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1
BATRA, T. R. "EFFECT OF COMPLETE DRY COW TREATMENT ON MASTITIS CONTROL IN DAIRY CATTLE." Canadian Journal of Animal Science 68, no. 2 (June 1, 1988): 553–56. http://dx.doi.org/10.4141/cjas88-061.
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Data on 297 cows which received complete dry cow treatment and subsequently calved were used in this study. At drying-off, 7.7% of the quarters were infected. Among the mastitic pathogens, Staphylococcus aureus was isolated in 75% of the infected quarters. About 92% of the infected quarters were cured with dry cow treatment and 6% of the quarters had new infections during dry period. It appears that dry cow treatment is an effective means of eliminating Staphylococcus aureus infections. Key words: Mastitis, dry cow treatment, dairy cattle
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Rajamanickam, Karthic, Jian Yang, Saravana Babu Chidambaram, and Meena Kishore Sakharkar. "Enhancing Drug Efficacy against Mastitis Pathogens—An In Vitro Pilot Study in Staphylococcus aureus and Staphylococcus epidermidis." Animals 10, no. 11 (November 15, 2020): 2117. http://dx.doi.org/10.3390/ani10112117.
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Background: Bovine mastitis is one of the major infectious diseases in dairy cattle, resulting in large economic loss due to decreased milk production and increased production cost to the dairy industry. Antibiotics are commonly used to prevent/treat bovine mastitis infections. However, increased antibiotic resistance and consumers’ concern regarding antibiotic overuse make it prudent and urgent to develop novel therapeutic protocols for this disease. Materials and methods: Potential druggable targets were found in 20 mastitis-causing pathogens and conserved and unique targets were identified. Bacterial strains Staphylococcus aureus (ATCC 29213, and two clinical isolates CI 1 and CI 2) and Staphylococcus epidermidis (ATCC 12228, and two clinical isolates CI 1 and CI 2) were used in the present study for validation of an effective drug combination. Results: In the current study, we identified the common and the unique druggable targets for twenty mastitis-causing pathogens using an integrative approach. Furthermore, we showed that phosphorylcholine, a drug for a unique target gamma-hemolysin component B in Staphylococcus aureus, and ceftiofur, the mostly used veterinary antibiotic that is FDA approved for treating mastitis infections, exhibit a synergistic effect against S. aureus and a strong additive effect against Staphylococcus epidermidis in vitro. Conclusion: Based on the data generated in this study, we propose that combination therapy with drugs that work synergistically against conserved and unique targets can help increase efficacy and lower the usage of antibiotics for treating bacterial infections. However, these data need further validations in animal models of infection.
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Vlkova, H., V. Babak, I. Vrtkova, D. Cervinkova, D. Marosevic, M. Moravkova, and Z. Jaglic. "Epidemiology of intramammary infections with Staphylococcus aureus and mastitis streptococci in a dairy cattle herd with a history of recurrent clinical mastitis." Polish Journal of Veterinary Sciences 20, no. 1 (March 28, 2017): 133–39. http://dx.doi.org/10.1515/pjvs-2017-0017.
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AbstractThe aim of the present work was to examine a dairy herd with an anamnesis of recurrent clinical mastitis and decreased milk production. A total of 239 individual cow milk samples originating from asymptomatic cows were collected at four-month intervals and examined mainly for the presence of Staphylococcus aureus and mastitis streptococci using standard cultivation methods. In total, 29.7% and 9.2% samples were positive for S. aureus and mastitis streptococci, respectively. Unlike for mastitis streptococci, the prevalence of animals positive for S. aureus had an increasing trend (p<0.05; Chi-squared test for trend) with rising parity. Despite in vitro susceptibility of S. aureus to potentiated penicillins and cephalosporins, the persistence of S. aureus was observed in cows undergoing intramammary treatment with amoxicillin/clavulanic acid (a potentiated penicillin antibiotic). All isolates of S. aureus were biofilm-positive and had the same macrorestriction pattern. Furthermore, no dependence was observed between the occurrence of S. aureus in milk and previous cases of clinical mastitis, reproductive and periparturient disorders and administration of antibiotics. In contrast to S. aureus, the occurrence of mastitis streptococci in milk was linked with previous cases of clinical mastitis and intramammary administration of antibiotics.
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Neamah, Ahmed Jasim, Hayder Naji Ayyez, Saba Falah Klaif, Yahia Ismail Khudhair, and Muthanna Hadi Hussain. "Molecular and phylogenetic study of Staphylococcus aureus isolated from human and cattle of Al-Qadisiyah Governorate, Iraq." Veterinary World 12, no. 9 (September 2019): 1378–82. http://dx.doi.org/10.14202/vetworld.2019.1378-1382.
