Dissertations / Theses on the topic 'Fungal cake'

Neem cake is a product of the cold pressing from the neem kernels to obtain neem oil. Bio-active substances from neem cake extracted solutions were evaluated for their potential to control the root knot nematodes and other pests of plants. In this study different concentrations of the solution extracted from neem cake was tested against the second stage juveniles of the plant parasitic nematode Meloidogyne spp. and four phytopathogenic fungi: Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytopthora capsici. Toxicity of neem cake extractions is represented by the EC50 value for the second-stage juvenile (J2) of Meloidogyne spp. and the four phytopathogenic fungi via Probit analysis. A 5% dilution of the solvent extracting from neem cake already caused 100% larval mortality after 24 hours exposure. Undiluted neem cake extraction effectively inhibited the growth of the four phytopathogenic fungi. The EC50 value of neem cake on J2-larvae of Meloidogyne nematode and on the fungi Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytophthora capsici was 0.51, 0.74, 0.30, 0.51 and 4.33%, respectively
Bánh dầu neem là sản phẩm của quá trình ép nhân hạt neem để lấy dầu. Các hoạt chất sinh học từ dịch chiết bánh dầu neem đã được đánh giá có tiềm năng lớn trong phòng trừ tuyến trùng nốt sưng và các loài dịch hại khác của nhiều loại cây trồng. Trong nghiên cứu này các nồng độ dịch chiết khác nhau của bánh dầu neem đã được thử nghiệm khả năng diệt tuyến trùng (ấu trùng tuổi 2 thuộc giống Meloidogyne spp.) và ức chế 4 loài nấm bệnh như: Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. và Phytopthora capsici. Độc tính của dịch chiết bánh dầu neem được biểu diễn bởi giá trị EC50 đối với ấu trùng tuổi 2 của tuyến trùng Meloidogyne spp. và các loài nấm bệnh thông qua phân tích Probit. Dịch chiết bánh dầu neem ở nồng độ 5% đã làm chết 100% cá thể IJ2 của Meloidogyne spp sau 24 giờ phơi nhiễm. Dịch nguyên chất bánh dầu neem ức chế cả 4 loài nấm bệnh. Giá trị EC50 của bánh dầu neem lên ấu trùng tuổi 2 của Meloidogyne spp và các loài nấm bệnh Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytophthora capsici tương ứng là 0.51, 0.74, 0.30, 0.51 và 4.33%

Neem cake is a product of the cold pressing from the neem kernels to obtain neem oil. Bio-active substances from neem cake extracted solutions were evaluated for their potential to control the root knot nematodes and other pests of plants. In this study different concentrations of the solution extracted from neem cake was tested against the second stage juveniles of the plant parasitic nematode Meloidogyne spp. and four phytopathogenic fungi: Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytopthora capsici. Toxicity of neem cake extractions is represented by the EC50 value for the second-stage juvenile (J2) of Meloidogyne spp. and the four phytopathogenic fungi via Probit analysis. A 5% dilution of the solvent extracting from neem cake already caused 100% larval mortality after 24 hours exposure. Undiluted neem cake extraction effectively inhibited the growth of the four phytopathogenic fungi. The EC50 value of neem cake on J2-larvae of Meloidogyne nematode and on the fungi Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytophthora capsici was 0.51, 0.74, 0.30, 0.51 and 4.33%, respectively.
Bánh dầu neem là sản phẩm của quá trình ép nhân hạt neem để lấy dầu. Các hoạt chất sinh học từ dịch chiết bánh dầu neem đã được đánh giá có tiềm năng lớn trong phòng trừ tuyến trùng nốt sưng và các loài dịch hại khác của nhiều loại cây trồng. Trong nghiên cứu này các nồng độ dịch chiết khác nhau của bánh dầu neem đã được thử nghiệm khả năng diệt tuyến trùng (ấu trùng tuổi 2 thuộc giống Meloidogyne spp.) và ức chế 4 loài nấm bệnh như: Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. và Phytopthora capsici. Độc tính của dịch chiết bánh dầu neem được biểu diễn bởi giá trị EC50 đối với ấu trùng tuổi 2 của tuyến trùng Meloidogyne spp. và các loài nấm bệnh thông qua phân tích Probit. Dịch chiết bánh dầu neem ở nồng độ 5% đã làm chết 100% cá thể IJ2 của Meloidogyne spp sau 24 giờ phơi nhiễm. Dịch nguyên chất bánh dầu neem ức chế cả 4 loài nấm bệnh. Giá trị EC50 của bánh dầu neem lên ấu trùng tuổi 2 của Meloidogyne spp và các loài nấm bệnh Rhizoctonia solani, Sclerotium rolfsii, Collectotrichum spp. and Phytophthora capsici tương ứng là 0.51, 0.74, 0.30, 0.51 và 4.33%.

Stale bread contributes to the biggest volume of food waste in Sweden. Current method on recovering bread waste is by producing biogas or bioethanol. Despite advantages in the energy sector, the bread which still has relatively high quality could be recovered into new products with higher value, such as food for human consumption. Development of a product, termed ‘fungal cake’ by solid state fermentation on bread waste using Neurospora intermedia in small scale petri dishes have previously been successfully conducted. This study aims to scale up the production of fungal cake into bench scale production. Two systems using different bioreactors were used in this study. The first system operated in batch mode using a tray bioreactor, in which the effect of particle size, mixing, and bread loading were evaluated. The fermentation was conducted during 5 days. Bread crumb with a larger particle size of 2 mm resulted in similar outcomes as bread crumb with a smaller particle size of 0.5 mm in terms of CO2 evolution rate, cumulative CO2 production, starch, and protein content of the final product. However, larger particle size resulted in a more homogeneous growth of the fungus throughout the product, which is preferred. The presence of daily mixing had no significant effect compared to static condition for all measured variables. Thus, mixing could be introduced to promote product homogeneity. Likewise, bread loading had no significant effect on the measured variables, which implies that a higher productivity can be achieved using a higher bread loading. The second system operated in continuous mode using a newly developed continuous tubular bioreactor with product recycle. Two experiments, in which the residence time (48h and 24h) and recycle ratio (10/65 and 20/55) were conducted. Both experiments yielded product with stable starch and protein content, indicated by a stable CO2 evolution rate over time. The performance using continuous tubular bioreactor was compared to batch fermentation in tray bioreactor using the same ratio of inoculum and both system yielded product with the same starch and protein content. Successful operation in continuous bioreactor certainly improved the productivity of fungal cake production.

Thesis (Master of Conservation Science)--Cape Peninsula University of Technology, 2018.
Introduction pathways of fungal pathogens in South Africa are far less quantified in the literature than those for plants, animals and human infectious diseases. Phytopathogens continue to be introduced to South Africa via several pathways at an unprecedented rate. A number of these species pose a significant threat to South African ecosystems and biodiversity. Despite this, fungal pathogens could also be beneficial when they are used as bio-control agents to control alien invasive plant species. Nevertheless, recent studies revealed pathogens are most likely to be studied after they have caused a detrimental impact on the environment. Invasive fungal pathogens, such as Phytophthora cinnamomi (Oomycota) do not only pose a threat to native species of the family Proteaceae but could also potentially be bio-control agents for emerging alien plant invaders. In this thesis, firstly, I review current knowledge of phytopathogenic fungi introduction pathways in South Africa; secondly, I aim to understand the importance of fungi in limiting plant invasions using Banksia as a case study in the Cape Floristic Region. In chapter two I investigate introduction pathways and dispersal vectors that facilitate the spread of fungal pathogens. I compiled comprehensive list of fungal pathogens in South Africa, and evaluated the dispersal vectors and introduction pathways for each species. I found fifty five casual species, three naturalised species, six invasive species and thirty six pathogens for which invasion status was not classified due to insufficient data. Agriculture is responsible for the introduction of most fungal pathogens in South Africa. Wind was identified to be the prominent dispersal vector facilitating the spread of pathogens. I conclude that knowing introduction pathways of pathogens and their dispersal vectors will assist in developing quarantine protocols that could improve bio-security. Lastly, I provide recommendations for the national invasive microbe species list. In chapter three the study investigates the variability in mortality rate of Banksia species in the Cape Floristic. Species abundance was calculated across known Banksia populations in the Cape Floristic Region to determine survival and mortality rates. Soil and leave samples were taken from Banksia plants to evaluate potential microbial pests that were present. Also, acetone leaf extracts of twelve Banksia species were screened for antimicrobial activity against P. cinnamomi (Oomycota). Lastly, a post-border risk assessment was conducted for 14 Banksia species− present in South Africa − using the Australian Weed Risk Assessment protocol, to evaluate potentially invasive species. The results indicated that survival and mortality rate varied across species; I found the two invasive species, B. integrifolia and B. ericifolia to have the highest survival rate. Phytophthora cinnamomi was the most prominent isolated fungal pathogen sampled from Banksia species roots. The detection of antifungal activities in the minimum inhibitory concentration (MIC) bioassay provided evidence that some Banksia species (B. ericifolia, B. integrifolia, B. hookeriana and B. formosa) have antimicrobial chemical constituents that could possibly inhibit infection and colonisation by P. cinnamomi. The weed risk assessments conducted on Banksia species showed five species pose a high risk of invasion while seven species required further evaluation. I conclude that P. cinnamomi could potentially regulate invasive Banksia species such as B. speciosa with minimal antimicrobial activity against the pathogen. I recommend an in-situ and ex-situ inoculation trials of Banksia species against P. cinnamomi to be conducted to evaluate pathogenicity, under different watering regimes since the pathogens proliferation is favoured by soils that are high in moisture. I present the main conclusions from this thesis in chapter four and provide recommendations for management and invasive species legislation.

BACKGROUND: Non malignant invasive tumors of the colon and rectum are very rare. Gastrointestinal Basidiobolomycosis can present as a mass lesion mimicking colorectal cancer. CASE PRESENTATION: A 56 year old Caucasian male was evaluated for abdominal and pelvic pain for 4 weeks complicated by acute urinary retention. Radiological evaluation showed him to have recto-sigmoid and cecal mass. Endoscopic examination and biopsies did not reveal a definite diagnosis. Computerized tomography guided biopsy of the mass showed fungal elements consistent with gastrointestinal basidiobolomycosis. He was treated with Itraconazole for 12 months with very good clinical and radiological response. CONCLUSION: Basidiobolomycosis of the gastrointestinal tract should be considered during evaluation of colorectal masses with atypical presentation. It is a rare entity seen more in endemic regions of the world for basidiobolomycosis including southwestern United States.

Research regarding the distribution and structure of fungal communities in caves is lacking. The current study examines fungal communities in Kartchner Caverns, a mineralogically diverse cave located in the Whetstone Mountains, Arizona, USA. The first study examines culturable fungal diversity from speleothem surfaces. Twenty-one fungal genera represented by 43 genotypes and 53 distinct morphological taxonomic units (MTU) were recovered from 15 speleothems. Analysis of DGGE profiles indicated a significant effect of sampling site on community structure. The second study examined fungal diversity from speleothem and rock wall surfaces using the 454 FLX Titanium sequencing platform using the rDNA internal transcribed spacer 1 (ITS1) as a genetic marker. Fungal diversity was estimated and compared between speleothem and rock wall surfaces and its variation with distance from the natural entrance of the cave was quantified. Effects of environmental factors and nutrient concentrations in speleothem drip water at different sample sites on fungal diversity were also examined. Sequencing revealed 2219 fungal operational taxonomic units (OTUs) at 95% similarity. Speleothems supported a higher fungal richness and diversity than rock walls, but community membership and the taxonomic distribution of fungal OTUs did not differ significantly. OTU richness and diversity were negatively correlated with distance from the natural cave entrance. Community membership and taxonomic distribution of fungal OTUs differed significantly between the front and back of the cave. There was no observed effect of drip water nutrient concentration on fungal community structure. The third study examined fungal community structure from bat guano over the course of a year. There was no significant difference in fungal OTU richness, diversity, or community membership and taxonomic affiliations among sampling times. There were no significant differences in nutrient concentrations of guano samples among sampling times. Nutrient concentration did have a significant effect on community structure, especially the level of nitrogen and calcium.

Magister Public Health - MPH
BACKGROUND AND RATIONALE: The Cape Town Metro District Health Service (MDHS) has introduced a Diabetic RetinopathyScreening (DRS) programme incorporating retinal fundal photography in diabetic services at primary health care (PHC) facilities. Hitherto, coverage of the DRS programme has been less than optimal in part due to volumes of diabetic patients attending PHC facilities. The aim of this study was to identify possible sub-groups of patients, attending the Cape Town DRS Programme, who are at most risk of diabetic retinopathy and might be prioritised for early diabetic retinopathy detection and subsequent sight-saving treatment. METHODOLOGY: A case-control study of risk factors for treatment-requiring diabetic retinopathy was conducted. This research sampled participants from the DRS programme provided by the MDHS eye care team to Type II diabetics attending public PHC facilities within the Klipfontein and Mitchells Plain Sub-Districts. Based on fundal images, cases were selected as those requiring ophthalmological treatment; and controls (three matched per case by area of residence) as those judged as not requiring ophthalmological treatment for diabetic retinopathy. Data on possible risk factors (clinical, laboratory) were extracted from the patients' folders. RESULT: The study included 453 participants, of whom 113 (24.9%) were cases and 340 (75.1%) were controls. Three factors were significantly associated with treatment-requiring diabetic retinopathy on multivariate analysis: Insulin dependency (OR of 2.96, 95% CI: 1.75 – 5.00); duration of diabetes of more than 10 years (OR of 3.44, 95% CI: 2.06 – 5.74) and sustained hyperglycaemia over the past six months (OR of 3.73, 95% CI: 1.69 – 8.22). A screening algorithm combining these criteria had a sensitivity of 61.2% (95% CI: 51.9 – 70.5). CONCLUSION: The findings indicate that a sub-set of patients attending the DRS programme in the Klipfontein and Mitchells Plain Sub-Districts have a greater likelihood of presenting with treatment-requiring diabetic retinopathy. Further research is required to develop a tool that is sufficiently sensitive to safely prioritise patients for fundal screening.
National Research Foundation (NRF)

Day care centers and schools are important environments for children, but few epidemiological studies exist from these environments. Mould, dampness, fungal DNA and allergens levels in these environments and respiratory health effects in school children were investigated in this thesis. In the day care centers studies, Allergen Avoidance Day care Centers (AADCs) and Ordinary Day care Centers were included. One third of the Swedish day care centers had a history of dampness or mould growth. Total fungal DNA levels were positively associated with risk construction buildings, reported dampness/moulds, rotating heat exchangers, linoleum floors and allergens (cat, dog, horse allergen) levels. The two school studies included secondary schools in Johor Bahru, Malaysia and elementary schools from five European countries (Italy, Denmark, Sweden, Norway, and France) (HESE-study). In Malaysia, 13 % of the pupils reported doctor-diagnosed asthma but only 4 % had asthma medication. The prevalence of wheeze in the last 12 months was 10 % in Malaysia and 13 % in the HESE-study. Cough and rhinitis were common among children in the HESE-study. There were associations between fungal DNA and reported dampness or mould growth. Fungal DNA levels and viable mould (VM) concentration in the classrooms were associated with respiratory symptoms (wheeze, rhinitis, cough, daytime breathlessness) in school children. In the HESE-study, associations were found between total fungal DNA, Aspergillus/Penicillium DNA and respiratory symptoms among children. Moreover, Aspergillus versicolor DNA and Streptomyces DNA were associated with respiratory symptoms in Malaysia and the HESE-study, as well as reduced lung function [forced vitality capacity (FVC) and forced expiratory volume in 1 second (FEV1)] among children in the HESE-study. In conclusion, fungal DNA and pet allergens were common in day care centers and schools and respiratory symptoms in school children were common. The associations between VM concentration and fungal DNA levels in the schools and respiratory health effects in school children indicated a need for improvement of these environments. Moreover, risk constructions should be avoided and buildings should be maintained to avoid dampness and microbial growth. Health relevance of microbial exposure and biodiversity needs to be further studied using molecular methods.

Thesis (MScAgric.)--Stellenbosch University, 2000.
ENGLISH ABSTRACT: Phomopsis cane and leaf spot disease of grapevine is an economically important disease in many of the vine-growing areas of the world. Four different Phomopsis spp. have previously been associated with this disease. The present study investigates the taxonomic significance of the different taxa found on grapevines in South Africa, as well as the endophytic growth and fungicide sensitivity of Phomopsis viticola isolates. The thesis is compiled of several different parts, which deal with specific, but related topics, and hence some duplication has been unavoidable. Understanding the epidemiology of a disease is important for the correct timing of disease control. To investigate the endophytic growth of P. viticola, asymptomatic shoots were collected at eight different growth stages. Nodes, internodes, leaf petioles, leaves, tendrils and bunch peduncles were investigated. Two Phomopsis spp., taxon 1 and 2 were identified in this study. The Phomopsis viticola-complex had a relative importance of 9% and accounted for 3% of the isolations. P. viticola (taxon 2) is mainly isolated from the nodes and internodes. Inoculations of healthy, young vine tissue confirmed taxon 2 to be a virulent pathogen, suggesting that it is a latent pathogen rather than an endophyte. In contrast, taxon 1 appeared to be a true endophyte, and did not seem to be an important pathogen on vines. The true identity of the causal organism of Phomopsis cane and leaf spot disease was investigated by collecting samples from 58 different vineyards in the grapevine growing areas of the Western Cape. P. viiicola occurred in grapevine material collected from Lutzville to Swellendam, but was not found in the Oudtshoorn and Orange River grapevine areas. Diaporthe perjuncta (taxon 1), P. vutcola (taxon 2), taxon 3 and a Phomopsis species commonly associated with shoot blight of peaches in the U.S.A., P. amygdali, were identified among the South African grapevine isolates. Examination of the Australian culture designated as taxon 4 found it to be a species of Libertella, thus excluding it from the P. viticola-complex. An Italian isolate was found to represent a species of Phomopsis not previously known from grapevines, and this was subsequently described as taxon 5. Species delimitation was based on morphological and cultural characteristics, stem inoculations and the formation of the teleomorph in vitro. The identity of each morphological taxon was confirmed by means of phylogenetic analyses of the nuclear ribosomal DNA internal transcribed spacers (ITS 1 and ITS2) and the 5' end partial sequence of the mitochondrial small subunit (mtSSU). P. amygdali, associated with peach shoot blight in the U.S.A., was isolated once only and appeared to be of lesser importance in this disease complex. Furthermore, taxa 1 (Diaporthe perjuncta) and 3 were also rarely encountered and proved to be non-pathogenic, indicating their non-functional role in Phomopsis cane and leaf spot disease. Taxon 2 (Phomopsis viticolas was common and widely distributed in diseased vineyards. This taxon was associated with the typical disease symptoms and proved to be pathogenic. Morphologically taxon 2 corresponded best with P. viticola, which was also neotypified in this study. Taxon 2 was mostly isolated from buds and nodes, indicating that these are important sites in which the fungus survives during winter. Molecular data indicated that taxon 3 and P. amygdali were not host specific to grapevine. The currently used foliar fungicides were compared to the new strobilurin fungicides. The effects of nine fungicides (azoxystrobin, flusilazole, folpet, fosetyl- Al+mancozeb, kresoxim-methyl, mancozeb, penconazole, spiroxamine and trifloxystrobin) were tested in vitro on inhibition of mycelial growth. The following EC50 (ug/ml) values were obtained: azoxystrobin (0.350), flusilazole (0.007), folpet (4.489), fosetyl-Al+mancozeb (3.925), kresoxim-methyl (1.665), mancozeb (2.891), penconazole (0.023), spiroxamine (0.321) and trifloxystrobin (0.051). Additionally, azoxystrobin, folpet, kresoxim-methyl, mancozeb, propineb and trifloxystrobin were tested for their ability to inhibit spore germination in vitro. The subsequent EC50 (ug/ml) values were obtained: azoxystrobin 0.123), folpet (0.510), kresoxim-methyl (0.0037), mancozeb (0.250), propineb (0.156) and trifloxystrobin (0.003). The results reported in part 4 showed that the strobilurin fungicides inhibited the mycelial growth and spore germination of P. viticola. However, further trials need to be conducted to verify these findings under field conditions. In the present study taxa 1, 3 and P. amygdali were infrequently isolated, suggesting that they played a less prominent role in the P. viticolacomplex.
AFRIKAANSE OPSOMMING: Streepvleksiekte van wingerd is 'n ekonomies belangrike siekte wat in die meeste wingerdproduserende gebiede van die wêreld voorkom. Vier Phomopsis spesies is in die verlede met dié siekte geassosieer. Hierdie studie ondersoek die taksonomiese belangrikheid van die verskillende taksa wat op wingerd in Suid Afrika gevind word, asook die endofietiese groei en fungisiedsensitiwiteit van die Phomopsis vitico/a isolate. Hierdie tesis bestaan uit verskeie dele met spesifieke, maar verwante onderwerpe wat tot onafwendbare duplisering lei. Dit is belangrik om die epidemiologie van 'n siekte te verstaan sodat korrekte en tydsberekende siektebeheer toegepas kan word. Die endofietiese groei van P. vitico/a is ondersoek deur simptoomlose lote by agt verskillende groei stadiums te versamel. Nodusse, internodusse, blaarstele, blare, rankies en trosstele is ondersoek. Twee Phomopsis spp., takson 1 en 2 is geïdentifiseer. Die Phomopsis vitico/a-kompleks het 3% van die isolasies uitgemaak en 'n relatiewe belangrikheid van 9% getoon. P. vitico/a (takson 2) is meestal uit die nodus en internodus geïsoleer. lnokulasies van gesonde, jong wingerdweefsel het bevestig dat takson 2 'n virulente patogeen is en dat die takson eerder 'n latente patogeen as 'n endofiet is. In teenstelling hiermee is takson 1 'n ware endofiet en 'n onbelangrike patogeen op wingerd. Die ware identiteit van die veroorsakende organisme van streepvlek is ondersoek deur plantmateriaal vanaf 58 verskillende wingerde in die wingerproduserende gebiede van die Wes-Kaap te versamel. P. vitico/a is in wingerdmateriaal vanaf Lutzville tot Swellendam aangetref, maar nie in die Oudtshoorn en Oranjerivier wingerd produserende gebiede nie. Diaporthe perjuncta (takson 1), P. vitico/a (takson 2), takson 3 en P. amygdali is in die Suid Afrikaanse wingerdisolate geïdentifiseer. P. amygdali word met lootverskroeiing van perske bome in die V.S.A. geassosieer. Die Australiese isolaat wat benoem is as takson 4, is met die huidige ondersoek gevind om 'n spesie van Libertella te wees. Takson 4 is daarvolgens uit die P. vitico/a-kompleks gelaat. 'n Italiaanse isolaat het 'n nuwe spesie van Phomopsis op wingerd verteenwoordig en is vervolgens as takson 5 beskryf. Spesie-onderskeiding is op morfologiese en kulturele eienskappe, staminokulasies en die vorming van die teleomorf in vitro gebaseer. Die identiteit vanelke morfologiese takson is met behulp van filogenetiese analises van die nukleêre ribosomale DNS intern transkriberende spasieerders (ITS 1 en ITS2) en die 5' punt gedeeltelike nukleotied volgorde van die mitochondriale klein subeenheid (mtSSU) bevestig. P. amygdali is slegs een keer geïsoleer en blyk van minder belang in die siektekompleks te wees. Takson 1 (Diaporthe perjuneta) en takson 3 het ook min voorgekom en is nie-patogenies, wat hul nie-funksionele rol in streepvleksiekte aandui. Takson 2 (P. viticola) is algemeen geïsoleer en kom wyd verspreid voor. Hierdie takson is geassosieer met die tipiese siektesimptome en is ook patogenies. Morfologies stem takson 2 met P. viiicola ooreen en is ook geneotipifiseer in hierdie studie. Takson 2 is meestal vanaf die ogies en nodusse geïsoleer, wat daarop dui dat hierdie belangrike setels is waar die swam tydens die winter oorleef. Die molekulêre data toon aan dat takson 3 en P. amygdali nie gasheerspesifiek tot wingerd is nie. Die swamdoders wat tans teen streepvlek gebruik word, is met die nuwe strobilurin swamdoders vergelyk. Die effek van nege swamdoders (azoksistrobin, flusilasool, folpet, fosetyl-Al + mancozeb, kresoxirn-metiel, mankozeb, penconasool, spiroksamien en trifloksistrobin) is in vitro op die inhibisie van miseliumgroei getoets. Die volgende EKso-waardes (g/ml) is verkry: azoxystrobin (0.350), flusilasool (0.007), folpet (4.489), fosetiel-Al + mankozeb (3.925), kresoxirn-metiel (l.665), mankozeb (2.891), penkonasool (0.023), spiroksamien (0.321) en trifloxystrobin (0.051). Azoxystrobin, folpet, kresoxim-rnetiel, mankozeb, propineb en trifloksistrobin is ook in vitro getoets vir hul inhibisie op spoorontkieming. Die volgende EKso-waardes is verkry: azoxystrobin (0.123), folpet (0.510), kresoxim-metiel (0.0037), mankozeb (0.250), propineb (0.156) en trifloxystrobin (0.003). Die resultate vervat in deel 4 toon dat die strobilurin swamdoders die miseliumgroei en spoorontkieming van P. viticola inhibeer. Toetsing in die veld word egter benodig om die effektiwiteit van die middels te bevestig. In hierdie studie is taksa I, 3 en P. amygdali selde geïsoleer, wat aangedui het dat hierdie taksa 'n minder belangrike rol in die P. viticola-kompleks speel.

