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E. coli is one of the major significant pathogens causing mastitis, the most complex and costly diseases in the dairy industry worldwide. Present study was undertaken to isolate, detect the virulence factors, phylogroup, antimicrobial susceptibility and antimicrobial resistance genes in E. coli from cows with clinical mastitis. A total of 68 milk samples comprising 53 from clinical mastitis and 15 from apparently healthy cattle were collected from four different established dairy farms in Bangladesh. E. coli was isolated from the milk samples and identified by PCR targeting malB gene and sequencing of 16S rRNA gene. E. coli isolates were screened by PCR for the detection of major virulence genes (stx, eae and cdt) of diarrheagenic E. coli followed by phylogenetic grouping. Antimicrobial susceptibility of the E. coli isolates was determined by disk diffusion test and E. coli showing resistance was further screened for the presence of antimicrobial resistance genes. E. coli was isolated from 35.8% of the mastitis milk samples but none from the apparently healthy cattle milk. All the E. coli isolates were negative for stx, eae and cdt genes and belonged to the phylogenetic groups A and B1 which comprising of commensal E. coli. Antibiotic sensitivity testing revealed 84.2% (16/19) of the isolates as multidrug resistant. Highest resistance was observed against amoxicillin (94.5%) followed by ampicillin (89.5%) and tetracycline (89.5%). E. coli were found resistant against all the classes of antimicrobials used at the farm level. Tetracycline resistance gene (tetA) was detected in 100% of the tetracycline resistant E. coli and blaTEM-1 was present in 38.9% of the E. coli isolates. Findings of this study indicate a potential threat of developing antimicrobial resistance in commensal E. coli and their association with clinical mastitis. Occurrence of multidrug resistant E. coli might be responsible for the failure of antibiotic therapies in clinical mastitis as well as pose potential threat of transmitting and development of antibiotic resistance in human.

Clinical and subclinical mastitis regarded as important diseases causes a reduction in reproductive performance in dairy cows in the UK during the last decade. This study was aimed to assess the association between clinical mastitis, subclinical mastitis and the reproductive performance of cows. Data from 184 multiparous Holstein dairy cows were collected. Binomial logistic regression used to determine the incidence rate of clinical mastitis and subclinical mastitis between parity, calving year and seasons of the year. Linear mixed model used to determine the effect of clinical and subclinical mastitis on reproductive performance. The association between clinical or subclinical mastitis and the probability of a cow to get pregnant at 1st, 2nd or 3rd service was evaluated using binomial logistic regression. The same model used to illustrate the probability of a cow getting pregnant from 30-60 days postpartum or 61-90 days postpartum. The incidence of clinical mastitis was different between calving years (P

Objective: The current investigation was designed to point out the prevalence of multidrug-resis¬tant Streptococcus spp. causing acute clinical mastitis and their pattern of antibiotic resistance in dairy cows. Materials and methods: Milk was sampled from 128 dairy cows with 191 infected quarters during the period from August 2017 to December 2018. Bacterial species were isolated from the milk samples and identified based on colony morphology and biochemical tests. Multiplex PCR was done for confirmatory detection of the Streptococcus spp. isolates. Results: The chief isolation percentages, from the sampled milk, were Escherichia coli (26%), then Staphylococcus aureus (23%), and Streptococcus dysagalactiae (23%), then Streptococcus agalac¬tiae (20.1%), and finally coagulase-negative Staphylococci (7.7%). In confirmed PCR streptococci isolates, the antibiotic resistance genes have been detected, including macrolides antibiotic resis¬tance genes (ermB and mefA genes), lincosamides antibiotic resistance genes (linB gene), and tetra¬cycline resistance genes (tetM and tetO genes). Age, parity number, cleaning of bedding materials, cleaning of milking facilities, and utensils and udder cleaning practice were significant risk factors for multidrug-resistant streptococcal mastitis in dairy cows. Conclusion: The results of this study explored the phenotypic and genotypic traits of Streptococcus spp. which constitute a usual cause of acute clinical mastitis in dairy cows. The ermB, mefA, tetM, and tetO antibiotic-resistant genes were identified in streptococci isolates from dairy cows milk with acute clinical mastitis, indicating a public health hazard. Thus, veterinary clinical breakpoints are needed to improve surveillance data, improve the hygiene regimen on the farms, and pro¬mote the wise use of antimicrobials. [J Adv Vet Anim Res 2020; 7(2.000): 186-197]

Ova studija predstavlja podatke prikupljene iz Veterinarske stanice “Zevet” d.o.o. Zenica, Bosna i Hercegovina o zastupljenosti bakterijskih patogena kod klinički prisutnih mastitisa mliječnih goveda i profilima njihove antimikrobne osjetljivosti. Za laboratorijsku izolaciju obrađena su 52 uzorka od kojih su 23 bila bakteriološki pozitivna. Najčešći izolati bili su Staphylococcus aureus u 21,74 % uzoraka i Streptococcus agalactiae u 17,39 % izolira- nih uzoraka, koagulaza-negativni stafilokoki (13,04 %), Klebsiella pneumoniae (13,04 %), Ente- rococcus spp. (8,70 %), Escherichia coli (8,70 %), Streptococcus spp., Enterobacter, Serratia spp. i Yersinia enterocolitica (4,35 %). Najveća antimikrobna otpornost među izolatima primjećena je na benzilpenicilin (56,25 %) i oksitetraciklin (46,15 %) s ukupnom otpornosti na uporabljena antimikrobna sredstava od 25,99 %. Na tri testirana antimikrobna sredstva (neomicin, klindamicin i cefotaksim) izolati nisu pokazali otpornost. Ostali antibiotici na koje je primijećena niska ukupna otpornost su ciprofloksacin (5,56 %) i ofloksacin (6,67 %). Samo jedan izolat nije pokazao nikakvu otpornost na testirane antimikrobike (S. aureus). Najveću otpornost među izolatima pokazao je Enterococcus spp., pokazujući senzibilitet samo za tetracikline. S. aureus je pokazao otpornost na benzilpenicilin i cefaleksin u 60 % slučajeva. Svi izolati S. agalactiae su bili otporni na tetracikline, oksitetracikline i gentamicin. Koagulaza negativni stafilokoki pokazali su otpornost na cefaleksin u dva slučaja i otpornost na benzilpenicilin, eri- tromicin i oksacilin u jednom izolatu. Klebsiella pneumoniae je pokazala otpornost na cefaleksin, amoksicilin i benzilpenicilin u svim ispitivanim izolatima, a pokazala je i otpornost na tetracikline u dva izolata. Svi ostali izolati pokazali su otpornost prema najmanje jednom testiranom antimikrobiku. U 46 slučajeva se odmah pristu- palo liječenju s 86,96 % uspjehom koji je procijenjen na osnovu izostanka kliničkog relapsa i smanjenja broja somatskih stanica u dopuštene granice za tri tjedna nakon provedene terapije., This study provides data on the bacterial pathogens present in clinical mastitis of dairy cattle and their antimicrobial susceptibility profiles, collected by the Zevet Veterinary Station in the Zenica region in Bosnia and Herzegovina. A total of 52 samples were taken for laboratory isolation and 23 bacterial pathogens were isolated. The most common isolates were Staphylococcus aureus in 21.74% of samples and Streptococcus agalactiae (17.39%), followed by coagulase negative staphylococci (13.04%), Klebsiella pneumoniae (13.04%), Enterococcus spp. (8.70%), Escherichia coli (8.70%), Streptococcus spp., Enterobacter, Serratia spp. and Yersinia enterocolitica (4.35%). The highest antimicrobial resistance among isolates was observed with tested benzylpenicillin (56.25%) and oxytetracyline (46.15%) with an overall resistance of used medical agents of 25.99%. Three of the tested antimicrobial agents (neomycin, clindamycin and cefotaxim) did not show resistance. Other antibiotics with low total resistance were ciprofloxacin (5.56%) and ofloxacin (6.67%). Only one isolate showed no resistance to the tested antimicrobials (S. aureus). The highest resistance between the isolates was shown by Enterococcus spp., showing sensitivity only to tetracycline. S. aureus showed resistance to benzylpenicillin and cephalexin in 60% of cases. All S. agalactiae isolates were resistant to tetracycline, oxytetracycline and gentamicin. Coagulase negative staphylococci showed resistance to cephalexin in two cases and resistance to benzylpenicillin, erythromycin and oxacillin in one isolate. Klebsiella pneumoniae showed resistance to cephalexin, amoxicillin and benzylpenicillin in all tested isolates, and showed resistance to tetracyclines in two isolates. All other isolates showed resistance to at least one of the antimicrobials tested. A total of 46 cases were treated immediately after clinical diagnosis with an 86.96% success rate, which was estimated based on the absence of clinical relapse and a decrease in the number of somatic cells within the permissible three week limit after therapy.