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Aim: This study was designed to detect the prevalence of Staphylococcus aureus, to estimate the frequency of methicillin resistance gene (mecA), femA (specific gene for S. aureus), and lukS gene, and the prevalence of urinary tract infection (UTI) in human and bovine mastitis caused by S. aureus. Materials and Methods: A total of 102 cases of S. aureus were included in this study; 72 specimens were isolated from human with UTIs and 30 specimens were isolated from milk of cattle with acute mastitis. Diagnosis was done by VITEK 2 Compact after subculture and purification. All isolates were examined for the presence of mecA, femA, and lukS (Panton- Valentine leukocidin) using multiplex polymerase chain reaction. Results: Culture and biochemical evaluation of the samples revealed the presence of S. aureus, among which the genes mecA, femA, and lukS were positively detected in 68 (94.4%), 36 (50%), and 20 (27.7%) of S. aureus isolates from methicillin-resistant humans, respectively. In the same manner, the genes mecA, femA, and lukS were positively detected in 27 (90%), 14 (46.7%), and 11 (36.7%) of S. aureus isolates from methicillin-resistant cattle. Sequencing of partial order of femA gene isolated from human isolate and from cattle with mecA isolated from human revealed high sequence identity with the National Center for Biotechnology Information (NCBI)-Basic Local Alignment Search Tool. S. aureus isolates and the phylogenetic analysis showed that there was a significant genetic similarity (0.5 genetic change) between human and animals isolates, and then, the gene sequences were deposited into NCBI-Genbank accession numbers MG696860.1 for mecA and femA from human, MG696861.1 for mecA and femA from cattle, MK474469.1 for mecA and femA gene from human, and MG696862.1 for mecA and femA gene from cattle. Conclusion: The study represents the first report of genetic relationship between S. aureus from humans and cattle of Iraq. Therefore, it is essential to define the role of animals as an important source of the distribution of pathogen related to public health. The continuous monitoring of methicillin susceptibility pattern of S. aureus isolates that have high standards of infections might prevent methicillin-resistant S. aureus transmission in either direction between human and cattle, the risk of dairy milk on humans, or self-direction between the same species.
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Hoekstra, Jurriaan, Victor P. M. G. Rutten, Theo J. G. M. Lam, Kok P. M. Van Kessel, Mirlin P. Spaninks, J. Arjan Stegeman, Lindert Benedictus, and Gerrit Koop. "Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent." Microorganisms 7, no. 12 (December 12, 2019): 688. http://dx.doi.org/10.3390/microorganisms7120688.
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Bovine mastitis is a costly disease to the dairy industry and intramammary infections (IMI) with Staphylococcus aureus are a major cause of mastitis. Staphylococcus aureus strains responsible for mastitis in cattle predominantly belong to ruminant-associated clonal complexes (CCs). Recognition of pathogens by bovine mammary epithelial cells (bMEC) plays a key role in activation of immune responsiveness during IMI. However, it is still largely unknown to what extent the bMEC response differs according to S. aureus CC. The aim of this study was to determine whether ruminant-associated S. aureus CCs differentially activate bMEC. For this purpose, the immortalized bMEC line PS was stimulated with S. aureus mastitis isolates belonging to four different clonal complexes (CCs; CC133, CC479, CC151 and CC425) and interleukin 8 (IL-8) release was measured as indicator of activation. To validate our bMEC model, we first stimulated PS cells with genetically modified S. aureus strains lacking (protein A, wall teichoic acid (WTA) synthesis) or expressing (capsular polysaccharide (CP) type 5 or type 8) factors expected to affect S. aureus recognition by bMEC. The absence of functional WTA synthesis increased IL-8 release by bMEC in response to bacterial stimulation compared to wildtype. In addition, bMEC released more IL-8 after stimulation with S. aureus expressing CP type 5 compared to CP type 8 or a strain lacking CP expression. Among the S. aureus lineages, isolates belonging to CC133 induced a significantly stronger IL-8 release from bMEC than isolates from the other CCs, and the IL-8 response to CC479 was higher compared to CC151 and CC425. Transcription levels of IL-8, tumor necrosis factor alpha (TNFα), serum amyloid A3 (SAA3), Toll-like receptor (TLR)-2 and nuclear factor κB (NF-κB) in bMEC after bacterial stimulation tended to follow a similar pattern as IL-8 release, but there were no significant differences between the CCs. This study demonstrates a differential activation of bMEC by ruminant-associated CCs of S. aureus, which may have implications for the severity of mastitis during IMI by S. aureus belonging to these lineages.
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Tesfaye, Kaleab, Zemichael Gizaw, and Aklilu Feleke Haile. "Prevalence of Mastitis and Phenotypic Characterization of Methicillin-Resistant Staphylococcus aureus in Lactating Dairy Cows of Selected Dairy Farms in and Around Adama Town, Central Ethiopia." Environmental Health Insights 15 (January 2021): 117863022110212. http://dx.doi.org/10.1177/11786302211021297.
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Background: The emergence of Methicillin resistant Staphylococcus aureus (MRSA) poses a serious public health threat. Strains of Staphylococcus aureus resistant to β-lactam antibiotics are known as MRSA. MRSA has gained attention as community pathogen. MRSA has been increasingly reported as emerging problem in veterinary medicine. However, little is known in Ethiopia. This study was, therefore, conducted to identify MRSA, to determine its drug susceptibility patterns, and mastitis infection in dairy cattle in and around Adama town, Central Ethiopia. Methods: A cross-sectional study was conducted to estimate the occurrence of MRSA in mastitic dairy cows in and around Adama town, central Ethiopia. A total of 384 lactating cows were included from the conveniently selected dairy farms in the study area. Approximately 10 ml of milk was aseptically collected from clinical and subclinical mastitic cows into sterile universal bottles after discarding the first 3 milking streams. Then, Staphylococcus aureus was isolated using the conventional bacteriological procedure. Resistance to methicillin was detected using the Kirby-Bauer disc diffusion antibiotic susceptibility method. Oxacillin disc was used to detect methicillin resistant Staphylococcus aureus strains. Antimicrobial susceptibility test was conducted against MRSA strains using streptomycin (S, 10 µg), amoxicillin (Am, 25 µg), kanamycin (k, 30 µg), nalidixic acid (NA, 30 µg), oxytetracycline (OT, 30 µg) sulphonamide (S, 300 µg) and ceftriazole (CRO, 30 µg). Results: The study found that the prevalence of mastitis was 121(31.5%). Among this 37(30.6%) were clinical mastitis and 84 (69.4%) of them were sub-clinical mastitis. Of 121 mastitis cases, Staphylococcus aureus was isolated in 37 (30.6%) of mastitic cow milk samples. The prevalence of mastitis was significantly affected by breed, age, floor type and hygienic status of the milkers ( P < .05). Moreover, 32.4% of Staphylococcus aureus isolates were resistant to oxacillin. A total of 75% percent of MRSA isolates were resistant to amoxicillin, 66.7% were resistant to oxytetracycline, and 50% were resistant to sulphonamide. However, 75% of MRSA isolates were susceptible to kanamycin, 58.3% were susceptible to streptomycin, and 50% were susceptible to nalidixic acid. Conclusion: The study revealed that relatively high number of strains are resistant to the antibiotics commonly used in the therapeutic protocol of many human and animal infections. Therefore, antimicrobial susceptibility test should be carried out at a regular basis and proper hygienic practices should be introduced at farm level. Creating public awareness about transmission, prevention and control of MRSA should also be considered.