Endophytic microorganisms were screened for hemicellulases production using plate assays and liquid cultivations. Two strains were selected and used in further studies. Aspergillus niger DR02 strain and Annulohypoxylon stigyum DR47. In A.niger fed-batch submerged cultivation approaches were developed using liquor from hydrothermal sugar cane pretreatment, and maximum xylanase activities obtained were 458.1 U/mL for constant fed-batch mode. For A. stygium DR47 media optimization and bioreactor cultivation using citrus bagasse and soybean bran were explored and revealed a maximum production of 6.26 U/mL of pectinase and 10.13 U/mL of β-glucosidase. Improved hemicellulase production was also done by genetic modifications at carbon catabolic repression levels in A. niger. Mass spectrometric studies were done in enzymatic extracts produced and contributed to understand the enzymatic supplementation of Celluclast 1.5L.In addition the extracts produced were tested for an enzyme formulation and that was able to increase sugar cane bagasse hydrolysis.
Microrganismos endofíticos foram selecionados para a produção de hemicelulases através de testes em placa e de cultivo líquido. Duas linhagens foram selecionadas para posteriores cultivos, Aspergillus niger DR02 e Annulohypoxylon stigyum DR47. Para o fungo A.niger batelada alimentada em cultivo submerso com licor do tratamento hidrotérmico do bagaço de cana de açúcar permitiram obtenção de máxima atividade de xilanase de 458,1 U/mL. Para a linhagem A. stygium DR47 otimização do cultivo utilizando bagaço cítrico e farelo de soja foram explorados e revelaram produção de 6,26 U/mL de pectinase e 10,13 U/mL de β-glucosidase. Melhoramento da produção de hemicelulase foi realizado com modificações genéticas ao nível de repressão catabólica de carbono em A. niger. Estudos de espectrometria de massas foram realizados nos extratos enzimáticos produzidos e contribuíram para entender a suplementação da Celluclast 1.5L. Além disso, os extratos produzidos foram testados para uma formulação que foi capaz de aumentar a hidrólise do bagaço de cana de açúcar.

Background The majority of invasive fungal disease in South African hospitals is HIV-related or associated with another secondary immunodeficiency e.g. haematopoietic stem cell transplant. After excluding secondary immunodeficiency, a detailed immune work-up can lead to a diagnosis of primary immunodeficiency. Objective To detail an appropriate step-wise immunological work-up for a series of patients with invasive fungal diseases and possible underlying primary immune deficiency. Methods Detailed review of all culture- or histologically confirmed cases of invasive fungal disease (IFD) at Groote Schuur Hospital between 2007-2017. Step-wise immunological work-up of IFD patients with no secondary immunodeficiency. Clinical characteristics and step-wise immunological profiles were evaluated. Results Sixty-seven adults with IFD were identified; 72% (48/67) were HIV-related. 8/19 HIVnegative cases were either deceased (4) or lost-to-follow-up (4). Work-up of the remaining 11 cases found five with non-HIV secondary immunodeficiencies (Lupus, liver transplant, endstage renal failure and haematological malignancy). A primary immunodeficiency was suspected in six cases, but 1 case of cutaneous sporotrichosis was excluded; with five cases (4 with disseminated Cryptococcus neoformans and 1 with cerebral aspergillosis) undergoing detailed immune work-up. A case of idiopathic CD4 lymphopenia was diagnosed; but all other cases had no evidence of neutrophil or a cell-mediated immune defect; including investigations of naïve and memory T-cell subsets and cytokine responses to PHA and candida. All cases were noted to have low baseline vaccine responses and Vitamin D deficiency. Conclusion Invasive fungal disease is predominantly associated with HIV and secondary immunodeficiency in South Africa. Known primary immunodeficiencies can be identified with basic immune work-up; but no obvious functional immune defect is evident in the majority of these cases.

Contamination and growth of fungi within buildings, has gained increasing public attention within the last decade. Indoor fungal growth can have many adverse effects on building materials and has been linked to many health effects. In an effort to better understand the different environmental conditions that lead to fungal growth on indoor materials of drywall, and observe the biodegradative capabilities of these organisms, a case study was conducted at the University of British Columbia-Okanagan campus. The results of this study showed that age, and type of rooms can affect the diversity and composition of fungal taxa, with rooms in older buildings and laboratories supporting a higher diversity of fungi. The type of flooring and presence of carpet was not an influencing factor on fungal growth and diversity observed on drywall samples. A higher degree of fungal growth also affected the physical and mechanical properties of these building materials, with more fungi increasing the dry weight loss and decreasing the tensile strength of drywall. Although there are several epidemiological studies on the association between indoor fungi and the development of health problems, much still needs to be learned about what factors in indoor environments lead to the growth of these microorganisms. With a better understanding of which factors and environmental conditions trigger fungal growth in built environments, we can eventually better design indoor living spaces occupied by humans and develop methods for dealing with such problems.
Irving K. Barber School of Arts and Sciences (Okanagan)
Graduate

Thesis (M Med.(Family Medicine))--University of Limpopo, 2010.
Aim: To understand the help seeking behaviour of parents having children with minor illnesses such as fungal skin infection and to respond appropriately to such behaviours. Design: A descriptive qualitative study using the free attitude interview technique Method: The study was conducted in Francistown City Council Clinics. Eight participants were interviewed. Using purposeful sampling chose the participants. Each respondent was asked the same exploratory question “How much do you know about this skin infection?” Exploratory question (Setswana) is “O itse go le kae ka bolwetsi jone jo jwa letlalo?” The probe follow up questions were used to encourage elaboration on the topic. The discussions were held in Setswana language. The interviews were audio – taped. The recordings were transcribed, and the ideas that emerged were developed into themes. Results: Most of the respondents believed that Skin fungal infection was common in the community and it was generally called skin rash. It was believed to be infectious that it may be associated with HIV infection. Some however believe that its cause was known and it was treatable The help seeking behaviour of parents having children with minor illness like skin fungal infection falls within these reasons: Availability of alternative treatment, Concern of the child especially if the child complains about the problem, 6 Concern of the parents when the parents consider such minor health concern as a health problem for the child, issues around difficult in management of the problem, If problem is tolerable. Conclusion: Residents of Francistown city council, which was where the study was carried out perceived skin fungal infection as a common infectious skin problem, that is treatable. There is adequate knowledge of the symptoms of skin fungal infection among the participants. Some of the participants believed that skin fungal infection is common in children because they see the skin lesion as a normal change in colour for children before adult life. It was believe that as they get older the change in colour will then disappear. Many of them believe that they do not know the cause and even the few that felt that they knew the cause could not give a good account of the exact cause of the skin fungal infection, only one said that it is caused by a germ. Most of the participants were aware that it is treatable but yet they were not keen seeking for help when they come to the clinics because of one or two of the following reasons: Availability of alternative treatment; Concern of the child especially if the child complains about the problem; Concern of the parents when the parents consider such minor health concern as a health on the child; Issues around difficult in management of the problem; Health problem being tolerable

O bagaço de cana-de-açúcar, um dos resíduos gerados durante a produção de açúcar, é composto basicamente por celulose (41 44 %), hemicelulose (25 27 %) e lignina (20 22 %); cerca de dois terços da energia existente na cana-de-açúcar está contida no bagaço e na palha. Diversos processos utilizam o bagaço excedente, mas principalmente para produção de energia. No entanto, devido à grande quantidade de bagaço e palha que ainda sobram no processo estes materiais podem ser utilizados como fonte de celulose para obtenção de etanol. Para tanto, se faz necessária a hidrólise do bagaço e da palha a fim de se obter um hidrolisado que possa ser fermentado por leveduras industriais. A hidrólise do bagaço pode ocorrer por explosão a vapor, hidrólise química com ácidos ou álcalis ou hidrólise enzimática entre outros processos; a hidrólise química pode gerar compostos que são inibidores do processo fermentativo enquanto a hidrólise enzimática é um processo bastante oneroso visto não existir no Brasil a produção de enzimas para este fim. Por outro lado, se a hidrólise for efetuada somente com o próprio fungo, o processo pode ser lento. Neste contexto, este trabalho visou utilizar bagaço de cana-de-açúcar, pré-tratado por explosão a vapor e pré-tratado com ácido como substrato para selecionar fungos capazes de hidrolisar o bagaço a açúcares potencialmente fermentáveis por leveduras industriais. Foram realizados experimentos em uma etapa preliminar para selecionar dentre seis linhagens de fungos o(s) mais apto(s) para produção de açúcares redutores. E, experimentos fatoriais 22 para otimização das condições de hidrólise para os fungos selecionados na etapa anterior. Os fungos Trichoderma reesei e Phanerochaete chrysosporium foram selecionados para hidrolisar bagaço moído pré-tratado por ácido em cinco dias de incubação. A análise de variância (ANOVA) possibilitou gerar modelos validados e, superfícies de resposta para liberação de açúcares redutores e hexoses na hidrólise pelos fungos selecionados. As superfícies de resposta indicaram faixas ótimas de temperatura e concentração de ácido (32,19°C/1,09%) para liberação de açúcares redutores por T. reesei e, (32,6°C/1,06%) para P. chrysosporium.
The bagasse from sugar cane, a waste generated during the production of sugar, is composed primarily of cellulose (41 - 44%), hemicellulose (25 - 27%) and lignin (20 - 22%), about two thirds of energy in existing sugar cane is contained in the bagasse and straw. Several processes using bagasse surplus, but mainly for energy production. However, due to the large amount of bagasse and straw still left in these materials can be used as a source of cellulose to obtain ethanol. Thus, it is necessary to hydrolysis of bagasse and straw in order to obtain a hydrolyzate that can be fermented by yeast industry. Hydrolysis of bagasse can occur for a steam explosion, hydrolysis with acid or alkali chemical or enzymatic hydrolysis and other processes, the hydrolysis may generate chemical compounds that are inhibitors of the fermentation process while the enzymatic hydrolysis process is very costly as there is in Brazil the production of enzymes for this purpose. Furthermore, if the hydrolysis is carried out only with the fungus itself, the process can be slow. In this context, this work aimed to use crushed sugar cane, pre-treated by steam explosion and pre-treated with acid as a substrate to select fungi able to hydrolyze the bagasse potentially fermentable sugars for the yeast industry. Experiments were carried out in a preliminary step to select among six strains of fungi as suitable for production of reducing sugars. And, 2² factorial experiments to optimize the conditions of hydrolysis for selected fungi in the previous step. The fungi Trichoderma reesei and Phanerochaete chrysosporium were selected to hydrolyze milled bagasse pre-treated by acid in five days of incubation. The analysis of variance (ANOVA) allowed generate validated models and response surfaces for the release of reducing sugars and hexose hydrolysis by the selected fungi. The areas of response indicated bands optimum temperature and concentration of acid (32.19°C/1.09%) for release of reducing sugars by T. reesei, and (32.6°C/1.06%) for P. chrysosporium.

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A cana-de-açúcar colhida sem queima é uma realidade em todo o Estado de São Paulo e os ataques da cigarrinha-da-raiz da cana estão cada vez mais freqüentes e intensos. O controle biológico desta praga com o fungo Metarhizium anisopliae também vem se desenvolvendo e adquirindo relevada importância. Com isso, este trabalho foi conduzido com os seguintes objetivos: avaliar a produção de conídios de diferentes isolados para o controle da cigarrinha-das-raízes, a partir de dois tipos de arroz; avaliar a eficiência dos isolados selecionados como mais produtivos em populações naturais de Mahanarva fimbriolata, na cultura da cana-de-açúcar colhida mecanicamente e; verificar o manejo da população de M. fimbriolata em áreas de cana-de-açúcar colhidas sem queima da palha, observando a influência da umidade sobre sua ocorrência. Foram avaliados 14 isolados, os quais foram produzidos em arroz tipo 1 e arroz parboilizado, em sacos de polipropileno, incubados em sala climatizada para desenvolvimento do fungo. Avaliou-se a concentração e viabilidade de cada isolado para os dois tipos de arroz. Os isolados que apresentaram maior produtividade foram aplicados em campo para avaliar a patogenicidade dos mesmos à cigarrinha-das-raízes, sendo eles: ESALQ 1037, IBCB 425, IBCB 353, IBCB 410, F 99 e IBCB 333. Foram pulverizados 2 kg/ha de arroz+fungo, contendo 1,0 x 1012 conídios/ha, além do tratamento com o inseticida tiametoxam 250 WG e a testemunha, sem aplicação. As avaliações foram realizadas aos 15, 30, 60, 90 e 120 dias após a aplicação (DAA), observando-se o número de ninfas e adultos de M. fimbriolata vivos, mortos, parasitados ou não, em cada parcela. A partir dos resultados, foi possível observar que os isolados IBCB 410 e F 99 causaram, respectivamente, mortalidades de 66,67 e 33,33% para ninfas, aos 15 DAA. Após 30 DAA, os isolados IBCB 425, IBCB 353 e IBCB 333...
The sugar cane is harvested without burning a reality throughout the state of Sao Paulo and the attacks of the leafhopper-root cane are increasingly frequent and intense. Biological control this pest with the Metarhizium anisopliae also has been developing and acquiring increasing importance. Therefore, this work was conducted to evaluate the production of conidial selected for the control of root speatlebug in two types of rice, evaluate the efficiency of the isolates selected as the most productive in natural populations of M. fimbriolata, the culture of cane sugar harvested mechanically, monitor the population of M. fimbriolata in areas of sugarcane harvested without burning the straw, and the influence of temperature and humidity on its occurrence. We evaluated 14 isolates, which were produced in rice type 1 and parboiled rice in polypropylene bags, incubated in a room for fungal growth and were evaluated the concentration and viability of each isolate for both types of rice. The isolates that had higher yields were applied in the field to assess the pathogenicity of the same root speatlebug, namely: ESALQ 1037, IBCB 425, IBCB 353, IBCB 410, F 99 and IBCB 333. Were sprayed 2 kg / ha of rice + fungus, containing 1,0 x 1012 conidia / ha, in addition to treatment with the insecticide tiametoxam 250 WG and the control. Evaluations were performed at 15, 30, 60, 90 and 120 days after the application, noting the number of nymphs and adults of M. fimbriolata alive in each plot, and the number of nymphs and adults dead, infected and non-parasitized. From the results, it was observed that isolates IBCB 410 and F 99 caused mortality to nymphs of 66.67 and 33.33%, respectively, at 15 days after application. After 30 days of spraying, the isolates IBCB 425, IBCB 353 and IBCB 333 had efficiencies... (Complete abstract click electronic access below)

