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HAYASHI, HISAO, TOSHIKUNI TAKlKAWA, KATSUMI KATO, SATOSHI TAKIYA, TSUNEAKI TAGAYA, JUNSUKE KURIKI, MOTOHIRO ARAO, and SHOSHI KATO. "BIOCHEMICAL IMPROVEMENT OF CHRONIC HEPATITIS C AFTER GASTROINTESTINAL BLEEDING." Nagoya University School of Medicine, 1994. http://hdl.handle.net/2237/16077.
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Ahonen, Laura. "An analysis of the perceived values to Northcentral Wisconsin phlebotomist of phlebotomy certification." Online version, 2009. http://www.uwstout.edu/lib/thesis/2009/2009ahonenl.pdf.
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Shiono, Yuhta, Hisao Hayashi, Shinnya Wakusawa, Fujiko Sanae, Toshikuni Takikawa, Motoyoshi Yano, Kenntaro Yoshioka, and Hiros Saito. "Body iron stores and Iron restoration rate in Japanese patients with chronic Hepatitis C as measured during therapeutic Iron removal revealed neither Increased body iron stores nor effects of C282y and H63d mutations on iron indices." Nagoya University School of Medicine, 2001. http://hdl.handle.net/2237/5367.
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Abbas, Mumtaz. "Development, implementation and impact of Phlebotomy training on blood sample rejection and Phlebotomy knowledge of primary health care workers at selected primary health care facilities in Cape Town: a quasi-experimental study design." Master's thesis, University of Cape Town, 2016. http://hdl.handle.net/11427/20331.
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Background: There is an increasing amount of blood sample rejection at primary health care facilities (PHCFs) impacting negatively on the staff, facility, patient and laboratory costs. Aim: The primary objective was to determine the rejection rate and reasons for blood sample rejection at four PHCFs pre and post phlebotomy training. The secondary objective was to determine whether phlebotomy training improved knowledge amongst primary health care providers (HCPs) and to develop a tool for blood sample acceptability. Study Setting: Two Community Health Centres (CHCs) and two Community Day Centres (CDCs) in Cape Town. Methods: A quasi-experimental study design. Results: The sample rejection rate was 0.79% (n= 60) at CHC A, 1.13% (n= 45) at CHC B, 1.64% (n= 38) at CDC C and 1.36% (n= 8) at CDC D pre training. The rejection rates remained approximately the same post training (p>0.05). The same phlebotomy questionnaire was administered pre and post training to HCPs. The average score increased from 6 3% (95% CI 6.97 - 17.03) to 96% (95% CI 16.91 - 20.09) at CHC A (p 0.039), 58% (95% CI 9.09 – 14.91) to 93% (95% CI 17.64 – 18.76) at CHC B (p 0.006), 60% (95% CI 8.84 – 13.13) to 97% (95% CI 16.14 – 19.29) at CDC C (p 0.001) and 63% (95% CI 9.81 – 13.33) to 97% (95% CI 18.08 – 19.07) at CDC D (p 0.001). Conclusion: There is no statistically significant improvement in the rejection rate of blood samples (p>0.05) post training despite knowledge improving in all HCPs (p <0.05).
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Cline, Patricia L. "The effects of modifying teaching methods to accommodate student learning styles." Online version, 1999. http://www.uwstout.edu/lib/thesis/1999/1999clinep.pdf.
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Scanlin, Matthew C. "The Effects of a Hydration Intervention on Cardiac Function, Autonomic Activity, and Cerebral Oxygenation during Phlebotomy." Ohio University / OhioLINK, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1428598371.
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Hormiga, Hernando Gonzalez [UNESP]. "Aplicação de diferentes pinças hemostáticas em veias de equinos: estudo morfológico." Universidade Estadual Paulista (UNESP), 2016. http://hdl.handle.net/11449/144581.
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Foi realizada a avaliação morfológica e morfométrica da veia cefálica submetida à pinçamento de cinco equinos hígidos. Foram testadas as pinças hemostáticas: Diffenbach bulldog, De Bakey bulldog, Rochester reta e De Bakey com cremalheira. Após 15 minutos da aplicação das referidas pinças, foi realizada a flebectomia parcial e coletadas as amostras referentes a cada segmento do vaso pinçado e do segmento controle sem pinçamento. Das peças procedeu-se as preparações histológicas dos segmentos da veia nas colorações de Hematoxilina-Eosina e Tricrômio de Masson, os cortes histológicos foram avaliados por microscopia óptica. Foi realizada análise morfológica das veias, de forma qualitativa, utilizando para isto uma escala de avaliação de lesões vasculares após pinçamento; a avaliação morfométrica, quantificando em micrometros o achatamento produzido pelas pinças nas diferentes camadas vasculares. Em ambos os estudos, morfológico e morfométrico, as pinças com serrilhamento transversal e fechamento tipo cremalheira causaram as maiores alterações, observou se marcada vacuolização das células musculares e desarranjo na túnica media com perda marcada das células endoteliais da túnica intima do vaso estudado.
Morphometric and morphologic evaluation of the cephalic vein of five healthy horses submitted to clamping was done. Hemostatic clamps tested were Dieffenbach bulldog, De Bakey bulldog, Rochester straight and De Bakey with ratchets. 15 minutes after mentioned clamps were applied partial phlebotomy was performed and histologic sections of the veins were prepared and stained with Hematoxylin-Eosin and Masson Trichrome, after the stained preparations were evaluated by light microscopy. A qualitative morphological analysis of the veins was performed using a rating scale of vascular lesions after clamping; the morphometric evaluation consisted in quantifying in micrometers the flattening produced by the hemostatic clamps in the different vascular beds. In both studies, morphologic and morphometric, hemostats with transverse serration and ratcheted mechanism caused major changes, pronounced vacuolization of the muscle cells, derangement of the medium tunic and marked loss of endothelial cells of the intima tunic was observed in the vessel studied.
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Eriksson, Mats. "Aspects of prevention and assessment of neonatal pain /." Linköping, 2003. http://www.bibl.liu.se/liupubl/disp/disp2004/med829s.pdf.
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Wells, Marjorie Joan. "The effects of pets on children's stress responses during medical procedures /." Thesis, Connect to this title online; UW restricted, 1998. http://hdl.handle.net/1773/7351.
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Hormiga, Hernando Gonzalez. "Aplicação de diferentes pinças hemostáticas em veias de equinos estudo morfológico /." Botucatu, 2016. http://hdl.handle.net/11449/144581.
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Orientador: Carlos Alberto HussniBanca: Marcos Jun Watanabe
Banca: Cláudia Helena Pellizzon
Resumo: Foi realizada a avaliação morfológica e morfométrica da veia cefálica submetida à pinçamento de cinco equinos hígidos. Foram testadas as pinças hemostáticas: Diffenbach bulldog, De Bakey bulldog, Rochester reta e De Bakey com cremalheira. Após 15 minutos da aplicação das referidas pinças, foi realizada a flebectomia parcial e coletadas as amostras referentes a cada segmento do vaso pinçado e do segmento controle sem pinçamento. Das peças procedeu-se as preparações histológicas dos segmentos da veia nas colorações de Hematoxilina-Eosina e Tricrômio de Masson, os cortes histológicos foram avaliados por microscopia óptica. Foi realizada análise morfológica das veias, de forma qualitativa, utilizando para isto uma escala de avaliação de lesões vasculares após pinçamento; a avaliação morfométrica, quantificando em micrometros o achatamento produzido pelas pinças nas diferentes camadas vasculares. Em ambos os estudos, morfológico e morfométrico, as pinças com serrilhamento transversal e fechamento tipo cremalheira causaram as maiores alterações, observou se marcada vacuolização das células musculares e desarranjo na túnica media com perda marcada das células endoteliais da túnica intima do vaso estudado.
Abstract: Morphometric and morphologic evaluation of the cephalic vein of five healthy horses submitted to clamping was done. Hemostatic clamps tested were Dieffenbach bulldog, De Bakey bulldog, Rochester straight and De Bakey with ratchets. 15 minutes after mentioned clamps were applied partial phlebotomy was performed and histologic sections of the veins were prepared and stained with Hematoxylin-Eosin and Masson Trichrome, after the stained preparations were evaluated by light microscopy. A qualitative morphological analysis of the veins was performed using a rating scale of vascular lesions after clamping; the morphometric evaluation consisted in quantifying in micrometers the flattening produced by the hemostatic clamps in the different vascular beds. In both studies, morphologic and morphometric, hemostats with transverse serration and ratcheted mechanism caused major changes, pronounced vacuolization of the muscle cells, derangement of the medium tunic and marked loss of endothelial cells of the intima tunic was observed in the vessel studied.
Mestre
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Thibeault, RaeAnn. "A comparison of the efficacy of three methods of antecubital fossa skin disinfection prior to phlebotomy in volunteer blood donors." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1998. http://www.collectionscanada.ca/obj/s4/f2/dsk2/tape17/PQDD_0013/MQ32263.pdf.
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Marchais, Régine. "Speciation et vicariance chez les larroussius du groupe perniciosus (diptera : psychodidae)." Reims, 1992. http://www.theses.fr/1992REIMP202.
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FLOER, ROGUET MARIE-THERESE. "Etude isoenzymatique d'une population de phlebotomus ariasi tonnoir, 1921." Strasbourg 1, 1991. http://www.theses.fr/1991STR15015.
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Gradin, Maria. "Procedural pain reducing methods and pain assessment in newborns /." Linköping : Univ, 2004. http://www.bibl.liu.se/liupubl/disp/disp2004/med868s.pdf.
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Silva, Nixon Ramos da. "Punção periférica da jugular externa para coleta de células progenitoras hematopoéticas de sangue periférico." Universidade de São Paulo, 2017. http://www.teses.usp.br/teses/disponiveis/17/17155/tde-08062017-094754/.
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Introdução: O transplante autólogo de células progenitoras hematopoéticas é indicado para tratamento de várias doenças neoplásicas e não-neoplásicas, tais como linfoma, mieloma múltiplo e doenças autoimunes. Para isso, é imprescindível coletar as células progenitoras hematopoéticas (CPHs), em geral mobilizadas para o sangue periférico, por meio de equipamentos de aférese e acesso venoso adequado, seja de veia periférica seja de veia central quando a primeira não for adequada. Usualmente, o acesso periférico se dá por veia da fossa cubital. Entretanto, a veia jugular externa também poderia ser uma opção em pacientes em que a veia cubital não proporcionar fluxo sanguíneo adequado, uma possibilidade ainda muito pouco explorada nesse contexto. Objetivos: Avaliar a viabilidade de coletar CPHs pela veia jugular externa em pacientes que não tem acesso por meio da veia da fossa cubital.Métodos: Trata-se de estudo observacional do tipo transversal, com coleta retrospectiva dos dados por meio da análise das fichas médicas arquivadas no Hemonúcleo do Hospital de Câncer de Barretos. A amostra foi composta de 26 indivíduos, sendo que 13 (50%) dos participantes tiveram a punção da veia jugular externa e os outros 13 (50%) foram extraídos aleatoriamente do conjunto de 913 pacientes que tiveram a punção da veia da fossa cubital para equilíbrio numérico das amostras, no período de 2007 a 2014. Resultados: Dos 26 participantes do estudo, 19 (76,9%) foram do sexo masculino e 7 (23,1%) do sexo feminino. Entre as doenças, o mieloma foi a mais prevalente, não houve diferença quanto ao diagnóstico entre os dois grupos fossa cubital e jugular. As médias de células CD34+ coletadas no grupo fossa cubital e no grupo jugular foram de 10,0 x 106/kg e 5,1 x 106/kg, respectivamente (p=0,20). As volemias processadas foram de 4,2 e 4,4 para os grupos fossa cubital de jugular (p=0,12). O fluxo médio de aspiração do sangue foi de 69±17 mL/minuto e 62±17 mL/minuto (p=0,29). Conclusão: Há viabilidade da coleta de CPH por via jugular externaIntroduction: Autologous hematopoietic progenitor cell transplantation is indicated for the treatment of various neoplastic and non-neoplastic diseases, such as lymphoma, multiple myeloma, and autoimmune diseases. For this, it is essential to collect the hematopoietic progenitor cells (CPHs), usually mobilized into the peripheral blood, by means of apheresis devices and adequate venous access, either peripheral vein or central vein when the former is not adequate. Usually, the peripheral access is by the ulnar fossavein. However, the external jugular vein could also be an option in patients where the ulnar vein does not provide adequate blood flow, a possibility still very little explored in this context. Objectives: To evaluate the feasibility of collecting CPHs through the external jugular vein in patients who do not have access through the ulnar fossa vein. Methods: It is an observational cross-sectional study, with retrospective data collection through the analysis of the medical records filed at the Barretos Cancer Hospital. The sample consisted of 26 individuals; 13 (50%) of the participants had external jugular vein puncture and the other 13 (50%) were randomly extracted from a set of 913 patients who had ulnar fossa vein puncture to numerical equilibrium of the samples from 2007 to 2014. Results: Of the 26 participants in the study, 19 (76.9%) were male and 7 (23.1%) were female. Among the diseases, myeloma was the most prevalent; there was no difference in diagnosis between the two groups of ulnar and jugular fossa. The mean CD34 + cells collected in the cubital fossa group and in the jugular group were 10.0 x 106/kg and 5.1 x 106/kg, respectively (p=0.20). The blood volumes processed were 4.2 and 4.4 for the jugular fossa group (p=0.12). The mean blood aspiration flow was 69 ± 17 mL/min and 62 ± 17 mL/min (p=0.29). Conclusion: there is viability of the collection of CPHs by external jugular vein
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Gillström, Alice, and Olivia Lundberg. "En kartläggning av effekten av olika distraktionsmetoder för att minska smärta vid venpunktion på barn 6-12 år : En litteraturstudie." Thesis, Umeå universitet, Institutionen för omvårdnad, 2021. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-178773.
