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There are several factors affecting global milk production leading to severe economic losses. Mastitis and leukosis are highly prevalent diseases especially in Brazilian cattle herds. The present study aimed to evaluate the correlation between mastitis and the bovine leukemia virus (BLV) infection in naturally infected cows belonging to dairy herds in Minas Gerais State. Additionally, the prevalence of both diseases, pathogens causing mastitis, and their association with BLV infection and persistent lymphocytosis were evaluated. The prevalence of BLV was determined using the agar gel immunodiffusion test (AGID) in 1,392 lactating cows from ten dairy herds. From the AGID results, 240 cows were selected and divided into two experimental groups: A case group composed of 160 BLV-positive cows (BLV-P) and the control group of 80 BLV-negative cows (BLV-N). Based on serological and leucocytic evaluations, the animals were classified into three subgroups: BLV-N, BLV-P without persistent lymphocytosis (BLV-PWPL), and BLV-positive with persistent lymphocytosis (BLV-PPL). The California Mastitis Test (CMT) was performed on days 0 (D0) and 60 (D60) in the subgroups and the milk from CMT-positive glands were analyzed. BLV infection was present in 100% of herds, with an overall prevalence of 80.9% in cows older than 24 months. Among BLV-P animals, 74% were classified as BLV-PWPL and 26% as BLV-PPL. Coagulase-negative Staphylococcus (73.30%), Corynebacterium spp. (12.04%), Streptococcus agalactiae (8.90%), and Staphylococcus aureus (5.76%) were predominant (61.41%). There were no statistical differences among the scores of clinical or subclinical mastitis with the pathogen type (contagious or environmental) in the BLV-PWPL or BLV-PPL subgroups. However, BLV-P animals showed higher CMT scores than BLV-N animals, suggesting a higher predisposition to subclinical mastitis in cows affected by leukosis. Tendo em vista a relevância econômica e social da produção de leite e a ampla ocorrência da leucose e mastite em rebanhos brasileiros, foi proposto o presente estudo com o objetivo de avaliar a correlação entre a mastite e a infecção pelo vírus da leucose bovina em vacas leiteiras naturalmente infectadas de 10 rebanhos leiteiros da região sul do estado de Minas Gerais. Além disso, foi avaliada a prevalência de ambas as doenças e dos patógenos causadores de mastite e sua associação com infecção por BLV e linfocitose persistente. A prevalência do VLB foi avaliada pelo teste de imunodifusão em gel de ágar (IDGA) em 1.392 vacas em lactação nos rebanhos estudados. De acordo com os resultados do IDGA, 240 vacas foram selecionadas para compor dois grupos experimentais: 1-caso: composto por 160 vacas BLV positivas (BLV-P) e 2-controle: composto por 80 vacas BLV negativas (BLV-N). De acordo com as avaliações sorológicas e leucocitárias, os animais foram classificados em três subgrupos: BLV-N; BLV-P sem linfocitose persistente (BLV-PWPL) e BLV positivos com linfocitose persistente (BLV-PPL). O California Mastitis Test (CMT) e a cultura do leite das glândulas CMT positivas foram realizados nos dias 0 (D0) e 60 (D60). A leucose foi detectada em 100% dos rebanhos com prevalência média de 80,9%. Entre os animais BLV-P, 74% foram classificados como BLV-PWPL e 26% como BLV-PPL. Staphylococcus coagulase negativos (73,30%), seguido por Corynebacterium spp. (12,04%), Streptococcus agalactiae (8,90%) e Staphylococcus aureus (5,76%) foram os agentes predominantes. Não houve diferença estatística entre os escores de mastite clínica ou subclínica e nem entre o tipo de patógeno (contagioso ou ambiental) nos subgrupos de animais BLV-PWPL ou BLV-PPL relacionados ao BLV-N. No entanto, animais BLV-P apresentaram escores mais elevados no CMT em relação aos animais BLV-N, sugerindo maior predisposição à mastite subclínica em vacas com leucose.

In the present study, subclinical mastitis (SCM) and clinical mastitis (CM) prevalence for various countries in the World were calculated by using online and offline databases. The SCM and CM prevalence studies reported during 1967-2019 were collected, reviewed, and a meta-analysis was done in R-Software. A total of 222 and 150 studies from the World and 103 and 37 studies from India on SCM and CM, respectively were included. The pooled prevalence of SCM and CM were 42% [Confidence Interval (CI) 38-45%, Prediction Interval (PI) 10-83%] and 15% [CI 12-19%, PI 1-81%] in the World respectively, 45% [CI 40-49%, PI 11-84%] and 18% [CI 14-23%, PI 3-60%] in India respectively. Continent-wise analysis indicated a higher prevalence of SCM in North America and CM in Europe and among the countries, a higher SCM prevalence in Uganda and CM in the United Kingdom was observed. Further, species-wise indicated a higher SCM and CM prevalence in buffaloes of the World than the cattle. Based on method-wise, SCM and CM prevalence were high in somatic cell count and clinical examination, respectively in the World. The SCM prevalence was higher than CM and indicated the importance of SCM in dairy cattle. This might result in low milk productivity in dairying and may set off losses to dairy farmers. Hence, there is an urgent need to reduce the SCM and CM prevalence by implementing scientific dairy management, good feeding practices, and timely therapeutic interventions for increasing the benefits from dairying to the farmers in the World.

Staphylococcus aureus memiliki kepentingan di dunia kedokteran hewan sebagai agen penyebab mastitis pada sapi perah. Bakteri S. aureus dapat menghasilkan bermacam faktor virulensi yang dapat membantu patogen ini dalam setiap tahap proses patogenesis mastitis, dimulai dari perlekatan S. aureus ke sel ambing hingga menimbulkan kerusakan yang menyebar ke jaringan ambing. Penelitian ini merupakan penelitian studi literatur yang dilakukan dengan mengumpulkan dan menelaah berbagai artikel ilmiah yang terkait dengan topik penelitian ini melalui mesin pencari Google Scholar (https://scholar.google.co.id). Penelitian ini bertujuan untuk mengulas karakteristik beberapa faktor virulensi S. aureus yang sering dilaporkan berperan dalam kejadian mastitis pada sapi perah. Faktor virulensi tersebut antara lain yaitu biofilm, clumping factor, fibronectin binding protein, hemolisin, koagulase, Panton-Valentine leukocidin, kapsul, dan superantigen seperti toksin toxic shock syndrome (TSST), serta enterotoksin (SE). Kombinasi produksi faktor virulensi oleh S. aureus dianggap dapat meningkatkan derajat keparahan mastitis.

There is ongoing debate regarding whether critically important antimicrobials (CIA) should be used to treat infections in food-producing animals. In this systematic review, we determined whether CIA and non-CIA have comparable efficacy to treat nonsevere bovine clinical mastitis caused by the most commonly reported bacteria that cause mastitis worldwide. We screened CAB Abstracts, Web of Science, MEDLINE, Scopus, and PubMed for original epidemiological studies that assessed pathogen-specific bacteriological cure rates of antimicrobials used to treat nonsevere clinical mastitis in lactating dairy cows. Network models were fit using risk ratios of bacteriological cure as outcome. A total of 30 studies met inclusion criteria. Comparisons of cure rates demonstrated that CIA and non-CIA had comparable efficacy for treatment of nonsevere clinical mastitis in dairy cattle. Additionally, for cows with nonsevere clinical mastitis caused by Escherichia coli and Klebsiella spp., bacteriological cure rates were comparable for treated versus untreated cows; therefore, there was no evidence to justify treatment of these cases with CIA. Our findings supported that CIA in general are not necessary for treating nonsevere clinical mastitis in dairy cattle, the disease that accounts for the majority of antimicrobial usage in dairy herds worldwide. Furthermore, our findings support initiatives to reduce or eliminate use of CIA in dairy herds.