Thesis (MSc)--University of Stellenbosch, 2002.
ENGLISH ABSTRACT: Five new species of ophiostomatoid fungi, colon ising the infructescences of serotinous Protea species, have recently been discovered in South Africa. Prior to this, ophostomatoid fungi were thought to be restricted to the Northern Hemisphere. The discovery of these five species thus extensively expanded the known geographical range of these fungi, now also to include the Southern Hemisphere. Since this discovery, few studies have focused on the interesting ecology of this group, which is uniquely adapted for spore dispersal by arthropods. Studies focussed on ophiostomatoid fungi provide an unique opportunity to study inter-organism interactions between fungi, their host plants and the arthropods responsible for their spore dispersal. Very few similar studies have been undertaken, particularly in the Fynbos Biome, to which most of our economically important Protea species are confined. The current study provides insight into some ecological aspects of these complex interactions. The seasonal distribution of the ophiostomatoid fungi associated with the infructescences of members of the Proteaceae was investigated. Definite seasonal patterns were observed, with peak fungal colonisation occurring during the wetter winter months. While determining the host specificity of these fungi, a new Protea host for Ophiostoma splendens was identified, and a new species of Rhyncomeliola was discovered. All ophiostomatoid species native to Fynbos in the Stellenbosch region are exclusively associated with the infructescences of species in the plant genus Protea. By employing a multiplex Polymerase Chain Reaction (PCR), a set of group specific primers was developed for the identification of South African Ophiostoma and Gondwanamyces fungal DNA. This newly developed method was used to scan arthropods collected from the infructescences of P. repens for the presence of 0. splendens and G. proteae spores. By using these group-specific primers, three insects possibly responsible for long range dispersal of fungal spores were identified. Three further insect species, possibly contributing to the short-range dispersal of ophiostomatoid spores, were also implicated through this method. All arthropods associated with the fruiting structures of selected members of the Proteaceae were also investigated. A total of 62 pseudospecies (ca. 7500 individuals), belonging to 45 different arthropod families, were collected. Proteaceae species with larger fruiting structures housed more arthropod species and higher numbers of individuals than Proteaceae taxa with smaller fruiting structures. Some plant species housed similar arthropod communities, while others housed unique suites of arthropods. Seasonal patterns in arthropod numbers were observed, and it was found that, in most instances, arthropod numbers peaked during the autumn and winter months. Twenty-five fungal taxa were isolated from various Proteaceae arthropods, many of which are genera known to include Proteaceae pathogenic species. Ophiostomatoid fungi have a saprophytic relationship with their Protea hosts, and may deter some of these potentially harmful fungi from colon ising the infrutescences. Some fungi showed a high degree of specificity towards potential vectors, while others were found on a diverse range of arthropods. This study highlights the existence of complex inter-organismal interactions within the Fynbos Biome, a study area where the interactions between plants, fungi and insects have been grossly neglected. The few studies that have been conducted in this field have mostly focussed on a limited number of organisms, and no wide-scale attempts, such as presented here, have been published. It is important to obtain a holistic view in any ecological study that focuses on interactions between different suites of organisms. Ultimately this will aid in the development of better conservation strategies. This study thus provides a muchneeded start in studies on multi-organismal interactions in the Fynbos Biome.
AFRIKAANSE OPSOMMING: Vyf nuwe ophiostoma-agtige fungus spesies is onlangs in die vrug-liggame van Suid Afrikaanse Protea plante ontdek. Voor hierdie ontdekking, was die algemene opvatting dat ophiostoma-agtige fungi beperk is tot die Noordelike Halfrond. Die ontdekking van hierdie vyf spesies het dus die bekende geografiese verspreiding van hierdie tipe fungi vergroot om nou ook die Suidelike Halfrond in te sluit. Sederdien het min studies gefokus op die interesante ekologie van hierdie groep, wat aangepas is vir spoorverspreiding deur geleedpotiges. Studies wat fokus op die ophiostoma-agtige fungi voorsien 'n unieke geleentheid om die inter-organismiese-interaksies tussen fungi, hul gasheer plante en die geleedpotiges wat hul spore versprei, te bestudeer. Baie min soortgelyke studies is al voorheen in die Fynbos Bioom, waar die meeste van ons ekonomies-belangrike Protea spesies voorkom, onderneem. Die huidige studie verleen meer inligting oor sommige ekologiese aspekte van hierdie komplekse interaksies. Seisoenale verspreidingspatrone van ophiostoma-agtige fungi geassosieer met die vrug-liggame van lede van die Proteaceae, is ondersoek. Daar is defnitiewe seisoenale patrone in die aanwesigheid van fungi gevind, met 'n piek in fungi kolonisasie-getalle gedurende die vogtiger wintermaande. 'n Nuwe Protea gasheer vir Ophiostoma splendens en 'n nuwe fungus spesie (Rhyncomeliola sp.), is ontdek. Alle Fynbos ophiostoma-agtige spesies in die Stellenbosch area, is beperk tot Protea spesies. 'n Multipleks Polimerase Ketting Reaksie is ontwikkel om die spesifieke DNS van ophiostoma-agtige fungi te herken. Hierdie metode is gebruik om te toets vir die aanwesigheid van spore van hierdie fungi op die liggame van geleedpotiges (geassosieerd met P. repens). Hierdeur is drie insekte wat waarskynlik verantwoordelik is vir die langafstand verspreiding van spore geidentifiseer. Drie verdere insekte wat moontlik verantwoordelik is vir die kortafstand verspreiding van ophiostoma-agtige fungi is ook aangewys. Alle geleedpotiges geassosieer met die vrugstrukture van geselekteerde lede van die Proteaceae is ondersoek. 'n Totaal van 62 pseudo-spesies (omtrent 7500 individue), wat tot 45 families behoort, is versamel. Proteaceae spesies met groter vrugstrukture het meer geleedpotige spesies en individue gehuisves as taksa met kleiner vrugstrukture. Sommige plant spesies het soortgelyke geleedpotige gemeenskappe gehuisves, terwyl unieke groepe geleedpotiges in ander plante voorgekom het. Alhoewel die geleedpotiges seisonale variasies in getalle geopenbaar het, het die meeste groepe 'n piek in getalle getoon gedurende die herfs- en wintermaande. Vyf-en-twintig fungi taksa is geïsoleer vanaf verskeie Proteaceae geleedpotiges. Sommige van hierdie is genera wat Proteaceae patogeniese spesies insluit. Sommige fungi het 'n hoë graad van spesifisiteit teenoor potensiële vektore getoon, terwyl ander weer algemeen op verskeie geleedpotige spesies gevind is. Hierdie studie dui op die bestaan van komplekse inter-organismiese-interaksies in die Fynbos Bioom, in 'n area waar die studie van interaksies tussen plante, fungi en insekte nog baie min aandag geniet het. Die beperkte aantal studies wat tot dusver in hierdie veld onderneem is, fokus gewoonlik slegs op 'n paar organismes. Geen grootskaalse studies, soos die een hier voorgelê, is al gepubliseer nie. In enige ekologiese studie wat fokus op interaksies tussen verskeie organismes, is dit belangrik om 'n holistiese siening te probeer verkry. Uiteindelik sal dit bydra tot die ontwikkeling van beter bewarings-strategië. Hierdie studie verleen dus 'n noodsaaklike begin tot studies op multiorganismiese- interaksies.

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Universidade Estadual Paulista (UNESP)
Nas usinas, no início da safra, a obtenção de matéria-prima de boa qualidade é maximizada com a aplicação de fitorreguladores, os quais aumentam o teor de sacarose da cana-de-açúcar. Em áreas onde eles são aplicados, tem se observado melhor desenvolvimento e perfilhamento das plantas. Avaliou-se aqui o efeito da aplicação dos fitorreguladores Ethrel e Moddus sobre o crescimento da cana-de-açúcar, de forma direta e indiretamente, através da modificação da microbiota rizosférica. Além disso, objetivou-se também avaliar o uso de rizobactérias, isoladas dos experimentos com fitorreguladores, para o biocontrole de doenças e seus possíveis mecanismos de ação. Os efeitos dos fitorreguladores sobre os microrganismos do solo foram avaliados em meios de cultura acrescidos de Ethrel e Moddus em concentrações de 0 a 1000 ppm. Estes fitorreguladores foram aplicados via foliar e via solo para análise do desenvolvimento da cana-de-açúcar (variedades RB72454, RB835486 e RB855156) em casa-de-vegetação, utilizando-se solo sem tratamento e tratado com brometo de metila. Após dez meses, foram avaliadas a brotação, altura e matéria seca da parte aérea e das raízes das plantas cultivadas. Rizobactérias foram isoladas dos solos contidos nos vasos e avaliadas in vitro quanto à capacidade de controle de fungos fitopatogênicos (Thielaviopsis paradoxa, Fusarium spp. e Hendersonina sacchari), e in vivo, quanto à capacidade de promoção de crescimento de plântulas de cana-de-açúcar. Alguns mecanismos de ação das rizobactérias foram também estudados, como produção de ácido indol acético, ácido cianídrico, sideróforos e solubilização de fosfato inorgânico. Constatou-se que as populações de fungos foram mais sensíveis à adição dos fitorreguladores do que outros grupos de microrganismos, com redução...
For sugar and alcohol industries, at the start of harvesting, to obtain good quality raw material is potentially possible with the application of plant regulators, which have a role in natural sugar cane maturation, increasing sucrose content. In areas where they have been applied, better plant development and shooting have been observed. The aim here was to evaluate the application of plant regulators Ethrel and Moddus on sugar cane growth, not only in a direct way, but also indirectly, through the modification of rhizosphere microorganisms. Besides, this work also aimed the evaluation of rhizobacteria isolated from the experiments using plant regulators upon the disease biocontrol and their action mechanisms in this respect. The effects of plant regulators upon the soil microorganisms were verified in culture media where Ethrel and Moddus were added in concentrations ranging from 0 to 1000 ppm, while the effects of these substances (applied in leaves and in soil) upon the sugar cane development (varieties RB72454, RB835486 and RB855156) were surveyed in greenhouse, using soil without treatment and treated with methyl bromide. After a ten-month period, the experiments were finished, and sprouting, height and aerial part and root dry matter were analyzed. Soil samples were taken from the pots for rhizobacteria isolation, which were evaluated initially in vitro regarding their ability to control plant pathogenic fungi (Thielaviopsis paradoxa, Fusarium spp. and Hendersonina sacchari), and in vivo, regarding their ability to promote sugar cane growth. Some action mechanisms were also studied, as indol acetic acid, cyanide acid and siderophore production and inorganic phosphate solubilization. It was verified that the fungi populations were more sensitive to the addition of plant regulators than other microorganisms, reducing their colony-forming unit (CFU)...(Complete abstract, click electronic access below)

The laboratory-derived calcite dissolution rate law of Plummer et al. (1978) is the most widely used and mechanistically detailed expression currently available for predicting dissolution rates as a function of water chemistry. Such rate expressions are of great use in understanding timescales associated with limestone karst development. Little work has gone into the field testing of the rate law under natural conditions. This work compared measured dissolution rates measured by a crystal weight loss experiment in Buffalo Creek within Fort’s Funnel Cave, which lies within a pristine, forested catchment of Mammoth Cave National Park. Continuous water chemistry sampling over the same period allowed a time-integrated prediction of the dissolution based on the Plummer et al (1978) expression. Results indicate that the rate law overpredicted dissolution by a factor of about ten. This concurs with earlier laboratory work suggesting that the law tends to overpredict rates in solutions close to equilibrium with respect to calcite, as were the waters in this study. Estimating dissolution rates with the expression under varying hydrologic conditions also allowed a prediction of storm scales change in cave forming processes. Neglecting effects of sediment masking on the bed, it was found that 78% of the work done in the dissolution of the cave passage during the study period occurred at or around baseflow conditions, with only a small amount during the effective but infrequent high flow conditions.

Thesis (MScAgric)--University of Stellenbosch, 2004.
ENGLISH ABSTRACT: Soilborne diseases of grapevines represent a complex problem with limited information available, both locally and internationally. Previous research in South Africa indicated that Phytophthora and Pythium spp. were the most widespread and devastating pathogens in grapevine nurseries and vineyards in the Western Cape province. The local grapevine industry is currently expanding; new cultivars, methods and agricultural chemicals are being used which can affect soilborne pathogens. It has therefore become necessary to reassess the status of soilborne pathogens in nurseries, since information in this regard is crucial for the development of disease management practices for the expanding local grapevine industry. Soilborne fungal genera associated with roots and crowns of declining nursery grapevines were assessed in surveys conducted at three different grapevine nurseries in the Western Cape province. Cylindrocarpon, Fusarium, Pythium, and Rhizoctonia spp. were consistently isolated from roots and crowns of declining nursery grapevines. Cylindrocladiella spp. and Phytophthora cinnamomi were infrequently isolated from diseased roots, crowns and soil whereas Pythium spp. were abundant in most of the soils. Results suggest that the status of soilborne fungal pathogens in grapevine nurseries in the Western Cape province has changed over the last 30 years. The DNA phylogeny and pathogenicity of the isolates of Cylindrocladiella were determined. Four species of Cylindrocladiella occur on grapevines in South Africa, namely C. lageniformis, C. parva, C. peruviana, as well as a new species, described in this study as C. viticola, which forms part of the C. infestans species complex. Pathogenicity trials were inconclusive. Ten Fusarium spp. were isolated from roots and crowns of declining nursery grapevines, namely F. acuminatum, F. anthophilum, F. chlamydosporum, F. equiseti, F. nygamai, F. oxysporum, F. proliferatum, F. scirpi, F. semitectum and F. solani. The dominant species was F. oxysporum, followed by F. proliferatum and F. solani. In pathogenicity trials F. oxysporum and F. solani significantly reduced root volume, root dry mass, length of new shoots, stem diameter and number of leaves, but increased the percentage of chlorotic leaves and root rot severity. Fusarium proliferatum also caused a significant reduction in new shoot growth, number of leaves and increased root rot severity compared to the controls. Fusarium so/ani seems to be more virulent than F. oxysporum, followed by F. pro/iferatum. This is the first report of F. oxysporum, F. pro/iferatum and F. so/ani as pathogens of grapevines in South Africa, and the first report of F. proliferatum as a pathogen of grapevines in the world. Phytophthora cinnamomi was isolated at low frequencies from declined grapevines, although present in the rhizosphere soil. It is possible that the extensive use of downy mildew chemicals in grapevine nurseries may protect grapevines from infection by P. cinnamomi. The effect of chemicals used to combat downy mildew on Phytophthora root rot of nursery grapevines was evaluated in a glasshouse. There was very little discernable effect of the chemicals tested relative to the control plants for the parameters measured and it was concluded that the inoculation technique needed refinement. However, plants treated with phosphorous acid tended to be taller and have more leaves, greater stem diameter and root volume than controls or plants treated with the other chemicals. The data obtained in this study are not conclusive, but indicated certain trends that more glasshouse trials and field trials would resolve. Results presented in this thesis indicate that a major shift has occurred in the status of soilborne fungi associated with roots and crowns of grapevines in nurseries in the Western Cape since the 1970s when Phytophthora and Pythium were predominant. The prevalence and role of soilborne fungi need to be determined so that new appropriate disease management strategies can be developed to limit losses in grapevine nurseries and ensure the sustainable production of healthy plants for the grapevine industry.
AFRIKAANSE OPSOMMING: 'N ONDERSOEK NA GRONDGEDRAAGDE SWAMME GEASSOSIEER MET WORTELS EN KRONE VAN WINGERD IN KWEKERYE Grondgedraagde siektes van wingerd is 'n komplekse probleem waaroor min inligting, beide plaaslik en internasionaal, beskikbaar is. Vorige navorsing in Suid-Afrika het aangedui dat swamme van die genera Phytophthora en Pythium die mees algemene en vernietigende grondgedraagde patogene in kwekerye en wingerde in die Wes-Kaap provinsie is. Die plaaslike wingerdbedryf brei huidiglik uit; nuwe kultivars, metodes en landbouchemikalieë word gebruik wat 'n invloed kan hê op grondgedraagde patogene. Gevolglik het dit noodsaaklik geword om die status van grondgedraagde patogene in wingerdkwekerye weer te bepaal, aangesien inligting in hierdie verband noodsaaklik is vir die ontwikkeling van siekte bestuurspraktyke vir die ontwikkelende plaaslike wingerdbedryf. Grondgedraagde swamgenera geassosieer met wortels en krone van terugsterwende wingerd in kwekerye is bepaal in opnames wat by drie verskillende wingerdkwekerye in die Wes-Kaap provinsie uitgevoer is. Cylindrocarpon, Fusarium, Pythium, en Rhizoctonia spp. is konstant vanuit wortels en krone van terugsterwende wingerdplante in kwekery geïsoleer, Cylindrocladiella spp. en Phytophthora cinnamomi is ongereeld vanuit siek wortels, krone en grond geïsoleer, terwyl Pythium spp. algemeen in meeste gronde voorgekom het. Resultate dui daarop dat die status van grondgedraagde swampatogene in wingerdkwekerye in die Wes- Kaap provinsie oor die laaste 30 jaar verander het. Die DNA filogenie en patogenisiteit van die isolate van Cylindrocladiella is bepaal. Vier spesies van Cylindrocladiella kom voor op wingerd in Suid-Afrika, naamlik C. lageniformis, C. parva, C. peruviana, sowel as 'n nuwe spesie, wat in hierdie studie as C. viticola aangedui is en wat deel is van die C. infestans spesie kompleks. Patogenisiteits proewe was onvoldoende om die patogeniese status van die swam me te bepaal. Tien Fusarium spp. is vanuit wortels en krone van terugsterwende wingerdplante in kwekery geïsoleer, naamlik F. acuminatum, F. anthophilum, F. chlamydosporum, F. equiseti, F. nygamai, F. oxysporum, F. proliferatum, F. scirpi, F. semitectum en F. solani. Die dominante spesies was F. oxysporum, gevolg deur F. proliferatum en F. solani. In pathogenisteitsproewe het F. oxysporum en F. solani gelei tot 'n betekenisvolle laer wortelvolume, droë massa van wortels, lengte en droë massa van nuwe groei en aantal blare, maar het die persentasie chlorotiese blare en graad van wortelvrot verhoog. Fusarium proliferatum het ook gelei tot 'n betekenisvolle afname in lengte en massa van nuwe groei, aantal blare en 'n verhoogde graad van wortelvrot in vergelyking met die kontrole behandelings. Dit wil voorkom asof Fusarium solani meer virulent is as F. oxysporum, gevolg deur F. proliferatum. Hierdie is die eerste aanmelding van F. oxysporum, F. proliferatum en F. solani as patogene van wingerd in Suid-Afrika, en die eerste aanmelding van F. proliferatum as 'n patogeen van wingerd in die wêreld. Phytophthora cinnamomi is konstant teen lae frekwensies vanuit terugsterwende wingerd in kwekerye geïsoleer, alhoewel dit in risosfeer gronde teenwoordig was. Dit is moontlik dat die ekstensiewe gebruik van chemikalieë teen donsskimmel in wingerdkwekerye die wingerdplante kan beskerm teen infeksie deur P. cinnamomi. Die effek van chemikalieë wat gebruik word teen donsskimmel op Phytophthora wortelverrotting van wingerd in kwekerye, is 'n glashuis geëvalueer. Die chemikalieë wat gestoets is, het vir die gemete parameters, tot baie min onderskeibare effek gelei relatief tot die kontrole plante, en daar is afgelei dat die inokulasie tegniek verbetering benodig. Plante wat met fosforiensuur behandel is, het egter geneig om langer te wees met meer blare, 'n groter stamdeursnee en wortelvolume as kontrole plante of plante behandel met ander chemikalieë. Data verkry vanuit die hierdie studie was onvoldoende, maar sekere neigings is aangedui wat deur verdere glashuis- en veldproewe verklaar sal word. Resultate wat in hierdie tesis weergegee is, het aangedui dat 'n algehele verskuiwing in die status van grondgedraagde swamme geassosieer met wortels en krone van wingerd in kwekerye vanaf die 1970s, toe Phytophthora en Pythium die dominante genera was, plaasgevind het. Die voorkoms en rol van grondgedraagde swamme moet bepaal word, sodat nuwe voldoende siektebestuurspraktyke ontwikkel kan word om verliese in wingerdkwekerye te beperk en sodoende die volhoubare produksie van gesonde plante vir die wingerdbedryf te verseker.

Thesis advisor: Melissa M. Kelley
Thesis advisor: Lisa Sowle Cahill
Thesis (STL) — Boston College, 2016
Submitted to: Boston College. School of Theology and Ministry
Discipline: Sacred Theology

Orientador: Samia Maria Tauk-Tornisielo
Banca: Sandra Regina Ceccato Antonini
Banca: Regina Teresa Rosim Monteiro
Banca: Antonio Ismael Bassinelo
Banca: Carlos Renato Corso
Resumo: Nas usinas, no início da safra, a obtenção de matéria-prima de boa qualidade é maximizada com a aplicação de fitorreguladores, os quais aumentam o teor de sacarose da cana-de-açúcar. Em áreas onde eles são aplicados, tem se observado melhor desenvolvimento e perfilhamento das plantas. Avaliou-se aqui o efeito da aplicação dos fitorreguladores Ethrel e Moddus sobre o crescimento da cana-de-açúcar, de forma direta e indiretamente, através da modificação da microbiota rizosférica. Além disso, objetivou-se também avaliar o uso de rizobactérias, isoladas dos experimentos com fitorreguladores, para o biocontrole de doenças e seus possíveis mecanismos de ação. Os efeitos dos fitorreguladores sobre os microrganismos do solo foram avaliados em meios de cultura acrescidos de Ethrel e Moddus em concentrações de 0 a 1000 ppm. Estes fitorreguladores foram aplicados via foliar e via solo para análise do desenvolvimento da cana-de-açúcar (variedades RB72454, RB835486 e RB855156) em casa-de-vegetação, utilizando-se solo sem tratamento e tratado com brometo de metila. Após dez meses, foram avaliadas a brotação, altura e matéria seca da parte aérea e das raízes das plantas cultivadas. Rizobactérias foram isoladas dos solos contidos nos vasos e avaliadas in vitro quanto à capacidade de controle de fungos fitopatogênicos (Thielaviopsis paradoxa, Fusarium spp. e Hendersonina sacchari), e in vivo, quanto à capacidade de promoção de crescimento de plântulas de cana-de-açúcar. Alguns mecanismos de ação das rizobactérias foram também estudados, como produção de ácido indol acético, ácido cianídrico, sideróforos e solubilização de fosfato inorgânico. Constatou-se que as populações de fungos foram mais sensíveis à adição dos fitorreguladores do que outros grupos de microrganismos, com redução...(Resumo completo, clicar acesso eletrônico abaixo)
Abstract: For sugar and alcohol industries, at the start of harvesting, to obtain good quality raw material is potentially possible with the application of plant regulators, which have a role in natural sugar cane maturation, increasing sucrose content. In areas where they have been applied, better plant development and shooting have been observed. The aim here was to evaluate the application of plant regulators Ethrel and Moddus on sugar cane growth, not only in a direct way, but also indirectly, through the modification of rhizosphere microorganisms. Besides, this work also aimed the evaluation of rhizobacteria isolated from the experiments using plant regulators upon the disease biocontrol and their action mechanisms in this respect. The effects of plant regulators upon the soil microorganisms were verified in culture media where Ethrel and Moddus were added in concentrations ranging from 0 to 1000 ppm, while the effects of these substances (applied in leaves and in soil) upon the sugar cane development (varieties RB72454, RB835486 and RB855156) were surveyed in greenhouse, using soil without treatment and treated with methyl bromide. After a ten-month period, the experiments were finished, and sprouting, height and aerial part and root dry matter were analyzed. Soil samples were taken from the pots for rhizobacteria isolation, which were evaluated initially in vitro regarding their ability to control plant pathogenic fungi (Thielaviopsis paradoxa, Fusarium spp. and Hendersonina sacchari), and in vivo, regarding their ability to promote sugar cane growth. Some action mechanisms were also studied, as indol acetic acid, cyanide acid and siderophore production and inorganic phosphate solubilization. It was verified that the fungi populations were more sensitive to the addition of plant regulators than other microorganisms, reducing their colony-forming unit (CFU)...(Complete abstract, click electronic access below)
Doutor

Background. As a result of the AIDS pandemic, many people areimmuno compromised andopportunistic fungal infections (OFIs) such as candidiasis are common. Despite the widespread use of medicinal plants in South Africa, there is a dearth of knowledge regarding the use of such plants in the management of these infections. This study evaluates three South African medicinal plants (Arctotis arctotoides, Pittosporum viridiflorum, and Gasteria bicolor) traditionally used in the treatment of OFIs in HIV/AIDS patients, in the Eastern Cape Province, South Africa. Materials and methods. A six-stage process of documentation, evaluation and analysis of results was conducted: (1) Selection of medicinal plants most frequently used in the treatment of OFIs through ethnomedical studies and the survey of specialised literature; (2) Collection and preparation of the extract of each plant; (3) Antifungal evaluation of the crude plant extracts. (4) Phytochemical and antioxidant evaluation of the active crude plant extracts; (5) Cytotoxicity evaluation of the bioactive extracts using the Chang liver cell line, and (6) Statistical analysis of the results. Ethnobotanical information was obtained through interviews with traditional healers and AIDS patients with the aid of semi-structured questionnaires, direct observations and by reviewing studies reported in the literature. Following the approval from the University of Fort Hare‘s Ethics Committee, 101 HIV/AIDS patients were recruited through convenience sampling into an anonymous cross-sectional questionnaire study. The agar diffusion and micro-dilution methods were used to determine the antifungal activities of the hexane, acetone and aqueous extracts of A. arctotoides, G. bicolor and P. viridiflorum against 10 opportunistic fungi.