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Bakgrund: Som grundutbildad sjuksköterska kan du möta barn inom olika verksamheter utanför den typiska barnsjukvården. Venpunktion är en smärtsam procedur och ett vanligt förekommande moment i sjukvården. Smärtan kan lindras och förebyggas med farmakologiska och icke-farmakologiska metoder, exempelvis aktiva och passiva distraktionsmetoder. Syfte: Syftet med studien var att kartlägga effekten av olika distraktionsmetoder för att minska smärta hos barn (6-12 år) vid venpunktion. Metod: En kvantitativ litteraturstudie med sammanställning av 11 interventionsstudier identifierade i PubMed och Cinahl. Resultat: Nio studier visade på statistiskt signifikant smärtlindrande effekt för åtta av elva distraktionsmetoder. Tre av sex studier som undersökte dess effekt på ångest visade på statistiskt signifikant effekt. Två studier presenterade i respektive studie en distraktionsmetod med bättre effekt, en av dessa presenterade att aktiv distraktion gav bättre effekt än passiv distraktion. Konklusion: Distraktionsmetoder kan antas ha både smärtlindrande och ångestdämpande effekt vid venpunktion på barn 6-12 år. Distraktionsmetoder bör vara aktuella att integrera i rutinen för venpunktion på barn inom svensk sjukvård. Det finns dock behov av vidare forskning inom området för att stärka dess evidens. Aspekter som distraktion som tillägg till topikal anestesi, effekt på ångest samt distraktion anpassat till barnets behov och förmågor bör studeras ytterligare.Background: A registered nurse can meet children in different healthcare departments besides the pediatric department. Phlebotomy is a common painful procedure in healthcare. This pain can be relieved and prevented with pharmacological and non-pharmacological methods, for example, active and passive distraction methods. Aim: The study aimed to collect an overview of the effects of different distraction methods to reduce pain during phlebotomy in children 6-12 years. Methods: A quantitative literature study identified and compared 11 intervention studies collected from PubMed and Cinahl. Results: Nine studies presented a statistically significant pain-relieving effect for eight of the total eleven distraction methods. Three out of six studies presented a statistically significant effect on anxiety. Two studies each presented one distraction method with better effect, one of these presented active distraction to be better than passive. Conclusion: Distraction methods present pain-relieving and anxiety-reducing effects during phlebotomy in children 6-12 years and could therefore be appropriate to integrate into the Swedish phlebotomy routine for children. There is further need for research in the field to strengthen its evidence. Aspects such as distraction in addition to topical anesthesia, the effect on anxiety and distraction based on the child's needs and abilities need further research.
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鄒惠娣. "井穴刺絡放血的臨床研究." HKBU Institutional Repository, 2010. http://repository.hkbu.edu.hk/etd_ra/1127.
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Günter, Christina Irene [Verfasser], Hans-Günther [Akademischer Betreuer] Machens, and Laszlo [Akademischer Betreuer] Kovacs-Hintz. "Retrospektive Evaluation zur Lebensqualität nach Seitenastexherese mittels Transilluminierter Powered Phlebotomy (TPP), unter Verwendung einer modifizierter Operationstechnik und des SQOR-V / Christina Irene Günter. Betreuer: Hans-Günther Machens. Gutachter: Hans-Günther Machens ; Laszlo Kovacs-Hintz." München : Universitätsbibliothek der TU München, 2015. http://d-nb.info/1081767987/34.
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Bölenius, Karin. "Improving venous blood specimen collection practices : method development and evaluation of an educational intervention program." Doctoral thesis, Umeå universitet, Institutionen för omvårdnad, 2014. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-87015.
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Background: About 60%–80% of decisions regarding diagnosis and treatment are based on laboratory test results. Low adherence to venous blood specimen collection (VBSC) guidelines may lead to erroneous or delayed test results, causing patient harm and high healthcare costs. Educational intervention programs (EIPs) to update, improve and sustain VBSC practices are seldom evaluated. After testing a self-reported venous blood sampling questionnaire, the overall aim of this thesis was to evaluate the impact of a large-scale EIP on healthcare personnel’s VBSC practices. Methods: The study settings were primary healthcare centres (PHCs) in northern Sweden. Participants were VBSC personnel. Data consisted of a VBSC questionnaire of self-reported practices, records of low-level haemolysis index in serum samples (specimen quality indicator), and interviews reflecting VBSC practices. First, experts on questionnaires and VBSC were consulted, and test-retest statistics were used when testing the VBSC questionnaire for validity and reliability. Thereafter, we evaluated the impact of a short, large-scale EIP with a before-after approach comparing self-reported VBSC questionnaire of two county councils. The personnel of the county councils (n = 61 PHCs) were divided into an intervention group (n = 84) and a corresponding control group (n = 79). In order to test changes in blood specimen quality we monitored haemolysis in serum samples (2008, n = 6652 samples and 2010, n = 6121 samples) from 11 PHCs. Finally, 30 VBSC personnel from 10 PHCs reported their experiences. The interview questions were open-ended with reflective elements and the interviews were analysed by qualitative content analysis. Results: The VBSC questionnaire was found to be valid and could be used to identify risk of errors (near misses) and evaluate the impact of an EIP emphasising VBSC guideline adherence. The intervention group demonstrated several significant improvements in self-reported practices after the EIP, such as information search, patient rest, test request management, patient identification, release of venous stasis, and test tube labelling. The control group showed no significant improvements. In total, PHCs showed minor differences in blood specimen quality. Interviews summarized VBSC personnel experiences in the overall theme: education opened up opportunities for reflection about safety. Conclusion: This thesis is, to our knowledge, the first to evaluate the impacts of a large-scale EIP on VBSC practices. The VBSC questionnaire and monitoring for low-level haemolysis reflected VBSC practices. The frequently occurring near-miss markers made it possible to compare and benchmark VBSC practices down to the healthcare unit and hospital ward. The short, general EIP opened up opportunities for reflection about safety and improved VBSC practices in PHCs with larger deviations from guidelines. EIPs that provide time for reflection and discussion could improve VBSC further. Directed EIPs focused on specific VBSC flaws might be more effective for some near misses in VBSC practices, while some near misses must be changed at a different level in the system. Clinical relevance: Our results indicate that monitoring and counteracting the near misses in VBSC practices is a well-functioning preventive action. We propose that the VBSC monitoring instruments (VBSC questionnaire & haemolysis index) we used and the EIP strategy proposed should be tested in additional countries with different healthcare settings. It is suggested that a national program intended to identify near misses and prevent VBSC errors be developed in the healthcare system. General e-learning programs may be cheaper than, and as effective as, the EIP program and may be performed everywhere and any time. Systematic planning, useful for reflection and with focus on the specific elements in a skill, together with VBSC guidelines, could probably increase improvements. Our studies have led to deeper and extended knowledge of the impact of an EIP on VBSC practices. Our results can be used when considering future VBSC practice interventions. Using a model for practical skills in nursing to describe VBSC in a more holistic and less technical way might highlight VBSC as a practical nursing skill.Bakgrund: Av kliniska beslut angående diagnostik och behandling baseras 60%–80% på laboratorieresultat. Därför är det helt nödvändigt att laboratorieresultat är tillförlitliga. Låg följsamhet till provtagnings anvisningar kan leda till felaktiga och fördröjda analysresultat, förorsaka skada och lidande för patienter och utgöra en stor kostnad för hälso- och sjukvården. Felaktiga provsvar beror till stor del på felaktig provtagning och provhantering och går oftast att undvika. Interventioner som avser att uppdatera och säkra korrekt venprovtagning kan leda till förbättringar men genomförda interventioner har sällan utvärderats. Efter att en enkät för självrapporterad venprovtagning testats för validitet och reliabilitet genomfördes ett omfattande interventionsprogram som utvärderades med hjälp av den testade enkäten och andra utvärderingsmått. Det övergripande syftet var att utvärdera i vilken utsträckning interventionsprogrammet påverkade provtagande personals praktiska utförande av venprovtagning. Metoder: Studierna i denna avhandling omfattade provtagande personal vid hälsocentraler i norra Sverige. För datainsamling användes en enkät som mäter självrapporterad venprovtagning, förekomst av låggradig hemolys (indikator på blodprovets kvalitet) och intervjuer. Initialt testades enkätens förmåga att mäta vad som avsetts (validitet) och testades enkätens förmåga att vid upprepade mätningar vara tillräckligt stabil (reliabilitet) för att användas i interventionsstudier. Därefter utvärderades ett kort men storskaligt interventionsprogram i preanalys inkluderande venprovtagning med före och efter mätningar. Vi jämförde provtagande personal från två landsting vid 61 hälsocentraler. Landstingens personal delades upp i en interventionsgrupp (n=84) och en motsvarande kontrollgrupp (n = 79). För att mäta kvaliteten av blodproverna extraherades uppgifter om hemolys i serumprover (2008, n = 6652 blodprov) och (2010, n = 6121 blodprov) från elva hälsocentraler i ett landsting. Slutligen, intervjuades 30 provtagande personal från 10 hälsocentraler efter att de deltagit i interventionsprogrammet. Intervjuerna var öppna och genererade korta berättelser och analyserades med innehållsanalys. Resultat: Venprovtagningsenkäten befanns vara valid och kan användas för att utvärdera personalens följsamhet till provtagningsanvisningar i venprovtagning och identifiera riskhändelser. Interventionsgruppen visade flera signifikanta förbättringar i självrapporterat utförande av venprovtagning såsom förbättrad informationssökning, vila inför provtagning, remissförfarande, kontroll av patientidentitet, användning av stas och etikettering av provrör. Kontrollgrupen visade inga signifikanta förbättringar. Blodprovskvaliteten visade små skillnader. Provtagande personals erfarenheter från intervjuerna sammanfattades i ett övergripande tema; utbildningsinsatsen öppnade upp möjligheter för reflektion om säkerhet. Slutsats: Avhandlingen är så vitt vi vet den första att utvärdera effekten av ett storskaligt interventionsprogram med hjälp av självrapporterat utförande av venprovtagning och blodprovers kvalitet (låggradig hemolys). Med dessa metoder ökar andelen riskhändelser så att jämförelser kunde göras även på enhetsnivå och avdelningsnivå. Utbildningsprogrammet öppnade upp för reflektioner om säkerhet och förbättrade utförande av venprovtagning vid enheter med större brister. Utbildningsprogram som öppnar upp för reflektion och diskussion kan leda till ökad patientsäkerhet i hälso- och sjukvården. Trots utfallet av resultaten, är riktade utbildningsinsatser för sjukvårdsenheter som uppvisar specifika brister troligtvis mer effektiva än breda utbildningsinsatser. Klinisk betydelse: Interventionsprogram avseende preanalys och venös provtagning förbättrade personalens praktiska utförande. Monitorering av och åtgärder för att minska riskhändelser är väl fungerande preventiva åtgärder. Instrumenten (självrapporterande enkät och hemolys) bör också testas i andra kontexter inom hälso- och sjukvården. Ett externt nationellt program för att identifiera och förebygga riskhändelser bör utvecklas i hälso- och sjukvården. Interventioner i form av e-lärande kan då vara ett alternativ som är billigt och effektivt. Dessutom kan systematisk planering och genomförande med fokus på reflektion av specifika delar i en färdighet vara effektivt för att uppnå förbättringar. Våra studier har bidragit till en djupare och utökad kunskap om effekten av ett interventionsprogram på utförande av venprovtagning. Resultaten kan användas vid framtida planering av utbildningsinsatser. Modeller för praktiskt färdighetsutövande inom omvårdnad kan beskriva venprovtagning ur ett helhetsperspektiv och synliggöra venprovtagning som en viktig praktisk färdighet inom omvårdnad.
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Nicholl, Analise. "Investigating cardiometabolic health and child-centred research in young children participating in The Milky Way Study: A randomised controlled dairy intervention." Thesis, Edith Cowan University, Research Online, Perth, Western Australia, 2022. https://ro.ecu.edu.au/theses/2585.
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Background
There is limited evidence to support the common public health guideline that children over 2 years should consume mostly reduced-fat dairy. Recruitment can be an issue in paediatric research, potentially due to confronting child assessments, and there is a growing international momentum to develop respectful research and healthcare protocols for children.
Aims
The primary aim of this thesis was to investigate the effects of whole-fat compared with reduced-fat dairy intake on measures of adiposity and cardiometabolic risk factors in healthy 4 to 6-year-old children. Secondary aims were to investigate: 1) differential changes in erythrocyte fatty acids (FAs) with changes in dairy fat consumption, and 2) to develop and test a child-centred approach in a paediatric clinical trial.
Methods
The Milky Way Study enrolled 49 children (mean ± SD age: 5.2 ± 0.9 years; 47% girls) who were habitual consumers of whole-fat dairy, then randomly assigned them in a double-blind fashion to remain on whole-fat dairy or change to reduced-fat products for 3 months. Primary endpoints included measures of adiposity, body composition, blood pressure, fasting serum lipids, blood glucose, glycated haemoglobin (HbA1c), and C-reactive protein (CRP); together with weighed 3-day food records and erythrocyte fatty acids, these were assessed at baseline and study end. Pre- and post-intervention results were compared using SPSS (version 28) linear mixed models, adjusted for growth, age, and sex. The Milky Way Study Respectful Approach to Child-centred Healthcare (ReACH) incorporated findings from pre-trial community consultation with parents. It was evaluated across all study clinics using mixed methods, with evaluation triangulated from 3 stakeholder perspectives.
Results
Dairy fat intake was significantly decreased in the reduced-fat compared with the whole-fat dairy group (p = 0.003), confirmed by significant changes in erythrocyte pentadecanoic acid, whereas dietary energy intakes remained similar (p = 0.936). We found no significant adjusted differential changes between dairy groups in any measure of adiposity, body composition, blood pressure, or fasting serum lipids, glucose, HbA1c, and CRP. Adjusted mixed models showed significant differential reductions over 3 months in the reduced-fat compared with the whole-fat dairy group for erythrocyte saturated fatty acids myristic acid (p = 0.043) and pentadecanoic acid (p < 0.001), but also for omega-3 fatty acids, EPA (p < 0.001) and DPA (p = 0.004). Children achieved high compliance rates for assessments, and adherence to study ReACH principles during clinic visits was positively associated with child compliance. Parents rated the Milky Way Study very highly, and qualitative feedback reflected an enjoyable study experience for parents and children.
Conclusions
Results do not support the guideline that reduced-fat dairy products are preferable over whole-fat for young children. Dairy fat intake can significantly impact erythrocyte fatty acid composition in young children. Adherence to the ReACH approach was associated with high child compliance rates and high participant satisfaction with the study.
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Söderberg, Johan. "Sources of preanalytical error in primary health care : implications for patient safety." Doctoral thesis, Umeå universitet, Klinisk kemi, 2009. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-21256.