Bovine mastitis continues to be a leading cause of heavy economic losses in the dairy industry and a public health hazard globally. This cross-sectional study investigated the prevalence, etiologies of clinical and subclinical mastitis, and associated predisposing factors in Embu and Kajiado counties in Kenya. A semistructured questionnaire was administered to 154 smallholder dairy farmers to collect data on management practices, animal factors, and disease history. A total of 395 dairy cows were initially screened for subclinical mastitis using the California mastitis test (CMT), and milk samples were aseptically collected. Both CMT positive and CMT negative samples were analyzed using conventional bacteriological isolation and identification procedures. In the present study, the overall prevalence of mastitis based on CMT and clinical examination was 80% (316/395), out of which 6.8% (27/395) was clinical mastitis, while 73.1% (289/395) was subclinical mastitis. Based on culture, the overall prevalence of clinical and subclinical mastitis was 51.6% (815/1580), 74.4% (294/395), and 76.6% (118/154) at the quarter, cow, and farm level, respectively. From the 1574 milk samples analyzed by cultured, 1016 bacteria were yielded. The predominant bacteria were coagulase-negative Staphylococcus (CNS), 42.8% (435/1016), and in decreasing order, Streptococcus species, 22.2% (226/1016), Staphylococcus aureus, 15.7% (160/1016), and Pseudomonas aeruginosa, 5.1% (52/1016), and the least was Enterobacter species, 0.7% (7/1016), while 23.7% of the sample yielded no bacterial growth. Risk factor analysis revealed that milking mastitic cows last (p=0.002), using a clean udder drying towel for each cow (p=0.033) and previous history of mastitis (p=0.046) were significantly associated with presence of mastitis. The current study has shown a relatively high prevalence of subclinical mastitis with CNS as predominant bacteria. Therefore, control measures are urgently warranted. Management factors such as milking mastitic cows last, using a clean towel for udder drying for each cow, and culling mastitic cows should be considered and included in the Kenyan mastitis control programs.

Mastitis is one of the most important health problems of dairy cattle as it causes physical, chemical and bacteriological changes in the produced milk, so the aim of this study is investigating the epidemiology of mastitis pathogens of dairy cattle and their associated risk factors. A total of 397 samples were gathered from mastitic cows and their surrounding environment including 127 milk samples from sub-clinically mastitic cows, 60 milk samples from clinically mastitic cows, 60 teat swabs, 50 milking machines swabs, 50 worker’s hand swabs and 50 bedding samples from different farms containing small and large dairy herds located in El Dakhalia and Menoufiya Governorate. These samples were examined bacteriologically and biochemically for isolation and biochemical identification of E. coli and S. aureus. Our results revealed that the prevalence of E.coli was 33.33% (11/33), 21.74% (20/92), 16.67% (4/24), 18.52% (5/27), 13.33% (2/15) and 13.33% (4/30) in milk samples from clinically mastitic cows, milk samples from subclinically mastitic cows, teat swabs, milking machines swabs, worker’s hand swabs and bedding samples, respectively, while the prevalence of Staphylococcus aureus was 23.08% (9/39), 17.43% (19/109), 11.9% (5/42), 10.53% (4/38), 17.07% (7/41) and 10.53% (4/38), in the same groups respectively. A significant relationship was found between of the number of these bacteria and cows age, herd size and hygienic condition of the farm. Moreover, PCR was done on 14 isolates of E. coli for detection of ESBL producing E. coli. The results of PCR revealed that the prevalence of blaTEM and blaSHV was 64.3% (9/14) and 92.9% (13/14), respectively, while none of the isolates carried blaCMY-2. We concluded that Staphylococcus aureus and E.coli are considered two main pathogens of mastitis and the prevalence of mastitis increases in old cattle, inadequate hygienic condition, lack of post milking teat dipping and absence of udder washing which are all predisposing risk factors of mastitis in the study area.

Mastitis in dairy cattle is the most common management disorder that causes higher economic losses by lowering production and quality of milk leads to substantial economical loss. The aim of this article was to review worldwide important advances in strategies to control mastitis for production augmentation in dairy cattle. Many scientists worked to identify effective strategies to control mastitis caused by Streptococcus agalactiae, Staphylococcus aureus, and others. It is necessary to identify mechanisms of infection, define clinical and subclinical states of disease, determine exposure time, and identify pathogen-specific characteristics. Evolvement of management strategies that incorporated hygienic procedures (animal, floor, and milkman), post milking standing period of animal and strategic use of antibiotic or herbal therapy at dry-off, nutritional supplementation, fly control, body condition score optimization, etc., resulted in widespread control of mastitis. The udder, teat of animal, scientific management of milking, automatic milking procedure, genetic selection are considered as important factors to control mastitis. As farm management changed, scientists were directed to redefine control of mastitis caused by opportunistic pathogens of environmental sources and have sought to explore management strategies which will maintain animal well-being in a judicial way. Although significant advances in mastitis management have been made changing herd structure, changing climatic scenario and more rigorous milk processing standards ensure that mastitis will remain important issue for future research.

Background: Mastitis is the most prevalent disease in dairy cattle and one of the most significant bovine pathologies affecting milk production, animal health, and reproduction. In addition, mastitis is the most common, expensive, and contagious infection in the dairy industry.Methods: A meta-analysis of microarray and RNA-seq data was conducted to identify candidate genes and functional modules associated with mastitis disease. The results were then applied to systems biology analysis via weighted gene coexpression network analysis (WGCNA), Gene Ontology, enrichment analysis for the Kyoto Encyclopedia of Genes and Genomes (KEGG), and modeling using machine-learning algorithms.Results: Microarray and RNA-seq datasets were generated for 2,089 and 2,794 meta-genes, respectively. Between microarray and RNA-seq datasets, a total of 360 meta-genes were found that were significantly enriched as “peroxisome,” “NOD-like receptor signaling pathway,” “IL-17 signaling pathway,” and “TNF signaling pathway” KEGG pathways. The turquoise module (n = 214 genes) and the brown module (n = 57 genes) were identified as critical functional modules associated with mastitis through WGCNA. PRDX5, RAB5C, ACTN4, SLC25A16, MAPK6, CD53, NCKAP1L, ARHGEF2, COL9A1, and PTPRC genes were detected as hub genes in identified functional modules. Finally, using attribute weighting and machine-learning methods, hub genes that are sufficiently informative in Escherichia coli mastitis were used to optimize predictive models. The constructed model proposed the optimal approach for the meta-genes and validated several high-ranked genes as biomarkers for E. coli mastitis using the decision tree (DT) method.Conclusion: The candidate genes and pathways proposed in this study may shed new light on the underlying molecular mechanisms of mastitis disease and suggest new approaches for diagnosing and treating E. coli mastitis in dairy cattle.

Streptococcus agalactiae, also known as group B Streptococcus (GBS), is a pathogen of humans and animals. It is an important cause of mastitis in dairy cattle, causing decreased milk quality and quantity. Denmark is the only country to have implemented a national surveillance and control campaign for GBS in dairy cattle. After a significant decline in the 20th century, prevalence has increased in the 21st century. Using a unique combination of national surveillance, cattle movement data and molecular typing, we tested the hypothesis that transmission mechanisms differ between GBS strains that are almost exclusive to cattle and those that affect humans as well as cattle, which would have implications for control recommendations. Three types of S. agalactiae, sequence type (ST) 1, ST23 and ST103 were consistently the most frequent strains among isolates obtained through the national surveillance programme from 2009 to 2011. Herds infected with ST103, which is common in cattle but rarely found in people in Europe, were spatially clustered throughout the study period and across spatial scales. By contrast, strains that are also commonly found in humans, ST1 and ST23, showed no spatial clustering in most or any years of the study, respectively. Introduction of cattle from a positive herd was associated with increased risk of infection by S. agalactiae in the next year (risk ratio of 2.9 and 4.7 for 2009–2010 and 2010–2011, respectively). Moreover, mean exposure to infection was significantly higher for newly infected herds and significantly lower for persistently susceptible herds, as compared to random simulated networks with the same properties, which suggests strong association between the cattle movement network and new infections. At strain-level, new infections with ST1 between 2009 and 2010 were significantly associated with cattle movements, while other strains showed only some degree of association. Sharing of veterinary services, which may serve as proxy for local or regional contacts at a range of scales, was not significantly associated with increased risk of introduction of S. agalactiae or one of the three predominant strains on a farm. Our findings support the reinstatement of restrictions on cattle movements from S. agalactiae positive herds, which came into effect in 2018, but provide insufficient evidence to support strain-specific control recommendations.