Thesis (MSc)--Stellenbosch University, 2012.
ENGLISH ABSTRACT: Only a limited number of systems involving anther-smut fungi have been studied, usually due to the economic significance of their crop plant hosts. A smut fungus of the genus Thecaphora has been discovered infecting Oxalis hosts in South Africa. This pathogenic fungus, Thecaphora capensis, produces dark-coloured spores in the anthers of host flowers, rendering it an anther-smut fungus. The host genus is the seventh largest plant genus in the Cape Floristic Region (CFR) and the largest geophytic genus of this region. Nine Oxalis species that host T. capensis have been identified across a wide distribution in the CFR of South Africa. A preliminary assessment of T. capensis infections of Oxalis was conducted in 2009, which provided a foundation for further research into the ecological and evolutionary consequences of hosting this fungus. In this study, a comprehensive host diversity assessment was conducted to determine the extent of infected Oxalis individuals within the CFR. Three new Oxalis host species for Thecaphora capensis were discovered. This brings the total number of known hosts to twelve. The morphological and reproductive effects of the fungus were assessed on two host species (O. incarnata and O. lanata) by comparing healthy and infected individuals of these species. Infection by Thecaphora capensis had a significantly negative effect on both of these factors. Host resources appear to be co-opted for fungal spore production, since floral morphological characters of infected individuals were reduced in size. Furthermore, infection by T. capensis ensured near-universal sterility in both hosts. Differences in floral characters and pollinator preferences for healthy Oxalis incarnata and O. lanata individuals from disease-free and diseased populations were compared to determine the evolutionary influence of Thecaphora capensis infections. It was shown that this pathogen can have a significant evolutionary influence on its hosts, showing its ability to shape flower size and pollinator activity in O. lanata, but not in O. incarnata. A need has therefore been identified to assess these evolutionary forces independently for each host and its pathogen before making erroneous assumptions for conservation practices. Plant pollinators play an integral role in plant fitness. Pollinator movements within a population are important when between-flower spore transfer by pollinators increases the likelihood of new infections. Pollinator movements may be influenced by host density and the frequency of diseased individuals, amongst other factors. Pollinators were found to mediate Thecaphora capensis spore transfers within diseased Oxalis populations. Host density and disease frequency affected the number of spores transferred under field and standardized conditions. More research is required to investigate confounding factors in these complex systems. This study highlighted the complexities of a fungal-plant-insect relationship, the evolutionary consequences of such fungal infections and the various factors influencing the likelihood of new infections. This research adds to the limited body of knowledge on multi-organismal interactions in the CFR and provides a base for more detailed future studies on this intriguing system.
AFRIKAANSE OPSOMMING: 'n Brandswam, wat deel is van die Thecaphora genus,is ontdek in 'n Oxalis blom waar dit die gasheer plant se blom gebruik om spore in te produseer. Die swam, Thecaphora capensis, produseer donker gekleurde spore in die helmknoppe van die blomme van gasheer plante, daarom word dit geklasifiseer as 'n brandswam van die helmknop. Die gasheer plante van die swam is deel van die genus Oxalis, die sewende grootste plant genus in die Kaapse Floristiese Streek (KFS) en die grootste geofitiese genus in die streek. Nege Oxalis species is al klaar identifiseer as gasheer plante van T. capensis. Hulle is versprei oor 'n groot area van die KFS van Suid Afrika. 'n Primêre ondersoek van T. capensis infeksies op Oxalis is in 2009 onderneem. Hierdie ondersoek het gelei tot meer vrae oor die sisteem en het 'n goeie fondasie geskep vir verdere navorsing rakende die ekologiese koste verbonde daaraan om as gasheer plant vir 'n swam op te tree. 'n Deeglike ondersoek is in die KFS aangepak om die Oxalis gasheer plante van die brand swam te identifiseer en om voort te bou op die basiskennis wat in die primêre ondersoek daargestel is. Drie nuwe Oxalis gasheer plante van Thecaphora capensis is ontdek. Die totale aantal gasheer plante staan nou op twaalf. Gesonde en geinfekteerde individuele gasheer plante is gebruik om die morfologiese en reproduktiewe effekte van die swam te toets in twee Oxalis spesies (O. incarnata en O. lanata). Die negatiewe gevolge om 'n gasheer plant van die brand swam te wees was duidelik toe gesonde en geinfekteerde individuele met mekaar vergelyk is. Dit kom voor asof gasheer plante se hulpbronne vir spoor produksie gebruik word, want hulle is morfologies kleiner en meestal steriel. Die evolusionêre effek van Thecaphora capensis op 'n populasie is getoets met gesonde individuele in populasies van twee Oxalis spesies. Blomkenmerke en insek bestuiwers van gesonde individue in gesonde en geinfekteerde populasies is ondersoek om die effekte van T. capensis op populasies te toets. Daar is suksesvol gedemonstreer dat swamme sterk evolusionêre kragte uitoefen, en die vermoë het om plantpopulasies te vorm en te verander, ofskoon nie in alle gevalle ewe sterk nie. Daarom is dit belangrik om die evolusionêre kragte vir elke gasheer plant en sy patogeen onafhanklik te assesseer, sonder om algemene aannames te maak in bewaringspraktyke. Plantbestuiwers speel 'n belangrike rol in die fiksheid van plante. Hulle kan hul fiksheid verbeter deur bestuiwers te lok met blomme en deur aspekte geassosieer met blomme. Die bewegingspatrone van plantbestuiwers is baie belangrik indien hulle helmknop-geproduseerde spore van brandswamme vervoer instede van stuifmeel, want dit vergroot die kanse vir nuwe infeksies. Die bewegingspatrone van plantbestuiwers word, onder andere, beinvloed deur die digtheid en frekwensie van geinfekteerde individue. Plantbestuiwers speel 'n belangrike rol in die vervoer van Thecaphora capensis spore in geinfekteerde Oxalis populasies. Die digtheid en frekwensie van geinfekteerde blomme het die vervoer van spore geaffekteer onder veld en gestandardiseerde kondisies, alhoewel baie veranderlikes so 'n komplekse natuurlike sisteem beinvloed. Hierdie studie beklemtoon die kompleksiteit van 'n fungus-plant-insek verhouding, die gevolge van so 'n interaksie en die verskeie faktore wat die waarskynlikheid van nuwe infeksies beinvloed. Tot dusver is daar 'n beperkte aantal sisteme soos hierdie bestudeer waarin 'n brandswam van die helmknop betrokke is, en die enkele beskikbare studies is onderneem meestal as gevolg van hulle ekonomiese effekte op landboukundig belangrike gasheer plante. Hierdie studie verteenwoordig 'n belangrike byvoeging tot die inter-organismiese studies in die KFS. 'n Holistiese ekologiese oorsig soos hierdie verskaf 'n belangrike basis vir toekomstige studies en bewarings- en bestuurspraktyke.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior
A cultura da mamoneira (Ricinus communis L.) à de grande importÃncia socioeconÃmica devido Ãs suas sementes, que sÃo utilizadas, principalmente, na produÃÃo do Ãleo que pode ser empregado para diversos fins industriais. A extraÃÃo do Ãleo gera um subproduto de alto valor nutricional, conhecido como torta da mamona. PorÃm, a presenÃa de compostos tÃxicos e alergÃnicos dificulta utilizaÃÃo desse resÃduo como fonte de alimentaÃÃo animal. Outra forma de agregar valor à torta da mamona seria buscar molÃculas bioativas que pudessem ter aplicaÃÃes na agricultura e saÃde humana. O presente trabalho teve como objetivos purificar e caracterizar um inibidor de tripsina da torta da mamona, bem como, testar a atividade deste in vitro contra fungos fitopatogÃnicos de importÃncia agrÃcola e proteases do intestino mÃdio de Aedes aegypti. O inibidor de tripsina da torta da mamona, nomeado RcTI, foi purificado atravÃs de tratamento tÃrmico, coluna de afinidade em matriz de anidrotripsina-Sepharose 4B e cromatografia de troca iÃnica em ResourceTM Q. Essa proteÃna apresentou massa molecular aparente de 14,0 kDa na ausÃncia ou presenÃa de agente redutor, pI 5,2 e nÃo à uma glicoproteÃna. A sequÃncia NH2-terminal do RcTI purificado mostrou similaridade de 83% com uma proteÃna de armazenamento rica em enxofre (albumina 2S) de R. communis e de 48% com uma napin-like (albumina 2S) de R. communis. O RcTI mostrou-se estÃvel apÃs tratamento tÃrmico a 98 ÂC durante duas horas. Da mesma forma, permaneceu com atividade inibitÃria estÃvel na faixa de pHs Ãcidos. No entanto, houve uma perda de, aproximadamente, 20% na capacidade inibitÃria em pHs alcalinos (pH 8-11). O RcTI (13 Âg) nÃo foi capaz de inibir a germinaÃÃo de esporos dos fungos Fusarium oxysporum, Fusarium solani e Rhizoctonia solani. No entanto, foi capaz de inibir a germinaÃÃo de esporos de Colletotrichum gloeosporioides. O RcTI nÃo foi efetivo na inibiÃÃo de crescimento vegetativo dos fungos mencionados acima. Esse inibidor foi efetivo contra as proteases intestinais de Aedes aegypti com 91% de inibiÃÃo. A partir dos resultados obtidos neste trabalho, conclui-se que o RcTI tem potencial biotecnolÃgico, podendo ser utilizado como uma alternativa para o combate do importante fungo fitopatogÃnico C. gloeosporioides e larvas de A. aegypti.
The castor bean (Ricinus communis L.) culture is of great socioeconomic importance because of its seeds, which are used mainly for production of oil that can be used for various industrial purposes. The oil extraction generates a by-product of high nutritional value known as the castor cake. However, the presence of toxic and allergenic compounds hinders the use of this residue to feeding source. Another way to add value to the castor cake would be to seek bioactive molecules that could have applications in agriculture and human health. This study aimed to purify and characterize a trypsin inhibitor of castor cake and, to test this in vitro activity against phytopathogenic fungi of agricultural importance and proteases from the midgut of Aedes aegypti. The trypsin inhibitor of castor cake, named RcTI, was purified by heat treatment, affinity column in anhydrotrypsin-Sepharose 4B and ion-exchange chromatography ResourceTM Q. RcTI has a apparent molecular mass of 14 kDa in the absence or presence of a reducing agent, pI 5.2 and isnât a glycoprotein. The NH2-terminal sequence of purified RcTI showed 83% similarity with sulfur-rich storage protein (2S albumin) R. communis and 48% with napin-like (2S albumin) R. communis. The RcTI was stable after heat treatment at 98 ÂC for two hours. The inhibition on trypsin was stable in the acid pH range. However, there was a loss of approximately 20% inhibitory effect on alkaline pH (pH 8-11). The RcTI (13 Âg) was unable to inhibit the germination of spores of the fungus Fusarium oxysporum, Rhizoctonia solani and Fusarium solani. However, it was able to inhibit the germination of spores of Colletotrichum gloeosporioides. RcTI 100 Âg was not effective in the inhibition of vegetative growth of the fungi mentioned above. This inhibitor was effective against proteases from the midgut of Aedes aegypti with 91% inhibition. The results of the present study indicate that RcTI has biotechnological potential, can be used as an alternative to combat the important plant pathogenic fungus C. gloeosporioides and larvae of A. aegypti.

Traditionally, funeral homes have been family-owned small businesses which pride themselves on their personal, caring service. Many are located in historic houses worthy of preservation. In the past few decades, however, many family-owned funeral homes have been bought out by large, national corporations in search of big profits. The future of oldhouse funeral homes is uncertain. An inventory of funeral homes was conducted in a ninecounty area of east-central Indiana, including Blackford, Delaware, Grant, Hancock, Henry, Jay, Madison, Randolph, and Wayne Counties. Findings show that a majority (59%) of the funeral homes in the nine-county area of east-central Indiana inventoried are located in former residences built before 1950, with varying degree of modification. While some have had minimal or moderate alterations, a large percentage (54%) of these have been extensively altered over the years. Case studies of four pre-1950 funeral homes in the inventory area revealed that alterations, many of which reflect the needs of the funeral business, have affected not only their integrity but also their ratings in the Indiana Historic Sites and Structures Inventory. Recommendations include greater communication between the funeral industry and preservationists, and the establishment of guidelines for sensitive additions that are addressed specifically to the needs of the funeral industry.
Department of Architecture

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The cave environments are oligotrophic and have specific characteristics that determine the native microbiota. Few studies have investigated the composition and function of soil microbiota of subterranean environments. This study aimed to expand the knowledge on the cave microbiota, discuss the functional role of the microbiota in these environments, isolate and select fungal strains with biotechnological potential for the degradation of cellulose. Soil epigean environment and two poins inside the cave Catão, São Desiderio BA, were sampled. The amount of total organic carbon, carbon and nitrogen in the microbial biomass and the microbial density was evaluated in the soil. All strains were evaluated for the activities of endoglucanase, β-glucosidase and total cellulase by submerged fermentation. Of these three strains of Aspergillus strains, one strain standard, and a Penicillium the process of solid state fermentation using wheat bran as substrate were selected. We observed a significant difference between epigeal and subterranean environment in relation to physical, chemical and biological soil parameters evaluated. We obtained 20 isolates belonging of the genus Aspergillus (SDC1.1, SDC 1.2, SDC 1.4, SDC 1.6, SDC 2.4, SDC 2.6, SDC 2.8, SDC 2.10, SDC 2.11, SDC 2.12), Penicillium (SDC 1.3, SDC 1.7, SDC 2.2, SDC 2.7, SDC2.9, SDC 2.13), Trichoderma (SDC 2.3), Scopulariopses (SDC 2.1) e Purpureocillium (SDC 2.5). Of all isolates showed 90% cellulolytic activity. During the submerged fermentation endoglucanase activity showed maximum values of 6.39 IU/mL in a strain of Aspergillus sp6 and maximum total cellulase activity was 1.7 IU/ml for Aspergillus sp8 strain. The strain Aspergillus sp6 showed maximum β- glucosidase activity of 13.67 IU/mL. Compared to solid-state fermentation, the Aspergillus sp6 and Aspergillus sp8 strains stood out compared to mutant strain Aspergillus niger 3T5B8, with values of endoglucanase and total cellulase activity higher whole with a lower protein content in their enzymatic extracts. From these resulados, we conclude that the cave environment presents singularities compared to epigean environment which probably allows you to select naturally microorganisms that use alternative sources for energy and nutrients.
Os ambientes cavernícolas são oligotróficos e possuem características específicas que determinam a microbiota local. Poucos estudos têm investigado a composição e funcionalidade da microbiota edáfica de ambientes subterrâneos. Este trabalho teve como objetivo ampliar o conhecimento sobre a microbiota cavernícola, discutir o papel funcional da microbiota nesses ambientes, isolar e selecionar linhagens fúngicas com potencial biotecnológico para a degradação de celulose. Foram coletadas amostras de solo do meio epígeo e de dois pontos no interior da caverna do Catão, São Desidério-BA. Nas amostras foram avaliadas as quantidades de carbono orgânico total, a densidade microbiana, carbono e nitrogênio da biomassa microbiana, bem como isoladas linhagens de fungos filamentosos. Todas as linhagens foram avaliadas quanto as atividades de endoglucanase, celulase total e β-glucosidases por meio de fermentação submersa. Dessas linhagens foram selecioandas três linhagens de Aspergillus, sendo uma linhagem padrão, e uma linhagem de Penicillium para o processo de fermentação em estado sólido, utilizando farelo de trigo como substrato. Observamos uma diferença significativa entre o meio epígeo e subterrâneo em relação aos parâmetros físicos, químicos e biológicos do solo avaliados. Obtivemos 20 isolados pertencentes aos gêneros Aspergillus (SDC1.1, SDC 1.2, SDC 1.4, SDC 1.6, SDC 2.4, SDC 2.6, SDC 2.8, SDC 2.10, SDC 2.11, SDC 2.12), Penicillium (SDC 1.3, SDC 1.7, SDC 2.2, SDC 2.7, SDC2.9, SDC 2.13), Trichoderma (SDC 2.3), Scopulariopses (SDC 2.1) e Purpureocillium (SDC 2.5). De todos os isolados 90% apresentaram atividade celulolítica. Durante a fermentação submersa a atividade endoglucanase apresentou valores máximos de 6,39 IU/mL em uma linhagem de Aspergillus sp6 e a atividade máxima de celulase total foi de 1,7 IU/mL para uma linhagem Aspergillus sp8. A linhagem Aspergillus sp6 apresentou atividade β-glucosidase máxima de 13,67 IU/mL. Em relação a fermentação em estado sólido, as linhagens Aspergillus sp6 e Aspergillus sp8 se destacaram em relação a linhagem padrão Aspergillus niger 3T5B8, com valores de atividade endoglucanase e celulase total superiores e com um menor conteúdo proteico em seus extratos enzimáticos. A partir desses resultados, concluímos que o ambiente cavernícola apresenta singularidades em relação ao meio epígeo e que provavelmente permite selecionar naturalmente microrganismos que utilizam fontes alternativas para obtenção de energia e nutrientes.