Aim: This study aimed to evaluate and quantify the different factors affecting the costs of mastitis in cattle, to quantify the annual and monthly financial losses attributed to mastitis, and to estimate production losses using average linear scores found on The Dairy Herd Improvement Association somatic cell count (SCC) sheets and bulk tank SCC per lactation number. Materials and Methods: All data (bovine mastitis-associated costs and expenditures in Egyptian dairy herds) were analyzed using SPSS/PCT, 2001. A partial budget technique using spreadsheet software and the general linear model procedure was used to analyze the productive and financial measures. Results: Mastitis was present in 57.1% of cows (240/420), while clinical mastitis (CM) was present in 19% of them (80/420). The geometric mean of SCC/ml in bulk tank milk samples of 150 cattle dairy farms was 556.3×103. The annual subclinical mastitis (SCM)-related economic loss was 21,933,258.6 LE, and the two most important cost components were the subsequent decrease in milk production and quality premium losses (93% and 7% of SCM costs, respectively). The quality premium loss was 1,369,602.1 LE. On the other hand, the annual economic loss due to decreased milk production as a result of SCM was 20,563,656.5 LE. The total cost of 80 CM cases, including the failure and preventive costs, was 1,196,871.4 LE, including 1,169,150.4 LE failure costs (106,336.0 LE in direct costs and 1,062,814.4 LE in indirect costs) and 27,721.0 LE preventive costs. The average cost per CM case was 28,760.9 LE, including veterinary time and consultation fees of 250.0 LE (1%), labor 562.5 LE (2%), premature culling 736,000.0 LE (77%), decreased milk production 4085.18 LE (13.7%), discarded milk 185.3 LE (1%), and drugs and treatments 328.9 LE (1%). The total costs of CM (expenditures) extra control and preventive measures, including the diagnosis of CM for 80 cows annually in 20 Egyptian dairy farms were 27,721.0 LE, representing 346.5 LE or 1% of the total cost of CM cases. The cost of monitoring and diagnostic measures was 8635.2 LE, representing 107.9 LE or 1% of the total cost of a case of CM. Conclusion: The method used for cost estimation, in this study, is highly adaptable to individual cattle farms and had a major role in assessing specific control and management measures. The concepts described in this paper help to improve our understanding of the full economic impact of clinical and subclinical mastitis in cattle in Egypt. Assessing the economic losses from mastitis to determine the economic costs and losses occurring in Egyptian dairy farms is critical for encouraging farmers to acknowledge the scale of the problem and implement effective management practices aimed at improving mastitis control and reducing the associated costs.

Cow mastitis, with high incidence rate and complex cause of disease, is one of the main diseases that affect the development of dairy industry in the world. Clinical mastitis and subclinical mastitis caused by Staphylococcus aureus, Escherichia coli, Streptococcus, and other pathogens have a huge potential safety hazard to food safety and the rapid development of animal husbandry. The economic loss caused by cow mastitis is billions of dollars every year in the world. In recent years, the omics technology has been widely used in animal husbandry with the continuous breakthrough of sequencing technology and the continuous reduction of sequencing cost. For dairy cow mastitis, the traditional diagnostic technique, such as histopathological screening, somatic cell count, milk pH test, milk conductivity test, enzyme activity test, and infrared thermography, are difficult to fully and comprehensively clarify its pathogenesis due to their own limitations. Metabolomics technology is an important part of system biology, which can simultaneously analyze all low molecular weight metabolites such as amino acids, lipids, carbohydrates under the action of complex factors including internal and external environment and in a specific physiological period accurately and efficiently, and then clarify the related metabolic pathways. Metabolomics, as the most downstream of gene expression, can amplify the small changes of gene and protein expression at the level of metabolites, which can more fully reflect the cell function. The application of metabolomics technology in cow mastitis can analyze the hetero metabolites, identify the related biomarkers, and reveal the physiological and pathological changes of cow mammary gland, so as to provide valuable reference for the prediction, diagnosis, and treatment of mastitis. The research progress of metabolomics technology in cow mastitis in recent years was reviewed, in order to provide guidance for the development of cow health and dairy industry safety in this manuscript.

Bedding can affect mammary health of dairy cows. The objectives of this study were to evaluate clinical mastitis incidence in cows housed on recycled manure solids bedding and, more specifically, to determine which pathogens were involved. We followed 26 recycled manure solids farms and 60 straw-bedded farms as a comparative group during 1 year (2018–2019). For each episode of clinical mastitis, defined as a visual alteration of the milk, with or without local or systemic signs of infection, producers sampled aseptically the affected quarter, provided some details about the animal, and sent the sample to the research team at the Université de Montréal. We received and analyzed 1,144 milk samples. The samples were cultured according to the National Mastitis Council guidelines and the different colony phenotypes were subsequently identified with mass spectrometry. In 54.6% of CM cases, a single phenotype of bacteria was cultured (pure culture), while two different phenotypes were found in 16.8% of the samples (mixed culture), and no growth was observed in 14.4% of the samples. Samples with three or more phenotypes were considered contaminated and were not included in the pathogen-specific analyses (14.3% of the submitted samples). The most frequently identified bacterial species in pure and mixed culture in farms using recycled manure solids were Streptococcus uberis (16.0%), Escherichia coli (13.8%), Klebsiella pneumoniae (13.2%), Streptococcus dysgalactiae (6.2%), and Staphylococcus aureus (3.4%). In straw farms, the most frequent species were S. aureus (16.6%), S. uberis (11.0%), E. coli (9.1%), S. dysgalactiae (8.0%), and K. pneumoniae (1.1%). The incidence of clinical mastitis (all cases together) was not higher in recycled manure solids farms (14.0 cases/100 cow-year; 95% CI: 8.3–23.7) compared with straw-bedded farms (16.3 cases/100 cow-year; 95% CI: 9.0–29.6). However, K. pneumoniae clinical mastitis episodes were 7.0 (95% CI: 2.0–24.6) times more frequent in recycled manure solids farms than in straw farms. Adjusted least square means estimates were 1.6 K. pneumoniae clinical mastitis cases/100 cow-year (95% CI: 0.8–3.4) in recycled manure solids farms vs. 0.2 cases/100 cow-year (95% CI: 0.1–0.6) in straw-bedded farms. Klebsiella pneumoniae clinical mastitis is in general severe. Producers interested in this bedding alternative need to be aware of this risk.

Background: Non-lactational infectious mastitis (NLIM) is an inflammatory breast disease with broad clinical presentation. Inadequate treatment can lead to chronic infections that cause breast deformities. NLIM information is limited, especially in the Americas. A systematic review and meta-analysis have been conducted here.Methods: Literature search was conducted in three databases (Lilacs, PubMed, and Scielo) on NLIM cases in the Americas. Demographic, epidemiological, clinical, radiological, and laboratory data were extracted. The main characteristics and results were also compared according to the country's gross national income.Results: A total of 47 articles were included, resulting in 93 cases. The etiological agent was described in 86 (92.5%) patients. Bacteria were the most prevalent etiology (73; 84.8%). Amongst bacterial diagnoses, more frequent cases were Mycobacterium tuberculosis (28; 38.4%); Corynebacterium spp. (15; 20.5%); non-tuberculous mycobacteria (13; 17.8%). The cases were reported in eight different countries, with the USA being the country with the highest number of cases (35; 37.6%). Patients from high-income countries group presented a shorter diagnostic time when compared to low, low-middle, and upper-middle-income countries. A greater number of radiographic studies with pathological findings were described in high-income countries.Conclusion: Non-lactational infectious mastitis is a complex public health problem with diagnostic and treatment challenges. Hence, multi-professional approach-based additional studies are recommended on its epidemiology, diagnosis, treatment, and control.