Group B Streptococcus (GBS; Streptococcus agalactiae ) is a major neonatal and opportunistic bacterial pathogen of humans and an important cause of mastitis in dairy cattle with significant impacts on food security. Following the introduction of mastitis control programmes in the 1950s, GBS was nearly eradicated from the dairy industry in northern Europe, followed by re-emergence in the 21st century. Here, we sought to explain this re-emergence based on short and long read sequencing of historical (1953–1978; n=44) and contemporary (1997–2012; n=76) bovine GBS isolates. Our data show that a globally distributed bovine-associated lineage of GBS was commonly detected among historical isolates but never among contemporary isolates. By contrast, tetracycline resistance, which is present in all major GBS clones adapted to humans, was commonly and uniquely detected in contemporary bovine isolates. These observations provide evidence for strain replacement and suggest a human origin of newly emerged strains. Three novel GBS plasmids were identified, including two showing >98 % sequence similarity with plasmids from Streptococcus pyogenes and Streptococcus dysgalactiae subsp. equisimilis , which co-exist with GBS in the human oropharynx. Our findings support introduction of GBS into the dairy population due to human-to-cattle jumps on multiple occasions and demonstrate that reverse zoonotic transmission can erase successes of animal disease control campaigns.

Made available in DSpace on 2022-04-28T19:42:49Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-08-01 This study aimed to obtain a better understanding of the regulatory genes and molecules involved in the development of mastitis. For this purpose, the transcription factors (TF) and MicroRNAs (miRNA) related to differentially expressed genes previously found in extracorporeal udders infected with Streptococcus agalactiae were investigated. The Gene-TF network highlighted LOC515333, SAA3, CD14, NFKBIA, APOC2 and LOC100335608 and genes that encode the most representative transcription factors STAT3, PPARG, EGR1 and NFKB1 for infected udders. In addition, it was possible to highlight, through the analysis of the gene-miRNA network, genes that could be post-transcriptionally regulated by miRNAs, such as the relationship between the CCL5 gene and the miRNA bta-miR-363. Overall, our data demonstrated genes and regulatory elements (TF and miRNA) that can play an important role in mastitis resistance. The results provide new insights into the first functional pathways and the network of genes that orchestrate the innate immune responses to infection by Streptococcus agalactiae. Our results will increase the general knowledge about the gene networks, transcription factors and miRNAs involved in fighting intramammary infection and maintaining tissue during infection and thus enable a better understanding of the pathophysiology of mastitis. Institute of Biological Sciences Universidade Federal de Juiz de Fora Zootechnics Department Universidade Federal Dos Vales Do Jequitinhonha e Mucuri Zootechnics Department Universidade Federal Do Espírito Santo Department of Exact Sciences Universidade Estadual Paulista Júlio de Mesquita Filho Molecular Genetics Laboratory Embrapa Gado de Leite Department of Exact Sciences Universidade Estadual Paulista Júlio de Mesquita Filho

Improving the production traits and resistance against mastitis in dairy cattle is a challenge for animal scientists across the globe. The present study was designed to investigate the genetic effects of single nucleotide polymorphisms (SNPs) in Janus kinase 2 (JAK2) and diacylglycerol acyltransferase (DGAT1) genes with production and mastitis-related traits. Four SNPs in JAK2 and one in DGAT1 were analyzed through Chinese Cow's SNPs Chip-I (CCSC-I) and genotyped in a population of 312 Chinese Holsteins. Our findings demonstrated that milk fat percentage, somatic cell count (SCC), somatic cell score (SCS), serum cytokines interleukin 6 (IL-6) and interferon gamma (IFN-γ) showed significant associations (P < 0.05) with at least one or more identified SNPs. Consequently, the analysis based on haplotypes amongst the SNPs in JAK2 revealed noteworthy (P < 0.05) association with SCC and IL-6. Collectively, our results verified the pleiotropic ability of detected SNPs in bovine JAK2 and DGAT1 for milk fat percentage as well as mastitis-related traits. The significant SNPs in both the genes could serve as powerful genetic markers to minimize mastitis risk. In addition, besides SCC and SCS, the IFN-γ and IL-6 could also be used as indicators of improved genetic resistance against mastitis.

Mastitis commonly occurs in dairy cattle during lactation. This disease has huge effects on the productivity of dairy cattle and the profits of stock farms. Enhancing the efficiency of mastitis treatment begins with the early detection of the disease. The purpose of this research is early detection of mastitis by monitoring the pathogenesis of mastitis through real-time measurements of the rumen temperature by utilizing an ingestible bio-sensor. The ingestible bio-sensor used for this research is orally administered and settles in the rumen. This bio-sensor can measure the cattle's body temperature accurately without any interference from the ambient environment. Also, this method requires less labor and the cattle experience less stress. The measured body temperature data is collected and transmitted to a server for analysis to detect any abnormalities. We administered bio-sensors to 50 Holstein dairy cattle and monitored their rumen temperatures for six months by measuring the temperatures in 10-min intervals. The farm manager could receive mastitis alert messages through a mobile application when a rise in a cow’s body temperature was detected. In addition, the manager conducted daily CMT to diagnose any mastitis incidence. The study results showed 15 subclinical mastitis incidences including recurrences from 9 cows, with 14 alert messages sent from the body temperature monitoring system. The average temperature of high fever cases was 41.10 °C. And the temperature difference between the average temperature of the experimental animals and the highest temperature of mastitis afflicted animals was on average 2.42 °C which shows that mastitis is accompanied with a high rise in body temperature.

Propolis is a substance produced by bees, especially Apis mellifera, from resins of plant shoots. It possesses numerous biological properties, such as antimicrobial and anti-inflammatory. Activities such as organic dairy farming show high demand for natural products with proprieties like those of propolis, since milk contamination and mastitis are major problems of the dairy industry. The objective of this study was to evaluate in vitro the antimicrobial activity of different concentrations of an ethanolic extract of propolis (EEP). Action of the EEP extracted from beehives in southern Brazil was tested against 13 different genera of bacterial agents that cause mastitis in dairy cattle. The experiment was carried out in the Laboratory of Parasitology and Microbiology at the Federal University of Pampa, Campus Dom Pedrito - RS, Brazil, during May and June 2016. Agar diffusion and microdilution plate methodologies were followed. Based on the results of the minimum inhibitory concentration test (MIC), the EEP had inhibitory activity on 100% of the bacteria tested at concentrations above 10% (w/v). These results show that propolis has antimicrobial potential against bacteria involved in the process of mastitis. A própolis é uma substância coletada por abelhas, especialmente pela Apis mellifera, a partir de resinas de brotos de plantas que possuem numerosas propriedades biológicas, como antimicrobiana e antiinflamatória. Atividades como a pecuária leiteira orgânica apresentam alta demanda por produtos naturais com propriedades como as presentes na própolis, uma vez que a contaminação do leite e a mastite são grandes problemas da indústria de laticínios. O objetivo deste trabalho foi avaliar a atividade antimicrobiana in vitro do extrato etanólico de própolis (EEP) em diferentes concentrações. Ação do EEP extraído de colmeias no sul do Brasil foi testada contra 13 diferentes gêneros de agentes bacterianos causadores de mastite em bovinos leiteiros. O experimento foi conduzido no laboratório de Parasitologia e Microbiologia da Universidade Federal do Pampa, Campus Dom Pedrito – RS, durante os meses de maio e junho de 2016. Seguiram-se as técnicas de difusão em ágar e microdiluição em placas. Através dos resultados do teste de concentração inibitória mínima (CIM), pode-se afirmar que o EEP possui atividade inibitória em 100% das bactérias testadas em concentrações acima de 10% (p/v). Tais resultados mostram que a própolis apresenta potencial antimicrobiano frente as principais bactérias envolvidas nos processos de mastite.

Mastitis in dairy cattle has high morbidity and mortality rates, having serious consequences in milk production and quality. The aim of this work was to evaluate the activity of metalloproteinase-2 (MMP-2) and metalloproteinase-9 (MMP-9) in milk from healthy dairy cattle and from cows with signs of clinical and subclinical mastitis. The levels of total proteins and proteases with caseinolytic activity were also evaluated. The highest caseinolytic activity was found in milk samples obtained from animals with mastitis, in which lower levels of casein, especially the κ-casein fraction, were found, as compared with healthy animals.