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Alternative methods that use a combination of nematophagous fungi associated with organic mineral fertilizer have been little used and shall be studied further, since they present potential for nematode control in addition to supplement fertility and improve soil biota. Moreover, there is a growing demand of the society, which has become more demanding on the use of management practices more environmentally appropriate. Thus, the objective of this work was to test the compatibility of an organic mineral fertilizer based on castor bean cake to Pochonia chlamydosporia for the control of Meloidogyne javanica in lettuce. Two tests were performed to study the viability of P. chlamydosporia in Rizomax (which contains fungus P. chlamydosporia), and UFV-TM100 (organic mineral fertilizer) with or without addition of water. One hundred grams of UFV-TM100 and 1 g of Rizomax were placed in a 3-L plastic bag with no addition of water. The bag was sealed and vigorously shaken until homogenization of the mixture and kept in a closed room. The determination of the CFU of P. chladoutmydosporia was carried out on days 0, 15, 30 and 45 both in pure Rizomax and Rizomax + UFV-TM100. The same methodology was used in the second test, but with the addition of water in the mixture and determination of CFU of P. chlamydosporia was carried out on days 0, 7 and 14. The number of CFU of P. chlamydosporia did not increase within the addition of water in Rizomax in the assessed period; however, the number of CFU was reduced by 27, 54 and 100% in the Rizomax + UFV-TM100 on days 0, 7 and 14, respectively, for the product Rizomax. The number of CFU of P. chlamydosporia in Rizomax did not decrease in the test with no addition of water, but the number of P. chlamydosporia in Rizomax + UFV-TM100 was reduced by 30% on day 45 in comparison to day 0, 15 and 30. A third test was conducted to study the colonization by P. chlamydosporia in a substrate in the presence of UFV-TM100. One litter of substrate was poured in a 5-L plastic bag containing 18 g of UFV- TM100 and 0.18 g of Rizomax. The bag was sealed and shaken vigorously for two minutes to homogenize the mixture, which was distributed into three 200 mL plastic glasses. A second treatment, with no UFV-TM100, was used to evaluate the development of P. chlamydosporia in the absence of UFV-TM100. The substrate in each of the plastic glass was kept moist and samples were taken on days 0 and 14 to determine the number of CFU of P. chlamydosporia. The CFU number of P. chlamydosporia did not differ between Rizomax and Rizomax + UFV-TM treatments in the evaluation periods but the CFU number for P. chlamydosporia was 61% higher on day 14 than on day 0. A forth test was carried out to study the effect of the doses of Rizomax on the growth of lettuce and on the development of M. javanica. Plastic pots containing 2 L of substrate received 36 g of UFV - TM100, and after 15 days, lettuce seedlings at 21 days of age were transplanted. Rizomax was added together with TM100-UFV at doses of 0, 0.36 and 0.45 g/kg substrate, which corresponded to 0, 1, and 1.25% Rizomax . The substrate in each pot was infested with 5,000 eggs of M. javanica on same day of incorporation of UFV- TM100 and Rizomax incorporation. The same doses of Rizomax , but in the absence of UFV - TM100 were tested as control treatment. Sixty days after transplanting, the diameter of the head of lettuce (cm) , the fresh matter weight , the dry matter weight, the fresh root system weight, the number of galls and eggs of M. javanica per gram of root system were evaluated. The diameter of the head of the lettuce, the weight of the shooting fresh matter and the weight of shooting dry matter increased by 41, 44 and 29% , respectively, in the presence of UFV-TM100. The number of gals and eggs per gram of root system was reduced up to 99% when UFV-TM100 + Rizomax were applied at the dose of 1.25%. The application of Rizomax in the absence of UFV-TM100pla root system and the numb also reduced the number of galls per root system and the number of galls per gram of root system, with reductions of up to 50 and 60%, respectively. Therefore, the use of organic mineral fertilizer with the addition of a biological control agent is an efficient alternative to the control of nematodes, in addition to improve plant development.
Métodos alternativos, utilizando a combinação de fungos nematófagos associados a um adubo organomineral têm sido pouco utilizados e devem ser mais estudados, visto que apresentam potencial de controle dos nematoides, além de suplementar a fertilidade e favorecer a biota do solo. Alia-se à isso a demanda crescente da sociedade, que está mais exigente quanto à utilização de práticas de manejo mais ecologicamente adequadas. Diante disto, o objetivo deste trabalho foi testar a compatibilidade de um fertilizante organomineral à base de torta de mamona com o fungo Pochonia chlamydosporia, visando o controle de Meloidogyne javanica em alface. Dois testes foram realizados para estudar a viabilidade de P. chlamydosporia nos produtos Rizomax (contém o fungo P. chlamydosporia), e UFV-TM100 (fertilizante organomineral), com e sem adição de água. Foram colocados em saco plástico de 3 L, 100 g de UFV-TM100 e 1 g de Rizomax, sem adição de água. O saco foi fechado e agitado vigorosamente até homogeneização da mistura e mantido em uma sala fechada. A determinação do número de UFC de P. chlamydosporia foi realizada aos 0, 15, 30 e 45 dias, tanto no produto Rizomax puro como na mistura Rizomax + UFV-TM100. Para o segundo teste foi utilizada a mesma metodologia, porém com adição de água e a determinação do número de UFC de P. chlamydosporia foi realizada aos 0, 7, e 14 dias. No teste com adição de água não houve decréscimo do número de UFC de P. chlamydosporia no produto Rizomax no período estudado, porém o número de UFC de P. chlamydosporia na mistura Rizomax + UFV-TM100 foi reduzido em de 27, 54 e 100%, respectivamente, aos 0, 7 e 14 dias em relação ao produto Rizomax. No teste sem adição de água não houve decréscimo do número de UFC de P. chlamydosporia no produto Rizomax no período estudado, porém o número de UFC de P. chlamydosporia na mistura Rizomax + UFV-TM100 foi reduzido em 30% aos 45 dias em relação aos 0, 15 e 30 dias. Um terceiro teste foi realizado com objetivo de estudar a colonização de um substrato por P. chlamydosporia na presença do UFV-TM100. Um litro de substrato foi colocado em um saco plástico de 5 L contendo 18 g de UFV-TM100 e 0,18 g de Rizomax. O saco foi fechado e agitado vigorosamente por dois minutos para homogeneizar a mistura, a qual foi distribuída em três copos plásticos de 200 mL de capacidade cada. Um segundo tratamento, sem UFV-TM100, foi utilizado para avaliar o desenvolvimento de P. chlamydosporia na ausência do UFV-TM100. O substrato em cada copo plástico foi mantido úmido e amostras foram retiradas aos 0 e 14 dias para determinar o número de UFC de P. chlamydosporia. Não houve diferença no número de UFC de P. chlamydosporia entre os tratamento Rizomax e Rizomax + UFV-TM no períodos de avaliação, porém o número de UFC de P. chlamydosporia foi 61% maior aos 14 dias com relação ao 0 dia. Um quarto teste foi realizado para estudar o efeito das doses do produto Rizomax nas variáveis de crescimento da alface e no desenvolvimento de M. javanica. Vasos plásticos contendo 2 L de substrato receberam 36 g de UFV- TM100 e, após 15 dias, mudas de alface com 21 dias de idade foram transplantadas. Juntamente com o UFV-TM100, adicionou-se o produto Rizomax nas doses de 0, 0,36 e 0,45 g/2 Kg de substrato, as quais corresponderam a 0, 1 e 1,25% de Rizomax. O substrato em cada vaso foi infestado com 5000 ovos de M. javanica no mesmo dia da incorporação do UFV-TM100 e Rizomax. Como tratamento testemunha, foram testadas as mesmas doses de Rizomax, porém na ausência do UFV-TM100. Após 60 dias do transplantio, avaliou-se o diâmetro da cabeça da alface (cm), o peso da matéria fresca, o peso da matéria seca, o peso do sistema radicular fresco, o número de galhas e de ovos de M. javanica por sistema radicular e o número de galhas e de ovos de M. javanica por grama de sistema radicular. Na presença do produto UFV-TM100 houve aumento de 41, 44 e 29% no diâmetro da cabeça da alface, no peso da matéria fresca da parte aérea e no peso da matéria seca da parte aérea, respectivamente. O número de galhas e ovos pos grama de sistema radicular sofreram reduções de até 99% quando aplicado o UFV- TM100 + Rizomax na dose de 1,25%. A aplicação do Rizomax na ausência do UFV- TM100 também reduziu o número de galhas por sistema radicular e número de galhas por grama de sistema radicular, com reduções de 50 e 60%, respectivamente. Assim, na aplicação do fertilizante organomineral com a adição de um agente de controle biológico é uma alternativa eficiente de controle de nematoides, além de aumentar o desenvolvimento das plantas.

Background and aims: Pollen and fungal spore concentrations in the atmosphere of Cape Town have been monitored since 1984 in two areas of Cape Town. Volumetric spore traps were used to monitor the air spora that trigger allergic disease in susceptible individuals. A pollen count was produced for the diagnosis and treatment of patients attending respiratory clinics at the academic hospitals but the findings of the different aerobiological monitoring areas have never been compared. We considered that more than one aerobiological area should be monitored to produce a representative pollen count for the most densely populated areas of Cape Town. Methods: The pollen taxa and fungal spore genera collected from the two aerobiological zones, now named the West Coast and Valkenberg Aerobiomes were defined. Eight of the air spora were selected for detailed comparison. The relative abundance and seasons of Poaceae, or grasses, the tree pollen taxa Cupressaceae, Platanus and Quercus and the fungal spores: Alternaria, Cladosporium, Epicoccum and Pleospora were evaluated and compared in each aerobiome. Differences in the annual distribution and seasonal limits of the air spora in the two aerobiomes were found using statistical techniques. Results: Significant differences were found between the tree pollen loads in the different aerobiomes. Spring pollinating trees were the most prevalent pollen taxa in the Valkenberg Aerobiome with short flowering seasons that spanned six weeks (August-September) for Platanus and Quercus but eight weeks (July-September) for Cupressaceae. The grass season was longer (September- March) in both aerobiomes and grasses flowered earlier at the inland site. Poaceae dominated the annual pollen catch at the coastal aerobiological zone. Parietaria was the only weed taxon with significant concentrations. Pleospora showed a seasonal trend peaking in mid to late winter at the West Coast. No comparable peaks for Pleospora were seen from the Valkenberg sites. Cladosporium concentrations were low and seldom breached the significant threshold of 3,000 spores/m-3 in either of the aerobiomes. The influence of meteorological parameters on Cladosporium and temperature on Poaceae was explored. A table was designed that clarified the ranges for Cladosporium and a formula was adapted for predicting the start of the grass season. Significant decreases in the Poaceae concentrations in both aerobiomes were observed and discussed with reference to Global Warming. Conclusions: The differences in the pollen spectra and seasonality of the selected allergenic air spora indicate that both aerobiomes should be monitored concurrently for patients who live and work in these different microclimates. Pollen profiles for skin, blood and specific IgE testing panels should be reassessed to include Cupressaceae, Parietaria, Myrica, Pleospora and basidiospores. When patients are recruited for clinical drug trials, their place of residence or work should be within the realm of the aerobiome that is being monitored. Current pollen monitoring programmes should be consulted for immunisation regimes to grass and tree pollen. These findings will be applied to the diagnosis and prescription of immunotherapy in clinical practice.

Background: The epidemiological studies investigating environmental risk factors associated with asthma among children living in informal settlements are scant as are studies on the independent and co-pollutant effect of short- and long-term exposures to ambient air pollutants as well as fungal spores on asthma-associated outcomes. This study systematically investigated these factors among schoolchildren residing in informal settlements in the Western Cape province of South Africa. Methods: A cohort study of grade-4 schoolchildren (n=590) recruited from six primary schools in four informal settlements was conducted over 12 months. In addition, a panel study, investigated the children for 2 consecutive school weeks in both summer and winter. Spirometry and fractional-exhaled nitric oxide (FeNO) measurements were conducted during the school day, while the International Study on Asthma and Allergy in Children (ISAAC) standardised questionnaire was administered to the parent or guardian at the child’s home at baseline and follow-up. The presence of atopy was determined based on a positive Phadiatop test on sera. In the cohort study, annual NO2 and PM2.5 levels were computed for each child’s address using a land-use regression model. Daily PM10 levels obtained from a stationary monitor near two of the study areas were used for the panel study. Airborne pollen and fungal spore measurements were obtained directly from a stationary monitor placed in each study area. Results: The prevalence of doctor-diagnosed asthma was 3.4% and only half of them were on asthma treatment. The prevalence of wheezing in the past 12 months (12.9%), airway obstruction (17.6%) and airway inflammation (10.2%) was much higher. The presence of damp conditions, visible mould growth, passive smoking as well as paraffin-use for cooking and heating were significant indoor risk factors for asthma. The estimated annual average NO2 level of 16.6 µg/m3 was below the WHO annual exposure standards, however more than a third of children were exposed to annual PM2.5 levels above the 10 µg/m3 WHO standard and the allergic symptom threshold level of 100 spores/m3 for Alternaria spores. In the panel study, daily exposure of schoolchildren to Alternaria and Cladosporium spores independently decreased FEV1 (-27.56 ml, 95% CI: -50.60 to -4.51 ml per 10 spores/m3 increase in Alternaria; and -86.19ml, 95% CI: -131.69 to -40.70 ml per 50 spores/m3 increase in Cladosporium respectively) from lag day-0 to lag day-5, especially in the winter monitoring period. In the cohort study, an interquartile range increase of 14.2 µg/m3 in annual NO2 was associated with an risk of new onset ocular-nasal symptoms (adjusted odds ratio – aOR: 1.63, 95% CI: 1.01 – 2.60), wheezing (aOR: 3.57, 95% CI: 1.18 – 10.92), more than two or more asthma symptom score (aOR: 1.71, 95% CI: 1.02 – 2.86), and airway inflammation defined as FeNO > 35ppb (aOR: 3.10, 95% CI: 1.10 – 8.71), independent of PM2.5 exposures. In addition, an interquartile increase of 83.1 spores/m3 in 24-hour annual Alternaria spore levels was associated with an increased risk of airway inflammation incidence and having a ≥ 10% increase in FeNO at follow-up both in the single-pollutant model and two-pollutant model. Conclusion: This study demonstrated a large proportion of undiagnosed and untreated asthma in schoolchildren living in informal settlements, with both indoor and outdoor mould exposures playing an important role in addition to ambient chemical pollutants. The incidence of new onset asthma symptoms and airway inflammation associated with NO2 at levels below the WHO Air Quality Standards raises the issue of the adequacy of these standards in protecting respiratory health. Raised long-term levels of airborne Alternaria spores contributing to increased airway inflammation is likely to form the basis for the increased risk of acute symptoms and airway effects observed in association with exposure peaks.

Thesis (PhDAgric)--University of Stellenbosch, 2000.
ENGLISH ABSTRACT: Barley leaf scald, caused by Rhynchosporium secalis, is the most important disease of barley (Hordeum vulgare) in the Western Cape province of South Africa. The disease was first reported from South Africa in 1937. The present study is the first attempt to characterise the South African R. secalis population. Topics such as pathogenesisrelated proteins, virulence spectra, variability of pathotypes, sources of variation, host resistance, breeding strategies, molecular characterisation and fungicide sensitivity are summarised in Part 1 of this dissertation. In succeeding Parts the focus is on the characteristics of the local R. secalis population regarding virulence spectrum, DNA polymorphisms, in vitro as well as in vivo fungicide sensitivity. These aspects are treated as separate entities, leading to some duplication which is unavoidable. In Part 2 the virulence spectra of 50 R. secalis isolates from a population in the. Western Cape province were determined. Twenty-one races were detected using 17 differential barley cultivars. The two most prevalent races, namely races 4 and 7 had three and four virulence genes respectively. Both race 4 and 7 were virulent on the most susceptible cultivars, namely West China, Steudelli, C.I.8618 and C.I.2226. Considering the resistance genes reported for cultivars Atlas 46, Turk, and C.I.3515 which showed no susceptible cultivar-pathogen interaction, it would appear that the Rh- Rh3-Rh4 complex is primarily involved in conferring resistance to the local R. secalis isolates. A total of 20 races (47 isolates) characterised in Part 2 were selected for further characterisation by means of DNA fingerprinting. In Part 3 an anonymous multilocus DNA probe was used to characterise the genotypic structure of these isolates by means of RFLP analysis. No correlation between any particular fingerprint pattern, race, district, field or lesion was found. The two most prevalent races, 4 and 7, did not share the same genotypes, even when isolated from the same field or lesion. The genotypic diversity of the isolates studied was 46.5% of the theoretical maximum diversity. The high level of genotypic variation observed in the South African R. secalis population resembled the genotypic diversity observed in other cereal pathogens with known sexual structures. Although no teleomorph has yet been observed, these data suggest that sexual recombination may operate within the local population of R. secalis. In South Africa barley scald is primarily controlled by means of fungicides. The continued use of fungicides on cereal crops results in the build-up of fungicide resistance in the population, which could lower the efficacy of these compounds. These aspects were investigated in Part 4, where isolates (collected during 1993 to 1995) were evaluated in vitro for sensitivity to triadimenol, tebuconazole, flusilazole and propiconazole. The sensitivity fluctuated but in 1995 isolates were significantly less sensitive towards triadimenol than in the previous two years. In a second experiment, isolates collected from two fields with a 5-6 year-history of triadimenol seed treatments and tebuconazole applications, were evaluated for their fungicide sensitivity. A significant positive correlation was observed between tebuconazole and triadimenol sensitivity among,R. secalis populations from these fields. However, such a correlation was not found within the R. secalis population collected during 1993-1995 where shorter crop rotation patterns and a range of fungicides was applied. In a third experiment, the fungicide sensitivity of local R. secalis isolates was evaluated towards two new triazole fungicides, namely bromuconazole and triticonazole. Correlation coefficients observed between these new triazoles and those previously applied in South Africa were not significantly positive. The lack of significant cross-resistance has important practical implications regarding the management of fungicide resistance. In Part 5, isolates with different minimum inhibitory concentration (MIC) towards tebuconazole in vitro (1, 3 and 10 ug/ml) were compared in vivo. The aim of this study was to determine how MIC values would influence virulence (leaf area affected) and sporulation. Results indicated that all isolates were equally fit to induce lesions and sporulate in the absence of tebuconazole. Thus no fitness cost was associated with the degree of tebuconazole sensitivity in the present study. All R. secalis isolates were able to induce lesions on tebuconazole treated leaves, but differed significantly with respect to the percentage leaf area affected. Isolates, least sensitive (MIC = 10 ug/rnl) towards tebuconazole were more adapted on tebuconazole treated leaves, being able to repeatedly cause larger lesions than sensitive R. secalis isolates (MIC = 1 ug/rnl), Sporulation was not significantly different between isolates on lesions of untreated or tebuconazole treated leaves. Larger leaf areas affected and adequate sporulation suggest that a less sensitive population would result in more disease in tebuconazole treated fields. In conclusion, this study revealed the variability associated with the South African R. secalis population regarding virulence spectrum and genotypic structure. The data in this study suggest that it is likely that the local population will easily adapt to newly introduced, single gene resistance. For more durable resistance, higher levels of quantitative resistance should be introduced. This type of resistance is, however, more difficult to identify and incorporate than single gene resistance. Consequently, barley scald control will remain dependent on the efficacy of fungicide applications. Furthermore, the lack of cross-resistance and low frequency of resistant isolates indicates a low risk for the development of fungicide resistance in the local R. secalis population. Other factors such as current crop rotation practices and the range of fungicides being ~pplied also contribute to this low risk level. However, the status of these factors can change over time. The in vivo tebuconazole sensitivity study has indicated that a resistant field population of R. secalis may be able to build-up. It is, therefore, necessary to monitor the fungicide sensitivity of R. secalis isolates at timely intervals with view to successful barley cultivation in the future.
AFRIKAANSE OPSOMMING: Blaarvlek op gars (Hordeum vulgare), veroorsaak deur Rhynchosporium secalis, is die belangrikste siekte van gars in die Wes-Kaap provinsie van Suid-Afrika. Die voorkoms van R. secalis op gars is in Suid-Afrika vir die eertse keer in 1937 gerapporteer. Hierdie studie is die eerste poging tot karakterisering van die plaaslike R. secalis-populasie. Aspekte soos proteïene betrokke by patogenese, virulensiespektra, variabiliteit van patotipes, bronne van variasie, gasheerweerstand, teeltprogramme, molekulêre karakterisering en swamdodersensitiwiteit word in Deel I van die tesis opgesom. In die daaropvolgende gedeelte is die fokus op die karakterisering van die R. secalis-populasie en behels DNA karakterisering, virulensiespektrum, en swamdodersensitiwiteit in vitro asook in vivo. .. In Deel 2 is die virulensiespektra van 50 R. secalis isolate van 'n populasie in die. Wes-Kaap geëvalueer teenoor 17 differensiëel weerstandbiedende gars kultivars en hieruit is 21 rasse geïdentifiseer. Die twee mees algemene rasse (rasse 4 en 7), met onderskeidelik drie en vier virulensie gene, het virulent vertoon teenoor die mees vatbare kultivars soos West China, Steudelli, C.I.8618 en C.I.2226. Geen vatbare kultivar-patogeen interaksies is met kultivars Atlas 46, Turk en C.I.3515, wat al drie die Rh-Rh3-Rh4 kompleks dra, gevind nie. Dit wil dus voorkom asof hierdie genekompleks effektiewe gasheerweerstand teen die plaaslike R. secalis isolate kan bied. 'n Totaal van 20 rasse (47 isolate), gekarakteriseer in Deel 2, is geselekteer vir verdere karakterisering met behulp van DNA bandpatrone. In Deel 3 is 'n anonieme multilokus DNA peiler gebruik om deur middel van RFLP analise die genotipiese struktuur van hierdie R. secalis-isolate te bepaal. Geen assosiasie is gevind tussen DNA bandpatroon en ras, distrik, garsland of letsel nie. Die twee rasse (4 en 7) wat mees algemeen voorkom, het nie dieselfde bandpatroon vertoon nie, ook nie dié afkomstig vanuit dieselfde garsland of letsel nie. Die genotipiese diversiteit van isolate was 46.5% van die teoretiese maksimum diversiteit. Die hoë vlak van variasie waargeneem in die R. secalis populasie is soortgelyk aan variasie waargeneem in ander graanpatogene wat oor 'n geslagtelike stadium in die lewenssiklus beskik. Alhoewel geen geslagtelike stadium tot dusver geidentifiseer is nie, dui die vlak van variasie daarop dat geslagtelike rekombinasie moontlik wel plaasvind binne die plaaslike R. secalis populasie. In Suid-Afrika word blaarvlek op gars primêr deur swamdoders beheer. Die toenemende gebruik van swamdoders op graangewasse veroorsaak moontlik 'n opbou van swamdoderweerstand in die populasie. Dit kan die effektiwiteit van swamdoders verlaag. Hierdie veronderstelling is in Deel 4 ondersoek, waar die sensitiwiteit van isolate in vitro teenoor triadimenol, tebukonasool, flusilasool en propikonasool geëvalueer is. Die triasooi sensitiwiteit van R. secalis isolate wat gedurende die 1993- 1995 seisoen versamel is het gewissel en slegs vir triadimenol was daar 'n tendens na meer weerstandbiedenheid. 'n Swamdoder-evaluasie is in 'n aparte eksperiment op isolate gedoen wat versamel is vanaf twee garslande met 'n 5-6 jaar geskiedenis van triadimenol saadbehandelings en tebukonasool bespuitings. 'n Betekenisvolle positiewe korrelasie is waaJ~geneem tussen tebukonasool en triadimenol sensitiwiteit in R. secalis isolate afkomstig vanaf hierdie twee garslande. 'n Soortgelyke korrelasie is egter nie gevind in die populasie wat gedurende die 1993-1995 seisoene versamel IS me. Laasgenoemde kan moontlik toegeskryf word aan korter wisselboupatrone en die toediening van 'n verskeidenheid van swamdoders. In 'n derde eksperiment is die sensitiwiteit van plaaslike R. secalis isolate teenoor twee nuwe triasole, naamlik bromukonasool en tritikonasool getoets. Die korrelasie waargeneem tussen die twee nuwe triasole en triasooi swamdoders reeds voorheen in gebruik in die Wes-Kaap was me betekenisvol positief me. Die gebrek aan betekenisvolle kruisweerstandbiedendheid het belangrike praktiese implikasies vir die bestuur van swamdoder -weerstandbiedendheid. In Deel 5 is isolate met wisselende minimum inhiberende konsentrasies (MIKs) teenoor tebukonasool in vitro (1, 3 en 10 ug/ml) en in vivo vergelyk. Die doel van hierdie studie was om te bepaal hoe wisselende MIK-waardes virulensie (blaaroppervlakte geïnfekteer) en sporulasie sal beïnvloed. Resultate dui daarop dat alle R. secalis isolate in hierdie studie ewe fiks was om, in die afwesigheid van tebukonasool, letsels te induseer en te sporuleer. Die bevinding is dat die verlies in fiksheid nie geassosieer is met die mate van tebukonasool weerstand nie. Alle R. secalis isolate het die vermoë gehad om letsels op tebukonasool-behandelde blare te veroorsaak maar het betekenisvol verskil ten opsigte van die blaaroppervlakte geaffekteer. Isolate wat minder sensitief (MIK = 10 ug/rnl) teenoor tebukonasool in vitro is, het meer aangepastheid op tebukonasool-behandelde blare getoon. Gevolglik het hierdie isolate herhaaldelik meer letsels veroorsaak as sensitiewe isolate (MIK = 1 ug/ml), Sporulasie het nie betekenisvol verskil tussen isolate vanaf letsels op ondehandelde of tebukonsoolbehandelde blare nie. Hierdie resultate dui egter daarop dat 'n minder sensitiewe populasie tot meer siektevoorkoms in tebukonasool-bespuite lande kan lei. Die studie het die veranderlike karakter van die Suid-Afrikaanse R. secalispopulasie aangaande virulensiespektrum en genotipiese struktuur blootgelê. Dit is dus baie moontlik dat die R. secalis-populasie maklik sal aanpas by teelmateriaal met nuwe enkelgeen-weerstand. Vir volgehoue gasheerweerstand is dit egter nodig dat hoër vlakke van kwantitatiewe weerstand ingeteel moet word. In die praktyk is hierdie tipe weerstand egter baie moeiliker om te identifiseer en by nuwe teelmateriaal in te sluit as in die geval van enkelgeen-weerstand, Dit bring mee dat blaarvlekbeheer afhanklik bly van swamdodertoedienings as beheermaatreël. Die resultate van hierdie studie dui daarop dat daar tans 'n lae risiko vir die ontwikkeling van swamdoderweerstand in die plaaslike populasie is, as gevolg van die afwesigheid van kruisweerstandbiedendheid en die lae voorkoms van weerstandbiediende isolate. Ander faktore soos die wisselboustelsels wat toegepas word en die verskeidenheid van swamdoders toegedien dra ook daartoe by. Ten spyte hiervan kan die status van hierdie faktore egter oor tyd verander. Die in vivo tebukonasool studie het daarop gedui dat 'n weerstandbiedende veldpopulasie van R. secalis die potensiaal het om te vermeerder. Gevolglik is die tydige monitering van swamdodersenisitiwiteit van R. secalis isolate noodsaaklik om 'n volhoubare garsproduksie te verseker.