Bovine paratuberculosis (PTB) is a chronic inflammatory disease caused by Mycobacterium avium susbp. paratuberculosis (MAP). Genome-wide association studies (GWAS) have identified single nucleotide polymorphisms (SNPs) significantly associated with susceptibility to bovine PTB. The main objective of this study was to identify quantitative trait loci (QTLs) associated with MAP infection in Spanish Holstein cows (N = 983) using combinations of diagnostic tests and imputed whole-genome sequence (WGS) data. The infection status of these animals was defined by three diagnostic methods including ELISA for MAP-antibodies detection, and tissue culture and PCR for MAP detection. The 983 cows included in this study were genotyped with the Bovine MD SNP50 Bead Chip, and the corresponding genotypes were imputed to WGS using the 1,000 Bull genomes reference population. In total, 33.77 million SNP variants per animal were identified across the genome. Linear mixed models were used to calculate the heritability (h2) estimates for each diagnostic test and test combinations. Next, we performed a case-control GWAS using the imputed WGS datasets and the phenotypes and combinations of phenotypes with h2 estimates > 0.080. After performing the GWAS, the test combinations that showed SNPs with a significant association (PFDR ≤ 0.05), were the ELISA-tissue PCR-tissue culture, ELISA-tissue culture, and ELISA-tissue PCR. A total of twelve quantitative trait loci (QTLs) highly associated with MAP infection status were identified on the Bos taurus autosomes (BTA) 4, BTA5, BTA11, BTA12, BTA14, BTA23, BTA24, and BTA28, and some of these QTLs were linked to immune-modulating genes. The identified QTLs on BTA23 spanning from 18.81 to 22.95 Mb of the Bos taurus genome overlapped with several QTLs previously found to be associated with PTB susceptibility, bovine tuberculosis susceptibility, and clinical mastitis. The results from this study provide more clues regarding the molecular mechanisms underlying susceptibility to PTB infection in cattle and might be used to develop national genetic evaluations for PTB in Spain. Financial support for this study was provided by a grant from the Spanish Ministry of Science, Innovation, and Universities (MICINN, project code: RTI2018-094192-R-C21) and by European Regional Development Funds (FEDER) to MAH. MC and GBB have been awarded fellowships from the National Institute for Agricultural Research (INIA) and MICINN, respectively

Mastitis-associated milk losses in dairy cows have a massive impact on farm profitability and sustainability. In this study, we analyzed milk losses from 4 553 treated mastitis cases as recorded via treatment registers at 41 AMS dairy farms. Milk losses were estimated based on the difference between the expected and the actual production. To estimate the unperturbed lactation curve, we applied an iterative procedure using the Wood model and a variance-dependent threshold on the milk yield residuals. We calculated milk losses both in a fixed window around the first treatment day of each mastitis case and in the perturbations corresponding to this day, at the cow level as well as at the quarter level. In a fixed time window of day -5 to 30 around the first treatment, the absolute median milk losses per case were 101.5 kg, highly dependent on the parity and the lactation stage with absolute milk losses being highest in multiparous cows and at peak lactation. Relative milk losses expressed in percentage were highest on the first treatment day, and full recovery was often not reached within 30 days from treatment onset. In 62 % of the cases, we found a perturbation in milk yield at the cow level at the time of treatment. On average, perturbations started 8.7 days before the first treatment and median absolute milk losses increased to 128 kg of milk per perturbation. Mastitis is not expected to have equal effects on the four quarters so this study additionally investigated losses in the individual udder quarters. We used a data-based method leveraging milk yield and electrical conductivity to project the presumably inflamed quarter. Next, we compared losses with the average of presumably non-inflamed quarters. Median absolute losses in a fixed 36-day window around treatment varied between 50.2 kg for front and 59.3 kg for hind inflamed quarters compared to respectively 24.7 and 26.3 kg for the median losses in the non-inflamed quarters. Also here, these losses differed between lactation stages and parities. Expressed proportionally to expected yield, the relative median milk losses in inflamed quarters on the treatment day were 20 % higher in inflamed quarters with a higher variability and slower recovery. In 86 % of the treated mastitis cases, at least one perturbation was found at the quarter level. This analysis confirms the high impact of mastitis on milk production, and the large variation between quarter losses illustrates the potential of quarter analysis for on-farm monitoring at farms with an automated milking system. ispartof: PREVENTIVE VETERINARY MEDICINE vol:194 ispartof: location:Netherlands status: published

Simple Summary Bovine mastitis is a complex disease with many potential risk factors. However, few studies have reported the risk factors for mastitis in Brazil. This prospective, repeated cross-sectional study was carried out between May 2018 and June 2019 in smallholder dairy farms located in the southeast of Brazil. The potential risk factors for clinical and subclinical mastitis in smallholder dairy farms in Brazil were investigated by a combination of a questionnaire, clinical udder examinations and tests for subclinical mastitis. Risk factors for mastitis were evaluated at the cow level and at the herd level. The risk factors evaluated at the herd level were related to milking management, environment and management practices. We identified some risk factors; increased parity, later stage of lactation, not milking clinical and subclinical cases last, lack of routine cleaning of the milking parlor, using the dry-off treatment and optimized feed before calving. Further research on more farms across more areas of Brazil is required to develop targeted control programs for mastitis on smallholder dairy farms. Abstract The aim of this study was to investigate the potential risk factors for clinical and subclinical mastitis in smallholder dairy farms in Brazil. A prospective, repeated cross-sectional study was carried out between May 2018 and June 2019 on 10 smallholder dairy farms. Potential risk factors for subclinical and clinical mastitis at the herd and cow level were recorded through interviewing the owner and by observation. A combination of clinical udder examination and the Tamis (screened mug with a dark base) test (Tadabras Indústria e Comércio de Produtos Agrovetereinário LTDA, Bragrança Paulista, SP, Brazil) were applied to observe clinical mastitis, and the California Mastitis Test (Tadabras Indústria e Comércio de Produtos Agrovetereinário LTDA, Bragrança Paulista, SP, Brazil) was used to determine subclinical mastitis. A total of 4567 quarters were tested, 107 (2.3%) had clinical mastitis, while 1519 (33.2%) had subclinical mastitis. At the cow level, clinical mastitis risk was highest in mid-lactation (50–150 days in milk) with OR 2.62 with 95% confidence interval (CI) of 1.03–6.67, while subclinical mastitis was highest in late lactation (> 150 days in milk) with OR 2.74 (95% CI 2.05–3.63) and lower in primiparous (OR 0.54, 95% CI 0.41–0.71) than multiparous cows. At the herd level, using dry-cow treatment (OR 4.23, 95% CI 1.42–12.62) was associated with an increased risk of clinical mastitis. Milking clinical (OR 0.37, 95% CI 0.24–0.56) and subclinical cases last (OR 0.21, 95% CI 0.09–0.47) and cleaning the milking parlor regularly (OR 0.27, 95% CI 0.15–0.46) had decreased odds for subclinical mastitis, while herds with optimized feed had greater odds (OR 9.11, 95% CI 2.59–31.9). Prevalence of clinical mastitis was at its lowest at the first visit in June/July and highest at the last visit in April/June (OR 3.81, 95% CI 1.93–7.52). Subclinical mastitis also presented increased odds in the last visit (OR 2.62, 95% CI 2.0–3.36). This study has identified some risk factors for mastitis on smallholder farms but further research on more farms across more areas of Brazil is required to develop a targeted mastitis control program for smallholder farms.

Background and Aim: Bovine mastitis is the costliest prevalent disease in the dairy sector due to the limitations of conventional treatments. Zinc oxide nanoparticles (ZnO-NPs) have been regarded as safe and economical antibacterial candidates against several microorganisms, but the tendency of these particles to aggregate is a major barrier to their application. This study aimed to enhance the antibacterial efficiency of ZnO-NPs against some bacterial agents, causing bovine mastitis. Materials and Methods: A total of 24 milk samples out of 300 cases from Nubaria farm, Beheira Governorate, Egypt, were collected from cows with clinical mastitis. ZnO-NPs were fabricated by a sonochemical method using starch as a capping agent and by an auto-combustion reaction using glycine as a fuel. The two preparations of synthesized ZnO-NPs at different concentrations were assessed for their antimicrobial activities in vitro against Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae isolated from milk of affected cows. Results: Sonochemically synthesized capped ZnO-NPs were dispersed and non-agglomerated in comparison with aggregated uncapped ZnO-NPs prepared by an auto-combustion reaction. Capped dispersed ZnO-NPs showed higher antibacterial activity against S. aureus, E. coli, and K. pneumoniae than particles synthesized by the auto-combustion reaction at same concentrations. However, the zone of inhibition for dispersed and agglomerated ZnO-NPs was concentration-dependent. In addition, Gram-positive S. aureus exhibited higher resistance to ZnO-NPs synthesized by both methods than Gram-negative E. coli and K. pneumoniae. Conclusion: Dispersed, non-agglomerated ZnO-NPs fabricated using starch as a capping agent under sonochemical irradiation could potentially be regarded as highly effective and inexpensive antimicrobial agents against S. aureus, E. coli, and K. pneumoniae for the management of bovine mastitis. Keywords: antibacterial activity, clinical mastitis, dairy cows, zinc oxide nanoparticles.