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

The objective of our study was to evaluate prevalence of selected bacterial and fungal pathogens of mastitis in dairy cattle in north-eastern Poland. Our study was conducted from 2013 to 2019 in 1,665 clinically and sub-clinically infected quarter milk samples (2013, n = 368; 2014, n = 350; 2015, n = 290; 2016, n = 170; 2017, n = 173; 2018, n = 224; and 2019, n = 90). The isolation and identification of the pathogens were performed in keeping with generally accepted microbiological procedures. In 2013, mastitis was most commonly caused by Staphylococcus aureus (24%), Streptococcus spp. (22%), Streptococcus agalactiae (12%) and coagulase-negative staphylococci (11%). In 2014, the most common pathogens were Streptococcus spp. (25%), Staphylococcus aureus (18%) and coagulase-negative staphylococci (10%); in 2015, 2016, 2017, 2018 and 2019, Streptococcus spp. (from 39–49%) were the most frequent strains isolated from the quarter milk samples. Other pathogens were isolated occasionally (below 15% in all years). In conclusion, the role of environmental bacteria has been gradually increasing in the Warmia Province. The importance of infectious pathogens has been decreasing, indicating the efficacy of the applied preventive programmes and a need for the development of new programmes targeting environmental pathogens.

BackgroundKlebsiella pneumoniae, an environmental pathogen causing mastitis in dairy cattle, is often resistant to antibiotics.K. pneumoniaewas used as the host bacteria to support bacteriophage replication; 2 bacteriophages, CM8-1 and SJT-2 were isolated and considered to have therapeutic potential. In the present study, we determined the ability of these 2 bacteriophages to mitigate cytotoxicity, pathomorphological changes, inflammatory responses and apoptosis induced byK. pneumoniae(bacteriophage toK. pneumoniaeMOI 1:10) in bovine mammary epithelial cells (bMECs) culturedin vitro.ResultsBacteriophages reduced bacterial adhesion and invasion and cytotoxicity (lactate dehydrogenase release). Morphological changes in bMECs, including swelling, shrinkage, necrosis and hematoxylin and eosin staining of cytoplasm, were apparent 4 to 8 h after infection withK. pneumoniae, but each bacteriophage significantly suppressed damage and decreased TNF-α and IL-1β concentrations.K. pneumoniaeenhanced mRNA expression of TLR4, NF-κB, TNF-α, IL-1β, IL-6, IL-8, caspase-3, caspase-9 and cyt-c in bMECs and increased apoptosis of bMECs, although these effects were mitigated by treatment with either bacteriophage for 8 h.ConclusionsBacteriophages CM8-1 and SJT-2 mitigatedK. pneumoniae-induced inflammation in bMECs cultured in vitro. Therefore, the potential of these bacteriophages for treating mastitis in cows should be determined in clinical trials.

Mycoplasma bovis is an important pathogen causing mostly pneumonia in calves and mastitis in dairy cattle. In the absence of an effective vaccine, antimicrobial therapy remains the main control measure. Antimicrobial use in veal calves is substantially higher than in conventional herds, but whether veal calves also harbor more resistant M. bovis strains is currently unknown. Therefore, we compared antimicrobial susceptibility test results of M. bovis isolates from different cattle sectors and genomic clusters. The minimum inhibitory concentration of nine antimicrobials was determined for 141 Belgian M. bovis isolates (29 dairy, 69 beef, 12 mixed, 31 veal farms), and was used to estimate the epidemiological cut-off. Acquired resistance was frequently observed for the macrolides, while no acquired resistance to oxytetracycline and doxycycline, minimal acquired resistance to florfenicol and tiamulin, and a limited acquired resistance to enrofloxacin was seen. M. bovis isolates from beef cattle or genomic cluster III had higher odds of being gamithromycin-resistant than those from dairy cattle or genomic clusters IV and V. In this study, no cattle industry could be identified as source of resistant M. bovis strains. A single guideline for antimicrobial use for M. bovis infections, with a small remark for gamithromycin, is likely sufficient.

Streptococcus uberis is a common cause of intramammary infection and mastitis in dairy cattle. Unlike other mammary pathogens, S. uberis evades detection by mammary epithelial cells, and the host&ndash, pathogen interactions during early colonisation are poorly understood. Intramammary challenge of dairy cows with S. uberis (strain 0140 J) or isogenic mutants lacking the surface-anchored serine protease, SUB1154, demonstrated that virulence was dependent on the presence and correct location of this protein. Unlike the wild-type strain, the mutant lacking SUB1154 failed to elicit IL-1&beta, from ex vivo CD14+ cells obtained from milk (bovine mammary macrophages, BMM), but this response was reinstated by complementation with recombinant SUB1154, the protein in isolation elicited no response. Production of IL-1&beta, was ablated in the presence of various inhibitors, indicating dependency on internalisation and activation of NLRP3 and caspase-1, consistent with inflammasome activation. Similar transcriptomic changes were detected in ex vivo BMM in response to the wild-type or the SUB1154 deletion mutant, consistent with S. uberis priming BMM, enabling the SUB1154 protein to activate inflammasome maturation in a transcriptionally independent manner. These data can be reconciled in a novel model of pathogenesis in which, paradoxically, early colonisation is dependent on the innate response to the initial infection.

Staphylococcus aureus is one of the main pathogens leading to both clinical and subclinical bovine mastitis in dairy cattle. Prediction of disease evolution based on the characteristics of Staph. aureus isolates that cause intramammary infections and understanding the host-pathogen interactions may improve management of mastitis in dairy herds. For this study, several strains were selected from each of the 6 major Canadian spa types associated with mastitis (t267, t359, t529, t605, t2445, and t13401). Adherence to host cells and intracellular persistence of these strains were studied using a bovine mammary gland epithelial cell line (MAC-T). Additionally, relative virulence and host response (cytokines production) were also studied in vivo using a mouse model of mastitis. Whole-genome sequencing was performed on all strains and associations between clonal complex, sequence type, and presence of certain virulence factors were also investigated. Results show that spa type t2445 was correlated with persistence in MAC-T cells. Strains from spa t359 and t529 showed better ability to colonize mouse mammary glands. The exception was strain sa3154 (spa t529), which showed less colonization of glands compared with other t359 and t529 strains but possessed the highest number of superantigen genes including tst. All strains possessed hemolysins, but spa types t529 and t2445 showed the largest diameter of β-hemolysis on blood agar plates. Although several spa types possessed 2 or 3 serine-aspartate rich proteins (Sdr) believed to be involved in many pathogenic processes, most t529 strains expressed only an allelic variant of sdrE. The spa types t605 (positive for the biofilm associated protein gene; bap+) and t13401 (bap-), that produced the largest amounts of biofilm in vitro, were the least virulent in vivo. Finally, strains from spa type t529 (ST151) elicited a cytokine expression profile (TNF-α, IL-1β and IL-12) that suggests a potential for severe inflammation. This study suggests that determination of the spa type may help predict the severity of the disease and the ability of the immune system to eliminate intramammary infections caused by Staph. aureus.

Simple Summary This literature review provides a synthesis and evaluation of the current knowledge on Staphylococcus agnetis (S. agnetis) and its implications in poultry pathology. Recent studies revealed that S. agnetis can cause bacterial chondronecrosis with osteomyelitis (BCO), endocarditis, and septicemia in broiler chickens. Lameness constitutes one of the major health and welfare problems causing huge economic losses in the poultry industry. To date, a range of infectious and non-infectious factors have been associated with lameness in poultry. Among bacteria of the genus Staphylococcus, Staphylococcus aureus is the main species associated with locomotor problems. This contrasts with S. agnetis, which until recently had not been considered as a poultry pathogen. Previously only reported in cattle, S. agnetis has expanded its host range to chickens, and due to its unique characteristics has become recognized as a new emerging pathogen. The genotypic and phenotypic similarities between S. agnetis and other two staphylococci (S. hyicus and S. chromogenes) make this pathogen capable of escaping recognition due to misidentification. Although a significant amount of research on S. agnetis has been conducted, many facts about this novel species are still unknown and further studies are required to understand its full significance in poultry pathology. Abstract This review aims to summarize recent discoveries and advancements regarding the characteristics of Staphylococcus agnetis (S. agnetis) and its role in poultry pathology. S. agnetis is an emerging pathogen that was primarily associated with mastitis in dairy cattle. After a presumed host jump from cattle to poultry, it was identified as a pathological agent in broiler chickens (Gallus gallus domesticus), causing lameness induced by bacterial chondronecrosis with osteomyelitis (BCO), septicemia, and valvular endocarditis. Economic and welfare losses caused by lameness are global problems in the poultry industry, and S. agnetis has been shown to have a potential to induce high incidences of lameness in broiler chickens. S. agnetis exhibits a distinct repertoire of virulence factors found in many different staphylococci. It is closely related to S. hyicus and S. chromogenes, hence infections caused by S. agnetis may be misdiagnosed or even undiagnosed. As there are very few reports on S. agnetis in poultry, many facts about its pathogenesis, epidemiology, routes of transmission, and the potential impacts on the poultry industry remain unknown.