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Background Venous blood tests constitute an important part in the diagnosis and treatment of patients. However, test results are often viewed as objective values rather than the end result of a complex process. This has clinical importance since most errors arise before the sample reaches the laboratory. Such preanalytical errors affect patient safety and are often due to human mistakes in the collection and handling of the sample. The preanalytical performance of venous blood testing in primary health care, where the majority of the patients contact with care occurs, has not previously been reported. Aims To investigate venous blood sampling practices and the prevalence of haemolysed blood samples in primary health care. Methods A questionnaire investigated the collection and handling of venous blood samples in primary health care centres in two county councils and in two hospital clinical laboratories. Haemolysis index was used to evaluate the prevalence of haemolysed blood samples sent from primary health care centres, nursing homes and a hospital emergency department. Results and discussion The results indicate that recommended preanalytical procedures were not always followed in the surveyed primary health care centres. For example, only 54% reported to always use name and Swedish identification number, and 5% to use photo-ID, the two recommended means for patient identification. Only 12% reported to always label the test tubes prior to blood collection. This increases the possibility of sample mix-up. As few as 6% reported to always allow the patient to rest at least 15 minutes before blood collection, desirable for a correct test result. Only 31% reported to have filed an incident report regarding venous blood sampling, indicating underreporting of incidents in the preanalytical phase. Major differences in the prevalence of haemolysed blood samples were found. For example, samples collected in the primary health care centre with the highest prevalence of haemolysed samples were six times (95% CI 4.0 to 9.2) more often haemolysed compared to the centre with the lowest prevalence. The significant variation in haemolysed samples is likely to reflect varying preanalytical conditions. Conclusions This thesis indicates that the preanalytical procedure in primary health care is associated with an increased risk of errors with consequences for patient safety and care. Monitoring of haemolysis index could be a valuable tool for estimating preanalytical sample quality. Further studies and interventions aimed at the preanalytical phase in primary health care are clearly needed.
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Le, Pont François. "Les phlebotomes et les leishmanioses en bolivie." Paris 11, 1990. http://www.theses.fr/1990PA112171.
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Des recherches sur les phlebotomes et les leishmanioses ont ete menees sur un transect s'etendant de la cordillere orientale des andes (2000 m) jusqu'a la plaine amazonienne (170 m). 1) 72 especes de phlebotomes ont ete inventoriees. 54 n'etaient pas connues de bolivie. Sept especes nouvelles ont ete decrites, dont deux, psychodopygus yucumensis et lutzomyia nuneztovari anglesi sont des vecteurs de leishmaniose tegumentaire. Le nombre d'especes anthropophiles diminue de la foret amazonienne aux contreforts andins, de 21 a 3, et la densite de piqures suit la meme tendance (de 40 h/h a 2,6 h/h). 2) leishmania (v. ) braziliensis, agent de la leishmaniose cutaneo-muqueuse, est present partout au-dessous de 2000 m. Dans les yungas, la transmission est principalement domestique et le vecteur presume est lu. N. Anglesi. Dans l'alto beni et le pando, l'homme entre dans le cycle sylvatique la journee, et les vecteurs sont ps. Yucumensis, ps. Llanosmartinsi, ps. C. Carrerai. Les formes evolutives n'apparaissent que chez 7% des sujets infectes dans les yungas, et 1,4% dans le pando, les autres guerissant spontanement. 3) la leishmaniose viscerale a le. (le. ) chagasi est cantonnee dans les yungas ou son vecteur est lu. Longipalpis. Rare chez l'homme, elle touche pratiquement tous les chiens. Le parasite est identique a le. (le. ) infantum. 4) dans les yungas des aspersions domiciliaires de deltamethrine a 0,025 g/m#2 ont donne de bons resultats concernant lu. Longipalpis, plus discutables concernant lu. N. Anglesi
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STOECKEL, GARCIA MARIA DE LA PAZ. "Contribution a l'etude des phlebotomes de l'ile de teneriffe (canaries)." Strasbourg 1, 1992. http://www.theses.fr/1992STR15083.
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BENABDENNBI, CHERKAOUI IHSANE. "Contribution a l'etude de la biodiversite chez les larroussius (diptera : psychodidae) du bassin mediterraneen : apports de l'analyse isoenzymatique." Université Louis Pasteur (Strasbourg) (1971-2008), 1998. http://www.theses.fr/1998STR10022.
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Doehl, Johannes. "Investigating the role of the Leishmania (Leishmania) major HASP and SHERP genes during metacyclogenesis in the sand fly vectors, Phlebotomus (Phlebotomus) papatasi and Ph. (Ph.) duboscqi." Thesis, University of York, 2013. http://etheses.whiterose.ac.uk/4807/.
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Leishmania parasites are the causative agents of a diverse spectrum of infectious diseases termed the leishmaniases. These digenetic parasites exist as intracellular, aflagellate amastigotes in a mammalian host and as extracellular flagellated promastigotes within phlebotomine sand fly vectors of the family Phlebotominae. Within the sand fly vector’s midgut, Leishmania has to undergo a complex differentiation process, termed metacyclogenesis, to transform from non-infective procyclic promastigotes into mammalian-infective metacyclics. Members of our research group have shown previously that parasites deleted for the L. (L.) major cDNA16 locus (a region of chromosome 23 that codes for the stage-regulated HASP and SHERP proteins) do not complete metacyclogenesis in the sand fly midgut, although metacyclic-like stages can be generated in in vitro culture (Sádlová et al. Cell. Micro.2010, 12, 1765-79). To determine the contribution of individual genes in the locus to this phenotype, I have generated a range of 17 mutants in which target HASP and SHERP genes are reintroduced either individually or in combination into their original genomic locations within the L. (L.) major cDNA16 double deletion mutant. All replacement strains have been characterized in vitro with respect to their gene copy number, correct gene integration and stage-regulated protein expression, prior to phenotypic analysis. HASPA1 was not detected in cultured promastigotes, but was expressed in mouse isolated amastigotes. Parasite mutant lines were passaged through susceptible BALB/c mice, during which HASPA2 gene containing mutant lines, in the absence of a HASPA1 gene, were shown not to develop lesions. Mouse-passaged parasites were used to infect the L. (L.) major specific sand fly vectors, Ph. (Ph.) papatasi and Ph. (Ph.) duboscqi. The progress of parasite metacyclogenesis was then monitored over twelve days, by midgut dissection and microscopy. Metacyclogenesis was not fully recovered in any of the replacement mutants tested. Surprisingly, HASPB protein expression could not be detected in the replacement mutants within the sand fly midgut, although HASPB protein was readily detected when the same parasite lines were cultured in vitro. The same was true for SHERP, although in situ expression was recovered in the presence of a HASPB gene, which itself did not expressed detectable HASPB protein levels. These observations suggest a requirement for one or multiple as-yet-unidentified regulatory component(s) for HASPB expression within the sand fly midgut and these are not required in culture. Quantitative PCR data suggested HASPB upregulation to be essential for metacyclogenesis completion, suggesting a sand fly specific function for HASPB.
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Sigle, Leah T. "Kazal-type serine proteinase inhibitors in the midgut of Phlebotomus papatasi." Thesis, Kansas State University, 2011. http://hdl.handle.net/2097/13140.
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Master of ScienceDepartment of Entomology
Marcelo Ramalho-Ortigao
Sand flies (Diptera:Psychodidae) are vectors of parasites of the genus Leishmania transmitted to suitable vertebrate host during blood feeding. For blood feeding arthropods, including sand flies, blood meal digestion requires the secretion of inhibitory molecules, such as Kazal-type serine proteinase inhibitors that are involved in preventing the blood from coagulating within the mouthparts and the midgut. Previous studies have identified such molecules in mosquitoes, ticks, and triatomine bugs. Following studies of the midgut transcriptome of Phlebotomus papatasi, the principal vector of Leishmania major, two non-classical Kazal-type serine proteinase inhibitors were identified (PpKzl1 and PpKzl2). We are interested in the role of these proteins as inhibitors of coagulation cascades, in addition to their potential effects on blood digestion in P. papatasi. Ppkzl1 is similar to thrombin and trypsin inhibitors in triatomines and mosquitoes and Ppkzl2 is similar to Kazal-type inhibitors in mosquitoes with unknown function. Analyses of expression profiles indicated that although both transcripts are expressed prior to blood feeding in the midgut of P. papatasi they are tightly regulated by the blood meal. Reverse genetics studies using RNAi-targeted knockdown of PpKzl1 and PpKzl2 by dsRNA injection did not result in a detectable effect on mRNA expression levels. Thus, we expressed a recombinant PpKzl2 in a mammalian expression system (CHO-S free style cells) that was applied to in vitro studies to assess activity against various serine proteinases. Recombinant PpKzl2 inhibited chymotrypsin at nanomolar levels and also inhibited thrombin and trypsin at micromolar levels, suggesting that native PpKzl2 is an active serine proteinase inhibitor and may regulate digestive enzymes and thrombin in the midgut. Leishmania development within the sand fly midgut is faced with several barriers that can severely impact the parasites. For transmission to occur, parasites must be able to overcome these barriers including digestive proteinases, escape from the peritrophic matrix, and midgut attachment. Early stages of Leishmania are susceptible to killing by digestive proteinases in the sand fly midgut. Thus, targeting serine proteinase inhibitors may provide a new strategy to prevent transmission of Leishmania.
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Haddad, Nabil. "Les phlebotomes du liban : inventaire et corollaire eco-epidemiologique (doctorat : parasitologie)." Reims, 1998. http://www.theses.fr/1998REIMP208.
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Marques, Maria Inês Lopes de Moura. "Leishmaniose canina." Bachelor's thesis, Universidade Técnica de Lisboa. Faculdade de Medicina Veterinária, 2008. http://hdl.handle.net/10400.5/950.
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Dissertação de Mestrado Integrado em Medicina VeterináriaA Leishmaniose canina é uma das principais zoonoses da actualidade, causada pelo protozoário Leishmania infantum, espécie que existe predominantemente na Bacia Mediterrânica. A forma infectante do parasita corresponde à forma promastigota desenvolvida no tracto intestinal do insecto género Phlebotomus, que é o vector biológico que transmite a doença ao canídeo. Pensa-se que a espécie mais predominante na Bacia Mediterrânea e consequentemente em Portugal seja o Phlebotomus perniciosus. Sendo a Leishmaniose uma doença sistémica de curso crónico e implicação viscero-cutânea, apresenta grande variabilidade clínica. O diagnóstico deve ser o mais precoce possível e a terapêutica instituída rigorosamente cumprida. É necessária monitorização laboratorial preferencialmente de seis em seis meses. No que concerne ao estudo retrospectivo, foram observados 35 canídeos com leishmaniose no período que compreendeu o estágio curricular no Hospital escolar da FMV. Apresentaram uma panóplia de sinais clínicos na consulta hospitalar, maioritariamente diminuição do apetite, prostração, linfoadenopatia generalizada e um conjunto de sintomatologia dermatológica, entre outros. Foi instituída terapêutica, na maioria dos casos alopurinol em monoterapia (44%) e associação medicamentosa de alopurinol com antimoniato de meglumina (47%), na maior parte dos casos houve resposta terapêutica eficaz, com melhoria da sintomatologia pré-existente e normalização das alterações laboratoriais.
ABSTRACT - Canine Leishmaniosis is one of the most important zoonosis nowadays, caused by the Leishmania infantum protozoan agent, which is the most common species of Leishmania in the Mediterranean basin. The promastigote organism is the infectious form of the parasite, contained in the intestinal tract of the Phlebotomus insect, the biological vector of the disease in dogs. It is thought that the Phlebotomus perniciosus is the most common species in the Mediterranean basin and Portugal. Leishmaniosis is a systemic disease with a chronic evolution and visceral-cutaneous involvement, having different clinical presentations. The diagnosis must be early defined and the therapeutics must be rigorously obeyed. It is necessary a laboratorial monitorization at each six months. In this study, 35 dogs with Leishmaniosis were observed during the curricular work accomplished at the School Hospital, Faculty of Veterinary Medicine. A large diversity of clinical signs was found in these animals, most of them having low appetite, prostration, generalized lymphadenopathy, dermatological manifestations and so on. In most cases was made a therapeutic approach with allopurinol only (44%) and an association of allopurinol and meglumine antimoniate (47%); most of the animals had a good therapeutic response, with the initial symptoms being eliminated and the laboratory parameters normalized.
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Yaman, Khatijah. "Semiochemical mediated oviposition and mating in Phlebotomus argentipes (Diptera: Psychodidae) sand flies." Thesis, Keele University, 2016. http://eprints.keele.ac.uk/2445/.
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Phlebotomus argentipes (Diptera: Psychodidae) is an important vector responsible for the transmission of Leishmania donovani that causes visceral leishmaniasis (VL) or kala-azar, in the sub-continent of India. The aims of this study were to investigate the semiochemicals that mediate oviposition and mating behaviour and also the courtship behaviours in P. argentipes. The result of ovipositional behaviour bioassays shows gravid P. argentipes females preferred to oviposit their eggs in the present of conspecific eggs and also eggs extract. This suggests the presence of an oviposition pheromone on the surface of the eggs which can be removed by washing with an organic solvent and transferred to an alternative surface. A Y-tube olfactometer was used to test an upwind anemotactic response of virgin females to male headspace volatiles and male extract, in the presence or absence of host odour. The results strongly suggest that a volatile maleproduced sex pheromone is present in P. argentipes. The results also suggest that under certain circumstances of the age of males and females and the presence of host odour, the females are attracted to live male and male extract of P. argentipes. Thus, presence of host odour might have a synergistic effect on the male-produced sex pheromone. Quantitative description and detailed of courtship behaviour(s) in both males and females of P. argentipes were observed. The results show that male behaviours during courtship are vital for the success of the mating. These predictor behaviours include approach wingflapping, abdomen bending and copulation attempt by male P. argentipes. Understanding of the biology, ecology and chemical mediated behaviour in P. argentipes will enhance and widen the knowledge leading to the improved of the efficiency and efficacy of the current sand fly control programmes.
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Cannet, Arnaud. "Insectes hématophages : terrains, élevages, infections : études chez Cimex lectularius et Phlebotomus perniciosus." Thesis, Aix-Marseille, 2016. http://www.theses.fr/2016AIXM5010.