Mastitis is a costly disease and in many areas of the world, these costs have been quantified to support farmers in their decision making with regard to prevention of mastitis. Although for subsaharan circumstances estimates have been made for the costs of subclinical mastitis (SCM), farm-specific cost estimations comprising both clinical mastitis (CM) and SCM are lacking. In this paper, we quantified failure costs of both CM and SCM on 150 Ethiopian market-oriented dairy farms keeping Holstein Friesian × Zebu breed cows. Data about CM were collected by face-to-face interviews and the prevalence of SCM was estimated for each farm using the California mastitis test. All other relevant information needed to calculate the failure costs, such as the consequences of mastitis and price levels, was collected during the farm visits, except for the parameter for milk production losses due to SCM, which was based on literature estimates and subjected to sensitivity analyses. The average total failure costs of mastitis was estimated to be 4 765 Ethiopian Birr (ETB) (1 ETB = 0.0449 USD) per farm per year of which SCM contributed 54% of the costs. The average total failure costs per lactating cow per farm per year were 1 961 ETB, with a large variation between farms (range 0 to 35 084 ETB). This large variation in failure costs between farms was mainly driven by variation in incidence of CM and prevalence of SCM. Milk production losses made the largest contribution (80%), whereas culling contributed 13% to 17% to the total failure costs. In our estimates, costs of veterinary services, drugs, discarded milk and labour made a minor contribution to the total failure costs of mastitis. Relative to the income of dairy farmers in North Western Ethiopia; the total failure costs of mastitis are high. In general, Ethiopian farmers are aware of the negative consequences of CM, but creating awareness of the high costs of SCM and showing large variation between farmers may be instrumental in motivating farmers to also take preventive measures for SCM.

Despite considerable efforts to control bovine mastitis and explain its causes, it remains the most costly and common disease of dairy cattle worldwide. The role and impact of non-aureus staphylococci (NAS) in udder health are not entirely understood. These Gram-positive bacteria have become the most frequently isolated group of bacteria in milk samples of dairy cows and are associated with (mild) clinical and subclinical mastitis. Different species and strains of NAS differ in their epidemiology, pathogenicity, virulence, ecology and host adaptation, and antimicrobial resistance profiles. They have distinct relationships with the microbiome composition of the udder and may also have protective effects against other mastitis pathogens. Some appear to persist on the skin and in the teat canal and udder, while others seem to be transient residents of the udder from the environment. Analyzing genotypic and phenotypic differences in individual species may also hold clues to why some appear more successful than others in colonizing the udder. Understanding species-level interactions within the microbiome and its interactions with host genetics will clarify the role of NAS in bovine mastitis and udder health.

Simple Summary Bovine clinical mastitis is an inflammatory disease of the mammary gland associated with visual changes in the milk and/or the udder. We show that the incidence of clinical mastitis in commercial dairy farms in Bangladesh is high but with large variation between farms. Streptococci and non-aureus Staphylococci were the most frequently isolated bacteria from quarter milk samples. Staphylococcus aureus and non-aureus Staphylococci were often resistant against penicillin and oxacillin. This work suggests an urgent need for improved udder health management and specifically a more prudent use of antimicrobial agents following a treatment protocol. Abstract Clinical mastitis (CM) is an important production disease in dairy cows, but much of the knowledge required to effectively control CM is lacking, specifically in low-income countries where most farms are small and have specific dairy management, such as regular udder cleaning and practicing hand milking. Therefore, we conducted a 6-month-long cohort study to (a) estimate the incidence rate of clinical mastitis (IRCM) at the cow and quarter level, (b) identify risk factors for the occurrence of CM, (c) describe the etiology of CM, and (d) quantify antimicrobial susceptibility (AMS) against commonly used antimicrobial agents in S. aureus and non-aureus Staphylococcus spp. (NAS) in dairy farms in the Chattogram region of Bangladesh. On 24 farms, all cows were monitored for CM during a 6-month period. Cases of CM were identified by trained farmers and milk samples were collected aseptically before administering any antimicrobial therapy. In total, 1383 lactating cows were enrolled, which totaled 446 cow-years at risk. During the study period, 196 new cases of CM occurred, resulting in an estimated crude IRCM of 43.9 cases per 100 cow-years, though this varied substantially between farms. Among the tested CM quarter samples, Streptococci (22.9%) followed by non-aureus staphylococci (20.3%) were the most frequently isolated pathogens and resistance of S. aureus and NAS against penicillin (2 out of 3 and 27 out of 39 isolates, respectively) and oxacillin (2 out of 3 and 38 out of 39 isolates, respectively) was common. The IRCM was associated with a high milk yield, 28 to 90 days in milk, and a higher body condition score. Our results show that there is substantial room for udder health improvement on most farms.

Made available in DSpace on 2022-04-29T08:45:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 It is unknown whether overuse of antimicrobials against clinical mastitis (CM) from Streptococcus uberis is associated with increased antimicrobial resistance (AMR). Therefore, the aim of this study was to evaluate the association between antimicrobial use (AMU) and AMR in relation to the Strep. uberis causing CM in dairy herds. A total of 83 Strep. uberis isolates were selected from a collection created during a previous study evaluating the epidemiology of CM in dairy herds (n = 17) of southeastern Brazil. For each case of CM identified on farm, the following information was recorded: cow's identification number, affected mammary quarter, date of CM diagnosis, antimicrobial commercial names, number of administrations, and descriptions of protocol changes during the treatment. Streptococcus uberis isolates were confirmed by conventional culture, MALDI-TOF mass spectrometry, and quantitative multiplex PCR analyses. Thus, a total of 8 antimicrobials commonly used for CM treatment were evaluated for antimicrobial activity against Strep. uberis isolates. The minimum inhibitory levels of antimicrobials were determined at the lowest concentrations able to inhibit 50 and 90%, respectively, of Strep. uberis isolates. Data related to the antibiotics used for treatment of CM was used to calculate the frequency of administered antimicrobials as the number of defined daily doses (DDD). The highest frequencies of resistant Strep. uberis were observed for erythromycin (80.7% resistant, R), tetracycline (R = 59%), and penicillin G (R = 57.8%), whereas against ceftiofur only 10.8% of Strep. uberis isolates were resistant, and only 1.2% of the Strep. uberis isolates were resistant to enrofloxacin. Regarding the evaluation of resistance for antimicrobial classes, the highest frequency was observed for macrolides (R = 80.7%; 19.3% susceptible, S). Additionally, a frequency of 18.7% of Strep. uberis isolates were resistant to cephalosporins (S = 81.3%), respectively. Further, 94% of Strep. uberis isolates were multiresistant; all these isolates presented resistance to at least 3 different antimicrobial classes. The overall monthly average of antimicrobial treatment incidence (ATI) among the 17 herds enrolled in the study was 23.7 DDD per 1,000 lactating dairy cows [standard deviation (SD) = 13.9], ranging from 5.0 to 55.4 DDD per 1,000 cows in lactation-day. Cephalosporins and penicillins were the most commonly used antimicrobial classes among the evaluated herds (n = 16; 94.1%), followed by tetracyclines (n = 15 herds; 88.2%), fluoroquinolones (n = 14; 82.3%), and sulfonamides (n = 14; 82.3%). The tetracycline class had the highest ATI mean (5.0 DDD per 1,000 lactating cow-days, SD = 5.8), followed by fluoroquinolones (4.7 DDD per 1,000 lactating cow-days, SD = 6.0) and cephalosporins (3.8 DDD per 1,000 lactating cow-days, SD = 6.0). The overall use of antimicrobials was associated with the resistance of Strep. uberis to the antimicrobial tetracycline. Department of Nutrition and Animal Production School of Veterinary Medicine and Animal Sciences University of São Paulo (USP) Department of Production Animal Health University of Calgary Department of Population Medicine and Diagnostic Sciences College of Veterinary Medicine Cornell University Department of Chemical and Biological Sciences Institute of Biosciences São Paulo State University Rua Dr. Plínio Pinto e Silva Department of Chemical and Biological Sciences Institute of Biosciences São Paulo State University Rua Dr. Plínio Pinto e Silva

To ensure milk quality and detect cows with signs of mastitis, visual inspection of milk by prestripping quarters before milking is recommended in many countries. An objective method to find milk changed in homogeneity (i.e., with clots) is to use commercially available inline filters to inspect the milk. Due to the required manual labor, this method is not applicable in automatic milking systems (AMS). We investigated the possibility of detecting and predicting changes in milk homogeneity using data generated by AMS. In total, 21,335 quarter-level milk inspections were performed on 5,424 milkings of 624 unique cows on 4 farms by applying visual inspection of inline filters that assembled clots from the separate quarters during milking. Images of the filters with clots were scored for density, resulting in 892 observations with signs of clots for analysis (77% traces or mild cases, 15% moderate cases, and 8% heavy cases). The quarter density scores were combined into 1 score indicating the presence of clots during a single cow milking and into 2 scores summarizing the density scores in cow milkings during a 30-h sampling period. Data generated from the AMS, such as milk yield, milk flow, conductivity, and online somatic cell counts, were used as input to 4 multilayer perceptron models to detect or predict single milkings with clots and to detect milking periods with clots. All models resulted in high specificity (98–100%), showing that the models correctly classified cow milkings or cow milking periods with no clots observed. The ability to successfully classify cow milkings or cow periods with observed clots had a low sensitivity. The highest sensitivity (26%) was obtained by the model that detected clots in a single milking. The prevalence of clots in the data was low (2.4%), which was reflected in the results. The positive predictive value depends on the prevalence and was relatively high, with the highest positive predictive value (72%) reached in the model that detected clots during the 30-h sampling periods. The misclassification rate for cow milkings that included higher-density scores was lower, indicating that the models that detected or predicted clots in a single milking could better distinguish the heavier cases of clots. Using data from AMS to detect and predict changes in milk homogeneity seems to be possible, although the prediction performance for the definitions of clots used in this study was poor.