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DALLA POZZA, MARIA CRISTINA, ANTONIA RICCI, and GADDO VICENZONI. "Protein A gene polymorphism analysis in Staphylococcus aureus strains isolated from bovine subclinical mastitis." Journal of Dairy Research 66, no. 3 (August 1999): 449–53. http://dx.doi.org/10.1017/s0022029999003672.
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Mammary infections caused by Staphylococcus aureus are still one of the most serious problems in dairy farms all over the world (Sischo et al. 1993), and the epidemiology of the infection has not yet been completely elucidated (Aarestrup et al. 1995). Any effective modern approach to this disease must therefore be based on more comprehensive epidemiological studies, conducted with valid microbiological typing tools.A technique for use in epidemiological studies should identify many types, and should be inexpensive, quick and easy to perform, but above all reproducible. Among the available methods, phage typing has up to now been widely and successfully used in differentiating strains of Staph. aureus isolated from cattle with mastitis (Mackie et al. 1987; Fox et al. 1991), but it has some limitations, being a technically demanding method subject to considerable experimental as well as biological variation (Maslow et al. 1993). Moreover, in some studies the number of strains that could not be typed with available bacteriophage panels has been high (Carroll & Francis, 1985; Farah et al. 1988).Alternative methods have been investigated, and of these molecular techniques have been the most intensely studied (Aarestrup et al. 1995). In studies of human infections caused by Staph. aureus, analysis of the so-called X region of protein A gene polymorphism has been a useful epidemiological marker (Frénay et al. 1994). This gene is ∼2150 bp and harbours some functionally distinct regions: an FC-binding region, the so-called X region and, at the C terminus, a sequence required for cell wall attachment (Guss et al. 1984; Frénay et al. 1994). The X region polymorphism depends on the presence, within the region itself, of a varying number of 24 bp repeats, highlighted by the amplification of this highly polymorphic DNA region and its subsequent restriction fragment length polymorphism (RFLP) analysis (Frénay et al. 1994).Human epidemic (MRSA) and non-epidemic methicillin-resistant Staph. aureus (non-MRSA) strains, which both cause infections but have completely different infection patterns, have been successfully distinguished by analysis of this polymorphism (Frénay et al. 1994). However, protein A has been identified in only 93% of Staph. aureus strains isolated from bovine intramammary infections (Poutrel & Ducelliez, 1979; Johne & Jarp, 1988).The aim of the present study was to determine whether the gene for protein A of Staph. aureus (Spa) was present in Staph. aureus strains isolated from cases of subclinical bovine mastitis. This was carried out using the polymerase chain reaction (PCR), as suggested by Frénay et al. (1994). In addition, we have investigated the genetic polymorphism related to the X region of the gene, by means of PCR amplification and subsequent RFLP analysis. Finally we verified the stability of this region after in vitro subculture.
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Kalashnikova, V. A. "Agr-typing of methicillin-susceptible Staphylococcus aureus (MSSA) isolated from non-human primates." Veterinary Science Today, no. 2 (June 16, 2020): 127–31. http://dx.doi.org/10.29326/2304-196x-2020-2-33-127-131.
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Staphylococcus aureus (S. aureus) is a microorganism that causes a great number of diseases in humans and animals, including sepsis, pneumonia, food toxicoinfections, wound abscess, etc. Numerous studies on genotyping S. aureus strains isolated from humans, food and mastitis in cattle and small ruminants have been carried out. The lack of information on the genotyping of methicillin-susceptible S. aureus detected in monkeys served as a stimulus to conduct a similar research, since staphylococcal infections in the primates are widespread. The present study is devoted to molecular genetic testing of S. aureus isolated from different biological samples taken from monkeys and is based on typing of agr polymorphic locus which acts as a regulator of pathogenic gene expression. As a result of PCR analysis of 301 S. aureus isolates it was established that most of S. aureus belonged to agr IV (55%), and agr I (34%) was the second most group. Data resulting from the study differ from the results of other researchers published in literary sources, who performed typing of salmonella isolated from people with agr I prevailing. In conducting the study, neither distinct correlation between microbial isolation source and agr complex groups, nor relationship between the diseases and S. aureus group specificity were detected. Prevalence ratio of each agr group is nearly similar in S. aureus isolated from rhesus macaques and crab-eating macaques. But in hamadryas baboons and green monkeys II and III groups of agr complex were not detected.
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JOHLER, SOPHIA, FRANZISKA LAYER, and ROGER STEPHAN. "Comparison of Virulence and Antibiotic Resistance Genes of Food Poisoning Outbreak Isolates of Staphylococcus aureus with Isolates Obtained from Bovine Mastitis Milk and Pig Carcasses." Journal of Food Protection 74, no. 11 (November 1, 2011): 1852–59. http://dx.doi.org/10.4315/0362-028x.jfp-11-192.