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Les insectes hématophages impactent sur la santé humaine et animale par la nuisance et l’allergie induite par la piqûre ou par leur risque de transmission d’agents pathogènes. Les étudier permet de mieux prévenir, gérer ou traiter l’ensemble des pathologies induites. Au cours de cette thèse, nous avons étudié deux insectes que sont la punaise de lit et le phlébotome.- Pour les punaises de lit une revue de la littérature sur les méthodes d'élevage a été rédigée. Nous avons déterminé deux aspects que sont les facteurs physiques (température, humidité relative, photopériode) et physiologiques (type, fréquence des repas) à prendre en considération. Un travail de terrain de recueil de spécimens a permis d’étudier la génétique des spécimens par localités et d’évaluer le portage de pathogènes notamment la bactérie Wolbachia sp.- Pour le phlébotome, un élevage de l’espèce Phlebotomus perniciosus a été mis en place permettant d'évaluer deux modèles infectieux expérimentaux à Leishmania infantum sur souris BALB/c et sur gorgeur artificiel. Des captures de terrain ont été réalisées et une étude de la génétique de population de Phlebotomus ariasi a été publiée. Une revue de la littérature sur les principaux événements historiques et évolutifs des Leishmanies, des phlébotomes et des réservoirs animaux associés a été publiée.- Dans le cadre de cette thèse nous avons collaboré à un nouveau mode d’identification des diptères hématophages (dépôt d’un brevet).- Au cours de ces 3 années de thèse, nos activités entomologiques hospitalières ont permis la description de cas entomologiques originaux.Cette thèse a fait l'objet d'une bourse CIFRE financée par les Laboratoires VIRBACBloodsucking insects have an impact on the human and animal health particularly for the risk of pathogens transmission and the nuisance caused by their bite. To have knowledge on these insects can help and apply to prevent, manage or treat the diseases. In this thesis, we mainly focused on two insects: bedbugs and sandflies.- For bedbug, a literature review was carried out to classify the published data demonstrating preferred bedbug colony conditions. We showed that physical factors (temperature, relative humidity, photoperiod), and physiological factors (type, frequency of blood meals) play important roles in the laboratory colonies. A huge number of Cimex lectularius species was sampled from infested regions to study the spatial genetic structure and restricted gene flow in bed bugs in France and to evaluate the symbiotic flora including Wolbachia bacteria.- For sandfly, Phlebotomus perniciosus species was colonized for experimental infections by Leishmania infantum using murine model and artificial feeder. A local sandfly sampling was carried out in the rural regions of Nice city and were served in a study targeting the genetic population structure of Phlebotomus ariasi. Moreover, a historical study was conducted on the classification, evolution and dispersion of Leishmania parasites, sandflies and animals reservoirs.- A new identification method of bloodsucking Diptera was invented (submitted patent).- Thanks to the hospital-entomological activities, an original case report of phthiriasis on an old person was reported.- During this thesis with a scholarship CIFRE funded by Virbac Laboratories, I succeeded tosubmit and publish 7 articles and 1 patent
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Souza, Cristian Ferreira de. "Estudo da leishmaniose tegumentar americana na região urbana do município de Timóteo e no Parque Estadual do Rio Doce, Minas Gerais, Brasil." reponame:Repositório Institucional da FIOCRUZ, 2015. https://www.arca.fiocruz.br/handle/icict/13820.
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Previous issue date: 2015Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil
O presente trabalho se caracterizou como estudo descritivo comparando duas áreas: uma área silvestre onde se encontra o Parque Estadual do Rio Doce (PERD) e outra área urbana, situadas no município de Timóteo-MG. Objetivando estudar o ciclo de transmissão dos agentes das leishmanioses nas duas áreas, verificando possíveis alterações no seu padrão de transmissão. Foram realizadas coletas de flebotomíneos com armadilhas luminosas HP, coleta de amostras biológicas de animais silvestres, sinantrópicos e cães, o período de coletas foi entre setembro de 2012 a fevereiro de 2014. Foram realizados estudos de infecção natural por Leishmania spp., utilizando o gene ITS1, em todas as amostras coletas. E por fim um estudo de fonte alimentar em fêmeas alimentadas naturalmente. As análises foram realizadas com técnicas moleculares (PCR/Sequenciamento). Ficou claro que no município de Timóteo há pelo menos três espécies de Leishmania: Leishmania braziliensis, Leishmania amazonensis e Leishmania infantum. Dando evidência a grande presença de Leishmania braziliensis, identificada em amostras biológica de flebotomíneos, roedores, marsupiais e cães. A participação de flebotomíneos e animais, possíveis reservatório, no ciclo da Leishmania spp. no município de Timóteo ficou mais evidente após as observações da presença infecção natural por Leishmania braziliensis. Nyssomyia whitmani foi a espécie mais capturada na área urbana, enquanto no PERD foi Pressatia choti ambas as espécies foram identificadas com infecção natural por Leishmania braziliensis. A presença de Rattus rattus e Mus musculus, com infecção natural, sugerem que sejam possíveis reservatórios de Leishmania spp. no município Preocupante também e a possibilidade da participação de cães, uma vez que tiveram amostras de sangue com infecção natural. E por fim, e não menos importante foi o registro da participação de Gallus gallus (galinha) e Sus scrofa (porco) enquanto fonte alimentar de flebotomíneos, o que ajuda a manter o ciclo de vida dos vetores na área urbana do município
This work was characterized as a descriptive study comparing two areas: a wild area where is located the Parque Estadual do Rio Doce (PERD) and other urban area, located in municipality of Timóteo-MG, in order to study the transmission\2019 agents of cycle of leishmaniasis in both areas suggested checking possible changes in their transmission\2019s pattern. Sandfly captures were conducted with HP light traps, collecting biologies samples of wild animals, synanthropic and dogs, the period of collections were between September 2012 to February 2014. A study of natural infection of Leishmania spp were performed using the ITS1 gene in all collected samples. And a food meals for the study in females fed naturally. Analyses were performed using molecular techniques (PCR/ Sequencing). It was clear that in Timóteo\2019s municipality exist at least three species of Leishmania: Leishmania braziliensis, Leishmania amazonensis and Leishmania infantum. Giving evidence of the strong presence of Leishmania braziliensis, identified in biological samples of sandflies, rodents, marsupials and dogs. Participation of sand flies and animals, possible reservoirs of the Leishmania spp cycle. In Timóteo became more evident after the observations with the presence of Leishmania braziliensis natural infection. Nyssomyia whitmani was the most captured species in urban areas, while in the PERD was Pressatia choti, both species were identified with Leishmania braziliensis natural infection
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Vasconcelos, Sílvia Alcântara. "Identificação dos vetores da Leishmaniose tegumentar americana em Altos, Estado do Piauí." reponame:Repositório Institucional da FIOCRUZ, 2017. https://www.arca.fiocruz.br/handle/icict/25149.
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silvia_vasconcelos_ioc_mest_2017.pdf: 3587086 bytes, checksum: 4d78bababf4956d9a42fb96d0a2d0446 (MD5)Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.
A leishmaniose tegumentar americana (LTA) é uma antropozoonose tropical negligenciada que acomete o homem e animais silvestres. A transmissão ocorre através da picada do flebótomo infectado com a Leishmania. A LTA tem apresentado elevada incidência em algumas regiões do estado do Piauí e tem despertado a atenção da vigilância epidemiológica e dos serviços de saúde por ser uma patologia relativamente recente no estado. A procura por potenciais vetores da leishmaniose tegumentar no Piauí é de grande importância entomológica e epidemiológica pela carência de estudos nessa área, o qual permitirá estratégias de controle destes insetos. O objetivo deste estudo foi identificar os potenciais vetores da LTA em áreas de surto na cidade de Altos, município este que tem apresentado altas taxas de detecção de casos da doença nos últimos dez anos. Para este estudo, foram listados os endereços com registros de notificação da doença por meio do Sistema de Informação de Agravos de Notificação. Foram instaladas armadilhas luminosas tipo CDC nas regiões com maior ocorrência de notificações, abrangendo zona urbana e zona rural. Oitenta e seis coletas foram realizadas em 12 bairros da cidade de Altos. As capturas ocorreram de outubro de 2016 a janeiro de 2017. O material capturado foi conduzido ao laboratório de entomologia para seleção dos flebótomos e preparo para identificação O método de preparo das espécies para identificação foi o de Young e Duncan (1994). Foram identificados 1.308 espécimes de flebótomos, sendo 1.300 de Lutzomyia longipalpis, 06 Lutzomyia whitmani, 01 Lutzomyia sordelli e 01 Lutzomyia carmelinoi. Destacou-se uma variação anatômica verificada em três espécimes de Lutzomyia longipalpis, o que demonstra o predomínio da referida espécie em Altos no período de captura. O presente trabalho traz uma contribuição ao identificar flebótomos presentes na cidade de Altos, entre eles Lutzomyia whitmani, flebótomo incriminado como transmissor da LTA no Brasil e nos ajuda a começar a entender os mecanismos de transmissão da LTA no Piauí
American Cutaneous Leishmaniasis (ACL) is a neglected tropical zoonosis that affects humans and wild animals. Transmission occurs through the bite of the sandfly infected with Leishmania. ACL has a high incidence in some regions of Piauí state and it has attracted the attention of epidemiological surveillance and a health service is a relatively new disease in the state. The study of potential vectors of cutaneous leishmaniasis in Piaui still unknown and is of great epidemiological and entomological importance in this area and will enable control strategies. Previous studies have only address vectors of visceral leishmaniasis. The aim of this study was to identify potential vectors of American Cutaneous Leishmaniasis (ACL) in an area of outbreak in the city of Altos. This municipality has shown a high case detection rate of ACL in the past decade. For this study, we initially accessed notification records of ACL through the Information System for Notifiable Diseases Later luminous type CDC traps were installed in regions with higher incidence of notifications, including urban and rural areas. A total of 86 sand fly captures were performed in 12 different neighborhoods in Altos, between October 2016 and January 2017. Captured sand flies were taken to an entomology laboratory for separation of sandflies and preparation for identification. Identification of sand fly species was performed according to Young and Duncan (1994). A total of 1.308 specimens were identified, including Lutzomyia longipalpis (n= 1.300), Lutzomyia whitmani (n= 06), Lutzomyia sordelli (n= 01) and Lutzomyia carmelinoi (n= 01). An anatomical variation was identified in three Lutzomyia longipalpis specimens, the predominant sand fly species in Altos. This work suggests that Lutzomyia whitmani, among the captured sandflies, might be responsible for transmitting ACL in Brasil contributing to the understanding of ACL transmission in Piauí
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Depaquit, Jérôme. "Revision du sous genre paraphlebotomus (phlebotomus, phlebotominae, psychodidae, diptera) ; approches morphologique et moleculaire." Reims, 1997. http://www.theses.fr/1997REIMP211.
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Barbosa, Sofia Diniz de Nazaré. "A leishmaniose canina e os condicionalismos determinados pelas respectivas alterações renais." Bachelor's thesis, Universidade Técnica de Lisboa. Faculdade de Medicina Veterinária, 2011. http://hdl.handle.net/10400.5/3548.
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Dissertação de Mestrado Integrado em Medicina VeterináriaA Leishmaniose é uma doença zoonótica causada pelo protozoário da espécie Leishmania infantum, que existe predominantemente na Bacia do Mediterrâneo. O ciclo de vida deste parasita requer a existência de um vector, um insecto do género Phlebotomus, que é responsável pela transmissão das formas infectantes, os promastigotas metacíclicos, aos hospedeiros vertebrados, nomeadamente os canídeos. Em Portugal, apenas as espécies P. ariasi e P. perniciosus são vectores comprovados de L. infantum. As características clínicas da Leishmaniose canina variam amplamente como consequência dos numerosos mecanismos patogénicos do protozoário, da diversidade de respostas imunológicas desenvolvidas nos hospedeiros e dos diferentes órgãos afectados. A doença renal pode ser a única manifestação clínica nos animais infectados, podendo progredir de proteinúria assintomática até síndrome nefrótico ou Insuficiência Renal Crónica, que é responsável pela degradação do estado geral e a principal causa de morte nos cães com Leishmaniose. A aplicação do tratamento adequado requer, previamente, um diagnóstico precoce da doença e a avaliação do estado sanitário e imunitário do animal, através do exame clínico e de exames complementares, que devem ser repetidos a cada 6 meses. A presença de Insuficiência Renal Crónica impõe algumas limitações no tratamento da Leishmaniose canina, sendo fundamental seleccionar cautelosamente os fármacos e respectivas doses a administrar a cada paciente. Neste estudo retrospectivo foram observados 19 canídeos com Leishmaniose, sendo a sua sintomatologia muito variável, da qual se destaca a linfadenopatia, o emagrecimento e as lesões dermatológicas. As alterações laboratoriais mais frequentes foram a anemia não-regenerativa, a trombocitopénia, a hiperproteinémia com hiperglobulinémia e a proteinúria. O diagnóstico etiológico foi realizado com base nas técnicas de imunocromatografia rápida e de imunofluorescência indirecta. Foi também diagnosticada Insuficiência Renal Crónica em 4 canídeos da amostra. Diferentes fármacos foram utilizados no tratamento etiológico e sintomático, porém foi mais frequente a administração de alopurinol em monoterapia ou em associação com o antimoniato de glucamina.
ABSTRACT - CANINE LEISHMANIASIS AND CONSTRAINTS DETERMINED BY THEIR RENAL CHANGES - Leishmaniasis is a zoonotic disease caused by the protozoan Leishmania infantum, which is the most common species of Leishmania in the Mediterranean basin. The life cycle of this parasite requires the existence of a vector, the insect of the genus Phlebotomus, which is responsible for the transmission of infectious form, the metacyclic promastigotes, to the vertebrate host, namely the dog. In Portugal, only the species P. ariasi and P. perniciosus are proven vectors of L. infantum. The clinical features of canine Leishmaniasis are highly variable as a consequence of the numerous pathogenic mechanisms, the diversity of immune responses of individual hosts and the different organs affected. Renal disease may be the only clinical manifestation in infected dogs and may progress from asymptomatic proteinuria to nephrotic syndrome or to Chronic Kidney Disease, which is responsible for the deterioration of general condition and leading cause of death in dogs with Leishmaniasis. The application of suitable treatment requires, previously, an early diagnosis and assessment of dog’s health and immunity status, by clinical examination and several routine diagnostic tests, which must be repeated every six months. The presence of Chronic Kidney Disease imposes some limitations in the treatment of canine Leishmaniasis and is necessary to select carefully the drugs and doses to be administered in each patient. In this retrospective study, 19 dogs with Leishmaniasis were observed and many clinical signs were found, mainly lymphadenopathy, weight loss and dermatologic lesions. The most frequent laboratory abnormalities were non-regenerative anemia, thrombocytopenia, hyperproteinemia with hyperglobulinemia and proteinuria. The definitive diagnosis was made by a rapid immunomigration test and indirect fluorescent antibody technique. Chronic Kidney Disease was also diagnosed in 4 dogs of sample. Different drugs were used in the etiological and symptomatic treatment, but the administration of alopurinol alone or in combination with glucamine antimoniate were the most frequent.
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Sackin, Jennifer Rose. "Characterization and evolution of the Pheline family of non-LTR retrotransposons in Phlebotomus sandflies." Thesis, Imperial College London, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.300511.
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Perrotey, Sylvie. "Etude critique des caracteres et de leurs etats utilises pour la diagnose des plebotomes femelles (diptera : psychodidae) (doctorat : parasitologie)." Reims, 1998. http://www.theses.fr/1998REIMP209.