Staphylococcus aureus (S. aureus) is an important pathogen that causes clinical mastitis in goats and produces infections difficult to cure. Different antimicrobials as fluoroquinolones have been used against S. aureus. However, the studies developed to evaluate the bacterial drug interaction only have used the MIC as a single reference point with artificial growth media. The aims of this study were to describe the effect of marbofloxacin on S. aureus isolated from mastitis goats’ milk by different approaches as the minimum inhibitory and bactericidal concentrations (MIC and MBC) in cation adjusted Mueller–Hinton broth (CAMHB), serum and milk of goats at two inoculum sizes of 105 and 108 CFU/mL, the determination and analysis of the time kill curves (TKC) by non-linear mixed effect models in each growth medium and inoculum size, as well as the estimation of their pharmacokinetics/pharmacodynamics (PK/PD) cutoff values. The results obtained indicate that MIC values were higher and increases 2,4-fold in serum and 3,6-fold in milk at high inoculum, as well as the EC50 values determined by each pharmacodynamics model. Finally, the PK/PD cutoff values defined as fAUC24/MIC ratios to achieve clinical efficacy were highly dependent on inoculum and growth medium, with median values of 60–180, especially at high inoculum in milk, suggesting that further studies are necessary to evaluate and optimize the best therapeutic strategies for treating S. aureus in lactating goats., Facultad de Ciencias Agropecuarias, Universidad Católica de Córdoba, Argentina, Universidad Complutense de Madrid

Mastitis is a prevalent disease in dairy cattle, and staphylococci are among the most common causative pathogens. Staphylococci can express resistance to a range of antimicrobials, of which methicillin resistance is of particular public health concern. Additionally, Staphylococcus aureus carries a variety of virulence factors, although less is understood about the virulence of non-aureus staphylococci (NAS). The aim of our study was to identify and characterize 3 collections of staphylococcal isolates from bovine milk samples regarding antimicrobial resistance, with emphasis on methicillin resistance, and their carriage of virulence genes typically displayed by Staph. aureus. A total of 272 staphylococcal isolates collected in Norway and Belgium in 2016 were included, distributed as follows: group 1, Norway, 100 isolates; group 2, Flanders, Belgium, 64 isolates; group 3, Wallonia, Belgium, 108 isolates. Species identification was performed by use of MALDI-TOF mass spectrometry. Phenotypic resistance was determined via disk diffusion, and PCR was used for detection of methicillin resistance genes, mecA and mecC, and virulence genes. Antimicrobial resistance was common in Staphylococcus epidermidis and Staphylococcus haemolyticus from all different groups, with resistance to trimethoprim-sulfonamide frequently occurring in Staph. epidermidis and Staph. haemolyticus as well as in Staph. aureus. Resistance to penicillin was most frequently observed in group 1. Ten Belgian isolates (1 from group 2, 9 from group 3) carried the methicillin resistance determinant mecA: 5 Staph. aureus from 2 different farms and 5 NAS from 3 different farms. Almost all Staph. aureus isolates were positive for at least 3 of the screened virulence genes, whereas, in total, only 8 NAS isolates harbored any of the same genes. Our study contributes to the continuous need for knowledge regarding staphylococci from food-producing animals as a basis for better understanding of occurrence of resistance and virulence traits in these bacteria.

International audience; Iron is an essential nutrient for bacterial growth. Therefore, bacteria have evolved chelation mechanisms to acquire iron for their survival. Enterobactin, a chelator with high affinity for ferric iron, is secreted by Escherichia coli and contributes to its improved bacterial fitness. In this preliminary study, we evaluated enterobactin deficiency both in vitro and in vivo in the context of E. coli mastitis. Firstly, we showed that expression of lipocalin 2, a protein produced by the host that is able to both bind and deplete enterobactin, is increased upon E. coli infection in the cow's mastitic mammary gland. Secondly, we demonstrated in vitro that enterobactin deficiency does not alter interleukin (IL)-8 expression in bovine mammary epithelial cells and its associated neutrophil recruitment. However, a significantly increased reactive oxygen species production of these neutrophils was observed. Thirdly, we showed there was no significant difference in bacterial in vitro growth between the enterobactin-deficient mutant and its wild-type counterpart. However, when further explored in a murine model for bovine mastitis, the enterobactin-deficient mutant vs. the wild-type strain revealed a significant reduction of the bacterial load and, consequently, a decrease in pro-inflammatory cytokines (IL-1α,-1β,-4,-6, and-8). A reduced neutrophilic influx was also observed immunohistochemically. These findings therefore identify interference of the enterobactin iron-scavenging mechanism as a potential measure to decrease the fitness of E. coli in the mastitic mammary gland.

Background Our aim was to investigate the incidence and prevalence of clinical mastitis, peracute mastitis, metabolic disorders, and peripartum disorders, and to examine factors affecting the prevalence of each disease in cows raised on a large dairy farm in a temperate climate in Japan. The present study was performed on a large commercial dairy farm with approximately 2500 Holstein cows. Data were collected from 2014 to 2018, and involved 9663 calving records for 4256 cows. Results The incidence rate on the farm was 21.9% for clinical mastitis, 10.4% for peracute mastitis, 2.9% for metabolic disorders, and 3.2% for peripartum disorders. The prevalence rates for clinical mastitis, peracute mastitis, metabolic disorders, and peripartum disorders were 28.0, 13.3, 3.7, and 4.0%, respectively. In all four diseases, the probability of time to occurrence for each disease was associated with parity and calving season (P Conclusions Our results showed that clinical mastitis occurred most often in this temperate zone, and that metabolic disorders and peripartum disorders occurred from calving to day 10 post-calving.

The incidence of milk leakage (ML) after dry-off (DO) and related risk factors was studied in 1,175 dairy cows from 41 commercial herds in 8 European countries: Belgium, Czech Republic, Denmark, France, Germany, Italy, the Netherlands, and Spain. Milk leakage was assessed twice for 30 s each during 3 visits at 20 to 24 h, 30 to 34 h, and 48 to 52 h after DO. Information related to dry-cow management and udder health was collected at herd and cow level, including individual somatic cell count (ISCC) from test-day controls and occurrence of clinical mastitis cases from DO until 30 d in lactation. Mixed-effect logistic regression analyses were used to identify possible risk factors for ML and to study the association between ML and new intramammary infections. Intramammary infections were defined as clinical mastitis cases during the dry period and in the first 30 d in lactation or a rise in ISCC from before to after the dry period (threshold: 200,000 cells/mL) or both. Milk leakage was observed in 24.5% of the cows between 20 and 52 h after DO, where the herd incidence varied between 0.0 and 77.8%. The reduction in number of milkings in the weeks before DO had statistically significant effect on the ML incidence. When the milking frequency was reduced from 3 times/d to 2 or maintained at twice a day, cows had 11 (95% CI = 3.43-35.46) or 9 (95% CI = 1.85-48.22) times higher odds of leaking milk, respectively, compared with cows where the milking frequency was reduced from twice to once a day. Also, the milk production 24 h before DO was associated with ML incidence. Hence, cows with a milk production between 13 and 21 L or above 21 L had 2.3 (95% CI = 1.48-3.53) and 3.1 (95% CI = 1.79-5.3) times higher odds of leaking milk, respectively, compared with cows with a milk production below 13 L. A higher ML incidence was present in the group of cows with an average ISCC in the last 3 mo before DO ≥200,000 cells/mL (odds ratio = 1.7; 95% CI = 1.13-2.41) compared with cows with an average ISCC100,000 cells/mL. Quarters with ML tended to have 2.0 times higher odds of developing clinical mastitis compared with quarters not leaking milk. Cows with ML tended to have 1.5 times higher odds of intramammary infections (i.e., an increase of ISCC or clinical mastitis) compared with cows without ML.