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Staphylococcus aureus is the etiological agent in a variety of infections in humans and livestock and produces enterotoxins leading to staphylococcal food poisoning (SFP), one of the most prevalent foodborne intoxication diseases worldwide. Pork and bovine milk are considered possible sources of SFP because pig skin is often colonized by S. aureus and bovine mastitis caused by S. aureus is common, but conclusive data are limited. The objective of the present study was to compare S. aureus isolates associated with cases of SFP with isolates obtained from bovine mastitis milk and pig carcasses. DNA microarray analysis and spa gene typing were performed with 100 S. aureus isolates: 20 isolates related to outbreaks of SFP in humans, 39 isolates obtained from pig carcasses, and 41 isolates collected from bovine mastitis milk. No overlap in spa types was observed for SFP isolates (t008, t015, t018, t024, t056, t084, t279, t377, t383, t648, t733, t912, t1239, t1270, t4802, and t6969) and isolates gathered from milk or pork. The porcine isolates were assigned to t034, t208, t337, t524, t899, t1939, t2922, t2971, t4475, and t7006, and the bovine isolates belonged to t267, t524, t529, t1403, t2953, t7007, t7008, and t7013. Comparison of microarray profiles revealed similar virulence gene patterns for isolates collected from the same host (pigs or cattle) but few similarities between SFP isolate profiles and the profiles of isolates obtained from bovine mastitis milk and pig carcasses. Although only some bovine and porcine isolates possessed the β-lactamase gene blaZ (milk, 24%; pork, 28%), significantly higher numbers of SFP isolates contained blaZ (90%). Investigations of these isolates provided no evidence that pork or bovine mastitis milk represent common sources of SFP.
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Donovan, David M., Shengli Dong, Wes Garrett, Geneviève M. Rousseau, Sylvain Moineau, and David G. Pritchard. "Peptidoglycan Hydrolase Fusions Maintain Their Parental Specificities." Applied and Environmental Microbiology 72, no. 4 (April 2006): 2988–96. http://dx.doi.org/10.1128/aem.72.4.2988-2996.2006.
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ABSTRACT The increased incidence of bacterial antibiotic resistance has led to a renewed search for novel antimicrobials. Avoiding the use of broad-range antimicrobials through the use of specific peptidoglycan hydrolases (endolysins) might reduce the incidence of antibiotic-resistant pathogens worldwide. Staphylococcus aureus and Streptococcus agalactiae are human pathogens and also cause mastitis in dairy cattle. The ultimate goal of this work is to create transgenic cattle that are resistant to mastitis through the expression of an antimicrobial protein(s) in their milk. Toward this end, two novel antimicrobials were produced. The (i) full-length and (ii) 182-amino-acid, C-terminally truncated S. agalactiae bacteriophage B30 endolysins were fused to the mature lysostaphin protein of Staphylococcus simulans. Both fusions display lytic specificity for streptococcal pathogens and S. aureus. The full lytic ability of the truncated B30 protein also suggests that the SH3b domain at the C terminus is dispensable. The fusions are active in a milk-like environment. They are also active against some lactic acid bacteria used to make cheese and yogurt, but their lytic activity is destroyed by pasteurization (63°C for 30 min). Immunohistochemical studies indicated that the fusion proteins can be expressed in cultured mammalian cells with no obvious deleterious effects on the cells, making it a strong candidate for use in future transgenic mice and cattle. Since the fusion peptidoglycan hydrolase also kills multiple human pathogens, it also may prove useful as a highly selective, multipathogen-targeting antimicrobial agent that could potentially reduce the use of broad-range antibiotics in fighting clinical infections.
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Thomas, Asha, Shubhada Chothe, Maurice Byukusenge, Tammy Mathews, Traci Pierre, Subhashinie Kariyawasam, Erin Luley, Suresh Kuchipudi, and Bhushan Jayarao. "Prevalence and distribution of multilocus sequence types of Staphylococcus aureus isolated from bulk tank milk and cows with mastitis in Pennsylvania." PLOS ONE 16, no. 3 (March 12, 2021): e0248528. http://dx.doi.org/10.1371/journal.pone.0248528.
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A total of 163 S. aureus isolates; 113 from mastitic milk (MM) and 50 from bulk tank milk (BTM) (2008, 2013–2015) submitted for bacteriologic analysis at the Penn State Animal Diagnostic Laboratory were examined for their phenotypic and genotypic characteristics. Multi-locus sequence typing (MLST) analysis identified 16 unique sequence types (STs) which belonged to eight clonal complexes (CCs). Majority of the isolates were variants of CC97 (68.7%) and CC151 (25.1%). CC97 comprised of seven STs, of which two were new STs (ST3273, ST3274), while CC151 comprised of three STs of which ST3272 was identified for the first time. Several farms had more than one ST type that were either members of the same clonal complex or unrelated STs. On one farm, six different STs of both categories were seen over the years within the farm. It was observed that ST352 and ST151 were the two main clonal populations in cattle not only in Pennsylvania but also globally. Most isolates were susceptible to all the antibiotics evaluated. 6.7% of isolates showed resistance to vancomycin and penicillin. Two isolates of ST398 showed multidrug resistance (>3 antibiotics) against clindamycin, erythromycin, tetracycline, and penicillin. It was noted that 59 of 163 (36.2%) isolates encoded for enterotoxigenic genes. Enterotoxin genes seg/sei accounted for ~85% of enterotoxin positive isolates. Toxic shock syndrome gene tsst-1 alone was positive in two isolates (ST352, ST 2187). 97.5% of CC151 isolates were enterotoxin seg/sei positive. Most isolates were positive for lukED (95%) and lukAB (96.3%) leukotoxin genes. Bovine specific bi-component leucocidin lukMF’ was present in 54% of isolates. A prominent observation of this study was the explicit association of lukMF’ with lineages ST151 and ST352. In conclusion, the findings of the study, suggest that small number of S. aureus STs types (ST352, ST2187, ST3028, and ST151) are associated with majority of cases of bovine mastitis in Pennsylvania dairy farms. It was observed that one ST of S. aureus predominated in the herd and this ST can coexist with several other ST types of S. aureus strains. When STs were interpreted along with virulence, leucocidin genes and antimicrobial resistance, ST-variants allowed better interpretation of the S. aureus molecular epidemiologic findings specifically for tracing recurrence or persistence of infections in cow over time, among cows in the herd, and between herds in Pennsylvania.