Mastitis, a disease with high incidence worldwide, is the most prevalent and costly disease in the dairy industry. Environmental mastitis pathogens, such as Escherichia coli (E. coli), are major etiological agents of bovine mastitis in well-managed dairy farms. However, there is still a need to develop more efficient, safe, and economical treatments for mastitis. In the current research, Heter-LP, a new system biology-based method of drug repositioning, was applied to potentially identify novel therapeutic avenues for the treatment of E. coli mastitis. On-line data repositories relevant to known diseases, drugs, and gene targets along with other specialized biological information for E. coli mastitis, including key genes with robust bio-signatures, drugs and related diseases were used as input data for analysis with the Heter-LP algorithm. Our analyses identified novel drugs such as Glibenclamide, Ipratropium, Salbutamol, and Carbidopa as possible therapeutics that could be used against E. coli mastitis. Predicted relationships can be used by pharmaceutical scientists or veterinarians to find commercially efficacious medicines or combination of two or more active compounds to treat mastitis.

Streptococcus agalactiae is a contagious pathogen that causes bovine mastitis worldwide, resulting in considerable economic losses. In this study, we isolated 42 S. agalactiae strains in 379 milk samples from cows with subclinical mastitis on 15 dairy farms in 12 Chinese provinces. Analysis based on capsular typing and multilocus sequence typing, combined with patterns of virulence gene scanning and antimicrobial resistance, identified the lineages and populations of the isolates. We grouped the 42 isolates into 7 sequence types belonging to 6 clonal complexes, mainly CC103 (31/42 isolates; 73.8%). We identified an ST-23 strain named Sa 129 for the first time on Chinese dairy farms-this strain is usually associated with human isolates. Capsular types Ia and II were predominant in capsular typing. The prevalence of virulence profile 1 (bibA, cfb, cspA, cylE, fbsA, fbsB, hylB, and pavA) was 64.3%, and represented the main trend in China. With respect to antimicrobial resistance, most isolates were susceptible to β-lactams, rifamycin, glycopeptides, and oxazolidone; resistance to several antimicrobial agents, including lincomycin, clindamycin, and doxycycline, varied in 4 different regions. Our research provides a profile for the molecular epidemiology, multilocus sequence typing, antimicrobial resistance, and virulence gene clustering of S. agalactiae, and may be beneficial for the clinical monitoring, prevention, and control of mastitis in dairy cattle.

Staphylococcus aureus can cause mastitis in dairy cattle. We report the genome sequence of a Staphylococcus aureus strain isolated from a dairy cow with a chronic case of mastitis. The infection with this strain of Staphylococcus aureus was not cleared from the animal with antibiotic treatment.

Mastitis is the most devastating economic disease of dairy cattle. Mastitis in dairy cattle frequently occurs during the dry period or in early lactation. E. coli. and S. aureus is the main causative agent of mastitis. S. aureus has an ability to form encapsulated focuses in the udder and develop subclinical form of mastitis. This property of bacteria hinders the effective cure during lactation period. Antibiotics used for treatments have a short half-life span at the site of action because of frequent milking, therefore unable to maintain desired drug concentration for effective clearance of bacteria. We demonstrated the potential of ciprofloxacin encapsulated nanocarriers which can improve availability of drugs and could provide effective means of the treatment of mastitis. These drug loaded nanoparticles show low toxicity & slow clearance from the site of action. Antimicrobial studied against clinical strain of E.coli. and Staphylococcus aureus zone of inhibition depend on dose 0.5 mg to 2 mg/ml nanoparticles solution from 11.6 to 14.5 mm and 15 to 18mm. These nanoparticles show good antimicrobial activity in broth culture as well against bacteria.

Mastitis is one of the major risks for public health and animal welfare in the dairy industry. Two of the most important pathogens to cause mastitis in dairy cattle are Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). While S. aureus generally induces a chronic and subclinical mastitis, E. coli is an important etiological pathogen resulting in an acute and clinical mastitis. The liver plays a central role in both, the metabolic and inflammatory physiology of the dairy cow, which is particularly challenged in the early lactation due to high metabolic and immunological demands. In the current study, we challenged the mammary glands of Holstein cows with S. aureus or E. coli, respectively, mimicking an early lactation infection. We compared the animals' liver transcriptomes with those of untreated controls to investigate the hepatic response of the individuals. Both, S. aureus and E. coli elicited systemic effects on the host after intramammary challenge and seemed to use pathogen-specific targeting strategies to bypass the innate immune system. The most striking result of our study is that we demonstrate for the first time that S. aureus intramammary challenge causes an immune response beyond the original local site of the mastitis. We found that in the peripheral liver tissue defined biological pathways are switched on in a coordinated manner to balance the immune response in the entire organism. TGFB1 signaling plays a crucial role in this context. Important pathways involving actin and integrin, key components of the cytoskeleton, were downregulated in the liver of S. aureus infected cows. In the hepatic transcriptome of E. coli infected cows, important components of the complement system were significantly lower expressed compared to the control cows. Notably, while S. aureus inhibits the cell signaling by Rho GTPases in the liver, E. coli switches the complement system off. Also, metabolic hepatic pathways (e.g., lipid metabolism) are affected after mammary gland challenge, demonstrating that the liver restricts metabolic tasks in favor of the predominant immune response after infection. Our results provide new insights for the infection-induced modifications of the dairy cow's hepatic transcriptome following mastitis.

S. agnetis has been previously associated with subclinical or clinically mild cases of mastitis in dairy cattle and is one of several Staphylococcal species that have been isolated from the bone and blood of lame broilers. We were the first to report that S. agnetis could be obtained frequently from bacterial chondronecrosis with osteomyelitis (BCO) lesions of lame broilers. Further, we showed that a particular isolate of S. agnetis, chicken isolate 908, can induce lameness in over 50% of exposed chickens, far exceeding normal BCO incidences in broiler operations. We have previously reported the assembly and annotation of the genome of isolate 908. To better understand the relationship between dairy cattle and broiler isolates, we assembled 11 additional genomes for S. agnetis isolates, including an additional chicken BCO strain, and ten isolates from milk, mammary gland secretions or udder skin, from the collection at the University of Missouri. To trace phylogenetic relationships, we constructed phylogenetic trees based on multi-locus sequence typing, and Genome-to-Genome Distance Comparisons. Chicken isolate 908 clustered with two of the cattle isolates along with three isolates from chickens in Denmark and an isolate of S. agnetis we isolated from a BCO lesion on a commercial broiler farm in Arkansas. We used a number of BLAST tools to compare the chicken isolates to those from cattle and identified 98 coding sequences distinguishing isolate 908 from the cattle isolates. None of the identified genes explain the differences in host or tissue tropism. These analyses are critical to understanding how Staphylococci colonize and infect different hosts and potentially how they can transition to alternative niches (bone vs dermis).ImportanceStaphylococcus agnetis has been recently recognized as associated with disease in dairy cattle and meat type chickens. The infections appear to be limited in cattle and systemic in broilers. This report details the molecular relationships between cattle and chicken isolates in order to understand how this recently recognized species infects different hosts with different disease manifestations. The data show the chicken and cattle isolates are very closely related but the chicken isolates all cluster together suggesting a single jump from cattle to chickens.