Bovine clinical mastitis (CM) is one of the most prevalent diseases caused by a wide range of resident microbes. The emergence of antimicrobial resistance in CM bacteria is well-known, however, the genomic resistance composition (the resistome) at the microbiome-level is not well characterized. In this study, we applied whole metagenome sequencing (WMS) to characterize the resistome of the CM microbiome, focusing on antibiotics and metals resistance, biofilm formation (BF), and quorum sensing (QS) along with in vitro resistance assays of six selected pathogens isolated from the same CM samples. The WMS generated an average of 21.13 million reads (post-processing) from 25 CM samples that mapped to 519 bacterial strains, of which 30.06% were previously unreported. We found a significant (P = 0.001) association between the resistomes and microbiome composition with no association with cattle breed, despite significant differences in microbiome diversity among breeds. The in vitro investigation determined that 76.2% of six selected pathogens considered "biofilm formers" actually formed biofilms and were also highly resistant to tetracycline, doxycycline, nalidixic acid, ampicillin, and chloramphenicol and remained sensitive to metals (Cr, Co, Ni, Cu, Zn) at varying concentrations. We also found bacterial flagellar movement and chemotaxis, regulation and cell signaling, and oxidative stress to be significantly associated with the pathophysiology of CM. Thus, identifying CM microbiomes, and analyzing their resistomes and genomic potentials will help improve the optimization of therapeutic schemes involving antibiotics and/or metals usage in the prevention and control of bovine CM.

The effect of an internal teat sealant (ITS) on subsequent infection of the mammary gland was evaluated on the following mammary gland health indicators: (a) bacteriological cure of preexisting intramammary infections at drying off, (b) risk of postpartum new intramammary infections (NIMI), (c) cure and risk of new cases of subclinical mastitis (SCM), and (d) risk of postpartum clinical mastitis (CM). A total of 553 cows during late gestation were randomly assigned into two treatment protocols at drying off: (a) Dry cow therapy with 0.25 g of intramammary anhydrous cefalonium (ADCT, Cepravin&reg, MSD Animal Health), or (b) ADCT combined with ITS (SDCT, 4 g bismuth subnitrate, Masti-Seal&reg, MSD Animal Health, S&atilde, o Paulo, Brazil). Mammary quarter (MQ) milk samples were collected for microbiological culture and somatic cell count (SCC) at drying off and early lactation, and data from 1756 MQ were used in the multivariate logistic regression. There was no effect on the risk of bacteriological cure, SCM cure, and new cases of postpartum SCM. Still, SDCT reduced the risk of CM up to 60 days postpartum (DPP), overall NIMI risk, and the NIMI caused by major pathogens compared to ADCT. Thus, the DCT combined with ITS at drying off is effective for preventing NIMI during the dry period and CM up to 60 DPP.

Understanding farmers’ mindsets is important to improve antimicrobial stewardship in the dairy industry. This cross-sectional study aimed to determine farmers’ knowledge, attitude, and practices with respect to lactational intramammary antimicrobial use (AMU) and antimicrobial resistance (AMR) in Swiss dairy herds. Based on their approach towards subclinical mastitis (SCM) and non-severe cases of clinical mastitis (CM), subgroups of farmers were identified and compared regarding their knowledge, attitude and practices towards AMU and AMR. After conducting qualitative interviews to develop a questionnaire, an online survey was sent to 1296 randomly selected Swiss dairy farmers. Information was gathered on demographic data and farmers’ knowledge, attitude, and practices towards AMU and AMR. A latent class analysis was performed to identify subgroups of farmers based on management of SCM and non-severe CM cases. Based on the results of 542 completed questionnaires, poor knowledge with respect to AMU and AMR was identified, as well as discrepancies between farmers’ perceptions and their actual practices. Farmers approached cows with SCM and non-severe CM similarly, indicating they perceived both mastitis states as the same disease. Intramammary antimicrobial products containing cefquinome, which is a highest priority critically important antimicrobial, were among the 3 most commonly applied intramammary antimicrobials. Five latent classes of farmers were identified based on their management towards SCM and non-severe CM. One group of farmers (18.5% of respondents) indicated that they did not treat those mastitis cases, one group only treated SCM cases (13.8% of respondents), one group only treated non-severe CM cases (28.6% of respondents) and the largest group treated both mastitis states (39.1% of respondents). The latter group was subdivided into a latent class of farmers following guidelines for AMU/AMR (25.5% of respondents) and a group of farmers that were not strictly following these guidelines (13.7% of respondents). Regional differences between farmers, according to altitude and language region, explained some of the variation in latent class membership. Latent class membership was associated with farmers’ attitude to use antimicrobials as little as possible and with using antimicrobials only after performing bacteriological and susceptibility testing. This study gave detailed insight into Swiss farmers’ knowledge, attitude, and practices regarding AMU and AMR and provides opportunities to improve antimicrobial stewardship in Swiss dairy herds. The identified groups of farmers, based on their management practices regarding SCM and non-severe CM, may help to design tailored intervention strategies for improving prudent AMU in the heterogeneous population of dairy farmers in Switzerland., Preventive Veterinary Medicine, 179, ISSN:0167-5877

Udder health problems are often associated with milk losses. These losses are different between quarters, as infected quarters are affected both by systemic and pathogen-specific local effects, whereas noninfected quarters are only subject to systemic effects. To gain insight in these losses and the milk yield dynamics during disease, it is essential to have a reliable reference for quarter-level milk yield in an unperturbed state, mimicking its potential yield. We developed a novel methodology to predict this quarter milk yield per milking session, using an historical data set of 504 lactations collected on a test farm by an automated milking system from DeLaval (Tumba, Sweden). Using a linear mixed model framework in which covariates associated with the linearized Wood model and the milking interval are included, we were able to describe quarter-level yield per milking session with a proportional error below 10%. Applying this model enables us to predict the milk yield of individual quarters 1 to 50 d ahead with a mean prediction error ranging between 8 and 20%, depending on the amount of historical data available to estimate the random effect covariates for the predicted lactation. The developed methodology was illustrated using 2 examples for which quarter-level milk losses are calculated during clinical mastitis. These showed that the quarter-level mixed model allows us to gain insight in quarter lactation dynamics and enables to calculate milk losses in different situations. ispartof: JOURNAL OF DAIRY SCIENCE vol:101 issue:11 pages:10327-10336 ispartof: location:United States status: published

International audience; Mastitis is a disease of major economic importance to the dairy cattle sector because of the high incidence of clinical mastitis and prevalence of subclinical mastitis and, consequently, the costs associated with treatment, production losses, and reduced animal welfare. Disease-recording systems compiling data from a large number of farms are still not widely implemented around the world; thus, selection for mastitis resistance is often based on genetically correlated indicator traits such as somatic cell count (SCC), udder depth, and fore udder attachment. However, in the past years, several countries have initiated collection systems of clinical mastitis, based on producers recording data in most cases. The large data sets generated have enabled researchers to assess incidence of this disease and to investigate the genetic background of clinical mastitis itself, as well as its relationships with other traits of interest to the dairy industry. The genetic correlations between clinical mastitis and its previous proxies were estimated more accurately and confirmed the strong relationship of clinical mastitis with SCC and udder depth. New traits deriving from SCC were also studied, with the most relevant findings being associated with mean somatic cell score (SCS) in early lactation, standard deviation of SCS, and excessive test-day SCC pattern. Genetic correlations between clinical mastitis and other economically important traits indicated that selection for mastitis resistance would also improve resistance against other diseases and enhance both fertility and longevity. However, milk yield remains negatively correlated with clinical mastitis, emphasizing the importance of including health traits in the breeding objectives to achieve genetic progress for all important traits. These studies enabled the establishment of new genetic and genomic evaluation models, which are more efficient for selection to mastitis resistance. Further studies that are potential keys for future improvement of mastitis resistance are deep investigation of the bacteriology of mastitis, identification of novel indicator traits and tools for selection, and development of a larger female reference population to improve reliability of genomic evaluations. These cutting-edge studies will result in a better understanding of the genetic background of mastitis resistance and enable a more accurate phenotyping and genetic selection to improve mastitis resistance, and consequently, animal welfare and industry profitability.