Thesis (MScAgric)--University of Stellenbosch, 2004.
ENGLISH ABSTRACT: Botrytis cinerea Pers.: Fr., a pathogen of grapevine (Vitis vinifera L.), moves mainly through conidia by air currents in vineyards which are deposited intermittently on the surfaces of leaves, inflorescences and bunches. Little is known about the relationship between the inoculum dosage in air and incidence of Botrytis bunch rot, and how the relationship is influenced by environmental and host factors. To better understand this relationship, information is needed on the period over which conidia have accumulated, the time they are able to survive and remain infectious, time of symptom expression in relation to conidium arrival at the infection court and host surface wetness. The aims of this study were (i) to estimate the amount of viable B. cinerea occurring in air in vineyards, and at different positions on leaves, inflorescences and bunches of grape at different phenological stages, (ii) to determine the relationships between the number of B. cinerea colonies recorded on spore traps placed in the bunch zone of vines and the incidence of B. cinerea recorded from the different tissues, and (iii) to compare the efficacy of fenhexamid on leaves and inflorescences carrying natural B. cinerea inoculum with those inoculated with dry, airborne conidia. Different techniques were used to detect viable Botrytis cinerea in air currents and on plant material obtained from table (cultivars Dauphine and Waltham Cross in Paarl- and Worcester-district) and wine grape (cultivars Chardonnay, Sauvignon Blanc and Merlot in Stellenbosch- and Malmesbury district) vineyards in the Western Cape province during 2001-02 and 2002-03. For four consecutive days during prebloorn, bloom, pea-size, bunch closure, veraison and harvest, sets of Petri dishes with freshly prepared Kerssies' B. cinerea selective medium (spore traps) were left overnight in the bunch zone of vines. Plant material was collected from the vines on the fourth day. Leaves, infloresence and bunches were treated with paraquat to terminate host resistance and to promote the development of the pathogen on the tissues. The B. cinerea inoculum dosage in air, and the incidence at which the pathogen was detected at various positions on leaves and in bunches normally differed between vineyards. However, the various tests revealed that the pathogen generally occurred in a consistent pattern in air in the bunch zone of vines, on leaves and in bunches from all vineyards. The inoculum dosage in air in the bunch zone of the vine was generally highest during prebioom or during bloom, it decreased at pea size and mostly remained at a very low level at the later growth stages. The estimations of viable B. cinerea residing naturally on leaves and in bunches, showed that their amounts depicted levels occurring in air in the bunch zone of the vine. Necrotic leaves occurring early season in vineyards were identified as an important source of secondary inoculum for dispersal to the developing bunches. Latent infections at the various positions in bunches were few at véraison and harvest. However, due to the necrotrophic ability of the pathogen, extensive berry rot (due to berry-to-berry contact) and thus severe bunch rot developed from a single berry that become symptomatic at the base of the pedicel/berry attachment zone. The B. cinerea occupation pattern explains why Botrytis bunch rot develops mostly from the inner bunch and why disease management strategies should concentrate on the bloom to pre-bunch closure stage and on inhibiting B. cinerea development in the inner bunch during the early part of the season. Thus, to effectively reduce B. cinerea in grapevine, preventative applications are recommended to reduce two primary infection events: (a) between budding and pre-bloom to counteract primary leaf infection; (b) during late bloom or early pea size stage, to reduce the amount of the pathogen on leaves and infloresences and to prevent colonisation of floral debris. A third spray can be applied at bunch closure to reduce the amount of B. cinerea at various positions of the inner bunch, especially for cultivars with tight bunches. The efficacy of fenhexamid on leaves and inflorescences carrying natural B. cinerea inoculum was compared with those inoculated with dry, airborne conidia. Shoots were obtained during late bloom from a vineyard (wine grape cultivar Merlot) in the Stellenbosch region. The shoots were divided into two main groups. One group of shoots was left uninoculated, the other shoots were inoculated by dusting with dry B. cinerea conidia in a settling tower. Before inoculation, equal numbers of shoots in each main group was sprayed with fenhexamid, or left unsprayed. Following inoculation and incubation, shoots of each treatment were divided in two equal groups. The one lot of shoots were rinsed in water. The other lot of shoots were immersed in paraquat solution to terminate host resistance and to promote the development of the pathogen from the tissues. For both uninoculated and inoculated shoots, irrespective of fungicide treatment, leaves remained asymptomatic at both the blade and petiole position for the water rinse treatment. No symptom of B. cinerea decay developed at any of the positions on leaves from shoots sprayed with fenhexamid. Spraying of shoots with fenhexamid completely suppressed B. cinerea infection and symptom expression on both uninoculated and inoculated inflorescens. For inoculated shoots, B. cinerea developed from approximately 50% of the laterals in the water rinse treatment. However, inflorescences rinsed in water remained asymptomatic. The laboratory studies showed that fungicides, if applied properly to shoots and bunches under controlled conditions, effectively reduced the amount of B. cinerea at the various positions on leaves and inflorescence, and prevented infection and symptom expression at bloom. However, these goals are not achieved in vineyards where the fungicides are applied by conventional spraying methods. Therefore, more work is needed to evaluate fungicide application techniques by conventional spraying methods for proper fungicide coverage, and the reduction of B. cinerea in bunches.
AFRIKAANSE OPSOMMING: Botrytis cinerea Pers.: Fr., 'n patogeen van druiwe (Vilis vinifera L.), beweeg hoofsaaklik deur middel van konidia in lugstrome deur die wingerd, en word dan afwisselend op die oppervlakte van die blare, bloeiwyses en trosse gedeponeer. Daar is nog min bekend oor die verhouding tussen die hoeveelheid inokulum in die lug en die voorkoms van Botrytis op die trosse, en hoe die verhouding deur omgewings- en gasheerfaktore beïnvloed word. Ten einde hierdie interaksie beter te verstaan, word inligting benodig oor die tydperk waarin die konidia akkumuleer, die tyd wat hulle oorleef en virulent bly, en die tyd van simptoom-uitdrukking in verhouding tot die verspreiding van die konidia by die infeksie-setel en benatbaarheid van die gasheer-oppervlakte. Die doel van hierdie studie was (i) om die hoeveelheid lewensvatbare B. cinerea wat in die lug voorkom, asook by verskeie posisies op blare, bloeiwyses en trosse by verskillende fenologiese stadiums te kwantifiseer, (ii) om die verhouding tussen die aantal aangetekende B. cinerea kolonies op spoorvangers wat in die trossone van die wingerd geplaas is, en die voorkoms van B. cinerea, aangeteken van verskeie weefsels, te bepaal, en (iii) om die effektiwiteit van fenhexamid op blare en bloeiwyses wat natuurlike B. cinerea inokulum dra, te vergelyk met dié wat met droë, luggedraagde konidia geïnokuleer is. Verskillende tegnieke is gebruik om lewensvatbare Botrytis cinerea in lugstrome en op plantmateriaal van tafeldruiwe (kultivars Dauphine en Waltham Cross In Paarl- en Worcester-distrik) en wyndruiwe (kultivar Chardonnay, Sauvignon Blanc en Merlot in Stellenbosch- en Malmesbury distrik) in wingerde van die Wes-Kaap provinsie gedurende 2001-02 en 2002-03 te kwantifiseer. Petri bakkies met vars voorbereide Kerssies medium, selektief vir B. cinerea (spoorvangers), is vir vier agtereenvolgende dae gedurende vóórblom, blom, ertjiekorrel, trostoemaak, kleurbreek en oes, oornag in die trossone van wingerdstokke in betrokke wingerde, gelaat. Plantmateriaal is op die vierde dag versamel. Blare, bloeiwyses en trosse is met paraquat behandel ten einde die gasheerweerstand af te breek en ontwikkeling van die patogeen op die weefsel te bevorder. B. cinerea inokulum in die lug, en die frekwensie waarby die patogeen op verskeie posisies op blare en in die trosse voorgekom het, het normaalweg tussen wingerde verskil. Die verskeie toetse het getoon dat die patogeen normaalweg in 'n vaste patroon in die lug en die trossones van wingerde, asook op blare en in trosse van alle wingerde voorkom. Die inokulumkonsentrasie in die lug in die trossones van wingerdstokke was normaalweg die hoogste gedurende vóórblom of gedurende blom. Die inokulumdruk het by ertjiekorrel verminder en meestal by 'n 'n baie lae vlak tydens die latere groeistadia gebly. Die bepaling van lewensvatbare B. cinerea wat natuurlik op blare en in trosse gedeponeer is, het getoon dat hul hoeveelhede ooreenstem met vlakke wat in die lug in die trossone van die wingerd voorkom. Nekrotiese blare vroeg in die seisoen is 'n belangrike bron van sekondêre inokulum en speel dus 'n belangrike rol by die verspreiding van Botrytis tussen die ontwikkelende trosse. Latente infeksies by die verskeie posisies in trosse was laag by kleurbreek en oes. Weens die saprofitiese vermoëns van die patogeen, kan uitgebreide korrelvrot (a.g.v. korrel-tot-korrel kontak) en dus ernstige trosvrot, ontwikkel. 'n Enkele korrel kan by die basis van die pedisel/korrel vashegtingsone simptomaties raak, en vandaar na aangrensende korrels versprei. Die B. cinerea kolonisasiepatroon verduidelik waarom Botrytis trosvrot meestal vanaf die binneste tros ontwikkel en waarom siektebeheerstrategieë op die vóórblom- tot blomstadium gekonsentreer moet word, en op die inhibering van B. cinerea ontwikkeling in die binneste tros gedurende die vroeë stadia van die seisoen. Dus, om B. cinerea effektief tydens die twee primêre infeksie stadiums in wingerde te verminder, kan voorkomende toedienings aanbeveel word: (a) tussen knopvorming en vóórblom om primêre blaarinfeksie te verhoed; (b) gedurende láátblom en vroeë ertjiekorrel om die hoeveelheid inokulum op die blare en bloeiwyses te verminder, en die kolonisasie van blomdebris te voorkom. 'n Derde toediening kan tydens trostoemaak aangewend word om B. cinerea by verskeie posisies in die binneste tros te verminder, veral by kultivars met digte trosse. Die effektiwitiet van fenhexamid op blare en bloeiwyses waarop natuurlike B. cinerea inokulum voorkom is vergelyk met dié wat met droë, luggedraagde konidia geïnokuleer is. Lote is vanaf 'n wingerd (wyndruif kultivar Merlot) in die Stellenbosch distrik tydens láátblom verkry en in twee hoofgroepe verdeel. Die een groep lote is geïnokuleer deur droë B. cinerea konidia in 'n afsettingstoring te strooi, terwyl die ander groep nie geïnokuleer is nie. Vóór inokulasie, is die helfte van die lote in elke groep met fenhexamid behandel, terwyl die ander helfte onbehandeld gelaat is. Ná inokulasie en inkubasie, is lote van elke behandeling verder in twee eweredige groepe verdeel. Die een groep lote is in water gespoel, terwyl die ander groep lote in 'n paraquatoplossing gedompel is om die gasheerweerstand te verwyder, en die ontwikkeling van die patogeen vanuit die weefsels te bevorder. Vir die waterspoelbehandeling van beide ongeïnokuleerde en geïnokuleerde lote, ongeag van die fungisiedbehandeling, het die blare asimptomaties by beide die bladoppervlakte en blaarsteelposisie gebly. Geen simptome van B. cinerea verrotting het by emge van die blaarposisies van die lote, met fenhexamid gespuit, ontwikkel nie. Die spuit van die lote met fenhexamid het die B. cinerea infeksie en die simptoomontwikkeling op beide die ongeïnokuleerde en geïnokuleerde bloeiwyses heeltemalonderdruk. By die geïnokuleerde lote, het B. cinerea vanaf ongeveer 50% van die laterale in die waterspoelbehandeling ontwikkel, alhoewel, bloeiwyses wat in water afgespoel is, heeltemal asimptomaties gebly het. Laboratoriumstudies het getoon dat fungisiedes, indien korrek toegedien op lote en trosse onder gekontroleerde toestande, tot effektiewe vermindering van B. cinerea getalle by die verskillende posisies op blare en bloeiwyses lei, en infeksie en simptoomuitdrukking tydens blom voorkom. Weens die feit dat die doelwitte nie behaal kan word in wingerde waar die fungisiede deur konvensionele spuitmetodes toegedien is nie, moet meer studies gedoen word om fungisied toedieningstegnieke, by konvensionele spuitmetodes, VIr deeglike fungisiedbedekking en die vermindering van B. cinerea in trosse, te evalueer.

Pectus excavatum é a deformidade congênita na parede torácica anterior mais comum e seu tratamento é essencialmente cirúrgico. Tratamentos cirúrgicos e repouso prolongado no leito levam a alterações metabólicas, músculoesqueléticas, cardiovasculares e respiratórias, que podem prolongar o tempo necessário para a recuperação funcional dos pacientes no pós-operatório, aumentar o tempo de internação e os custos em saúde. A reabilitação precoce após cirurgias, com a realização de exercícios físicos e respiratórios, reduzir estes efeitos adversos, garante melhor recuperação pós-operatória, com aumento da independência dos pacientes, maior bem-estar psicológico e melhores resultados funcionais. O objetivo deste estudo foi avaliar se os pacientes submetidos a reabilitação precoce após correção cirúrgica de Pectus excavatum pela técnica de Nuss apresentam melhor capacidade funcional de exercício e função pulmonar no dia da alta hospitalar quando comparados com os pacientes submetidos a cuidados convencionais. Os Candidatos a correção cirúrgica foram alocados aleatoriamente em 2 grupos: o grupo reabilitação precoce (GRP), que iniciou a intervenção logo após a cirurgia e o grupo de cuidados convencionais (GC), que recebeu os cuidados de rotina da instituição. O teste de função pulmonar por meio da espirometria simples e a avaliação da capacidade funcional de exercício pelo teste de caminhada de seis minutos (TC6), foram realizados antes da cirurgia (pré-operatório) e no dia da alta hospitalar (pós-operatório). Quarenta pacientes foram analisados, 20 em cada grupo. No teste de função pulmonar, todos os pacientes apresentaram redução significativa dos valores de CVF, VEF1 e PFE no pós-operatório, sem diferença entre os grupos. Todos os pacientes apresentaram redução significativa da distância percorrida no TC6 pós-operatório comparada com a distância percorrida no pré-operatório (p < 0,005). Houve diferença estatisticamente significante entre o GRP e GC na avaliação pós-operatória (506.26 ± 66.54 vs 431.11 ± 75.61, p=0.02), e a diferença entre as distâncias percorridas no pré-operatório e no dia da alta hospitalar foi significativamente menor no GRP em comparação com o GC (76.57 ± 49.41 vs 166.82 ± 70.13, p < 0.001). Concluímos que os pacientes submetidos a reabilitação precoce após técnica de Nuss apresentam melhor capacidade funcional de exercício no dia da alta hospitalar em comparação com os pacientes do grupo convencional, sem diferença da função pulmonar entre os grupos
Pectus excavatum is the most common congenital chest wall deformity and its treatment is essentially surgical. Surgical treatments and postoperative bed rest lead to metabolic, musculoskeletal, cardiovascular and respiratory alterations, with the possibility of prolonging the time required for postoperative patient recovery, increasing hospitalization time and health expenditure. Early rehabilitation after surgeries, which involves physical and breathing exercises, reduces these adverse effects and ensure better postoperative recovery, with increased independence of patients, greater psychological well-being and better functional outcomes. The objective of this study was to assess whether patients undergoing early rehabilitation after Pectus excavatum repair using the Nuss procedure have better functional exercise capacity and lung function on hospital discharge day compared with patients undergoing conventional care. Patients were randomly allocated into two groups: the early rehabilitation group (ERG) which started rehabilitation after surgery and the group of conventional care (CG) which received routine care of the institution. The lung function was assessed by simple spirometry and the functional exercise capacity by the 6-minute walk test (6MWT) were performed before surgery (preoperative) and in hospital discharge day (postoperative). Forty patients were evaluated, 20 in each group. All patients presented a significant reduction in FVC, FEV1 and PEF in the postoperative lung function test and there was no statistically significant difference between groups. All patients showed significant reduction in postoperative distance walked in 6MWT compared with the preoperative distance (p < 0.005). There was statistically significant different in functional exercise capacity between the ERG and CG in the postoperative evaluation (506.26 ± 66.54 vs 431.11 ± 75.61, p=0.02) and the difference between distance walked in the preoperative and postoperative period was significantly lower in the ERC compared to the CG (76.57 ± 49.41 vs 166.82 ± 70.13, p < 0.001). We conclude that patients undergoing early rehabilitation after Nuss procedure presented better postoperative functional exercise capacity in hospital discharge day compared to patients in the conventional group, with no difference in lung function between groups

Orientador: Rosemeire Cristina Linhari Rodrigues Pietro
Resumo: O complexo celulolítico é composto principalmente por endoglucanases, exoglucanases e β-glicosidases. As endoglucanases atuam clivando a celulose nas ligações internas da fibra, enquanto as exoglucanases hidrolisam as ligações externas. A necessidade de novas pesquisas de enzimas celulolíticas capazes de degradar eficientemente resíduos industriais como bagaço de cana-de-açúcar para diminuir o custo da hidrólise enzimática e favorecer a produção de etanol de segunda geração. Neste trabalho foram estudadas as melhores condições dos fungos filamentosos em agitação orbital, além de caracterizar os parâmetros bioquímicos como pH e temperatura ótimos e determinar o melhor produtor do complexo celulolítico e analisar a capacidade quanto à biodegradação de bagaço de cana-de-açúcar comparado com enzima comercial. Através da análise de atividade enzimática, o Aspergillus terreus mostrou-se o fungo com a maior produção nos diferentes meios de cultivo, atingindo a maior atividade enzimática no meio contendo CMC, apresentando 230,65 UI.mg proteína-1 de endoglucanase, 10,79 UI.mg proteína-1 de β-glicosidase, 57,79 UI.mg proteína-1 de exoglucanase e 68,60 UI.mg proteína-1 de atividade celulolítica total (FPAse). A análise realizada através do delineamento experimental de Plackett & Burman realizada no meio contendo glicose e celulose cristalina não mostrou influencia da composição do meio de cultura na produção das enzimas do complexo celulítico. O estudo de estabilidade ao pH revelou que... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The cellulolytic complex is composed mainly of endoglucanases, exoglucanases and β-glycosidases. Endoglucanases act by cleaving the cellulose in the internal bonds of the fiber, while the exoglucanases hydrolyze the outer bonds. The need for new research on cellulolytic enzymes capable of efficiently degrading industrial residues such as sugarcane bagasse to reduce the cost of enzymatic hydrolysis and favor the production of second generation ethanol. In this work the best conditions of the filamentous fungi were studied in orbital agitation, besides characterizing the biochemical parameters as pH and temperature optimum and to determine the best producer of the cellulolítico complex and to analyze the capacity regarding the biodegradation of sugarcane bagasse compared to Commercial enzyme. Through enzymatic activity analysis, Aspergillus terreus showed the fungus with the largest output in the different culture media, achieving higher production in D medium, showing 230,65 65 μmol.min-1.mg protein-1 for endoglucanase, 10788,34 nmol.min-1.mg protein-1 for β-glycosidase, 57.79 μmol.min-1.mg protein-1 for exoglucanase and 68,60 μmol.min-1.mg protein-1 for Total Cellulolytic Activity - Filter Papel Activity (FPAse). The analysis made with the Plackett & Burman design for the medium content glucose and crystal cellulose showed that the medium composition not show an influence on the enzyme production of cellulolytic complex. The enzymatic stability of pH showed that pH 3 has a negative influence for all enzymatic complex. The endoglucanase and FPAse tend to 0% of residual activity after 24 hours of incubation in all studied pHs for all fungi. The partial purification showed best results with the acetone precipitation and, also, the exoglucanase enzyme was the only... (Complete abstract click electronic access below)
Mestre