The aim of this study was to employ the principal component technique to physiological data and environmental thermohygrometric variables correlated with detection of clinical and subclinical mastitis in dairy cattle. A total of 24 lactating Girolando cows with different clinical conditions were selected (healthy, and with clinical or subclinical mastitis). The following physiological variables were recorded: udder surface temperature, ST (°C); eyeball temperature, ET (°C); rectum temperature, RT (°C); respiratory frequency, RF (mov. min-1). Thermohygrometric variables included air temperature, AirT (°C), and relative humidity, RU (%). ST was determined by means of thermal images, with four images per animal, on these quarters: front left side (FL), front right side (FR), rear right side (RR) and rear left side (RL), totaling 96 images. Exploratory data analysis was run through multivariate statistical technique with the employment of principal components, comprehending nine variables: ST on the FL, FR, RL and RR quarters; ET, RT; RF, AirT and RU. The representative quarters of the animals with clinical and subclinical mastitis showed udder temperatures 8.55 and 2.46° C higher than those of healthy animals, respectively. The ETs of the animals with subclinical and clinical mastitis were, respectively, 7.9 and 8.0% higher than those of healthy animals. Rectum temperatures were 2.9% (subclinical mastitis) and 5.5% (clinical mastitis) higher compared to those of healthy animals. Respiratory frequencies were 40.3% (subclinical mastitis) and 61.6% (clinical mastitis) higher compared to those of healthy animals. The first component explained 91% of the total variance for the variables analyzed. The principal component technique allowed verifying the variables correlated with the animals' clinical condition and the degree of dependence between the study variables.

This work presents a novel handheld mastitis detection device for detecting mastitis in dairy cattle, as well as a cattle management information system for managing cattle health information. The management information system comprises of three major subsystems—a mastitis detection device, an information display device, and a cloud database. The mastitis detection device detects and evaluates the degree of mastitis immediately before milking operations are carried out. The information display device shows the health information of the dairy cattle obtained from a cloud database and the database manages the milk production of a dairy farm. The combination of these two devices helps the dairy farmer to verify the mastitis of cattle effectively and find its health information rapidly.

Sub-clinical mastitis (SCM) affects milk composition. In this study, we hypothesise that large-scale mining of milk composition features by pattern recognition models can identify the best predictors of SCM within the milk composition features. To this end, using data mining algorithms, we conducted a large-scale and longitudinal study to evaluate the ability of various milk production parameters as indicators of SCM. SCM is the most prevalent disease of dairy cattle, causing substantial economic loss for the dairy industry. Developing new techniques to diagnose SCM in its early stages improves herd health and is of great importance. Test-day Somatic Cell Count (SCC) is the most common indicator of SCM and the primary mastitis surveillance approach worldwide. However, test-day SCC fluctuates widely between days, causing major concerns for its reliability. Consequently, there would be great benefit to identifying additional efficient indicators from large-scale and longitudinal studies. With this intent, data was collected at every milking (twice per day) for a period of 2 months from a single farm using in-line electronic equipment (346 248 records in total). The following data were analysed: milk volume, protein concentration, lactose concentration, electrical conductivity (EC), milking time and peak flow. Three SCC cut-offs were used to estimate the prevalence of SCM: Australian > 250 000 cells/ml, European >= 200 000 cells/ml and New Zealand >= 150 000 cells/ml. At first, 10 different Attribute Weighting Algorithms (AWM) were applied to the data. In the absence of SCC, lactose concentration featured as the most important variable, followed by EC. For the first time, using attribute weighted modelling, we showed that the concentration of lactose in milk can be used as a strong indicator of SCM. The development of machine-learning expert systems using two or more milk variables (such as lactose concentration and EC) may produce a predictive pattern for early SCM detection. Refereed/Peer-reviewed

Mycoplasma bovis is an important cause of mastitis in dairy cattle, and pneumonia, arthritis, and otitis in calves. Milk and colostrum are considered important sources of infection for calves. knowledge on the effect of on-farm freezing (-18°C) and thawing methods on the recovery of M. bovis from colostrum samples is missing. In this study, 2 separate experiments were performed. The first experiment consisted of a longitudinal study examining the survival [as measured by log(10) reduction] of 2 M. bovis strains in frozen colostrum over 14 wk. The second experiment examined the effect of different thawing temperatures (45 and 20°C), thawing frequencies (once or twice), and initial colostrum titer (104 or 106 cfu/mL) on M. bovis survival. A single freeze-thaw cycle led to an approximate 1 log reduction of M. bovis titer, independent of the thawing temperature. Freezing for 14 wk did not significantly further reduce the titer of bacteria compared with freezing for 2 wk. A second freeze-thaw cycle further reduced the M. bovis count by approximately 0.5 log compared with a single freeze-thaw cycle. Thawing temperature and initial bacterial concentration did not significantly affect M. bovis reduction. In conclusion, storage of colostrum samples in the freezer at -18°C during epidemiological studies, herd monitoring, or test and cull programs will probably have little influence on qualitative bacteriological test results for M. bovis. The epidemiological or clinical relevance of an approximate 1 log reduction of M. bovis in colostrum is currently unclear.

This study described the occurrence of clinical and subclinical forms of mastitis in 250 cattle from 5 dairy farms around the cities of Santa Rosa and Machala, El Oro Province, Ecuador. Clinical mastitis (CM) was determined based on obvious changes in milk (mild), signs of inflammation in the udder (moderate), and/or generalized clinical symptoms (severe). Subclinical mastitis (SCM) was assessed using the California mastitis test. CM and SCM were detected in 30 (12.0%) and 150 (60%) of the 250 tested cattle, respectively. Prevalence at the udder quarter level was 57.7% (577/1,000), which was higher among forequarters (369/577; 63.9%) than hindquarters. Of the 577 mastitic milk samples subjected to microbiological analysis, 35 were excluded due to contamination and 20 tested negative. Identification of bacterial isolates revealed that 33.3% of the 93 CM samples contained coliforms, 25.8% coagulase-positive staphylococci, 20.4% coagulase-negative staphylococci (CNS), 9.7% streptococci, 7.5% Bacillus spp., and 3.2% Klebsiella spp. Bacterial profiling of the 429 SCM milk samples showed that 55.4% contained CNS, 22.1% Bacillus spp., 9.3% streptococci, and 6.1% coagulase-positive staphylococci. In vitro antibiotic susceptibility testing of the obtained isolates indicated that all were susceptible to amoxicillin, ampicillin, cefotaxime, enrofloxacin, sulfamethoxazole-trimethoprim, gentamicin, and neomycin. No multidrug-resistant strains were observed.

Background: Staphylococcus aureus is a leading cause of contagious mastitis in dairy cattle. Internalization of S. aureus by bovine mammary gland epithelial cells is thought to be responsible for persistent and chronic intramammary infection, but the underlying mechanisms are not fully understood. Methods: In the present study, we evaluated the role of Annexin A2 (AnxA2), a membrane-binding protein, in S. aureus invasion into bovine mammary epithelial cell line (MAC-T). In vitro binding assays were performed to co-immunoprecipitate the binding proteins of AnxA2 in the lysates of S. aureus. Results: AnxA2 mediated the internalization but not adherence of S. aureus. Engagement of AnxA2 stimulated an integrin-linked protein kinase (ILK)/p38 MAPK cascade to induce S. aureus invasion. One of the AnxA2-precipitated proteins was identified as S. aureus clumping factor B (ClfB) through use of mass spectrometry. Direct binding of ClfB to AnxA2 was further confirmed by using a pull-down assay. Pre-incubation with recombinant ClfB protein enhanced S. aureus internalization, an effect that was specially blocked by anti-AnxA2 antibody. Conclusion: Our results demonstrate that binding of ClfB to AnxA2 has a function in promoting S. aureus internalization. Targeting the interaction of ClfB and AnxA2 may confer protection against S. aureus mastitis.

E-coli mastitis is one of the most frequent causes of environmental mastitis in the dairy cattle worldwide. The purpose of this field study was to compare the efficacy of ceftiofur (HCL) in conjunction with supportive measures versus supportive measures alone for treatment of dairy cows affected with naturally occurring acute form of E. coli mastitis. From January 2014 to December 2016 a total number of 100 cows naturally affected by acute E-coli mastitis randomly were allocated into two groups. A milk sample from the affected quarter was collected for bacteriological tests on the first day of treatment. In group A (control), fifty cows received ceftiofur (HCL) 1mg/5kg/BW, flunixin meglumine 2.2mg/kg, calcium borogluconate 40%, 250ml and hypertonic saline (Nacl 7.2 %,) 5ml/kg. In group B (treatment, n=50), cows received the same drugs mentioned for group A, except ceftiofur (HCL) which replaced by placebo. In the group A, 41cows (82%) and in the group B, 2 cows (4%) were survived respectively. The rates of quarter health recovery in the groups A and B were 31.7% and 0% respectively. The differences between two groups were significant (P≤0.01). In conclusion our results indicated that treatment of cows affected with naturally occurring acute form of E. coli mastitis without application of effective antibiotic(s) such as ceftiofur (HCL) and fluid therapy almost impossible.