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FIGUEIRÊDO, JÚNIOR Carlos Alberto Santiago. "Análise do polimorfismo do gene que codifica a proteína salivar SP15 em três populações do Oriente Médio de Phlebotomus papatasi (Diptera: Psychodidae), vetor da Leishmania major." Universidade Federal de Pernambuco, 2010. https://repositorio.ufpe.br/handle/123456789/6797.
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Previous issue date: 2010Conselho Nacional de Desenvolvimento Científico e Tecnológico
As proteínas presentes na saliva dos flebotomíneos possuem uma grande importância na proteção contra parasitas do gênero Leishmania. Uma das proteínas identificadas, denominada de SP15, demonstrou ser responsável pela proteção contra a progressão da doença e aumento do tamanho da lesão em modelos animais. A elaboração de vacinas de múltiplos componentes com proteínas salivares pode ser uma forma viável para o desenvolvimento de vacinas anti-Leishmania. Partindo desta estratégia adotada para elaboração de vacinas, é necessário entender a variabilidade dos genes que codificam proteínas salivares e suas implicações nas respostas imunes dos humanos. Neste trabalho, investigamos a variabilidade genética de SP15 de populações naturais de Phlebotomus papatasi do Oriente Médio. Adicionalmente, associamos a variabilidade genética observada com a predição da estrutura secundária da proteína e possíveis epítopos de ligação a MHC classe II. Os resultados obtidos indicaram um baixo nível de variabilidade genética entre as populações oriundas do Oriente Médio, apesar da ocorrência de um grande número de haplótipos, onde alguns deles são compartilhados entre as populações distintas. Em conjunto, estas observações evidenciam a existência de fluxo gênico ou retenção de polimorfismo ancestral entre as populações e a ausência da seleção purificadora atuando sobre este gene. Para os preditos epítopos de MHC classe II, muitos foram identificados como possuindo sítios de mutação, mas pelo menos três epítopos identificados não apresentaram qualquer tipo de mutação, sugerindo que esta molécula apresenta-se moderadamente conservada e deve induzir respostas imunes uniformes nos humanos
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Nascimento, Bruno Warlley Leandro. "Estudo de Flebotomíneos (Diptera: Psychodidae) no município de Divinópolis, Minas Gerais, Brasil." reponame:Repositório Institucional da FIOCRUZ, 2013. https://www.arca.fiocruz.br/handle/icict/6520.
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A transmissão das espécies de Leishmania aos hospedeiros vertebrados envolve várias espécies de flebotomíneos no Brasil. Um estudo sobre a composição da fauna de flebotomíneos foi realizado no município de Divinópolis, Minas Gerais, Brasil, área endêmica para leishmaniose tegumentar (LT), e de transmissão moderada para leishmaniose visceral (LV). Foram realizadas coletas sistematizadas mensais para captura de flebotomíneos durante um ano, de setembro de 2010 a agosto de 2011, com armadilhas luminosas HP instaladas em área urbana, em 15 peridomicílios de casas onde se registrou pelo menos um caso de LT ou LV, e em 5 fragmentos de mata. Também foram realizadas coletas não sistematizadas utilizando armadilhas de Shannon nos fragmentos de mata. A detecção de DNA de Leishmania sp. nos espécimes de flebotomíneos foi realizada por meio da Reação em Cadeia da Polimerase (PCR) e para a identificação da amostra positiva foi utilizada a técnica de sequenciamento genético de fragmentos de DNA. Um total de 1.088 espécimes de flebotomíneos foram coletados pertencentes ao gênero Brumptomyia e Lutzomyia, e 18 espécies. Lutzomyia longipalpis, o principal vetor da Leishmania infatum no Brasil, foi a espécie mais freqüente, sendo encontrada em 14 das 20 localidades. As espécies Lu. longipalpis e Lu. whitmani foram as únicas coletadas em todos os meses de estudo. Nenhuma das espécies foi encontrada em todas as localidades de estudo. As cinco espécies mais frequentes foram: Lu. longipalpis (76,9%), Lu. lenti (8,3%), Lu. whitmani (5,0%), Lu. sallesi (2,8%) e Lu. aragaoi (2,2%). Não foi observada correlação estatisticamente significativa entre densidade total de flebotomíneos e variáveis climáticas durante o período de estudo. As análises moleculares detectaram DNA de Leishmania infantum em um exemplar de Lu. longipalpis. Os dados apontam para a necessidade de medidas de controle da população de flebotomíneos no município de Divinópolis e adoção de estratégias de vigilância entomológica.
The transmission of Leishmania species to vertebrate hosts involves several species of sand flies in Brazil. A study of the sand flies fauna composition was carried out in the city of Divinópolis, Minas Gerais state, Brazil, an endemic area for cutaneous leishmaniasis (CL), and of moderated transmission of visceral leishmaniasis (VL). Monthly systematic collections were made during one year, from September 2010 to August 2011. The HP light traps were installed inperidomicile of 15 houses where at least one case of CL or VL has been recorded and in five forested areas. Systematized collections were carried out using Shannon traps. The detection of DNA of Leishmania sp. in the sand fly specimens was performed by Polymerase Chain Reaction (PCR) and the technique for identification of positive samples genetic sequencing of DNA fragments technique. A total of 1,088 specimens of sand flies were collected, belonging to the genera Brumptomyia e Lutzomyia and 18 species. Lutzomyia longipalpis, the main vector of Leishmania infatum in Brazil, was the most frequent species, being collected in 14 of the 20 locations. The species Lu. longipalpis and Lu. whitmani were the only ones collected in every month studied. None of the collected species were found in all localities sampled. The five most abundant species were: Lu. longipalpis (76.9%), Lu. lenti (8.3%), Lu. whitmani (5.0%), Lu. sallesi (2.8%) and Lu. aragaoi (2.2%). No significant statistical correlation between the total sand fly density and the climatic variables was found. Molecular analysis identified natural infection by Leishmania infantum in one specimen of Lu. longipalpis. The data points to for measures to control the sand fly populations and adoption of strategies for entomological surveillance in Divinópolis municipality.
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Delinger, David. "Understanding the Mechanisms of Insecticide Resistance in Phlebotomus papatasi and Lutzoymia longipalpis Sand Flies (Diptera: Psychodidae: Phlebotominae)." DigitalCommons@USU, 2017. https://digitalcommons.usu.edu/etd/5730.
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The prevalence of insecticide resistance in vector species around the world is a continuous threat for any success at mitigating the spread of vector-borne diseases. With a limited arsenal of new insecticides, it is crucial for public health programs to understand the geographic range and the genetic mechanisms of resistance to best approach controlling insect vectors. Insecticide resistance is being increasingly observed in phlebotomine sand fly (Diptera: Psychodidae) populations in both the Old World and New World. Sand flies transmit the protozoans that cause leishmaniasis, a disfiguring disease that kills tens of thousands of people each year. The goal of this dissertation was to have both an applied and basic research focus towards understanding resistance in phlebotomines. I began by comparing in vivo and in vitro methods for blood-feeding two species of sand flies, Phlebotomus papatasi and Lutzomyia longipalpis, in the laboratory, both of which are important leishmaniasis vectors. I investigated the susceptibility of both species to ten different insecticides by calculating lethal concentrations that caused varying levels of mortality. Based on these results, I determined diagnostic doses and diagnostic times for both species to the same ten insecticides using an accepted, but novel, assay for sand flies. Finally, I tested for known mechanisms of insecticide resistance in four artificially resistant-selected colonies of sand flies, as well as tested for novel resistance mechanisms. Through applied research, I developed methods for efficient sand fly rearing and for determination of population resistance to insecticides, tools that have worldwide applicability.
Through basic research, I determined that laboratory populations of sand flies have sufficient standing genetic variation needed to survive sublethal doses of insecticides; however, I was unable to develop artificially-selected colonies resistant to these insecticides. My research has generated information to provide new insights into the evolution of insecticide resistance in natural sand fly populations. My results support that resistance development may be possible, but evolutionary challenging, an encouraging finding that may be exploited by vector biologists and public health officials to prevent or slow the development of resistance in sand flies to insecticides
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MARTIN, ELENA. "SYMBIOSIS BETWEEN ARTHROPODS AND FUNGI: THE CASE OF PHLEBOTOMUS PERNICIOSUS, THE VECTOR OF VISCERAL AND CANINE LEISHMANIASIS." Doctoral thesis, Università degli Studi di Milano, 2018. http://hdl.handle.net/2434/577035.
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Introduction
Vector borne diseases (VBDs) are one of the greatest public health problems worldwide and their control represents a key global public health challenge. In particular, leishmaniases are parasitic VBDs endemic in 98 countries from five continents distributed in tropical, subtropical and temperate zones. During the last twenty years, their distribution have undergone an expansion, also due to climate and environmental changes, the reason why they are considered re-emerging diseases. Italy is traditionally endemic for cutaneous and visceral leishmaniasis caused by the protozoan parasite Leishmania infantum, whose reservoir is the dog. In the Old World, the natural vectors of the protozoa Leishmania spp. are several species of sand flies belonging to the genus Phlebotomus. To control VBDs, the World Health Organization promotes the rational use of insecticides and integrated strategies for vector control through chemical and biological strategies. The "symbiotic control" is one of the biological control strategies that exploits microorganisms that live in symbiosis with the vector, in order to reduce or interfere with the pathogen transmission.
The aim of my thesis project was to increase the knowledge on killer yeasts, in general, and in the arthropod Phlebotomus, in particular, in order to open the way toward their development as tools for the biological/integrated control of vector-borne diseases.
Article 1
My first work, here presented, reports the investigation of the fungal community of the sand fly Phlebotomus perniciosus, the main vector of leishmaniasis in the western Mediterranean area and the most widespread sand fly in Italy, with a focus on the dominant yeast species Meyerozyma guilliermondii. The yeast displayed higher prevalence in the reared population than in the analyzed wild individuals, letting us hypothesize a contribution of this yeast to the diet of reared sand flies with nutrients that are on the other side easily available in nature. The localization of the yeast, found in the midgut of adults of both sexes and larvae and in Malpighian tubules of female sand flies, in addition to the genome analysis, let us speculate that the yeast may confer the additional advantage of contributing to the removal of the excess of nitrogenous wastes after the blood meal of the insect host.
Article 2
Whole-genome sequencing, de novo assemby and phylogenomic analysis of one strain of M. guilliermondii isolated from P. perniciosus and of other yeast strains of the M. guilliermondii complex isolated from arthropods were the goals of another research paper here presented. This work led to publish the first four genomes of isolates of Meyerozyma caribbica and to better understand evolutionary relationships within the clade.
Articles 3-5
The other works presented in this document were focused on the yeast Wickerhamomyces anomalus, here studied as a yeast candidate for symbiotic VBDs control.
In one of these papers (Article 3) the isolation and phylogenetic characterization of a killer strain of W. anomalus from the sand fly P. perniciosus is reported. The association between the yeast and sand flies seems to be facultative.
The killer toxin produced by the isolated W. anomalus strain was purified, characterized and tested against fungal pathogens in another work (Article 4). This toxin is a β-1,3-glucanase that possesses in vitro activity against Candida glabrata isolates, independently from their drug-resistance phenotypes. The obtained results suggest a potential application of this toxin against fungal infections at skin and mucosal membrane levels provoked by C. glabrata and may be useful for the design of new therapeutic molecules and the study of yeast resistance mechanisms.
Finally, W. anomalus is widely distuributed in nature and is considered by EFSA as a non-pathogenic agent, completely safe for healthy individuals (level of bio-security 1/QPS). Nevertheless, in the last paper I investigated the circulation of W. anomalus by qPCR screening examining a large panel of immunocompromised and / or intensive care patients, as well as healthy donors as controls (Article 5).
The screening revealed only one qPCR-positive subject, out of over 500 analyzed samples. Considering the few published cases of clinical infection by this yeast, in immunocompromised patients, we can conclude that this microorganism is a rather uncommon opportunistic pathogen.
In conclusion, the study of the mycobiota associated with sand flies could contribute to the understanding of the biology these insects, and offers the potential of discovering novel microrganisms for future application in the symbiotic control of leishmaniasis.
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Menezes, Júlia Alves. "Leishmanioses : estudos epidemiológicos e o conhecimento da população de Formiga, Minas Gerais." s.n, 2014. https://www.arca.fiocruz.br/handle/icict/9933.
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Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Belo Horizonte, MG, Brasil
No Novo Mundo, as leishmanioses constituem um grupo de doenças negligenciadas, com alta prevalência mundial e que podem assumir formas graves. Elas são transmitidas ao homem pela picada de flebotomíneos, existindo uma gama de reservatórios, sendo o cão o principal hospedeiro urbano. Neste contexto, as leishmanioses, principalmente a forma visceral zoonótica, tem se urbanizado no Brasil e em várias partes do mundo. O objetivo do presente trabalho foi estudar alguns aspectos epidemiológicos das leishmanioses e o conhecimento da população sobre a doença no município de Formiga, MG. Estes aspectos se referem principalmente à fauna flebotomínica e à infecção canina. Para se conhecer a fauna flebotomínica, foram realizadas coletas sistemáticas mensais entre maio 2012 e abril 2013 utilizando armadilhas luminosas HP em 24 peridomicílios. Os exemplares foram identificados e sua densidade relacionada a fatores climáticos e ambientais. O estudo da frequência da leishmaniose canina foi realizado através da coleta de sangue de 570 cães e posterior análise sorológica (DPP® e ELISA) e molecular (LnPCR). As amostras positivas no LnPCR foram submetidas ao sequenciamento e PCR-RFLP a fim de determinar a espécie de Leishmania. Todos os locais de captura entomológica e os endereços dos cães positivos foram georreferenciados sobre a malha urbana do município e correlacionados aos casos humanos e a fatores ambientais locais. A população foi entrevistada a partir de um questionário quantitativo validado que abordou conceitos relevantes sobre a doença e sobre as condições do peridomicílio. Foram capturados 224 flebotomíneos de 10 espécies diferentes, sendo Lutzomyia longipalpis (35,3%), Lutzomyia cortelezzii (33,5%) e Lutzomyia whitmani (18,3%) as mais abundantes. Foi observada correlação estatisticamente significativa (p<0,05) entre a densidade total de flebotomíneos e a precipitação e temperatura. Em relação aos cães, a taxa de infecção encontrada, considerando os testes sorológicos e o molecular, foi de 19,1%. O sequenciamento e a PCR-RFLP permitiram identificar as espécies Leishmania infantum e Leishmania braziliensis infectando os cães. As análises espaciais demonstraram que L. whitmani (99,8%) se concentra nas proximidades de extensas áreas verdes e em distâncias de até 200m da vegetação. Lutzomyia longipalpis está mais associado à rede hidrográfica, se concentrando, principalmente, nas faixas distantes até 400m de um curso d’água. Para o município, flebotomíneos, cães positivos e casos humanos estão geograficamente relacionados, embora com pouca sobreposição de suas áreas de ocorrência. O estudo sobre o conhecimento da população revelou que os moradores de Formiga desconhecem completamente a doença (93%) e que 95% deles estão sob algum fator de risco. A existência de flebotomíneos comprovadamente vetores de leishmaniose visceral e tegumentar associado ao fato de existirem cães infectados com espécies, tanto dermotrópicas quanto viscerotrópicas do parasito, sugerem que no município está ocorrendo um ciclo de transmissão de leishmanioses visceral. Além disso, pode ocorrer um possível surgimento de casos humanos de leishmaniose tegumentar. Dessa forma, fica evidente que medidas de vigilância e controle precisam ser urgentemente estabelecidas na cidade para se evitar a dispersão e os óbitos por leishmaniose na região.