SILVA, Rodolpho Glauber Guedes. Purificação, caracterização bioquímica e atividade biológica de um inibidor de tripsina da torta da mamona (Ricinus communis L.). 2012. 94 f. : Dissertação (Mestrado em Bioquímica)-Universidade Federal do Ceará, Fortaleza-CE, 2012.
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The castor bean (Ricinus communis L.) culture is of great socioeconomic importance because of its seeds, which are used mainly for production of oil that can be used for various industrial purposes. The oil extraction generates a by-product of high nutritional value known as the castor cake. However, the presence of toxic and allergenic compounds hinders the use of this residue to feeding source. Another way to add value to the castor cake would be to seek bioactive molecules that could have applications in agriculture and human health. This study aimed to purify and characterize a trypsin inhibitor of castor cake and, to test this in vitro activity against phytopathogenic fungi of agricultural importance and proteases from the midgut of Aedes aegypti. The trypsin inhibitor of castor cake, named RcTI, was purified by heat treatment, affinity column in anhydrotrypsin-Sepharose® 4B and ion-exchange chromatography ResourceTM Q. RcTI has a apparent molecular mass of 14 kDa in the absence or presence of a reducing agent, pI 5.2 and isn’t a glycoprotein. The NH2-terminal sequence of purified RcTI showed 83% similarity with sulfur-rich storage protein (2S albumin) R. communis and 48% with napin-like (2S albumin) R. communis. The RcTI was stable after heat treatment at 98 °C for two hours. The inhibition on trypsin was stable in the acid pH range. However, there was a loss of approximately 20% inhibitory effect on alkaline pH (pH 8-11). The RcTI (13 µg) was unable to inhibit the germination of spores of the fungus Fusarium oxysporum, Rhizoctonia solani and Fusarium solani. However, it was able to inhibit the germination of spores of Colletotrichum gloeosporioides. RcTI 100 µg was not effective in the inhibition of vegetative growth of the fungi mentioned above. This inhibitor was effective against proteases from the midgut of Aedes aegypti with 91% inhibition. The results of the present study indicate that RcTI has biotechnological potential, can be used as an alternative to combat the important plant pathogenic fungus C. gloeosporioides and larvae of A. aegypti.
A cultura da mamoneira (Ricinus communis L.) é de grande importância socioeconômica devido às suas sementes, que são utilizadas, principalmente, na produção do óleo que pode ser empregado para diversos fins industriais. A extração do óleo gera um subproduto de alto valor nutricional, conhecido como torta da mamona. Porém, a presença de compostos tóxicos e alergênicos dificulta utilização desse resíduo como fonte de alimentação animal. Outra forma de agregar valor à torta da mamona seria buscar moléculas bioativas que pudessem ter aplicações na agricultura e saúde humana. O presente trabalho teve como objetivos purificar e caracterizar um inibidor de tripsina da torta da mamona, bem como, testar a atividade deste in vitro contra fungos fitopatogênicos de importância agrícola e proteases do intestino médio de Aedes aegypti. O inibidor de tripsina da torta da mamona, nomeado RcTI, foi purificado através de tratamento térmico, coluna de afinidade em matriz de anidrotripsina-Sepharose® 4B e cromatografia de troca iônica em ResourceTM Q. Essa proteína apresentou massa molecular aparente de 14,0 kDa na ausência ou presença de agente redutor, pI 5,2 e não é uma glicoproteína. A sequência NH2-terminal do RcTI purificado mostrou similaridade de 83% com uma proteína de armazenamento rica em enxofre (albumina 2S) de R. communis e de 48% com uma napin-like (albumina 2S) de R. communis. O RcTI mostrou-se estável após tratamento térmico a 98 ºC durante duas horas. Da mesma forma, permaneceu com atividade inibitória estável na faixa de pHs ácidos. No entanto, houve uma perda de, aproximadamente, 20% na capacidade inibitória em pHs alcalinos (pH 8-11). O RcTI (13 µg) não foi capaz de inibir a germinação de esporos dos fungos Fusarium oxysporum, Fusarium solani e Rhizoctonia solani. No entanto, foi capaz de inibir a germinação de esporos de Colletotrichum gloeosporioides. O RcTI não foi efetivo na inibição de crescimento vegetativo dos fungos mencionados acima. Esse inibidor foi efetivo contra as proteases intestinais de Aedes aegypti com 91% de inibição. A partir dos resultados obtidos neste trabalho, conclui-se que o RcTI tem potencial biotecnológico, podendo ser utilizado como uma alternativa para o combate do importante fungo fitopatogênico C. gloeosporioides e larvas de A. aegypti.

False codling moth (FCM), Thaumatotibia leucotreta is an extremely important pest of citrus in South Africa and with the shift away from the use of chemicals, alternate control options are needed. One avenue of control which has only recently been investigated against the soil-borne life stages of FCM is the use of entomopathogenic fungi (EPF). In 2009, 12 entomopathogenic fungal isolates collected from South African citrus orchards showed good control potential during laboratory conducted bioassays. The aim of this study was to further analyse the potential of these isolates through concentration-dose and exposure-time response bioassays. After initial re-screening, concentration-dose response and exposure-time response sandconidial bioassays, three isolates were identified as exhibiting the greatest control potential against FCM in soil, Metarhizium anisopliae var. anisopliae (G 11 3 L6 and FCM Ar 23 B3) and Beauveria bassiana (G Ar 17 B3). Percentage mycosis was found to be directly related to fungal concentration as well as the amount of time FCM 5th instar larvae were exposed to the fungal conidia. LC50 values for the three isolates were not greater than 1.92 x 10⁶ conidia.ml⁻ₑ and at the LC₅₀, FCM 5th instar larvae would need to be exposed to the fungus for a maximum of 13 days to ensure a high mortality level. These isolates along with two commercially available EPF products were subjected to field persistence trials whereby net bags filled with a mixture of autoclaved sand and formulated fungal product were buried in an Eastern Cape citrus orchard. The viability of each isolate was measured on a monthly basis for a period of six months. All isolates were capable of persisting in the soil for six months with the collected isolates persisting far better than the commercially used isolates. Two of the isolates, G 11 3 L6 and G Ar 17 B3, were subjected to small scale laboratory application trials. Two formulations were investigated at two concentrations. For each isolate, each formulation and each concentration, FCM 5th instar larvae were applied and allowed to burrow into the soil to pupate before fungal application or after fungal application. Contact between fungi and FCM host is essential as, in contrast to pre-larval treatments, percentage mortality in post-larval treatments was low for both formulations and both isolates. For isolate G Ar 17 B3, a conidial suspension applied as a spray at a concentration of 1 x 10⁷ conidia.ml⁻ₑ obtained the highest percentage mortality (80 %). For isolate G 11 3 L6 however, both formulations performed equally well at a high, 1 x10⁷ conidia.ml⁻ₑ concentration (conidial suspension: 60 %; granular: 65 %) The results obtained thus far are promising for the control of FCM in citrus, but if these EPFs are to successfully integrate into current FCM control practices more research, some of which is discussed, is essential

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
O termo cancro é utilizado para designar lesões localizadas na casca do caule de plantas lenhosas que podem resultar no anelamento do caule e no secamento da parte superior da copa e a morte da planta. A melhor opção para o controle da doença é a resistência genética dos indivíduos quanto ao patógeno utilizado. Com o objetivo de estudar o comportamento de resistência e mecanismos moleculares de clones dos parentais e das progênies de eucalipto quanto ao cancro causado por Chrysoporthe cubensis e Botryosphaeria sp. foram executados ensaios em casa de vegetação e em campo. Os isolados utilizados foram Chrysoporthe cubensis e Botryosphaeria sp. sendo produzidos em meio BDA e inoculados nos indivíduos pelo método do disco. A avaliação foi realizada 60 dias após a inoculação, medindo a área da lesão de cada material. Para o patógeno C. cubensis inoculados nos parentais foi encontrado uma média de 3,20 e 6,14 cm2 de área de lesão do tronco causado pelos isolados provenientes de Piracicaba e Bofete, respectivamente, enquanto que para Botryosphaeria sp. a média variou de 7,29 cm ² a 9,72cm ² da área da lesão. Dos indivíduos testados, com C. cubensis, foram encontrados 46,66% de plantas classificadas como resistentes; 6,67% plantas moderadamente resistente, 13,33% planta moderadamente suscetíveis e 33,34% plantas suscetíveis. Já para o teste de clones com Botryosphaeria sp. verificou-se 91,67% resistentes, 5% moderadamente resistentes, 1,67% moderadamente suscetível e 1,67% suscetíveis. As variâncias fenotípicas encontradas foram de 41,32 a 0,23 inoculados com C. cubensis e 0,285 a 156,88, quando inoculados com Bothyosphaeria sp. Os coeficientes de variação foram de 125% a 15,05% para C. cubensis e 29,86% a 101,13% para Bothyosphaeria sp. Os resultados indicam que os materiais genéticos testados possuem boa fonte de resistência para os dois fitopatógenos...
The term canker is used to assign to injuries located in the bark of stems of woody plants that can result in the anelamento of stems and drying of the superior part of the crown and the death of the plant. The best option for the control of the illness is the resistance genetic of the individuals concerning to the used patógeno. The objective of this study the molecular behavior of resistance and mechanisms of clonals of parental and the lineages the eucalyptus concerning the canker caused for Chrysoporthe cubensis and Dothiorella sp. The essays executed in the greenhouse and field. The isolated ones used concerning Chrysoporthe cubensis and Dothiorella sp. being produced in inoculated medium BDA and in the individuals for the method of the disk. The evaluation was concluded 60 days after the inoculation, measuring the area of the injury of each material. For pathogeno C. cubensis inoculated in the parental ones a 6,14 was found average of 3,20 and cm² of area of isolated injury of the trunk caused for the proceeding ones from Piracicaba and Bofete, respectively. The average of the area of the injury for the studied lineages varied of 7,29 cm ² 9,72cm ² for Dothiorella sp. The clonals inoculated C. cubensis were grouped into the categories HS (highly susceptible – 33,34%), MS (moderately susceptible -13,3%), MR (moderately resistant- 6,67%), and HR (highly resistant -46,66%). The clonal tested, with Botryosphaeria sp., verified resistant 91.67%, 5% moderately resistant, 1.67% moderately susceptible and 5,26% susceptible. The phenotypic variation found was from 41,32 to 0,23, inoculated with C. cubensis while Botryosphaeria sp from 0,285 to156,88. The coefficient of variation from 125% to 15,95% for C. cubensis and 29,86% to 101,13% for Botryosphaeria sp. The results indicate that the tested genetic materials possess good source of resistance for the two fitopatógenos tested... (Complete abstract click electronic access below)

Mealybugs (Planococcus citri) and thrips (Scirtothrips aurantii) are common and extremely damaging citrus crop pests which have proven difficult to control via conventional methods, such as chemical pesticides and insect growth regulators. The objective of this study was to determine the efficacy of entomopathogenic fungi against these pests in laboratory bioassays. Isolates of Metarhizium anisopliae and Beauveria bassiana from citrus orchards in the Eastern Cape, South Africa were maintained on Sabouraud Dextrose 4% Agar supplemented with Dodine, chloramphenicol and rifampicin at 25°C. Infectivity of the fungal isolates was initially assessed using 5th instar false codling moth, Thaumatotibia leucotreta, larvae. Mealybug bioassays were performed in 24 well plates using 1 x 107 ml-1 conidial suspensions and kept at 26°C for 5 days with a photoperiod of 12 L:12 D. A Beauveria commercial product and an un-inoculated control were also screened for comparison. Isolates GAR 17 B3 (B. bassiana) and FCM AR 23 B3 (M. anisopliae) both resulted in 67.5% mealybug crawler mortality and GB AR 23 13 3 (B. bassiana) resulted in 64% crawler mortality. These 3 isolates were further tested in dose-dependent assays. Probit analyses were conducted on the dose-dependent assays data using PROBAN to determine LC₅₀ values. For both the mealybug adult and crawlers FCM AR 23 B3 required the lowest concentration to achieve LC₅₀ at 4.96 x 10⁶ conidia ml-1 and 5.29 x 10⁵ conidia ml-1, respectively. Bioassays on adult thrips were conducted in munger cells with leaf buds inoculated with the conidial suspensions. Isolate GAR 17 B3 had the highest mortality rate at 70% on thrips while FCM AR 23 B3 resulted in 60% mortality. Identification of the isolates, FCM AR 23 B3, GAR 17 B3 and GB AR 23 13 3, were confirmed to be correct using both microscopic and molecularly techniques. ITS sequences were compared to other sequences from GenBank and confirmed phylogenetically using MEGA6. Mealybug infection was investigated using scanning electron microscopy, mycosis was confirmed but the infection process could not be followed due to the extensive waxy cuticle. These results indicate that there is potential for the isolates FCM AR 23 B3 and GAR 17 B3 to be developed as biological control agents for the control of citrus mealybug and thrips. Further research would be required to determine their ability to perform under field conditions.

Early in 2003, African Muslims in Uitenhage’s township, Kwa-Nobuhle, learnt that Muslim women, led by Sheikh Nceba Salamntu, in South Africa’s Port Elizabeth New Brighton township, were allowed, contrary to previous practice, to follow a funeral procession right up to the graveyard. The resultant discomfort on the one hand, and excitement on the other caused by this event among Muslims in the township, forms the basis of this research. It gives focus to Muslim women, the ones most affected by their customary restriction from the gravesites. The researchi exposes the basis for women’s exclusion from funeral processions in the Muslim community. It was established that many of these Muslim women who challenged the practice were converts from Christianity to Islam. One of the bases for their action was that they were passive recipients of Islam. Furthermore, it was found that the exclusion of women from the funeral procession has no basis in Islamic writings.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Dos 27 microrganismos recém isolados, A. fumigatus M.7.1 e Myceliophthora sp M.7.7 foram os melhores produtores de FPase (0,8 e 2,0 U/g de substrato) e xilanase (1040 e 1292 U/g de substrato), quando cultivados por fermentação em estado sólido (FES) em mistura de bagaço de cana e farelo de trigo (1:1). As produções máximas de xilanase (7237,9 U/g de substrato) e endoglucanase (46,2 U/g de substrato) por A. fumigatus M.7.1 foram observadas pelo cultivo do fungo em palha de milho e farelo de trigo (9:1 p/p) e bagaço de cana e farelo de trigo (9:1 p/p), respectivamente. Em relação ao isolado Myceliophthora sp M.7.7, os picos de produção de ambas as enzimas (xilanase: 1044,6 U/g de substrato; endoglucanase 53,7 U/g de substrato) foram obtidos quando este foi cultivado em mistura de bagaço de cana e farelo de trigo (9:1 p/p). A endoglucanase e xilanase produzida por A. fumigatus M.7.1 apresentaram atividade ótima em pH 4,5, a 70 e 60 ºC, respectivamente. Nos ensaios com a linhagem Myceliophthora sp M.7.7, ambas as enzimas apresentaram maior atividade em pH 5,0 a 65-70 ºC. Ambas as enzimas de Myceliophthora sp M.7.7 e a endoglucanase de A. fumigatus M.7.1 mantiveram aproximadamente 70-100% da atividade inicial na faixa de pH entre 3,5 a 9,0 e nas temperaturas de 35 a 65 ºC. A xilanase de A. fumigatus M.7.1 manteve-se estável em pH entre 5,5 e 10,5 e 40 e 50 ºC, respectivamente. Dos tratamentos ao qual o bagaço foi submetido, o que mostrou maior eficiência na liberação de açúcares redutores (0,09%) e compostos fenólicos (0,74%), foi a solução de glicerol em microondas por 5 min. O bagaço de cana pré tratado com glicerol foi incubado com o preparado enzimático de A. fumigatus M.7.1 a 55 ºC. Após 24 h de incubação, foram liberados 0,7 mg/mL de açúcar redutor. As mesmas condições foram utilizadas na sacarificação do bagaço utilizando o extrato enzimático...
The microorganisms A. fumigatus M.7.1 and Myceliophthora SP M.7.7 were the best producers of FPase (0,8 and 2,0 U/g of substrate) and xylanase (1040 and 1292 U/g of substrate) of the 27 isolated microorganisms, when cultivated by solid state fermentation (SSF) in a mixture of cane bagasse and wheat bran (1:1). The greatest production of xylanase (7237,9 U/g of substrate) and endoglucanase (46,2 U/g of substrate) by A. fumigatus M.7.1 were noted by the fungi cultivation in corn straw and wheat bran (9:1 p/p) and cane bagasse and wheat bran (9:1 p/p), respectively. In the isolated Myceliophthora sp M.7.7, the peak production of the both enzymes (xylanase: 1044,6 U/g of substrate; endoglucanase 53,7 U/g of substrate) were obtained when the microorganism were cultivated in a mixture of cane bagasse and wheat bran (9:1 p/p). The endoglucanase and the xylanase produced by A. fumigatus M.7.1 showed higher activity in pH 4,5, at 70 and 60 oC, respectively. In the essays with the Myceliophthora sp M.7.7 strains, the both enzymes showed higher activity in pH 5,0, at 65-70 oC. The both enzymes of Myceliophthora sp M.7.7 and the endoglucanase of A. fumigatus M.7.1 maintained approximately 70-100% of the initial activity between pH 3,5 and 9,0 and between 35 and 65 oC. The xylanase of A. fumigatus M.7.1 was stable between the pH 5,5 and 10,5 and the temperature 40 and 50 oC. About the treatments of the bagasse, the most efficient in the liberation of reducing sugars (0,09%) and phenolic compounds (0,74%) was the glycerol solution in microwave for 5 minutes. The cane bagasse pretreated with glycerol was incubated with the enzymatic solution of A. fumigatus M.7.1 at 55 oC. After 24 hours of incubation, 0,7mg/mL of reducing sugar was liberated. The same conditions were used in the saccharification of the bagasse using the enzymatic extract produced by Myceliophthora sp. M.7.7, with a reducing sugar... (Complete abstract click electronic access below)

Resumo: Dos 27 microrganismos recém isolados, A. fumigatus M.7.1 e Myceliophthora sp M.7.7 foram os melhores produtores de FPase (0,8 e 2,0 U/g de substrato) e xilanase (1040 e 1292 U/g de substrato), quando cultivados por fermentação em estado sólido (FES) em mistura de bagaço de cana e farelo de trigo (1:1). As produções máximas de xilanase (7237,9 U/g de substrato) e endoglucanase (46,2 U/g de substrato) por A. fumigatus M.7.1 foram observadas pelo cultivo do fungo em palha de milho e farelo de trigo (9:1 p/p) e bagaço de cana e farelo de trigo (9:1 p/p), respectivamente. Em relação ao isolado Myceliophthora sp M.7.7, os picos de produção de ambas as enzimas (xilanase: 1044,6 U/g de substrato; endoglucanase 53,7 U/g de substrato) foram obtidos quando este foi cultivado em mistura de bagaço de cana e farelo de trigo (9:1 p/p). A endoglucanase e xilanase produzida por A. fumigatus M.7.1 apresentaram atividade ótima em pH 4,5, a 70 e 60 ºC, respectivamente. Nos ensaios com a linhagem Myceliophthora sp M.7.7, ambas as enzimas apresentaram maior atividade em pH 5,0 a 65-70 ºC. Ambas as enzimas de Myceliophthora sp M.7.7 e a endoglucanase de A. fumigatus M.7.1 mantiveram aproximadamente 70-100% da atividade inicial na faixa de pH entre 3,5 a 9,0 e nas temperaturas de 35 a 65 ºC. A xilanase de A. fumigatus M.7.1 manteve-se estável em pH entre 5,5 e 10,5 e 40 e 50 ºC, respectivamente. Dos tratamentos ao qual o bagaço foi submetido, o que mostrou maior eficiência na liberação de açúcares redutores (0,09%) e compostos fenólicos (0,74%), foi a solução de glicerol em microondas por 5 min. O bagaço de cana pré tratado com glicerol foi incubado com o preparado enzimático de A. fumigatus M.7.1 a 55 ºC. Após 24 h de incubação, foram liberados 0,7 mg/mL de açúcar redutor. As mesmas condições foram utilizadas na sacarificação do bagaço utilizando o extrato enzimático... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The microorganisms A. fumigatus M.7.1 and Myceliophthora SP M.7.7 were the best producers of FPase (0,8 and 2,0 U/g of substrate) and xylanase (1040 and 1292 U/g of substrate) of the 27 isolated microorganisms, when cultivated by solid state fermentation (SSF) in a mixture of cane bagasse and wheat bran (1:1). The greatest production of xylanase (7237,9 U/g of substrate) and endoglucanase (46,2 U/g of substrate) by A. fumigatus M.7.1 were noted by the fungi cultivation in corn straw and wheat bran (9:1 p/p) and cane bagasse and wheat bran (9:1 p/p), respectively. In the isolated Myceliophthora sp M.7.7, the peak production of the both enzymes (xylanase: 1044,6 U/g of substrate; endoglucanase 53,7 U/g of substrate) were obtained when the microorganism were cultivated in a mixture of cane bagasse and wheat bran (9:1 p/p). The endoglucanase and the xylanase produced by A. fumigatus M.7.1 showed higher activity in pH 4,5, at 70 and 60 oC, respectively. In the essays with the Myceliophthora sp M.7.7 strains, the both enzymes showed higher activity in pH 5,0, at 65-70 oC. The both enzymes of Myceliophthora sp M.7.7 and the endoglucanase of A. fumigatus M.7.1 maintained approximately 70-100% of the initial activity between pH 3,5 and 9,0 and between 35 and 65 oC. The xylanase of A. fumigatus M.7.1 was stable between the pH 5,5 and 10,5 and the temperature 40 and 50 oC. About the treatments of the bagasse, the most efficient in the liberation of reducing sugars (0,09%) and phenolic compounds (0,74%) was the glycerol solution in microwave for 5 minutes. The cane bagasse pretreated with glycerol was incubated with the enzymatic solution of A. fumigatus M.7.1 at 55 oC. After 24 hours of incubation, 0,7mg/mL of reducing sugar was liberated. The same conditions were used in the saccharification of the bagasse using the enzymatic extract produced by Myceliophthora sp. M.7.7, with a reducing sugar... (Complete abstract click electronic access below)
Orientador: Eleni Gomes
Coorientador: Daniela A. Bocchini-Martins
Banca: Beatriz Vahan Kilikian
Banca: Mauricio Boscolo
Mestre