Whole blood stimulation assay (WBA) has been widely used to study production of the pro-inflammatory cytokine, tumor necrosis factor-alpha (TNF-alpha) and is considered a relatively good predictor for the in vivo release of TNF-a during endotoximia in dairy cattle. However, it requires a standardized and optimized dose of lipopolysaccharides (LPS) to simulate the in vivo cytokine response during WBA when using killed bacteria. Moreover, if an association between genome (e. g., Quantitative trait loci) and ex vivo TNF-alpha responsiveness can be established; it helps to reduce mastitis in dairy cattle using selective breeding. Three experiment scenarios for pathogen dose, stimulation time and estimation of genetic variables effect in ex vivo TNF-alpha production were carried out to investigate differences of TNF-alpha responsiveness in early lactation in 4, 38 and 40 Danish Holstein Friesian dairy cows respectively. Statistical analysis done using ANOVA, t.test and lm functions in R software. Results showed that, ex vivo TNF-alpha response to E. coli is dose and time dependent and a dose of 2.5 x 10(6) CFU/mL for 3.5 h is optimal for observing TNF-alpha response after ex vivo stimulation.Relatively consistent inter-individual cow differences in the TNF-alpha response ex vivo were observed from week 3 to week 8. No statistically significant association were found between the ex vivo TNF-alpha responsiveness and the two E. call associated mastitis QTLs which may be due to the low number of animals (n =) in the study.

Mycoplasma bovis (M. bovis) is a small bacterium that lacks a cell wall. M. bovis infection results in chronic pneumonia and polyarthritis syndrome (CPPS), otitis media, conjunctivitis, and meningitis in feedlot cattle and mastitis in dairy cattle. Numerous studies of peripheral mononuclear cells (PBMC) indicate that M. bovis evades host immunity through targeted effects on immune cell activity, including inhibition of effector function and simultaneous aberrant activation of immune cell activity that has no effect on protection against the bacterium. Few studies have addressed the interaction between M. bovis and neutrophils, one of the most important cell subsets of innate immunity. We hypothesized that M. bovis modifies specific neutrophil activities to support its persistence and systemic dissemination. In this study, we demonstrate that M. bovis enhances neutrophil apoptosis, stimulates production of pro-inflammatory cytokines, IL-12 and TNF-α, inhibits production of nitric oxide (NO) but augments elastase release. We also show that IL-17 an inflammatory cytokine produced by Th-17 cells does not enhance the capacity of neutrophils to destroy M. bovis. These findings present novel mechanisms of mycoplasma evasion of host innate immunity and provide potential opportunities for immuno-therapeutic interventions.

Streptococcus uberis (S. uberis ) is an important environmental pathogen causing mastitis in dairy cattle. Mastitis or mammary gland infection is the most common disease in milking cows and cause significant economic burden to farmers due to the reduction of the amount of milk and its quality. The development of rapid analytical methods for the determination of mastitis-causing pathogens in milk is of utmost importance for the early identification of the causes of mastitis and the beginning of timely treatment of cows. Combining in silico bioinformatic analysis and solid phase peptide synthesis using Fmoc chemistry, we have made two different peptides to mimic the adhesion protein of S. uberis, which is promoting the attachment of bacteria to epithelial cells. After purification with RP-HPLC, the peptides were conjugated with a larger carrier protein (KLH) and used for immunization of rabbits to produce specific antibodies. The separation of anti- S. uberis antibodies from rabbit blood antisera was carried out with affinity chromatography, using the synthetic peptides as affinity ligands. The purified antibodies showed high affinity and specificity towards S. uberis and were used for rapid bio-recognition and identification of S. uberis with an immunobiosensing system.

he objective of this study was to identify and quantify the most representative factor affecting the economic impact of mastitis in dairy cattle herds in order to establish those that should receive greater attention from farmers and technicians to reduce the economic impact of this important disease. A simulation study was conducted with the CU$TO MASTITE program, considering 324 different herds and using combinations of the following factors: somatic cell count (250,000; 500,000; 750,000 and 1,000,000 somatic cells/mL milk); production scale (50; 100 and 150 lactating cows); productivity per animal (10; 20 and 30 L/cow/day); average annual incidence of clinical mastitis (1%; 7% and 15%), and involuntary culling rate due to mastitis (2%; 4% and 6%). Preventive measures included expenses with monitoring [culture and antibiogram, bulk tank somatic cell count (BTSCC), and individual somatic cell counts], pre- and post-dipping, vaccination, treatment of dry cows, and maintenance of the milking machine. Only treatments of clinical cases were considered as curative measures. The impact of mastitis was estimated as total losses plus expenses with prevention and treatment of clinical cases. The mean incidence of clinical mastitis (MIM) and BTSCC were significant (P

Mastitis in dairy cattle is an inflammatory reaction of the udder tissue. Mastitis increases plasmin levels, leading to an increased proteolysis of milk proteins such as casein, resulting in a significant decrease in milk quality and related dairy products. Due to its key-role in mastitis, we used plasmin proteolytic activity as a biomarker for the detection of mastitis in bovine mastitic milk. Inspired by earlier studies on protease activity using mastitic milk samples, we developed a simple colorimetric assay to distinguish mastitic milk from milk derived from healthy animals. The plasmin substrate coupled to magnetic nanoparticles form a black self-assembled monolayer on a gold sensor surface. In the presence of increased levels of plasmin, the substrate is cleaved and the peptide fragment attached to the magnetic beads, will be attracted by the magnet which is present under the sensor strips revealing the golden surface. We found the area of the golden color surface proportional to plasmin activity. The sensitivity of this method was determined to be 1 ng/ml of plasmin in vitro. Next, we tested the biosensor using mastitis positive milk of which infection is confirmed by bacterial cultures. This newly developed colorimetric biosensor has high potential in applications for the diagnosis of mastitis with potential spin offs to health, food and environmental sectors.

The study was aimed at assessing heat load-related risk of clinical mastitis (CM) in dairy cows. Records of CM for the years 2014 and 2015 were obtained from a large conventional dairy farm milking about 1,200 Holstein cows in central Italy. A case of CM was defined by the presence of clinical signs and veterinary confirmation. Quarter milk samples were collected and bacteriological investigated for each CM. Etiological agents were identified and classified as environmental or contagious pathogens. Hourly weather data from the nearest weather station were used to calculate heat load index (HLI). Upper and lower thresholds of HLI, at which the animal accumulates or dissipates heat, were settled and used to measure heat load balance through the accumulated heat load (AHL) model. Zero and positive values of AHL indicate periods of thermo-neutral and heat accumulation, respectively. Each case of CM was associated with HLI-AHL values recorded 5 d before the event. The risk of CM was evaluated using a case-crossover design. A conditional logistic regression model was used to calculate the odds ratio and 95% confidence intervals of CM recorded in thermo-neutral (AHL = 0) or heat load (AHL0) days, pooled or stratified for pathogen type (environmental or contagious). Classes of AHL as low (6.5), medium (6.6-34.9), and high (35) were included in the model. Other variables included in the model were milk yield as liters (20, 20-30, and30), days in milk (60, 60-150, and150), and parity (1, 2-3, and3). A total of 1,086 CM cases were identified from 677 cows. Escherichia coli, Streptococcus spp., and Streptococcus uberis were the environmental pathogens isolated with the highest frequency; Staphylococcus aureus prevailed within contagious species. The analysis of pooled data indicated a significant effect of heat load on the occurrence of CM in the contagious pathogen stratum. Higher milk yield, middle and late stage of lactation, and older parity increased the risk of CM under heat load conditions. However, the association between pathogen type and these factors was not clear because the model provided significant odds ratios within all pathogen categories. The present study provided the first evidence of an association between HLI and CM in dairy cattle and suggested the ability of the AHL model to assess the risk of mastitis associated with heat load.