Leishmaniases in the New World are neglected diseases with high prevalence worldwide and may assume severe forms. They are transmitted to humans by the bite of sand flies and exhibit a range of wild and domestic reservoirs including the dog as the main urban host. In this context, especially zoonotic visceral leishmaniasis form has urbanized in Brazil and around the world. The aim of this work was to study some epidemiological aspects of leishmaniasis and the population’s knowledge about the disease in Formiga, Minas Gerais State. Those aspects are mainly related to the phlebotomine fauna and canine infection. To determine the sand fly fauna, monthly systematic collections were performed between May 2012 and April 2013 using HP light traps in 24 peridomicile. Specimens were identified and their density related to climatic and environmental conditions. The study of the frequency of canine leishmaniasis was performed by collecting blood samples from 570 dogs and subsequent serologic (DPP and ELISA) and molecular (LnPCR) analysis. The LnPCR positive samples were subjected to sequencing and PCR-RFLP to determine the Leishmania species. All the entomological capture sites and addresses of positive dogs were georeferenced on the urban grid of the city and correlated to human cases and local environmental factors. The population was interviewed using a validated quantitative questionnaire addressing relevant concepts about the disease and the conditions of the peridomicily. Two hundred twenty-four phlebotomines from 10 different species were captured, where Lutzomyia longipalpis (35.3%), Lutzomyia cortelezzii (33.5%) and Lutzomyia whitmani (18.3%) were the most abundant. Statistically significant correlation (p <0.05) was observed between total sandfly density and precipitation and temperature. Regarding the dogs, the rate of infection found by considering the serologic and molecular assays was 19.1%. Sequencing and PCR-RFLP allowed the identification of the species Leishmania infantum and Leishmania braziliensis infecting dogs. The spatial analysis showed that L. whitmani (99.8%) is concentrated close to green areas and up to a distance of 200m from vegetation. Lutzomyia longipalpis is most associated with the hydrographic network, concentrating mainly on the distances up to 400m alongside the water. For the Formiga city, phlebotomines, positive dogs and human cases are geographically related, although poor overlap of their occurrence areas has been observed. The study on the knowledge of the population revealed that the residents of Formiga are completely unaware of the disease (93%) and that 95% of them are under some risk factor. The existence of proven sandflies vectors of visceral and tegumentary leishmaniasis and the existence of infected dogs - with both dermotrophic as visceral species - suggest that the city is experiencing a cycle of transmission of visceral leishmaniasis. Besides, the city may be also iin risk of an outbreak of tegumentary human cases. It is clear that surveillance and control measures need to be urgently established in the town to avoid dispersion and deaths per leishmaniasis in the region.
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Zulfiqar, Bilal. "Identification and Characterization of Novel Anti-leishmanial Compounds." Thesis, Griffith University, 2017. http://hdl.handle.net/10072/379283.
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Leishmaniasis is characterized as a parasitic disease caused by the trypanosomatid
protozoan termed Leishmania. Leishmaniasis is endemic in 98 countries around the
globe with increased cases of morbidity and mortality emerging each day. The mode of
transmission of this disease is via the bite of a sand fly, genus Phlebotomus (Old World)
and Lutzomyia (New World). The life cycle of Leishmania parasite exists between the
sand fly (promastigote form) and the mammalian host (amastigote form). Leishmaniasis
can be characterized as cutaneous, muco-cutaneous or visceral leishmaniasis based on
clinical manifestations exhibited in infected individuals. Although leishmaniasis is
treatable, it faces challenges largely due to emerging resistance and extensive toxicity
for current drugs. Therapeutic efficacy varies depending upon the species, symptoms
and geographical regions of the Leishmania parasite.
The drug discovery pipeline for neglected trypanosomatid diseases remains sparse. In
particular, the field of leishmaniasis drug discovery has had limited success in
translating potential drug candidates into viable therapies. Currently there are few
compounds that are clinical candidates for leishmaniasis, it is therefore essential that
new compounds that are active against Leishmania are identified and evaluated for their
potential to progress through the drug discovery pipeline. In order to identify new
therapeutics, it is imperative that robust, biologically relevant assays be developed for
the screening of anti-leishmanial compounds. The currently available assays have low
predictive compatibility and high attrition rate in the identification of the compounds.
In an aim to identify compounds with activity against L. donovani (MHOM/IN/80/DD8)
promastigotes, a phenotypic, high-throughput, resazurin based 384-well viability based
assay was successfully developed to estimate the effect of compound treatment on L.
donovani DD8 parasites. Complementary cytotoxicity assays were also established to
access the toxicological profile of the compounds. The assays developed are robust and
reproducible, using standard statistical parameters to assess assay quality and suitability
for use in high throughput screening (HTS) based drug discovery.
To identify compounds active against L. donovani DD8 parasites, two compound
collections were evaluated, a synthetic scaffold and a natural product (NP) based
library. A primary screen of the open scaffold library constituting 5560 structurally diverse synthetic compounds was undertaken. Screening incorporated a promastigote
viability assay (extracellular form) and a high-content intracellular amastigote imaging
assay. Confirmation of activity was performed together with cytotoxicity studies against
both THP-1 (host cell) and HEK-293 cell lines. The second library, Davis open access
natural product-based (DOANP) library was tested against kinetoplastids (Leishmania
donovani DD8, Trypanosoma brucei brucei and Trypanosoma cruzi) using highthroughput
phenotypic assays. This library currently consists of 472 distinct
compounds, the majority of which are natural products that have been obtained from
Australian natural sources, such as endophytic fungi, plants, macrofungi and marine
invertebrates.
The confirmed hits were prioritized based on structure activity relationships to identify
potential analogues. Then the activities of these analogues were evaluated to access the
in vitro effect of these compounds. Based on the anti-leishmanial activity and
selectivity, two compounds identified from the synthetic scaffold library were selected
to be taken further for characterization and biological profiling. Of these two
compounds, the first compound BZ1 exhibited an IC50 value of 0.59 ± 0.13 μM against
the intracellular form (amastigote) of the parasite and IC50 value of 2.37 ± 0.85 μM
against the extracellular form (promastigote) of L. donovani DD8 (Old World - Indian
strain) parasites. The second compound BZ1-I demonstrated to have an IC50 of 0.57 ±
0.17 μM against intracellular amastigotes and an IC50 value of 0.60 ± 1.12 μM against
promastigotes.
The two compounds were subjected to additional assays which included cidal/static
effect, time to kill and pre-incubation studies. The mechanism of action studies were
also conducted to assess whether the compounds induce apoptosis in parasites and their
effect on mitochondrial morphology. Resistance was generated and confirmed for both
the compounds in order to determine the target protein using whole genome sequencing
technique. Compounds cytotoxicity was also determined against a cytotoxicity panel
constituting of THP-1, HEK-293, HepG2, J774.1, Raw-264.1 cell lines to establish the
safety profile. The lead molecules were subsequently progressed to Drug Metabolism
and Pharmacokinetics (DMPK) and pH stability studies. In addition to the activity
against the L. donovani DD8 (Old World - Indian strain) parasite, compounds also
showed activity against other species and strains of the Old World and New World parasites, namely L. infantum (Old World), L. donovani (Old World - Sudanese strain)
and L. infantum (New World) amastigotes. The compounds were also shown to have in
vitro activity against T. brucei species, T. cruzi and Plasmodium falciparum strains.
Screening against the extracellular and intracellular forms of the parasite, as well as
across a panel of leishmanial species, provides a unique activity profile for prioritizing
the identified hits. These hit compounds provide urgently needed starting points for the
development of novel lead series for future anti-leishmanial therapeutics.Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
School of Environment and Sc
Science, Environment, Engineering and Technology
Full Text
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Mahamdallie, Shazia Sophie. "Effects of environmental change on genetic diversity and distribution pf Phlebotomus ariasi, a vector of visceral leishmaniasis in Southwest Europe." Thesis, London School of Hygiene and Tropical Medicine (University of London), 2010. http://researchonline.lshtm.ac.uk/768498/.
Full textAbstract:
Leishmania infantum is the causative agent of zoonotic visceral leishmaniasis (ZVL) in the Mediterranean region, with the domestic dog as the main reservoir host. Phlebotomus (Larroussius) ariasi is the principal vector in cooler, forested ecotopes in southwest Europe, which suggests that it might be subject to environmental and geographical isolation. However, the population genetics of P. ariasi had been little studied before this thesis, which investigated how the population differentiation of this vector might affect its ability to spread northwards, or persist in the Mediterranean region, in response to climate and habitat change. Thirty-six spatio-temporal populations of P. ariasi were molecularly characterized across its range, predominantly from southwest France but including geographical outgroups from Spain, Portugal and North Africa. Phylogenetic and population genetic assessments were made based on five DNA sequences: mitochondrial cytochrome b, nuclear elongation factor-la and apyrase, plus two anonymous nuclear loci, AAm20 and AAm24. The results demonstrated the absence of cryptic sibling species of P. ariasi and the selective neutrality of each locus. Mitochondrial DNA revealed a historical phylogeographic structure, which was consistent with Pleistocene climate change driving multiple haplogroup divergences within glacial refuges and phalanx-like population expansions in interglacial periods. Nuclear loci mostly showed isolation by distance, but some supported restricted gene flow between the Pyrenees and the Massif Central, France, as indicated by cytochrome b. A glacial refuge may have existed north of the Pyrenees. The genetic diversity observed in the northeast Pyrenees, France, permitted an assessment of the effects of broadleaf forest fragmentation on the differentiation of P. ariasi. No conclusive evidence was found to support contemporary genetic substructuring or impoverishment associated with a recent increase in forest fragmentation. The salivary peptide apyrase revealed a geographical pattern of polymorphism consistent with the other selectively neutral loci. A range of selection tests indicated that apyrase was not evolving under positive directional or balancing selection and, therefore, a genetic arms race with the mammalian host and/or Leishmania parasite was not supported. The approach taken provides a proof of principle for helping to assess apyrase and other salivary peptides as vaccine candidates against leishmaniasis.
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Saraiva, Lara. "Estudo populacional de flebotomíneos (Diptera: Psychodidae) no Município de Belo Horizonte e no Parque Estadual do Sumidouro – Minas Gerais - Brasil." s.n, 2015. https://www.arca.fiocruz.br/handle/icict/13672.
Full textAbstract:
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Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil
O município de Belo Horizonte e sua região metropolitana ilustram o mais sério exemplo de expansão da leishmaniose visceral no sudest e do Brasil. O município e o parque Estadual do Sumidouro localizam-se no bioma cerrado, segundo ma ior bioma terrestre da América do Sul. Os objetivos deste projeto foram: estudar a variação sazonal da fauna flebotomínica no DS Venda Nova (Belo Horizonte) e no PES; descrever os padrões de riqueza e diversidade fauna flebotomínica nas formações vegetais do PES e no DS Venda Nova; de terminar a taxa de infecção natural de fêmeas capturadas; est udar a variação morfológica das populações de Lu. longipalpis coletadas nos diferentes ambientes; aval iar os impactos das ações de controle realizadas no DS Venda Nova nas taxas de oc orrência sazonal e dens idade das populações de flebotomíneos. Foram realizadas coletadas mensais no pe ríodo de agosto de 2011 a agosto de 2013. Os insetos foram identificados de acordo com Galati 2003. As fêmeas coletadas não ingurgitadas foram submetidas à extração de DNA e reações de PCR e de PCR-RFLP para a averiguação da infecção natural por espécies de Leishmania. Para comparação das localidades de estudo foi utilizada análise descritiva e índices ecológicos. Para a comparação morfológica trinta casais (quinze proveni ente do DS Venda Nova e quinze do PES) da espécie Lu. longipalpis foram medidos, e foram comparados tantos as médias como as variâncias dos caracteres entre os grupos. As ações de contro le da LV no DS Venda Nova realizadas nos período de janeiro de 2011 a dezembro de 2013 foram analisadas descritivamente e relacionadas à curva sazonal de Lu. longipalpis . A riqueza, a diversidade e a equitabilidade da fauna de flebotomíneos foram marcadamente dife rentes entre o DS Venda Nova e o PES. Na área urbana foram coletados 2.247 espécimes pertencentes a cinco gênero s e oito espécies, a curva de acumulação de espécies atingiu a satu ração na 19º amostragem. No DS Venda Nova a curva de variação sazonal foi delineada principalmente por Lu. longipalpis e 95,3% dos espécimes pertenciam a esta espécie. Lu. longipalpis apresentou taxas de infecção natural de 1,01% para Leishmania infantum e 1,77% para Leishmania braziliensis . No PES foram coletados 4.675 espécimes pertencentes a ci nco gêneros e 25 esp écies e a curva de acumulação atingiu a saturação na 16º amostragem. No PES não houve uma única espécie que apresentasse padrão tão pronunciado de dominância. Lu. longipalpis correspondeu a 5,35% dos espécimes. Diversas espécies com envolvi mento (suspeito ou comprovado) nos ciclos de LTA foram registradas. Uma fêmea do comple xo cortellezzi apresent ou detecção positiva para Le. braziliensis. A comparação morfométrica e de co mposição de feromônios indicou grande similaridade entre as duas populações de Lu. longipalpis avaliadas. A análise realizada para co rrelação das ações de controle com a variação sazonal de Lu. longipalpis indicam a necessidade de estudos pormenorizados para inferências mais robustas. Os resultados evidenciam a dominância da espécie Lu. longipalpis no ambiente urbano, e indicam que as mudanças antrópicas no cerrado podem a lterar a composição das populações de flebotomineos e os ciclos de tran smissão de patógenos aos humanos.