Aim: This study was conducted to evaluate the incidence of clinical mastitis in bovines of Jammu region, to identify the infectious organisms responsible for it, and the antimicrobial sensitivity of isolated pathogens. Materials and Methods: The study was conducted on cases that were presented to the Medicine Division of Teaching Veterinary Clinical Complex, Faculty of Veterinary Sciences and Animal Husbandry, R.S. Pura, Jammu, Jammu and Kashmir. A total of 260 cases of bovines were presented from June 30, 2012, to July 01, 2013, out of which 30 cases were of clinical mastitis. The diagnosis of clinical mastitis was made on the basis of history and clinical examination of affected animals. Results: Animal and quarter-wise incidence of clinical mastitis were found to be 11.5% and 5.76%, respectively. Of the 23 isolates obtained, Staphylococcus aureus (60.87%) was the most frequently isolated organism, followed by coagulase negative Staphylococci (13.04%), Streptococcus uberis (4.35%), Streptococcus dysgalactiae (8.69%), and Escherichia coli (13.04%). The antimicrobial sensitivity of isolates revealed maximum sensitivity to enrofloxacin, gentamicin, amoxicillin/ sulbactam, ceftriaxone/tazobactam, ceftizoxime, ampicillin/sulbactam and least sensitivity for oxytetracycline and penicillin. Conclusion: Staphylococcus spp. is the major causative agent of clinical mastitis in bovines of Jammu region. The causative agents of the clinical mastitis were most sensitive to enrofloxacin and gentamicin.

Aim: Evaluation of the effect of clinical mastitis (CM) and its treatment outcome on the reproductive performance in crossbred cows retrospectively. Materials and Methods: Datasets of 835 lactating cows affected with CM during a period of 12 years (2001-2012) were considered for this study. Mastitis treatment related data and reproductive parameters such as days to first detected heat (DTFDH), days to first insemination (DTFI), days open (DO), and number of services per conception (SC) were collected from mastitis treatment and artificial insemination registers, respectively. Data were analyzed by ANOVA using SPSS 20 software. The means were compared with the Duncan's multiple comparison post-hoc test. Results: CM affected cows had significantly (p

The aim of this study was to determine risk factors for bovine intramammary infection (IMI) associated with the most common bacterial species in Finland. Large databases of the Finnish milk-recording system and results of microbiological analyses of mastitic milk samples from Valio Ltd. (Helsinki, Finland) were analyzed. The study group comprised 29,969 cows with IMI from 4,173 dairy herds. A cow with a quarter milk sample in which DNA of target species was detected in the PathoProof Mastitis PCR Assay (Thermo Fisher Scientific, Waltham, MA) was determined to have IMI. Only cows with IMI caused by the 6 most common pathogens or groups of pathogens, coagulase-negative staphylococci (CNS), Staphylococcus aureus, Streptococcus uberis, Streptococcus dysgalactiae, Corynebacterium bovis, and Escherichia coli, were included. The control group comprised 160,176 IMI-free cows from the same herds as the study group. A multilevel logistic regression model was used to study herd- and cow-specific risk factors for incidence of IMI. Pathogen-specific results confirmed those of earlier studies, specifically that increasing parity increases prevalence of IMI regardless of causative pathogen. Holsteins were more susceptible to IMI than Nordic Reds except when the causative pathogen was CNS. Occurrence of IMI caused by C. bovis was not related to milk yield, in contrast to IMI caused by all other pathogens investigated. Organic milk production was associated with IMI only when the causative pathogen of IMI was Staph. aureus; Staph. aureus IMI was more likely to occur in conventional than in organic production. Cows in older freestall barns with parlor milking had an increased probability of contracting an IMI compared with cows in tiestall barns or in new freestall barns with automatic milking. This was the case for all IMI, except those caused by CNS, the prevalence of which was not associated with the milking system, and IMI caused by Staph. aureus, which was most common in cows housed in tiestall barns. A better breeding index for milk somatic cell count was associated with decreased occurrence of IMI, indicating that breeding for improved udder health has been successful in reducing the incidence of IMI caused by the most common pathogens in Finland. In the Finnish dairy sector, the importance of other measures to control IMI will increase as the Holstein breed progressively takes the place of the Nordic Red breed. Attention should be paid to hygiene and cleanliness, especially in old freestall barns. Based on our results, the increasing prevalence of automatic milking is not a reason for special concern.

Background and Aim: Staphylococcus aureus is argued as one of the principal organisms responsible for mammary gland infection in lactating goats, causing both clinical and subclinical mastitis. Being highly zoonotic potential, pathogen emergence of methicillin-resistant S. aureus (MRSA) has a significant clinical impact on treatment and management of clinical mastitis. We conducted a cross-sectional study to investigate the prevalence of coagulase-positive S. aureus (CoPS), antimicrobial resistance profile of Staphylococcus spp., prevalence of MRSA, and association between different clinical parameters with CoPS. Materials and Methods: A total of 67 clinical mastitic goats were sampled based on clinical examination and California mastitis test. Standard bacteriological methods were performed to isolate and identify Staphylococcus spp. CoPS were confirmed by nuc gene using polymerase chain reaction (PCR). All staphylococcal isolates were further examined for antimicrobial susceptibility testing by disk diffusion method. MRSA was confirmed based on mecA gene-based PCR. Results: Here, 49 (73.13%; 95% confidence interval [CI], 61.41-82.35) samples were positive for Staphylococcus spp., of which 17 (34.69%; 95% CI, 22.88-48.73) isolates were CoPS and rest of the isolates (32; 65.30%; 95% CI, 51.27-77.12) were identified as coagulase-negative Staphylococcus spp. (coagulase-negative staphylococci [CNS]). Both, CoPS and CNS isolates displayed the highest resistance against tetracycline (76.47% and 75%, respectively) and oxacillin (70.58% and 68.75%, respectively). Notably, all staphylococcal isolates were multidrug-resistant (showed resistance to ≥3 classes of antimicrobials). mecA gene was found in 6 (8.96%; 95% CI, 3.84-18.52) CoPS isolates indicating MRSA strains. Among different clinical parameters, presence of high body temperature (p

The main objective of this study was to compare the performance of different ‘nonlinear quantile regression’ models evaluated at the τth quantile (0·25, 0·50, and 0·75) of milk production traits and somatic cell score (SCS) in Iranian Holstein dairy cows. Data were collected by the Animal Breeding Center of Iran from 1991 to 2011, comprising 101 051 monthly milk production traits and SCS records of 13 977 cows in 183 herds. Incomplete gamma (Wood), exponential (Wilmink), Dijkstra and polynomial (Ali & Schaeffer) functions were implemented in the quantile regression. Residual mean square, Akaike information criterion and log-likelihood from different models and quantiles indicated that in the same quantile, the best models were Wilmink for milk yield, Dijkstra for fat percentage and Ali & Schaeffer for protein percentage. Over all models the best model fit occurred at quantile 0·50 for milk yield, fat and protein percentage, whereas, for SCS the 0·25th quantile was best. The best model to describe SCS was Dijkstra at quantiles 0·25 and 0·50, and Ali & Schaeffer at quantile 0·75. Wood function had the worst performance amongst all traits. Quantile regression is specifically appropriate for SCS which has a mixed multimodal distribution. The main objective of this study was to compare the performance of different 'nonlinear quantile regression' models evaluated at the τth quantile (0·25, 0·50, and 0·75) of milk production traits and somatic cell score (SCS) in Iranian Holstein dairy cows. Data were collected by the Animal Breeding Center of Iran from 1991 to 2011, comprising 101 051 monthly milk production traits and SCS records of 13 977 cows in 183 herds. Incomplete gamma (Wood), exponential (Wilmink), Dijkstra and polynomial (Ali & Schaeffer) functions were implemented in the quantile regression. Residual mean square, Akaike information criterion and log-likelihood from different models and quantiles indicated that in the same quantile, the best models were Wilmink for milk yield, Dijkstra for fat percentage and Ali & Schaeffer for protein percentage. Over all models the best model fit occurred at quantile 0·50 for milk yield, fat and protein percentage, whereas, for SCS the 0·25th quantile was best. The best model to describe SCS was Dijkstra at quantiles 0·25 and 0·50, and Ali & Schaeffer at quantile 0·75. Wood function had the worst performance amongst all traits. Quantile regression is specifically appropriate for SCS which has a mixed multimodal distribution.