Thesis (MScAgric)--University of Stellenbosch, 2002.
ENGLISH ABSTRACT: Historically root diseases have been a production-limiting problem for the strawberry industry worldwide. In the Western Cape Province of South Africa the most serious root disease is black root rot, which causes losses of up to 30%, annually. The aims of this study were to investigate aspects of the etiology and epidemiology of this disease in the Western Cape, and to provide information that can be incorporated in an integrated disease management strategy. In Chapter I a summary of published information on this disease is presented. Disease symptoms include severe stunting of plants, which have black, rotted, reduced root systems. Even though this disease is of great economic importance, the etiology remains unresolved. However, soilborne fungal root pathogens, particularly Pythium and Rhizoctonia spp. have been implicated as major role players. Control of this disease, as well as the other root diseases affecting strawberries, has relied on soil fumigation with broad spectrum chemical fumigants, in particular methyl bromide. However, due to the ozone depleting action of methyl bromide it was decided at the signing of the Montreal Protocol to remove this chemical from the market. This action has caused great demand for alternative measures to control root diseases on many crops including strawberries. Development of integrated disease management strategies is dependent upon a more complete understanding of the etiology, biology and ecology of the disease complex. In Chapter 2 fungal pathogens associated with diseased plants were isolated and Koch's postulates were carried out. The most frequently isolated fungal pathogens were Pythium irregulare, Rhizoctonia spp. and Cylindrocarpon destructans. Two morphotypes of Rhizoctonia were isolated viz. a brown and a white type. Pythium irregulare was isolated more frequently in July than in September, and was not isolated at all in November. Rhizoctonia spp. were present at all sampling dates but were more frequently isolated in November than at the other times. All the fungi that were tested were pathogenic and caused root lesions. Cylindrocarpon destructans and Coniella fragariae did not have a stunting effect on the plants. These results confirm a major role for Pythium and Rhizoctonia in the black root rot complex and suggest that there is a complimentary seasonal variation in occurrence between these two pathogens. The Rhizoctonia species and anastomosis groups of isolates obtained from diseased strawberries in the Western Cape Province were determined, and their pathogenicity and relative virulence assessed. Both binucleate and multinucleate types were recovered from diseased roots and identified as R. fragariae and R. so/ani, respectively. All isolates of R. solani were members of anastomosis group (AG) 6, whereas three AG types were identified among isolates of R. fragariae, viz. AG-A, AG-G and AG-I at a relative occurrence of 69%, 25%, 6% respectively. All Rhizoctonia isolates were pathogenic to strawberry, but R. solani (AG 6) was the most virulent causing severe stunting of plants. This is the first species confirmation and AG type identification of Rhizoctonia taxa causing root rot of strawberries in South Africa. An assessment of the presence and quantity of black root rot pathogens associated with soils prior to fumigation and post fumigation with methyl bromide, was made in Chapter 4. Isolations were also made from nursery plants to determine whether any black root rot pathogens were in the plants before transplanting. Results demonstrated that after fumigation the soil was free of all pathogenic fungi associated with the disease. However, the main pathogens involved in black root rot, viz. Rhizoctonia fragariae, R. solani and Pythium spp. were isolated from nursery plants. The fact that the plants are already infected with these pathogens renders the prospects for control of this disease difficult. Further studies are urgently required to develop production practices that can be included in disease management programmes. In vitro studies were carried out to determine the ECso values of different fungicides for isolates of Pythium irregulare, Rhizoctonia fragariae AG-A, AG-G and AG-I and R. solani AG 6. Benomyl, fludioxonil and tolc1ofos-methyl were used in these tests. Field trials were also conducted using these fungicides. In Chapter 5 it is shown that in general application of fungicides improved the yield and did not affect the survival rate of strawberry plants. Fludioxonil showed potential for short-term use. Applications of fungicides that targeted specific fungal genera were not sufficient to control the disease. Seasonal fluctuation of Pythium and Rhizoctonia spp. became apparent with the occurrence of Pythium being relatively high early in the season but low late in the season. Conversely, the occurrence of Rhizoctonia was low at the beginning of the season but high late in the season. In the short-term there is potential for fungicide applications as part of an integrated disease management strategy, but the economic feasibility of this practice needs to be assessed. In this study the major pathogens causing black root rot were identified in the Western Cape Province of South Africa, and important information regarding the epidemiology of the disease was reported. These results can be incorporated in an integrated management strategy to reduce losses of strawberry production, attJibutable to black root rot.
AFRIKAANSE OPSOMMING: Wortelsiektes is wêreldwyd 'n produksie-beperkende probleem vir die aarbeibedryf. . Swartwortelvrot, wat jaarliks verliese van tot 30% veroorsaak, is die belangrikste wortelsiekte in die Wes-Kaap Provinsie van Suid-Afrika. Die doelwitte van hierdie studie was om aspekte van die etiologie en epidemiologie van die siekte in die Wes- Kaap te ondersoek en inligting wat in geïntegreerde siektebestuurstrategië ingesluit kan word, te voorsien. In Hoofstuk 1 word 'n opsomming van gepubliseerde inligting aangaande die siekte uiteengesit. Siektesimptome sluit ernstige verdwerging van plante met swart verotte en verkleinde wortelstelsels in. Alhoewel die siekte van groot ekonomiese belang is, is die etiologie grootliks onbekend. Grondgedraagde wortelpatogene swamme, spesifiek Pythium en Rhizoctonia spp., is egter as belangrike rolspelers geïdentifiseer. Tot dusver het die beheer van hierdie siekte sowel as ander wortelsiektes van aarbeie berus op grondberoking met breë spektrum chemiese berokingsmiddels, spesifiek metielbromied. As gevolg van die osoonafbrekende aksie van metielbromied is daar egter tydens die ondertekening van die Montreal Protocol besluit om dié middel van die mark te verwyder. Hierdie besluit het 'n groot aanvraag na alternatiewe beheermaatreëls vir wortelsiektes van verskeie gewasse, insluitende aarbeie, veroorsaak. Die ontwikkeling van geïntegreerd siektebestuurstrategieë is egter afhanklik van 'n meer volledige begrip van die etiologie, biologie en ekologie van die siektekompleks. In Hoofstuk 2 is die patogene swamme wat met die siekte geassosieer word, geïsoleer, en is Koch se postulate uitgevoer. Die mees algemeen geïsoleerde patogene swamme was Pythium irregulare, Rhizoctonia spp. en Cylindrocarpon destructans. Twee morfotipes van Rhizoctonia is geïsoleer, nl. 'n bruin tipe en 'n wit tipe. Pythium irregulare is meer dikwels in Julie as in September geïsoleer, maar glad nie in November nie. Rhizoetonia het tydens alle monstertye voorgekom, maar is meer dikwels in November geïsoleer. Al die swamme wat getoets is, was patogenies en het letsels op die wortels veroorsaak. Cylindroearpon des true tans en Coniella fragariae het nie'n verdwergingseffek op plante gehad nie. Hierdie resultate bevestig die dominante rol van Pythium en Rhizoctonia in die swartwortelvrot kompleks en dui op 'n komplementêre seisoenale variasie in die voorkoms van hierdie twee patogene. Die Rhizoctonia spesies en anastomose groepe (AG) van die isolate geisoleer vanaf siek aarbeiplante in die Wes-Kaap Provinsie is bepaal, en die patogenisiteit en relatiewe virulensie is beraam. Sowel tweekernige as multikernige tipes is vanaf siek wortels geïsoleer en respektiewelik as R. fragariae en R. so/ani geïdentifiseer. Alle isolate van R. so/ani was lede van anastomose groep 6, terwyl drie AG tipes, nl. AGA, AG-G en AG-I onder die R. fragariae isolate geïdentifiseer is met relatiewe voorkomste van 69%, 25%, 6% respektiewelik. Alle Rhizoctonia isolate was patogenies op aarbeie, maar R. so/ani (AG 6) was die mees virulente en het ernstige verdwerging van plante veroorsaak. Hierdie is die eerste bevestiging van spesies en identifisering van AG tipes van Rhizoctonia taksa wat wortelvrot van aarbeie in Suid Afrika veroorsaak. In Hoofstuk 4 is 'n beraming van die voorkoms en hoeveelheid swartwortelvrot patogene geassosieer met grond voor, en na beroking met metielbromied, gemaak. Isolasies is ook vanaf kwekeryplante gemaak om te bepaal of enige swartwortelvrot patogene voor oorplanting in die plante teenwoordig was. Die resultate het getoon dat grond na beroking vry was van alle patogeniese swamme geassosieër met die siekte. Die hoof patogene betrokke in die swartwortelvrot kompleks, nl. Rhizoctonia fragariae, R. so/ani en Pythium spp. was egter in die kwekery plante teenwoordig. Die feit dat plante reeds met hierdie patogene geïnfekteer is, maak die vooruitsigte vir die beheer van hierdie siekte moeilik. Verdere studies word dringend benodig vir die ontwikkeling van produksiepraktyke wat by siektebestuursprogramme ingesluit kan word. In vitro studies om die ECso waardes van die isolate van Pythium irregulare, Rhizoctonia fragariae AG-A, AG-G en AG-I en R. so/ani AG 6 vir die fungisiedes benomyl, fludioxonil en tolclofos-metiel te bepaal, is uitgevoer. Hierdie fungisiedes is ook in veldproewe getoets. In Hoofstuk 5 is getoon dat aanwending van fungisiedes die opbrengs verbeter het en nie die oorlewing van aarbeiplante beïnvloed het nie. Fludioxonil het potensiaal vir korttermyn gebruik getoon. Die aanwending van fungisiedes wat spesifieke swamgenera teiken, was nie voldoende om die siekte te beheer nie. Seisoenale fluktuasies van Pythium en Rhizoctonia spp. het duidelik geword met die relatief hoë voorkoms van Pythium vroeg in die seisoen, maar lae voorkoms laat in die seisoen, terwyl die voorkoms van Rhizoctonia laag was aan die begin van die seisoen, maar hoog later in die seisoen. In die korttermyn is daar potensiaal vir fungisiedtoedienings as deel van 'n geïntegreerde siektebestuurstrategie, maar die ekonomiese haalbaarheid van hierdie praktyk moet bepaal word. In hierdie studie is die hoof patogene wat swartwortelvrot van aarbeie in die Wes-Kaap Provinsie van Suid-Afrika veroorsaak geïdentifiseer, en belangrike inligting rakende die epidemiologie van die siekte is aangeteken. Hierdie resultate kan in 'n geïntegreerde bestuurstrategie geïnkorporeer word om verliese van aarbeiproduksie, toeskryfbaar aan swartwortelvrot te, verminder.

Thesis (PhD)--Stellenbosch University, 2014.
ENGLISH ABSTRACT: Very little is known about the diversity of fungi associated with Afromontane forests of the Cape Floristic Region (CFR) of South Africa. The ophiostomatoid fungi include many species, some known as pathogens in the CFR, while others are well-known saprophytes important in wood degradation. This study focused on the biodiversity and ecology of tree-associated ophiostomatoid fungi (Ophiostomatales) in the CFR. In addition to this, mites and subcortical beetles associated with the CFR trees were collected, regardless of whether they were associated with ophiostomatoid fungi or not. A relatively high diversity of ophiostomatoid fungi were collected from native trees, ten of which were newly described here. Three further fungal species, two of which are probably new to science, were also collected from exotic Pinus species growing in these forests. Four Ophiostomatales species (including three newly described species) were associated with subcortical beetles on Rapanea melanophloeos and Olea capensis ssp. macrocarpa. These were Sporothrix pallida, Sporothrix aemuluphilus, Raffaelea scabbardiae and Raffaelea rapaneae, associated with the beetles Lanurgus sp. 1, Ctonoxylon sp. 1, Xyleborinus aemuluphilus and a Platypodinae species. This represents a first study to explore the associations between subcortical beetles and ophiostomatoid fungi on native trees in the CFR. In addition to fungi associated with subcortical beetles, several members of the Ophiostomatales associated with wounds on Rapanea melanophloes trees were also collected. These included Ophiostoma stenoceras, Sporothrix reniformis, S. rapaneae, S. lunateae and S. noisomeae. All but O. stenoceras were new to science, and were formally described here. All of these wound-associated species from R. melanophloeos belong to the Sporothrix schenckii – O. stenoceras complex, except for S. noisomeae that was provisionally placed in the S. lignivora complex. Besides fungal taxa collected from wounds on Rapanea melanophloeos, other fungi were also collected from wounds on other host trees species. Three more previously undescribed ophiostomatoid fungal species were collected from this niche. They included Sporothix capensis collected from O. capensis ssp. macrocarpa, Graphilbum roseus collected from many different, unrelated host trees and Graphium ilexiense (Microascales), isolated from wounds on Ilex mitis. The latter represented the first isolation of an ophiostomatoid fungus from this host tree species. Two possibly new fungal species (Sporothrix sp. 1, Ceratocystiopsis sp. 1) and Ophiostoma ips, associated with three bark beetles (Orthotomicus erosus, Hylurgus ligniperda and Hylastes angustatus), were collected from Pinus. Several fungal species were collected from both native trees and non-native trees. These included Sporothrix fusiforme from Brabejum stellatifolium and Acacia mearnsii, O. quercus and O. pluriannulatum-like fungus from several native trees and from A. mearnsii. This suggests a possibility for host shifting of some of these fungi between native and non-native hosts or even between different native hosts. Eight non-ophiostomatoid fungi associated subcortical beetles taxa were found also to infest native trees in the Afromontane forests and in total more than 4500 beetle individuals were collected. Some species of ophiostomatoid fungi collected in this study were found to be associated with other arthropods such as mites. Four phoretic mites species associated with ophiostomatoid fungi (Dendrolaelaps quadrisetus, Histiogaster sp. 3, Elattoma sp. 1 & 2) were collected. In addition, sixteen species of tree wound-associated mites were collected from 12 native trees. Of these, nine were associated with several ophiostomatoid fungi (Graphilbum roseus, O. pluriannulatum-like, O. quercus) that were isolated from several different host trees. This suggests that they may aid in the transport of these fungi from one host species to another. The possible consequences of transfers of Ophiostomatales species between hosts were tested using pathogenicity tests, which highlighted that some fungi are pathogenic on several different trees. Transfers seemed most likely in fungal species isolated from wounds, especially those associated with mites, because the mites may aid in the vectoring of these. When phoretic mites were tested for their specificity to their vector beetles, they proved to be highly specific. Although some of the fungi associated with these mites and their sub-cortical beetles were also pathogenic, it is less likely for these fungi to be transferred to other host tree species due to the high specificity of their arthropod associates. This study represents one of a few studies that focused on ophiostomatoid fungi, subcortical beetles and mites associated with trees in the Afromontane forests of South Africa. Although we collected a high diversity of Ophiostomatales members, many more still await discovery. It is recommended that future studies focus on the complex inter-organismal interactions in many of the systems uncovered in this study.
AFRIKAANSE OPSOMMING: Baie min is bekend oor die diversiteit van fungi wat met die Afromontane woude van die Kaapse Floristiese Streek (KFS) van Suid Afrika geassosieer is. Die ophiostomatoïde fungi sluit baie spesies in, sommiges bekend as patogene in die KFS, terwyl ander bekende en belangrike saprofiete in houtdegradasie is. Hierdie studie het op die biodiversiteit en ekologie van die boom-geassosieerde ophiostomatoïde fungi (Ophiostomatales) in die KFS gefokus. Daarbenewens is myte en subkortikale kewers wat met die KFS bome geassosieer word ook versamel, ongeag of hulle geassosieerd was met ophiostomatoïde fungi of nie. „n Relatief hoë diversiteit van ophiostomatoïde fungi is van inheemse bome versamel, tien waarvan hier nuut beskryf is. Drie verdere fungi spesies, twee waarvan ook waarskynlik nuut is tot die wetenskap, is ook vanaf Pinus spesies versamel wat in hierdie woude gegroei het. Vier Ophiostomatales spesies (insluitend drie nuut beskryfde spesies) wat met subkortikale kewers op Rapanea melanophloeos en Olea capensis L. ssp. macrocarpa geassosieer is, is ook versamel. Hulle was Sporothrix pallida, Sporothrix aemuluphilus, Raffaelea scabbardiae en Raffaelea rapaneae, geassosieer met die kewers Lanurgus sp. 1, Ctonoxylon sp. 1, Xyleborinus aemuluphilus en „n Platypodinae spesie. Hierdie verteenwoordig die eerste studie wat die assosiasies tussen subkortikale kewers en ophiostomatoïde fungi op inheemse bome in die KFS ondersoek. Addisioneel tot fungi geassosieer met die subkortikale kewers, is verskeie lede van die Ophiostomatales vanaf wonde op Rapanea melanophloes bome versamel. Hulle sluit in Ophiostoma stenoceras, Sporothrix reniformis, S. rapaneae, S. lunateae en S. noisomeae. Almal behalwe O. stenoceras was nuut tot die wetenskap, en is hier formeel beskryf. Al hierdie wond-geassosieerde spesies vanaf R. melanophloeos behoort aan die Sporothrix schenckii – O. stenoceras kompleks, behalwe vir S. noisomeae wat voorlopig in die S. lignivora kompleks geplaas is. Benewens fungi taxa wat van die wonde op Rapanea melanophloes versamel is, is ander fungi ook vanaf die wonde op ander gasheer boom spesies versamel. Drie verdere ophiostomatoïde fungus spesies is in hierdie nis versamel. Hulle sluit in Sporothix capensis wat vanaf O. capensis ssp. macrocarpa versamel is, Graphilbum roseus wat vanaf baie verskillende, onverwante gasheer bome versamel is en Graphium ilexiense (Microascales), wat vanaf wonde op Ilex mitis versamel is. Laasgenoemde verteenwoordig die eerste isolasie van „n ophiostomatoïde fungus vanaf hierdie gasheer boom spesie. Twee moontlik nuwe fungus spesies (Sporothrix sp. 1, Ceratocystiopsis sp. 1) en Ophiostoma ips, geassosieer met drie baskewers (Orthotomicus erosus, Hylurgus ligniperda en Hylastes angustatus) is vanaf Pinus versamel. Verskeie fungi spesies is van beide inheemse en nie-inheemse bome versamel. Hulle het Sporothrix fusiforme vanaf Brabejum stellatifolium en Acacia mearnsii, O. quercus en O. pluriannulatum-like fungus vanaf verskeie inheemse bome en vanaf A. mearnsii ingesluit. Dit suggereer die moontlikheid van gasheer-skuiwing van sommige van hierdie fungi tussen inheemse en uitheemse gashere of selfs tussen verskillende inheemse gashere. Agt nie- ophiostomatoïde geassosieerde subkortikale kewers was ook versamel en in totaal is meer as 4500 kewer indiwidue versamel. Sommige ophiostomatoïde fungus spesies wat in hierdie studie versamel is, was met ander geleedpotiges soos myte geassosieer. Vier foretiese myt spesies wat met ophiostomatoïde fungi geassosieer is (Dendrolaelaps quadrisetus, Histiogaster sp. 3, Elattoma sp. 1 & 2), is versamel. Nege addisioneële myt spesies was met verskeie ophiostomatoïde spesies vanaf verskeie boomspesies geassosieer (Graphilbum roseus, O. pluriannulatum-like, O. quercus). Dit suggereer dat myte die vervoer van hierdie fungi van een gasheer spesie na die ander mag bewerkstellig. Die moontlike gevolge van die oordrag van Ophiostomatales spesies tussen gashere is getoets deur patogeniteitstoetse. Dit het beklemtoon dat sommige fungi patogenies is op verskeie onverwante boomspesies. Oordraag van spesies is mees waarskynlik in fungi spesies wat vanaf wonde geisoleer is, veral dié wat met myte geassosieer is, want die myte mag hierdie fungi help vervoer. Toe foretiese myte getoets is vir hulle spesifisiteit tot hulle vektore, is hulle hoogs spesifiek bevind. Alhoewel sommige fungi wat met hierdie myte en hulle geassosieerde kewers geassosieer word wel patogenies is, is dit minder waarskylik dat hulle na ander gasheer bome sal verskuif as gevolg van die hoë spesifisiteit van hulle geleedpotige assosiate. Hierdie studie verteenwoordig een van net enkele studies gefokus op ophiostomatoïde fungi, subkortikale kewers en myte wat met bome van die Afromontane woude van Suid-Afrika geassosieer is. Alhoewel ons „n hoë diversiteit van Ophiotomatale lede versamel het, wag baie meer fungi spesies waarskynlik nog op ontdekking. Daar word voorgestel dat toekomstige studies fokus op die komplekse inter-organismiese interaksies in baie van die sisteme wat in hierdie studie blootgelê is.