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Paduch, Jan-Hendrik, Elmar Mohr, and Volker Krömker. "The association between bedding material and the bacterial counts of Staphylococcus aureus, Streptococcus uberis and coliform bacteria on teat skin and in teat canals in lactating dairy cattle." Journal of Dairy Research 80, no. 2 (February 28, 2013): 159–64. http://dx.doi.org/10.1017/s0022029913000046.
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Several mastitis-causing pathogens are able to colonize the bovine teat canal. The objective of this study was to investigate the association between the treatment of sawdust bedding with a commercial alkaline conditioner and the bacterial counts on teat skin and in the teat canal. The study used a crossover design. Ten lactating Holstein cows that were free of udder infections and mastitis were included in the study. The animals were bedded on either untreated sawdust or sawdust that had been treated with a hydrated lime-based conditioner. Once a day, fresh bedding material was added. After 3 weeks, the bedding material was removed from the cubicles, fresh bedding material was provided, and the cows were rotated between the two bedding material groups. Teat skin and teat canals were sampled using the wet and dry swab technique after weeks 1, 2, 3, 4, 5 and 6. Staphylococcus aureus, Streptococcus uberis, Escherichia coli and other coliform bacteria were detected in the resulting agar plate cultures. The treatment of the bedding material was associated with the teat skin bacterial counts of Str. uberis, Esch. coli and other coliform bacteria. An association was also found between the bedding material and the teat canal bacterial counts of coliform bacteria other than Esch. coli. For Staph. aureus, no associations with the bedding material were found. In general, the addition of a hydrated lime-based conditioner to sawdust reduces the population sizes of environmental pathogens on teat skin and in teat canals.
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Goldammer, T., H. Zerbe, A. Molenaar, H. J. Schuberth, R. M. Brunner, S. R. Kata, and H. M. Seyfert. "Mastitis Increases Mammary mRNA Abundance of β-Defensin 5, Toll-Like-Receptor 2 (TLR2), and TLR4 but Not TLR9 in Cattle." Clinical Diagnostic Laboratory Immunology 11, no. 1 (January 2004): 174–85. http://dx.doi.org/10.1128/cdli.11.1.174-185.2004.
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ABSTRACT Coordination of the primary defense mechanisms against pathogens relies on the appropriate expression of pathogen recognition receptors (PRRs) triggering the early release of effector molecules of the innate immune system. To analyze the impact of this system on the counteraction of infections of the mammary gland (mastitis), we characterized the bovine gene encoding the key PRR Toll-like receptor 9 (TLR9) and mapped its precise position on chromosome BTA22. The sequence information was used to establish real-time PCR quantification assays to measure the mRNA abundances of TLR9, TLR2, and TLR4 together with those of β-defensin 5 (BNBD5), an early bactericidal effector molecule of the innate system, in healthy and infected mammary glands. Mastitis strongly increased (4- to 13-fold) the mRNA abundances of all of these genes except TLR9. Slight subclinical infections already caused a substantial increase in the copy numbers, though they did so the least for TLR9. Induction was not systemic, since mRNA abundance was low in uninfected control quarters of the udder but high in the severely infected quarters of the same animal. The number of TLR2 copies correlated well with those of TLR4, indicating coordinated regulation of these two PRRs during infection of the udder. Their coordinated regulation explains our unexpected observation that pure Staphylococcus aureus infections caused a strong increase also in TLR4 mRNA abundance. In situ hybridizations revealed that BNBD5 is expressed predominantly in the mammary epithelial cells (MEC) of the infected gland. Our data therefore suggest a significant contribution of the innate immune system to counteract mastitis and attribute a prominent effector function to the MEC.
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El-Deeb, Wael, Mahmoud Fayez, Naser Alhumam, Ibrahim Elsohaby, Sayed A. Quadri, and Hermine Mkrtchyan. "The effect of staphylococcal mastitis including resistant strains on serum procalcitonin, neopterin, acute phase response and stress biomarkers in Holstein dairy cows." PeerJ 9 (May 31, 2021): e11511. http://dx.doi.org/10.7717/peerj.11511.
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Staphylococcal mastitis (SM) is a frequent disease in the dairy cattle that is costly to treat. This study aimed to investigate the alterations in the levels of procalcitonin (PCT), neopterin (NPT), haptoglobin (HP), serum amyloid A (SAA), proinflammatory cytokines (IL-1β, IL-8, TNF-α, IF-γ) and oxidative stress (OS) biomarkers in Holstein dairy cows with SM under field conditions. In addition, we also evaluated the role of examined biomarkers in disease pathogenesis and their use as diagnostic biomarkers for the disease in dairy cows. Fifty-three dairy cows with SM, including those with infections caused by Staphylococcus aureus (n = 42) and methicillin resistant S. aureus (MRSA) (n = 11) were selected for this study. In addition, 20 healthy dairy cows were enrolled as a control group. Higher serum levels of PCT, NP, IL-1β, IL-8, TNF-α, IF-γ, HP and SAA and a state of OS was detected in SM group in comparison with the controls. Moreover, the levels of all examined biomarkers in mastitic cows with S. aureus when compared with those infected with MRSA was not significantly different. All examined biomarkers demonstrated a significant degree of discrimination between SM cows and healthy controls (the area under the curve (AUC) ranged from 83.6 for SAA to 100 for PCT). Our study showed that SM in dairy cows was associated with substantial changes in serum PCT, NPT, Acute phase proteins (APPs), proinflammatory cytokines, and OS levels. This study demonstrates that clinical examination in tandem with quantification of PCT, NPT, APPs and cytokines, OS biomarkers could be a useful assessment tool for SM in dairy cows.