Mastitis in dairy cattle is extremely costly both in economic and welfare terms and is one of the most significant drivers of antimicrobial usage in dairy cattle. A critical step in the prevention of mastitis is the diagnosis of the predominant route of transmission of pathogens into either contagious (CONT) or environmental (ENV), with environmental being further subdivided as transmission during either the nonlactating “dry” period (EDP) or lactating period (EL). Using data from 1000 farms, random forest algorithms were able to replicate the complex herd level diagnoses made by specialist veterinary clinicians with a high degree of accuracy. An accuracy of 98%, positive predictive value (PPV) of 86% and negative predictive value (NPV) of 99% was achieved for the diagnosis of CONT vs ENV (with CONT as a “positive” diagnosis), and an accuracy of 78%, PPV of 76% and NPV of 81% for the diagnosis of EDP vs EL (with EDP as a “positive” diagnosis). An accurate, automated mastitis diagnosis tool has great potential to aid non-specialist veterinary clinicians to make a rapid herd level diagnosis and promptly implement appropriate control measures for an extremely damaging disease in terms of animal health, productivity, welfare and antimicrobial use.

Escherichia coli is a normal inhabitant of the enteric microflora of human and animal. Intestinal and extra-intestinal infections caused by E. coli in mammals are characterized by the presence of diversity of virulence factors. In addition it can be isolated from environment surrounding human and animal farms. E. coli is the main environmental pathogen causing clinical mastitis in dairy cattle. It causes a wide range of disease severity, from changes seen exclusively in milk to severe systemic signs. The severity of clinical mastitis has been conventionally classified into three levels: mild (grade 1), moderate (score 2), and severe (score 3). Recently, reports of cases of bovine mastitis caused by environmental agents have been on the rise, in particular in countries that have succeeded in controlling contagious microorganisms. Unlike enteric and certain extra-enteric conditions in domestic animals and humans, the impact of virulence factors on the occurrence of bovine mastitis due to E. coli, as well as the clinical severity of the cases, is not fully understood. In this regard, the present study reviewed the most relevant virulence factors of E. coli in human and animals, with emphasis in bovine mastitis. Escherichia coli (E. coli) é um micro-organismo que pertence a microbiota entérica de humanos e animais. As afecções entéricas e extra-entéricas causadas por E.coli em mamíferos são caracterizadas pela presença de ampla diversidade de fatores de virulência. Além disso, este micro-organismo também pode ser isolado em ambientes de produção animal. E. coli é o principal agente ambiental que causa mastite clínica em vacas leiteiras, provocando sinais clínicos que podem variar desde mudanças observadas exclusivamente no leite até sinais sistêmicos graves. A gravidade da mastite clínica é convencionalmente classificada em três níveis: leve (grau 1), moderada (grau 2), e grave (grau 3). Atualmente, os relatos de casos de mastite bovina causada por agentes ambientais têm aumentando, principalmente nos países que conseguiram controlar os micro-organismos contagiosos. Diferentemente de infecções entéricas e certas extra-entéricas em humanos e animais, o impacto dos fatores de virulência de E. coli na ocorrência de mastite bovina não é totalmente compreendido, assim como na gravidade clínica dos casos. Neste contexto, o presente estudo revisou os fatores de virulência de E. coli mais relevantes para humanos e animais, com ênfase na mastite bovina.

Aims This study was conducted to determine the occurrence of staphylococci from cows with subclinical mastitis from independent herds in Algeria, and to characterize Staphylococcus aureus isolates. Methods and results Quarter milk samples were collected separately, somatic cells were counted and samples with more than 200 000 somatic cells per ml were cultured on blood agar. Staphylococci isolates were identified by routine diagnostics, and S. aureus isolates were tested for antibiotic susceptibility by disk diffusion and microdilution. Congo red agar was used to detect biofilm formation and capsule synthesis was detected on serum soft agar (SSA). The S. aureus isolates were characterized by spa typing. DNA microarray analysis was performed to detect resistance and virulence genes. Overall, 40·0% (167/418) of the cows suffered from mastitis. In 63·5% (106/167) of the cows staphylococci were identified. Nine of the 106 Staphylococcus isolates (8·5%) were S. aureus. The coagulase-negative staphylococci belonged to 14 species. All S. aureus isolates were multiresistant and biofilm forming, with 66·67% of them showing diffuse colonies on SSA and belonged to CC97-agrI-cap5. Biofilm genes (icaA/C/D), 13 genes encoding for adhesion, six genes encoding proteases, 11 genes encoding superantigen like toxins were found. Genes conferring resistance to tetracycline (tet(K)), penicillin (blaZ/I/R) and macrolide-lincosamide-streptogramin B (erm(B), erm(A)) were also detected in the S. aureus from this study. Conclusions The current investigation provides a detailed molecular and biofilm formation ability of S. aureus involved in subclinical mastitis in Algeria and shows the wide distribution of adhesion and enterotoxin(-like) genes among S. aureus responsible for causing subclinical bovine mastitis. Significance and impact of the study These findings are valuable in tracking the evolution and genomic variation of S. aureus from bovine origin.

Mastitis in dairy cattle is a highly prevalent infectious disease, causing considerable economic loss worldwide. In this study, we used Angelica dahurica and Rheum officinale extracts (designated as Yi-Xiong-Tang, YXT) for mastitis treatment. California mastitis test (CMT) was performed and 67 mastitis udder quarters were identified among 179 lactating dairy cows. These 67 mastitis udder quarters were subjected to treatments by intramammary infusion of YXT twice a day for three consecutive days. The mastitis indicators including clots, lactate dehydrogenase (LDH), TNF-α, IL-6, IL-8, and total viable count of bacteria (TVC) in milk were examined before and after the YXT treatment to evaluate its effectiveness. Levels of mastitis indicators from mastitis udder quarters were elevated. After YXT treatment, normal levels of these indicators were restored: TVC, 2.10 × 104 – 9.20 × 106 CFU/mL; clots, 6.56 ± 0.43 mg/mL; LDH, 181.0 ± 18.55 U/L; TNF-α, 0.02 ± 0.02 ng/mL; IL-6, 41.4 ± 11.46 pg/mL; and IL-8, 1.85 ± 0.60 pg/mL. Compared with the antibiotic therapy, YXT treatment has a shorter treatment course and might have lower probability for the causative agents to develop drug resistance because YXT is in fact a cocktail containing multiple active ingredients.

This study evaluated the efficacy of a recombinant lysostaphin fused to a protein transduction domain (rLYS-PTD) as a dry-cow therapy for the treatment of experimentally induced chronic, subclinical Staphylococcus aureus mastitis. Twenty-two Holstein dairy cows were experimentally infected with Staph. aureus in a single pair of diagonal mammary quarters approximately 45d before dry off. Staphylococcus aureus-infected mammary quarters of cows were randomly assigned to 1 of 2 treatment groups at dry off: (1) 279mg of rLYS-PTD in 50mL of vehicle (n=11 cows; 22 quarters) or (2) 50mL of vehicle solution (n=11 cows; 22 quarters) by intramammary infusion. All cows were followed for 30d postpartum to determine cure rates using bacteriologic culture, somatic cell counts, and clinical mastitis scores. No cures were recorded in either the treatment or control groups. Milk somatic cell count, bacterial colony counts, and mastitis scores did not significantly differ between treatment groups. In conclusion, rLYS-PTD was not an effective dry-cow therapeutic for chronic, subclinical Staph. aureus mastitis at the tested dose and formulation.

Staphylococcus aureus is known as a major causative agent of mastitis in dairy cattle. In the presentstudy, 104 isolates of Staphylococcus originated from subclinical mastitis cattle characterized for thephenotypic properties and the presence of Staphylococcal protein A (Spa). Some bacteria were resistancesagainst several antibiotics were also studied, such as erythromycin, streptomycin, tetracycline, cefepime,nitrofurantoin, amikacin, chloramphenicol, and ciprofloxacin. About 78% of the isolated were moderatelysensitive to nitrofurantoin, while 89% were highly resistant to cefepime and ciprofloxacin. Using thevarious mammals’ sera, seven isolates out of 104 revealed the presence of Spa.