The municipality of Belo Horizonte and its me tropolitan region illustrate the most serious example of the expansion of leishmaniasis in southeast Brazil. The m unicipality of Belo Horizonte, its metropolitan region, and th e Sumidouro State Park, are in the Brazilian Savannah biome (Cerrado). This is the second la rgest terrestrial biome in South America. The aims of this project were: to study the seasona l variation of sand flies in SD Venda Nova (Belo Horizonte) and in the PES; to describe the ric hness and diversity pa tterns of sand flies in the vegetable formations of PES and SD Venda Nova; to determine the natural infection rate of females captured; to study the morphological variation of Lu. longipalpis populations from different environments; to evaluate the im pacts of LV control actio ns carried out in DS Venda Nova in seasonal occurr ence of sandflies. Sampling was performed every month over three consecutive days using HP light trap s form August 2011 to August 2013. The insects were identified according to Galati 2003. Th e non-engorged females were subjected to DNA extraction and PCR reactions and the PCR-RFLP for the investigation of natural infection with Leishmania species. To compare the study sites was used descriptive analysis and ecological indexes. In the morphological comp arison thirty couples (fifteen from the DS Venda Nova and fifteen PES) of Lu. longipalpis species were measured and the means and variances were compared. The LV control actions in DS Venda Nova conducted in the period from January 2011 to December 2013 were analyzed descriptively and related to seasonal curve Lu. longipalpis. Richness, diversity and evenness in the study areas, urban and park protected area, were distinct. In Venda Nova SD 2.427 sand flies specimen s belonging to five genera and eight species were collected and the species accumulation curve reached saturation in the 19º sampling. In Sumidouro State Park a total of 4,675 sand flie s specimens belonging to nine genera and 25 species were collected and the species accumulation curve reached the saturation in the 16º sampling. In Venda Nova SD the species seasonal variation curve is predominantly determined by Lu. longipalpis . In Venda Nova 95.3% of the collected specimens belonged to the species Lu. longipalpis and this species exhibited a natural infection rate of 1.01% for Le. infantum and 1.77% for Le. braziliensis . In the Sumidouro State Park a total of 4,675 sand fly specimens of 25 species bel onging to nine genera were collected. The seasonal curve is not delineated by a unique species. In the park area, Lu. longipalpis accounted for only 5.35% of the collect ed specimens. The main vector of Le. infantum , Lu. longipalpis, accounted for only 5.35% of the specimens collected. Proven or suspected vectors of Le. braziliensis were recorded, and one female of the cortellezzii complex tested positive for Le. braziliensis DNA. The results make evident the Lu. longipalpis dominance in the urban environment. Our d ata demonstrate that anthropic modifications in the savannah studied area sign ificantly altered the sandflies populations and the Leishmania transmission cycles.
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Cruz, Malpica Cristhopher Donat's. "Estandarización de la prueba de ensayo inmunoenzimático para la detección de anticuerpos IgG en sueros de humanos para el virus Phlebotomus fever." Bachelor's thesis, Universidad Nacional Mayor de San Marcos, 2001. https://hdl.handle.net/20.500.12672/1276.
Full textAbstract:
El virus Phlebotomus fever es un arbovirus cuya transmisión al hombre ocurre por la picadura de moscas hematófagas de vuelo limitado, pertenecientes a los géneros Phlebotomus, Sergentomyia y Lutzomyia. La infección en humanos ocurre principalmente en personas que trabajan o viven en la selva.
En el presente trabajo se estandarizó una prueba inmunoenzimática indirecta (ELISA) para detectar anticuerpos IgG contra el virus en sueros humanos. También se aplicó esta prueba en la determinación de la seroprevalencia de anticuerpos contra dicho virus en personas procedentes de Iquitos. La confirmación de los casos positivos se realizó mediante la Prueba de Neutralización por Reducción de Placa (PRNT).
El ELISA se realizó en cuatro pasos: fijación del antígeno, reacción con el suero, adición del conjugado y reacción con el sustrato. En la producción del antígeno se utilizó la línea celular Vero E-6, siendo la dilución de trabajo 1:1500. El conjugado utilizado es un Anti IgG humano de cabra purificado y marcado con peroxidasa, siendo su dilución de trabajo 1:8 000.
Se analizó con la prueba de ELISA 400 sueros procedentes de Iquitos, encontrándose una seroprevalencia de 16.25%. Al confirmar los resultados con la prueba de PRNT, el ELISA obtuvo una especificidad de 83.02% y una sensibilidad de 98.24%. ELISA desarrollado para el virus Phlebotomus fever es aplicable para realizar estudios de prevalencia.-- The Phlebotomus fever virus is an arbovirus transmitted to humans by the bits of hematofagous flies with limited flight range, which belong to the genus Phlebotomus, Sergentomyia and Lutzomyia. The human infection mainly occurs among people who work and live in the jungle. An inmunoenzimatic indirect assay (ELISA) was standardized in this thesis to detect antibodies IgG against the virus in human serum. This test was applied to determine the antibody seroprevalence against this virus in people coming from Iquitos. The positive case confirmation was performed using the Plaque Reduction for Neutralization Test (PRNT). The ELISA test was performed in four stages: the antigen fixation, reaction to the serum, addition to the conjugate and reaction to the substrate. During the antigen production, the cell line Vero E-6 was used, obtaining a work dilution of 1:1500. The conjugated used is a purified antibody peroxidase labeled goat antihuman IgG, obtaining a work dilution of 1:8000. Using the ELISA test, 400 sera from Iquitos were tested, resulting with a seroprevalence of 16.25%. When confirming the results with the PRNT trial, the ELISA test showed a specificity of 83.02% and a sensitivity of 98.24%. I concluded that the ELISA test developed for the Phlebotomus fever virus is applicable to perform prevalence studies.
Tesis
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Azevedo, Nuno Miguel Pereira. "Leishmaniose cutânea na Beira Interior." Master's thesis, Universidade da Beira Interior, 2009. http://hdl.handle.net/10400.6/888.
Full textAbstract:
Este trabalho expõe o levantamento estatístico e a caracterização clínica da
leishmaniose cutânea na Beira Interior. Pretende-se, assim, tentar definir a importância que
esta infecção assume neste meio e identificar os principais problemas epidemiológicos e
clínicos com ela relacionados, bem como propor algumas medidas a tomar para a sua
resolução.
Foi efectuado um estudo descritivo tipo série de casos exploratório, tendo por base os
doentes observados nas consultas de Dermatologia do Centro Hospitalar da Cova da Beira e
da Unidade Local de Saúde da Guarda, entre Abril de 2004 e Maio de 2009, com diagnóstico
de leishmaniose cutânea confirmado por biópsia cutânea.
Os dados submetidos à análise estatística provêm das informações recolhidas nos
processos clínicos e junto dos doentes, através de um inquérito epidemiológico. Para
caracterização epidemiológica foi inquirido o local de residência, a proximidade a zonas
verdes e a recursos hídricos, o contacto com animais domésticos, nomeadamente cães, mas
também selvagens ! raposas e roedores. Para além da ocupação profissional e de
passatempos, foram também registadas viagens para fora de Portugal, assim como o momento
em que ocorreu a picada de flebótomo potencialmente infectante. Na caracterização da doença
tentou-se obter a descrição da lesão, questionando-se igualmente a ocorrência de leishmaniose
mucocutânea ou visceral, e o tempo decorrido até à procura de ajuda médica e tratamentos
prévios realizados. Recolheram-se dados sobre a abordagem diagnóstica, a resposta ao
tratamento e a evolução depois deste. Para avaliar o grau de informação sobre a leishmaniose,
os doentes foram inquiridos quanto à ocorrência de história familiar e quanto ao seu
conhecimento prévio acerca da leishmaniose cutânea.
Foram seleccionados 13 doentes. Todos viviam na região da Beira Interior. A maior
ocorrência foi no sexo feminino (77%). Cinquenta porcento dos doentes tinham idade inferior
a quarenta e quatro anos. O maior número de diagnósticos confirmados por biópsia ! seis !,
ocorreu no ano de 2006. A maioria vivia perto de espaços verdes (92%), numa zona rural
(69%) e próximo de recursos hídricos (62%). A apresentação típica foi a lesão única na face
(85%), mais frequentemente um nódulo (69%), sem eritema, prurido ou dor. O tempo médio
de espera até procurar assistência médica foi de 22 semanas. A maioria dos doentes foi
primeiro ao Médico de Família (92%). Em nenhum caso se observou associação com a forma
mucosa ou visceral da doença. Todos os doentes evoluíram para a cura, não tendo ocorrido
recidivas nem reinfecções.
Em conclusão, o presente estudo permite-nos sugerir que a leishmaniose cutânea deve
ser considerada no diagnóstico diferencial das lesões cutâneas inflamatórias crónicas na Beira
Interior, principalmente em doentes com factores de risco adequados associados.This paper presents a survey of statistical and clinical characterization of cutaneous leishmaniasis in Beira Interior. The aim is to try to define the importance of this infection in this region, as well as identify key epidemiological and clinical problems related with it, as well as some measures to resolve them. A descriptive exploratory case-series study was conducted, based on patients seen in the Dermatology department of the Hospital Center of Cova da Beira and the Local Health Unit of Guarda, between April 2004 and May 2009, with diagnosis of cutaneous leishmaniasis confirmed by skin biopsy. "#$% &'('% )*+,-(($&% (.% )('(-)(-/'0% '1'02)-)% 3')% /.00$/($&% 45.,% (#$% 6'(-$1()7% ,$&-/'0% records and through field work. For the epidemiological characterization, besides the place of residence, it was asked the proximity to green areas and water resources, contact with household animals, mainly dogs but also wildlife ! foxes and rodents. In addition to occupation and hobbies, the trips out of Portugal and the time when the bite occurred was inquired. For the characterization of the disease, we tried to get a description of the injury, and we also questioned the occurrence of mucocutaneous or visceral leishmaniasis, the weeks that elapsed until the patient seeked medical advice and any treatments he made before. We also analyzed the diagnostic approach, the response to treatment and the evolution after this. To assess the degree of information about leishmaniasis, the patient was questioned about previous family history and their prior knowledge of the disease. Thirteen patients were selected. All lived in the region of Beira Interior. Most occurrences were in females (77%). Fifty percent of patients had less than forty-four years. The highest number of cases confirmed by biopsy, 6 patients, was registered in 2006. Most patients lived close to green areas (92%), in a rural zone (69%), near water resources (62%). The typical presentation was a single lesion on the face (85%), most often a nodule (69%) without erythema, itching or pain. The average time elapsed before seeking medical advice was 22 weeks. Most patients first went to the family practitioner (92%). In no case the disease was associated with the mucosal or visceral form. All patients progressed to a cure. There were no relapses or reinfections. In conclusion, this study allows us to suggest that cutaneous leishmaniasis should be considered in the differential diagnosis of chronic inflammatory skin lesions in Beira Interior, mainly in patients with known risk factors.
47
Vieira, Coutinho Abreu Gomes Iliano. "Molecular aspects of sand-fly-based vaccine development." Diss., Kansas State University, 2011. http://hdl.handle.net/2097/9224.
Full textAbstract:
Doctor of PhilosophyDepartment of Entomology
Marcelo Ramalho-Ortigao
The emergence and reemergence of vector-borne diseases pose significant threats to humans and other animals worldwide. Although vector control relies mostly on insecticides, the emergence of insecticide resistance urges for the development of new strategies to control the spread of such diseases. For sand fly-transmitted leishmaniasis, Transmission Blocking Vaccines (TBV) may constitute a feasible strategy to impair Leishmania transmission from infected to uninfected vertebrate hosts. Moreover, sand fly saliva-based vaccines represent an alternative or complementary approach as these vaccines protect different mammalian hosts against Leishmania. Based on the potential use of sand fly molecules as vaccines against leishmaniasis, we assessed the potential of Phlebotomus papatasi midgut secreted proteins as TBV candidates and the expression variability of sand fly salivary gland genes. Regarding the TBV approach, we took advantage of the RNA interference (RNAi) technique to evaluate the effects of knocking down P. papatasi midgut-specific genes on Leishmania major development within the sand fly midgut. Whereas peritrophin 1 (PpPer1) knock down led to increased Le. major load by 39%, knocking down chitinase 1 (PpChit1) reduced Le. major load in P. papatasi midguts by 63%. Thus, our data strongly suggest that PpChit1 constitutes a potential target for TBV approaches against Leishmania transmission in endemic areas. Concerning protective vaccines based on salivary gland secreted proteins, we searched for expression polymorphism in selected salivary gland genes in natural and colonized populations of P. papatasi. Significant differences in salivary gland gene expression were not only exhibited in P. papatasi specimens collected in different geographic habitats but also seasonal difference in gene expression was displayed by specimens belonging to the same population. As antigen dose is an important component of immune responses, different doses of salivary protein inoculated into host skin may interfere with vaccine protection. Thus, the efficacy of sand fly saliva-based vaccine upon exposure to different salivary protein doses must be evaluated before deployment in endemic areas. Our data also ruled out some biotic factors as responsible for fine-tuning the expression of such genes. Overall, this dissertation makes significant contribution to the development of sand fly-based vaccines against leishmaniasis.
48
Bron, Magali. "Etude écologique et éthologique de Phlebotomus perniciosus (vecteur de leihmaniose canine et humaine) et de Sergentomya minuta dans le Sud Est de la France (région marseillaise)." Aix-Marseille 3, 1994. http://www.theses.fr/1994AIX30039.
Full textAbstract:
Les investigations portent sur les phlebotomes de la banlieue marseillaise. Elle ont permis de recencer deux especes: sergentomyia minuta, herpetophile et p. Perniciosus, vecteur de leishmaniose. Les differences interstationnelles ont permis de degager une typologie basee sur leur mode de fonctionnement. Elles sont en parties regies par des facteurs physiographiques biologiques dont certains ont ete utilises comme descripteurs lors de l'elaboration de la carte ecologique. La dynamique des populations est conditionnee par les facteurs climatiques. L'influence des facteurs biologiques semble se determiner par une concurrence interspecifique exerce entre s. Minuta et p. Perniciosus au detriment de cette derniere. Elle se traduit par des phenomenes adaptatifs tels que la diminution de la taille de p. Perniciosus et la derive du sex ratio en faveur des males de cette espece. L'etude des facteurs lithologiques en laboratoire et sur le terrain montre le role essentiel de la matiere organique dans la regulation des facteurs hygrothermiques et du relargage des infrarouges thermiques. Associee aux sables, la matiere organique intervient dans le degre de fertilite des gites, et dans le choix d'un gite de ponte par les femelles des deux especes
49
Signorini, Manuela. "Analisi ecologica dei flebotomi vettori della leishmaniosi canina in Italia nord-orientale attraverso l'utilizzo di Sistemi Informativi Geografici (GIS) e Modelli di Distribuzione di Specie." Doctoral thesis, Università degli studi di Padova, 2014. http://hdl.handle.net/11577/3423675.