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Mdegela, Robinson H., Elibariki R. Mwakapeje, Bachana Rubegwa, Daniel T. Gebeyehu, Solange Niyigena, Victoria Msambichaka, Hezron E. Nonga, Nicolas Antoine-Moussiaux, and Folorunso O. Fasina. "Antimicrobial Use, Residues, Resistance and Governance in the Food and Agriculture Sectors, Tanzania." Antibiotics 10, no. 4 (April 16, 2021): 454. http://dx.doi.org/10.3390/antibiotics10040454.
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All infections are potentially curable as long as the etiological agents are susceptible to antimicrobials. The increased rate at which antimicrobials are becoming ineffective is a global health risk of increasing concern that threatens withdrawal of beneficial antimicrobials for disease control. The increased demand for food of animal origin, in particular eggs, meat and milk has led to intensification and commercial production systems where excessive use and misuse of antimicrobials may prevail. Antimicrobials, handled and used by farmers and animal attendants with no formal education, may be predisposed to incorrect dosages, misuse, incorrect applications and non-adherence to withdrawal periods. This study was conducted to assess the regulatory roles and governance of antimicrobials, establish the pattern and extent of their use, evaluate the antimicrobial residues and resistance in the food animals and crop agriculture value chains, and relate these findings to existing strategies in place for combating the emergence of antimicrobial resistance in Tanzania. A multimethod approach (desk review, field study and interviews) was used. Relevant establishments were also visited. High levels of resistance to penicillin G, chloramphenicol, streptomycin and oxytetracycline have been reported, especially for Actinobacter pyogenes, Staphylococcus hyicus, Staphylococcus intermedius and Staphylococcus aureus from dairy cattle with mastitis and in humans. Similar trends were found in poultry where eggs and meat are contaminated with Escherichia coli strains resistant to amoxicillin + clavulanate, sulphamethoxazole and neomycin. An increasing trend of emerging multidrug resistant E. coli, Klebsiella pneumoniae, Staphylococcus aureus and Salmonella was also found in food animals. An increase in methicillin resistant Staphlococcus aureus (MRSA) and extended-spectrum beta-lactamase (ESBL) in the livestock sector in Tanzania have been reported. The pathogens isolated in animals were resistant to ampicillin, augmentin, gentamicin, co-trimoxazole, tetracycline, amoxicillin, streptomycin, nalidixic acid, azithromycin, chloramphenicol, tylosin, erythromycin, cefuroxime, norfloxacin and ciprofloxacin. An increased usage of antimicrobials for prophylaxis, and therapeutics against pathogens and for growth promotion in livestock, aquaculture and crop production were observed. A One Health strategic approach is advocated to combat antimicrobial resistance (AMR) in the food and agriculture sectors in Tanzania. Practical recommendations include (a) legislation review and implementation; (b) antimicrobial use (AMU), AMR and antimicrobial residue (AR) awareness and advocacy among stakeholders along the value chain; (c) strengthening of surveillance and monitoring programs for AMU, AMR and AR; (d) enhanced development and use of rapid and innovative diagnostic tests and the promotion of biosecurity principles; and (e) good husbandry practices. The utilization of this information to improve public health policies and reduce the burden of AMR will be beneficial.
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Mahmmod, Yasser S., Nils Toft, Jørgen Katholm, Carsten Grønbæk, and Ilka C. Klaas. "Bayesian estimation of test characteristics of real-time PCR, bacteriological culture and California mastitis test for diagnosis of intramammary infections with Staphylococcus aureus in dairy cattle at routine milk recordings." Preventive Veterinary Medicine 112, no. 3-4 (November 2013): 309–17. http://dx.doi.org/10.1016/j.prevetmed.2013.07.021.
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Saidi, Radhwane, Djamel Khelef, and Rachid Kaidi. "Subclinical mastitis in cattle in Algeria: Frequency of occurrence and bacteriological isolates." Journal of the South African Veterinary Association 84, no. 1 (February 12, 2013). http://dx.doi.org/10.4102/jsava.v84i1.929.
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The present study was carried out to determine the prevalence of subclinical mastitis in cattle in eighteen herds in the center region of Algeria. Milk samples were collected from 560 quarters of 140 cows free of clinical mastitis. The samples were subjected to California Mastitis Test (CMT) and the positive samples were analysed by bacteriological culture and Speed Mam® Color. The overall quarter prevalence was 28.77% whilst animal prevalence was 28.57%.Bacteriological analysis showed that there was a wide range of bacteria that cause these infections. Staphylococcus aureus (40%) was found to be the most prevalent organism followed by Streptococcus spp. (12.5%), Enterobacteriaceae (2.5%), Pseudomonas spp. (2.5%), Staphylococcusaureus + Streptococcus spp. (12.5%), Streptococcus spp.+ Escherichia coli (7.5%), S. aureus + Mycoplasma spp.(7.5%), and S. aureus +Streptococcus spp.+ E. coli (5%).
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Hoekstra, Jurriaan, Aldert L. Zomer, Victor P. M. G. Rutten, Lindert Benedictus, Arjan Stegeman, Mirlin P. Spaninks, Torben W. Bennedsgaard, et al. "Genomic analysis of European bovine Staphylococcus aureus from clinical versus subclinical mastitis." Scientific Reports 10, no. 1 (October 23, 2020). http://dx.doi.org/10.1038/s41598-020-75179-2.
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Abstract Intramammary infections (IMI) with Staphylococcus aureus are a common cause of bovine mastitis and can result in both clinical (CM) or subclinical mastitis (SCM). Although bacterial isolates of S. aureus differ in their virulence potential it is largely unclear which bacterial virulence factors are responsible for increased clinical severity. We performed a genome wide association study and used a generalized linear mixed model to investigate the correlation between gene carriage, lineage and clinical outcome of IMI in a collection of S. aureus isolates from cattle with CM (n = 125) and SCM (n = 151) from 11 European countries. An additional aim was to describe the genetic variation of bovine S. aureus in Europa. The dominant lineages in our collection were clonal complex (CC) 151 (81/276, 29.3%), CC97 (54/276, 19.6%), CC479 (32/276, 11.6%) and CC398 (19/276, 6.9%). Virulence and antimicrobial resistance (AMR) gene carriage was highly associated with CC. Among a selection of nine virulence and AMR genes, CC151, CC479 and CC133 carried more virulence genes than other CCs, and CC398 was associated with AMR gene carriage. Whereas CC151, CC97 were widespread in Europe, CC479, CC398 and CC8 were only found in specific countries. Compared to CC151, CC479 was associated with CM rather than SCM (OR 3.62; 95% CI 1.38–9.50) and the other CCs were not. Multiple genes were associated with CM, but due to the clustering within CC of carriage of these genes, it was not possible to differentiate between the effect of gene carriage and CC on clinical outcome of IMI. Nevertheless, this study demonstrates that characterization of S. aureus CC and virulence genes helps to predict the likelihood of the occurrence of CM following S. aureus IMI and highlights the potential benefit of diagnostics tools to identify S. aureus CC during bovine mastitis.
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Barasuol, Bibiana Martins, Valessa Lunkes Ely, Antônio Francisco Igor Magalhães de Matos, Luis Antônio Sangioni, Agueda Castagna de Vargas, Daniela Isabel Brayer Pereira, Amanda Carneiro Martini, Luciana Pötter, and Sônia de Avila Botton. "In vitro lytic efficiency of Staphylococcus aureus bacteriophages in bacteria from bovine mastitis: a meta-analysis." Ciência Rural 51, no. 10 (2021). http://dx.doi.org/10.1590/0103-8478cr20200839.
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ABSTRACT: Bacteriophages have been investigated as alternative to the treatment of bacterial infections, including bovine mastitis, in production animals. In this meta-analysis, we evaluated in vitro efficiency of phages of Staphylococcus aureus against S. aureus, which is involved in the etiology of bovine mastitis. Seventeen studies were included and the bacterial lytic activity was extracted using proportion analysis. The lytic efficiency of phages was obtained in this meta-analysis using a random-effects model [significant difference (P<0.05)]. Forest plots were used to graphically represent the efficiency of phages on bacterial isolates. Most phages (e.g., CS1, DW2, ΦSA011, ΦSA012, ΦSA022, ΦSA023, ΦSA024, ΦSA025, ΦSA037, ΦSA038, ΦSA039, ΦSA041, ΦSA042, ΦSA043, ΦSA044, MSA6, Ufv-aur2 to Ufv-aur11, SAH-1, SPW, vB_SauM_JS25, SaPh1 to SaPh6, SA, SANF, SA2, ΦSA012, ΦSA039, phi11, phiIPLA88, phiIPLA35, phiIPLA-RODI, phiIPLA-C1C, SAJK-IND, vBSP-A1, vBSP-A2, STA1.ST29, EB1.ST11, EB1.ST27, Remus, and ISP) were efficiently lytics or infected most S. aureus isolates, demonstrating 80% (P<0.05) lytic efficiency. The phages SA, SANF and SA2, also demonstrated lytic activity or infected the non-Staphylococcus aureus and Macrococcus caseolyticus isolates. In this meta-analysis, we compared and demonstrated the in vitro efficiency and host range of S. aureus phages. Additionally, the phages represent an alternative to be researched to treat bovine mastitis in dairy cattle caused by the prevalent microorganism, S. aureus.
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Vrieling, M., K. J. Koymans, D. A. C. Heesterbeek, P. C. Aerts, V. P. M. G. Rutten, C. J. C. de Haas, K. P. M. van Kessel, A. P. Koets, R. Nijland, and J. A. G. van Strijp. "Bovine Staphylococcus aureus Secretes the Leukocidin LukMF′ To Kill Migrating Neutrophils through CCR1." mBio 6, no. 3 (June 4, 2015). http://dx.doi.org/10.1128/mbio.00335-15.
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ABSTRACTAlthoughStaphylococcus aureusis best known for infecting humans, bovine-specific strains are a major cause of mastitis in dairy cattle. The bicomponent leukocidin LukMF′, exclusively harbored byS. aureusof ruminant origin, is a virulence factor associated with bovine infections. In this study, the molecular basis of the host specificity of LukMF′ is elucidated by identification of chemokine receptor CCR1 as its target. Bovine neutrophils, the major effector cells in the defense against staphylococci, express significant cell surface levels of CCR1, whereas human neutrophils do not. This causes the particular susceptibility of bovine neutrophils to pore formation induced by LukMF′. BovineS. aureusstrains produce high levels of LukMF′in vitro. In culture supernatant of the mastitis field isolate S1444, LukMF′ was the most important cytotoxic agent for bovine neutrophils. In a fibrin gel matrix, the effects of thein situsecreted toxins on neutrophils migrating towardS. aureuswere visualized. Under these physiologicalex vivoconditions, bovineS. aureusS1444 efficiently killed approaching neutrophils at a distance through secretion of LukMF′. Altogether, our findings illustrate the coevolution of pathogen and host, provide new targets for therapeutic and vaccine approaches to treat staphylococcal diseases in the cow, and emphasize the importance of staphylococcal toxins in general.IMPORTANCEThis study explains the mechanism of action of LukMF′, a bicomponent toxin found in bovine lineages ofS. aureusthat is associated with mastitis in cattle. At a molecular level, we describe how LukMF′ can specifically kill bovine neutrophils. Here, we demonstrate the contribution of toxins in the determination of host specificity and contribute to the understanding of mechanisms of coevolution of pathogen and host. Our study provides new targets that can be used in therapeutic and vaccine approaches to treat staphylococcal diseases in the cow. We also demonstrate the importance of toxins in specific elimination of immune cells, which has broader implications, especially in human infections.