Full textAbstract:
Since the 1990s, new foci of Canine Leishmaniosis have being detected in northern Italy, previously regarded as non-endemic. The disease has increased gradually in the regions of the Alps and entomological surveys demonstrated the established presence of two vectors of the disease, P. perniciosus and P. neglectus, in several localities of the area. The changing pattern of the disease is mainly due to movement of infected dogs from endemic areas, to the increase in human and animal travels and goods trade and to climatic changes, critical to the establishment of the vectors.
The study describes the results of the 12-years entomological survey in Veneto, Friuli Venezia Giulia and Trentino Alto Adige Regions, analysed through the use of GIS tools and Species Distribution Models (SDMs), based on remote sensing environmental data. Several climatic and environmental features were explored in regard to the entomological data, in order to better understand the ecology of the vectors and the epidemiology of the disease, to create a tool in support of surveillance activities.
Sandfly trapping was conducted from 2001 to 2012 in 175 sites, using sticky traps (n=114 sites), CDC light traps (n=53) and CO2 traps (n=66). GPS coordinates were acquired for all sites and potential risk factors (altitude, number and species of domestic bait animals, structural characteristics of site and level of urbanization) were identified and registered. The presence/absence of sandflies were compared with the risk factors considered and environmental variables, such as MODIS data (Normalized Difference Vegetation Index and Land Surface Temperature), a land cover map (Corine Land Cover 2006), a Digital Elevation Model (GTOPO30) and a bioclimatic variable taken from the database WorldClim (BIO 18, precipitation of Warmest Quarter). The environmental features resulted more relevant, were used to built a predictive model of presence of P. perniciosus and sandflies in Veneto, Friuli Venezia Giulia and Trentino Alto Adige Regions, using the software MaxEnt (Maximum Entropy Modeling System).
Overall, 6.144 sandflies were collected and identified and P. perniciosus was the most abundant species (3.797, 61,8%), with density values comparable to endemic areas of southern and central Italy. The ecological analyses of risk factors identified the altitudinal range between 100 and 300 m as the optimal environment for sandflies. The analyses of eco-climatic variables showed that the species P. perniciosus and the sandflies prefer hilly areas, characterized by temperate climate, high vegetation cover and moderate rainfall.
The SDMs developed showed a high predictive power and demonstrated to be realistic, since areas highly suitable for sandflies overlap with Canine Leishmaniosis foci in the study area.
Visualisation of patterns of distribution of vector species in ecological space, using SDMs, was a useful tool for the understanding of the ecological requirements of the sandfly vectors. The used approach may be considered a new resource for the proper identification of the surveillance actions for the control of Canine Leishmaniosis.A partire dagli anni '90, sono stati evidenziati nuovi focolai autoctoni di leishmaniosi canina in Italia settentrionale, area fino agli anni precedenti considerata indenne. Gradualmente, la malattia si è diffusa nelle Regioni dell’arco alpino e i monitoraggi entomologici condotti, hanno evidenziato la presenza stabile di due vettori della malattia, P. perniciosus e P. neglectus, in diverse aree del Territorio. Il cambiamento della distribuzione della malattia sembra legato principalmente al movimento dei cani infetti dalle aree endemiche, all’aumento degli spostamenti di merci, animali e persone ed ai cambiamenti climatici, determinanti per la sopravvivenza dei vettori. Lo studio descrive i risultati di 12 anni di campionamenti entomologici, condotti in Veneto, Friuli Venezia Giulia e Trentino Alto Adige, analizzati attraverso l’utilizzo di strumenti GIS e modelli di distribuzione di specie (SDMs) basati su dati ambientali ottenuti tramite remote sensing. Sono state analizzate diverse caratteristiche climatiche ed ambientali, relazionandole con i dati relativi ai monitoraggi, con la finalità di comprendere meglio l’ecologia dei vettori e l’epidemiologia della malattia, per creare uno strumento di supporto alle attività di sorveglianza. Il monitoraggio è stato condotto dal 2001 al 2012 in 175 siti, utilizzando sticky traps (n=114 siti), CDC light traps (n=53 siti) e CO2 traps (n=66 siti). I siti sono stati geo-referenziati ed alcuni possibili fattori di rischio (altitudine, numero e specie di animali esca, tipologia di sito e livello di urbanizzazione) sono stati individuati e registrati sul campo. I dati di presenza/assenza dei flebotomi sono stati analizzati in funzione dei fattori di rischio considerati, di due variabili ambientali ottenute dal sensore MODIS (Normalized Difference Vegetation Index e Land Surface Temperature), di una mappa di uso del suolo (Corine Land Cover 2006), di un modello digitale del terreno (GTOPO30) e di una variabile climatica ottenuta dal data set WorldClim (BIO 18), relativa alle precipitazioni medie registrate nei trimestri più caldi. Le variabili risultate più rilevanti sono state utilizzate per elaborare un modello predittivo della presenza di P. perniciosus e dei flebotomi in Triveneto, utilizzando il software MaxEnt (Maximum Entropy Modeling System). In totale sono stati raccolti e identificati 6.144 flebotomi e P. perniciosus si è dimostrato essere la specie più rappresentata nel territorio (3.797 esemplari, 61,8%), con densità paragonabili a quelle registrate nelle aree endemiche del centro-sud Italia. L’analisi ecologica dei fattori di rischio ha individuato la fascia altitudinale tra i 100 e i 300 m s.l.m. come quella più idonea ai flebotomi. L’analisi delle variabili ambientali ha evidenziato che i flebotomi e P. perniciosus prediligono ambienti collinari, caratterizzati da un clima mite, elevata copertura vegetazionale e precipitazioni non abbondanti. Il modello di distribuzione di specie elaborato, ha mostrato un buon potere predittivo e si è dimostrato realistico, in quanto l’area risultata più idonea alla presenza dei flebotomi nel territorio si sovrappone in modo chiaro con le aree interessate da focolai di leishmaniosi canina. La visualizzazione dei pattern di distribuzione dei flebotomi vettori nello spazio ecologico, attraverso l’utilizzo di modelli di distribuzione di specie, si è dimostrata essere un valido strumento per comprendere quali fattori condizionano maggiormente la presenza degli stessi. Gli strumenti utilizzati possono essere considerati una nuova risorsa da integrare alle azioni di sorveglianza per la prevenzione della leishmaniosi canina, con il fine di destinare le risorse correttamente e di sviluppare strategie sempre più appropriate.
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Malta, Juliana. "Efeitos de antissoros específicos para proteínas associadas a matriz peritrófica, silenciamento gênico da quitinase 1 e morfologia do intestino médio durante a metamorfose de flebotomíneos." Universidade Federal de Viçosa, 2016. http://www.locus.ufv.br/handle/123456789/9222.
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Flebotomíneos (Diptera: Psychodidae: Phlebotominae) são importantes vetores das leishmanioses, doenças causadas por protozoários do gênero Leishmania, distribuídos em dois grandes gêneros de importância médica: Phlebotomus no Velho Mundo e Lutzomyia no Novo Mundo. Após a ingestão de sangue o bolo alimentar é envolto por uma matriz quitino-proteica, chamada matriz peritrófica (MP). Em uma infecção por Leishmania, o intestino do vetor tem papel crucial, pois, para se estabelecer, o protozoário deve escapar do espaço endoperitrófico e se fixar na parede do intestino para evitar sua eliminação durante a excreção. Nesse sentido, a MP funciona como barreira ao desenvolvimento do parasito, sendo um componente importante na competência vetorial de flebotomíneos. Neste trabalho foi estudado o efeito da alimentação com células sanguíneas reconstituídas com anti-soros específicos para duas proteínas associadas à MP, a quitinase PpChit1 e a peritrofina PpPer2, na morfologia da MP de fêmeas de Phebotomus papatasi. A MP foi avaliada por microscopia de luz (ML) e microscopia eletrônica de transmissão (MET) (24, 42–46, 48 e 72 h após a alimentação), microscopia de força atômica (MFA) (30 h após a alimentação) e microscopia confocal (WGA-FITC) (72 horas após a alimentação). Nesta mesma espécie, também foi estudado a inibição da expressão de PpChit1 pela técnica de RNA de interferência (RNAi) após a injeção de dsPpChit1 (24, 48, 72 e 96 h após a alimentação sanguínea). Adicionalmente, o desenvolvimento pós embrionário do intestino médio foi investigado nas seguintes fases/estágios: larvas de 4o instar com três dias (L4-3) e com cinco dias (L4-5) após a ecdise, pré-pupa, pupa 24 horas e 72 horas após início da metamorfose e adulto recém-emergido, nos flebotomíneos Lutzomyia longipalpis e P. papatasi. Amostras de intestinos médios dissecados de cada fase foram avaliados por microscopias de luz (ML), eletrônica de transmissão (MET) e fluorescência. Verificamos que a alimentação de fêmeas de P. papatasi com antisoros específicos para PpChit1 e PpPer1, levou a um aumento na espessura da MP 72 h após a alimentação, bem como um aumento na amplitude da rugosidade na superfície da MP 30 h após a alimentação. A detecção de quitina com WGA-FITC, identificou que 72 h após a alimentação com anti-PpChit1, o conteúdo de quitina associada a MP no intestino médio do inseto era maior que nos insetos alimentados com soro naïve. A alimentação com antisoros específicos contra as proteínas associadas a MP (PpChit1 e PpPer2) afetam a cinética de maturação e degradação da MP, evidenciando o papel dessas proteínas na estruturação da MP de P. papatasi. A injeção de dsPpChit1 levou a uma reducão nos níveis de transcritos em todos os horários analisados, sendo esses resultados o primeiro passo para contribuir futuramente para o entendimento do papel de PpChit1 na MP P. papatasi. As mudanças morfológicas no intestino médio das duas espécies tiveram início no quarto instar larval, no entanto, em P. papatasi o processo degenerativo das células epiteliais iniciou um pouco antes em L4-3 enquanto que em L. longipalpis em L4-5. Durante a metamorfose, células regerativas foram vistas na base do epitélio, nas duas espécies. Além disso, as marcações positivas para a histona fosforilada H3, em ambas, sugerem que as células regenerativas se dividem durante o processo de remodelamento do intestino médio em flebotomíneos. A histólise do epitélio intestinal larval se dá possivelmente por autofagia, pela presença de numerosos vacúolos autofágicos, bem como por marcações positivas para a proteína LC3, entretanto, a detecção de caspase-3 sugere que a apoptose possa acontecer durante o processo de troca do epitélio larval pelo do adulto. Finalmente, o estudo do remodelamento do intestino médio em P. papatasi e L. longipalpis mostrou de forma inédita que o processo é conservado nas duas espécies, se diferenciando apenas no tempo do início do processo degenerativo entre as duas espécies. Os conhecimentos relacionados as proteínas da MP, bem como ao desenvolvimento pós-embrionário do intestino médio em flebotomíneos, o qual tem papel fundamental na transmissão de Leishmania, são importantes para uma melhor compreensão do inseto vetor.
Sand flies (Diptera: Psychodidae: Phlebotominae) are vectors of Leishmaniasis, a disease caused by parasitic protozoa of the genus Leishmania. They are distributed in two large medical importance genus: Phlebotomus from the Old World, and Lutzomyia from the New World. Leishmania suprapilarian life cycle in the vector midgut begins when insect females intake infected blood with amastigotes forms from the vertebrate host. After the blood meal, the food bolus is surrounded by a chitin-protein layer, called peritrophic matrix (PM). Sand fly midgut plays a crucial role during a Leishmania infection. In order to survive and develop, Leishmania parasites must escape from endoperitrophic space and attach themselves in the intestinal epithelium, preventing excretion with the fecal pellets. The PM can act as a barrier to parasite development, working as a relevant component in the vector competence. This study investigated the effects of reconstituted blood cells feeding with specific antisera targeting two PM associated proteins, chitinase PpChit1 and peritrophin PpPer2 in the PM formation. The PM was studied under light (LM) and transmission electron (TEM) microscopies (24, 42-46 , 48 and 72 h after blood meal), under atomic force microscopy (AFM) (30 h after blood meal) and under confocal microscopy (WGA–FITC) (72 hours after blood meal) in Phlebotomus papatasi. PpChit1 knockdown was performed in P. papatasi by means RNA interference technique (RNAi) after dsPpChit1 injection (24, 48, 72 and 96 h after blood meal). Additionally, the post-embryonic development of the midgut was investigated in the following life-stages: 4th instar larvae three days (L4-3) and five days (L4-5) after molting, pré-pupae, pupae 24 hours and 72 hours, and newly emerged adult in Lutzomyia longipalpis and P. papatasi. Midgut samples from each stage were dissected and assessed by LM, TEM and immunofluorescence. P. papatasi females feeding with anti-PpChit1 and anti-PpPer1 had the PM thickness increased at 72 h after blood meal, as well as a PM roughnes’s amplitude increase at 30 hr after feeding. WGA-FITC staining indicates that PM chitin content on insect midgut was higher in treated individuals than those treated with naïve serum. The feeding of P. papatasi females with red blood cells reconstituted with antisera targeting PM associated proteins (PpChit1 and PpPer2) affected the PM maturation and degradation, indicating the role of these proteins on PM structure. Injection of dsPpChit1 led to significant decrease in corresponding mRNA levels. These results are the first step on contribution to understand PpChit1 role in P. papatasi PM. The midgut metamorphosis in of the two species begins in the 4th instar, however, in P. papatasi, epithelial cells degeneration started shortly, in L4-3, while in L. longipalpis it began in L4-5. Larval gut epithelium degeneration was intensified in pré-pupa in both species by the presence of numerous autophagic vacuoles. During metamorphosis, midgut remodeling occurs by differentiation of stem or regenerative cells to replace larval digestive cells. Regenerative cells were seen at the epithelium basal region in both species. Furthermore, the detection of phosphohistone H3- positive cells suggested that the stem cells can divide during the remodeling process of the midgut. Stem cells in proliferation and differentiation were seen forming the new digestive epithelium in the pupae. Larval midgut replacement possibly occurs by autophagy by the presence of numerous autophagic vacuoles, as well as by the detection of LC3-positive cells. Additionally, cells positive for caspase-3 suggested that the apoptosis may occur during the elimination of larval epithelium. Finally, the study of midgut remodeling in P. papatasi and L. longipalpis was showed for the first time, and this process is conserved in these species, differing only in the time of th beginning of the degenerative processof the midgut epithelium. The study of MP formation as well as the post-embryonic development of the midgut of sandflies represent important steps for a better vector biology understanding.