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Marbach, Helene, Katharina Mayer, Claus Vogl, Jean Y. H. Lee, Ian R. Monk, Daniel O. Sordelli, Fernanda R. Buzzola, Monika Ehling-Schulz, and Tom Grunert. "Within-host evolution of bovine Staphylococcus aureus selects for a SigB-deficient pathotype characterized by reduced virulence but enhanced proteolytic activity and biofilm formation." Scientific Reports 9, no. 1 (September 17, 2019). http://dx.doi.org/10.1038/s41598-019-49981-6.
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Abstract Staphylococcus aureus is a major cause of bovine mastitis, commonly leading to long-lasting, persistent and recurrent infections. Thereby, S. aureus constantly refines and permanently adapts to the bovine udder environment. In this work, we followed S. aureus within-host adaptation over the course of three months in a naturally infected dairy cattle with chronic, subclinical mastitis. Whole genome sequence analysis revealed a complete replacement of the initial predominant variant by another isogenic variant. We report for the first time within-host evolution towards a sigma factor SigB-deficient pathotype in S. aureus bovine mastitis, associated with a single nucleotide polymorphism in rsbU (G368A → G122D), a contributor to SigB-functionality. The emerged SigB-deficient pathotype exhibits a substantial shift to new phenotypic traits comprising strong proteolytic activity and poly-N-acetylglucosamine (PNAG)-based biofilm production. This possibly unlocks new nutritional resources and promotes immune evasion, presumably facilitating extracellular persistence within the host. Moreover, we observed an adaptation towards attenuated virulence using a mouse infection model. This study extends the role of sigma factor SigB in S. aureus pathogenesis, so far described to be required for intracellular persistence during chronic infections. Our findings suggest that S. aureus SigB-deficiency is an alternative mechanism for persistence and underpin the clinical relevance of staphylococcal SigB-deficient variants which are consistently isolated during human chronic infections.
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Naushad, Sohail, Diego B. Nobrega, S. Ali Naqvi, Herman W. Barkema, and Jeroen De Buck. "Genomic Analysis of Bovine Staphylococcus aureus Isolates from Milk To Elucidate Diversity and Determine the Distributions of Antimicrobial and Virulence Genes and Their Association with Mastitis." mSystems 5, no. 4 (July 7, 2020). http://dx.doi.org/10.1128/msystems.00063-20.
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ABSTRACT Staphylococcus aureus causes persistent clinical and subclinical bovine intramammary infections (IMI) worldwide. However, there is a lack of comprehensive information regarding genetic diversity, the presence of antimicrobial resistance (AMR), and virulence genes for S. aureus in bovine milk in Canada. Here, we performed whole-genome sequencing (WGS) of 119 Canadian bovine milk S. aureus isolates and determined they belonged to 8 sequence types (ST151, ST352, ST351, ST2187, ST2270, ST126, ST133, and ST8), 5 clonal complexes (CC151, CC97, CC126, CC133, and CC8), and 18 distinct Spa types. Pan-, core, and accessory genomes were composed of 6,340, 1,279, and 2,431 genes, respectively. Based on phenotypic screening for AMR, resistance was common against beta-lactams (19% of isolates) and sulfonamides (7% of isolates), whereas resistance against pirlimycin, tetracycline, ceftiofur, and erythromycin and to the combination of penicillin and novobiocin was uncommon (3, 3, 3, 2, and 2% of all isolates, respectively). We also determined distributions of 191 virulence factors (VFs) in 119 S. aureus isolates after classifying them into 5 functional categories (adherence [n = 28], exoenzymes [n = 21], immune evasion [n = 20], iron metabolism [n = 29], and toxins [n = 93]). Additionally, we calculated the pathogenic potential of distinct CCs and STs and determined that CC151 (ST151 and ST351) had the highest pathogenic potential (calculated by subtracting core-VFs from total VFs), followed by CC97 (ST352 and ST2187) and CC126 (ST126 and ST2270), potentially linked to their higher prevalence in bovine IMI worldwide. However, there was no statistically significant link between the presence of VF genes and mastitis. IMPORTANCE Staphylococcus aureus is a major cause of bovine intramammary infections, leading to significant economic losses to dairy industry in Canada and worldwide. There is a lack of knowledge regarding genetic diversity, the presence of antimicrobial resistance (AMR), and virulence genes for S. aureus isolated from bovine milk in Canada. Based on whole-genome sequencing and genomic analysis, we have determined the phylogeny and diversity of S. aureus in bovine milk and concluded that it had a large accessory genome, limited distribution of AMR genes, variable VF gene profiles and sequence types (ST), and clonal complex (CC)-specific pathogenic potentials. Comprehensive information on the population structure, as well as the virulence and resistance characteristics of S. aureus from bovine milk, will allow for source attribution, risk assessment, and improved therapeutic approaches in cattle.