Aim of study: We have classified into homogenous groups a wide spectrum of mammary pathogens according to their frequency of isolation in clinical mastitis and their somatic cell counts in non-clinical mastitis. Area of study: The study was conducted in Galicia (NW Spain) Material and methods: 163,741 dairy cattle quarter milk samples were analyzed. We identified mastitis pathogens to the species level and performed a cluster analysis to classify these microorganisms by their median of Linear Score (mLS), percentage of isolation in clinical mastitis samples (%ICS) and percentage of isolation in samples with somatic cell counts under 100,000 cells/mL (%ISU100). Main results: Forty-three different species were isolated. Cluster analysis identified 4 groups of pathogens; mLS and %ICS progressively increased from Group I to Group IV and %ISU100 decreased. However, several pathogens included in groups II and III showed %ISU100 values higher than 2% and 3%. Minor pathogens were mainly clustered in Group I (e.g., Corynebacterium spp. and most of Staphylococcus spp.), while known major pathogens were included in Groups II, III y IV. Species of the same family, genus or microbiological group like Enterobacteriaceae or Enterococcus spp. were frequently separated into different groups, thus showing heterogeneity among the members of these groups. Research highlights: Results obtained here may aid in assessing the pathogenicity of sporadic pathogens in relation to more well-known pathogens and suggest that the traditional classification between minor and major pathogens is an oversimplification of the reality, especially for the latter category. Xunta de Galicia, Program for consolidating and structuring competitive research groups: GRC2015/003 SI

Mastitis remains the most expensive disease of dairy cows, and antibiotic dry cow therapy (DCT) at dry-off is an important part of mastitis control. Regardless of the infection status, blanket DCT is administered to all quarters of all cows, which is controversial due to the worldwide problem of antimicrobial resistance. Even though selective DCT of only infected cows is a more sustainable approach, choosing animals for treatment is not always straightforward. Our aim was to evaluate whether the herd-level DCT approach is associated with early lactation udder health problems, taking into account the cow characteristics. The information source was 2015?2017 Dairy Herd Improvement data with 7461 multiparous cows from 241 Finnish dairy herds. Information on the herd-level DCT approach was obtained from farmers? questionnaire responses in 2017, and the three different approaches were selective DCT, blanket DCT, and no DCT. The statistical tool for the data analysis was a generalized linear mixed model with a random herd effect for binary outcomes and a linear mixed model with a random herd effect for a continuous outcome. The two binary outcomes were the odds of having high milk somatic cell count (SCC ? 200 000 cells/mL) on the first test-day within 5?45 days in milk (DIM) and the odds of mastitis treatment in early lactation up to 45 DIM. The third outcome was the mean milk lnSCC (? 1000 cells/ mL) within 120 DIM. Selective DCT was the prevailing treatment practice in our data. Blanket DCT was associated with lower SCC after calving. Cows more likely to have high SCC after calving were older cows, cows with high average SCC during the previous lactation, and cows with high milk yield near dry-off. A mastitis treatment in the early lactation was more likely if, during the previous lactation, the cow had high average SCC, high peak milk production, or high milk yield near dry-off. Our findings indicate that DCT is still effective in mastitis control. Cows with high milk yield, especially near dry-off, and cows with persistently high SCC require attention when considering next lactation udder health. Mastitis remains the most expensive disease of dairy cows, and antibiotic dry cow therapy (DCT) at dry-off is an important part of mastitis control. Regardless of the infection status, blanket DCT is administered to all quarters of all cows, which is controversial due to the worldwide problem of antimicrobial resistance. Even though selective DCT of only infected cows is a more sustainable approach, choosing animals for treatment is not always straightforward. Our aim was to evaluate whether the herd-level DCT approach is associated with early lactation udder health problems, taking into account the cow characteristics. The information source was 2015?2017 Dairy Herd Improvement data with 7461 multiparous cows from 241 Finnish dairy herds. Information on the herd-level DCT approach was obtained from farmers? questionnaire responses in 2017, and the three different approaches were selective DCT, blanket DCT, and no DCT. The statistical tool for the data analysis was a generalized linear mixed model with a random herd effect for binary outcomes and a linear mixed model with a random herd effect for a continuous outcome. The two binary outcomes were the odds of having high milk somatic cell count (SCC ? 200 000 cells/mL) on the first test-day within 5?45 days in milk (DIM) and the odds of mastitis treatment in early lactation up to 45 DIM. The third outcome was the mean milk lnSCC (? 1000 cells/ mL) within 120 DIM. Selective DCT was the prevailing treatment practice in our data. Blanket DCT was associated with lower SCC after calving. Cows more likely to have high SCC after calving were older cows, cows with high average SCC during the previous lactation, and cows with high milk yield near dry-off. A mastitis treatment in the early lactation was more likely if, during the previous lactation, the cow had high average SCC, high peak milk production, or high milk yield near dry-off. Our findings indicate that DCT is still effective in mastitis control. Cows with high milk yield, especially near dry-off, and cows with persistently high SCC require attention when considering next lactation udder health.

The objective of this study was to determine cow characteristics and farm management factors during the dry period associated with early postparturient intramammary infection (IMI) and subsequent clinical mastitis (CM). Data were collected three times: before drying off (P1), during the dry period (P2), and 5 to 14 days after calving (P3), using questionnaires and farm investigation. Milk samples were aseptically collected for bacterial identification at P1 and P3. Factors associated with IMI and CM were analyzed using multiple logistic regression models. The final model showed that IMI in early postpartum was significantly associated with full insertion of dry cow antibiotic, dry cows in barns with a combination of tie and free stalls, body condition score (BCS) in dry period and after calving, and milk yield before drying off. For IMI cows, factors significantly associated with clinical expression of mastitis were having daily barn cleaning, teat disinfected with alcohol before administration of dry cow therapy, BCS before drying off, milk yield before drying off, and days in milk at drying off. In conclusion, both cow and farm management factors are associated with the IMI rate and subsequent expression of clinical signs of mastitis in early postpartum cows.

Bovine mastitis, an infection of the cow's mammary gland, is frequently caused by Streptococcus uberis and causes major economic losses in the dairy industry. The intramammary administration of antibiotics currently remains the predominant preventive and therapeutic measure. These antimicrobial compounds, of which some are considered critical in human health care, are frequently applied as dry therapy resulting in their consistent overuse. Therefore, the use of antibiotics in the dairy sector is being questioned. We here identified two endolysins, i.e., PlySs2 and PlySs9, respectively derived from Streptococcus suis serotype-2 and -9 prophages, with lytic activity against S. uberis in an in vitro setting. Both endolysins gave clear lysis zones in spot-on-plate assays and caused a reduction of the optical density in a turbidity reduction assay. In depth characterization identified PlySs9 as the more potent endolysin over PlySs2 with a lower MIC value and about one additional log of killing. PlySs2 and PlySs9 were challenged to a panel of subclinical and clinical S. uberis milk isolates and were both able to lyse all strains tested. Molecular dissection of these endolysins in catalytic and cell wall binding subdomains resulted in major loss of killing and binding activity, respectively. Taken together, we here propose PlySs2 and PlySs9 as candidate compounds to the current antimicrobial arsenal known against bovine mastitis-causing S. uberis as future add-on or replacement strategy to the currently used intramammary antibiotics. ispartof: ANTIBIOTICS-BASEL vol:9 issue:9 ispartof: location:Switzerland status: published

The study was conducted to know the factor that potentially affect the incidence of subclinical and clinical mastitis in dairy cows in the area Agribusiness Cooperative Dana Mulya Pacet, Mojokerto. Data retrieval were done by examining the incidence of subclinical and clinical mastitis in 100 lactation dairy cows. Milk samples were taken from milking each quarter of dairy cows in the afternoon, then tested using CMT reagent (California Mastitis Test). In addition data was obtained from farmers through structured questionnaires. The data was processed and presented in tabular form and analyzed statistically by the method of regression trees. The conclusion of this research showed that dairy cows in the area of Agribusiness Cooperative Dana Mulya Pacet, Mojokerto affected by subclinical mastitis reached 76% and clinical mastitis 11%. This was influenced by the milking sequence factor of the nipple which was not distinguish between sick and healthy nipples, the lack of sanitation before milking, and the location of livestock disposal is too close to the cowshed.

Aims: The objective of this study was to compare qualitatively and quantitatively the results of identification of the bacteria present in milk samples from cows with subclinical mastitis using multiplex qPCR assay and matrix-assisted laser desorption-ionization time of flight mass spectrometry (MALDI-TOF MS (R)) after bacteriological growth. Methods and Results: A total of 182 samples were aseptically collected from 119 cows with high somatic cell counts (>2.10(5) SCC per ml) on 11 farms in Belgium in 2014. The mutiplex qPCR assay was carried out on 350 mu l of milk with the PathoProof (R) Complete-16kit. Ten microlitre of milk was streaked on Columbia blood agar and three selective agar plates. Growing colonies were identified by MALDI-TOF MS. Of the 182 samples, 90 gave positive results with either or both tests for one or two bacterial species/genera. Total qualitative agreement of the bacteria identified was observed in 41 mono- or bi-bacterial samples (46%) and partial agreement in 19 bi-bacterial samples at both or either tests (21%). The results of both tests on those mono- and bi-bacterial samples were not significantly different (McNemar test; P = 0.395) with a fair agreement (Cohen's kappa test; k = 0.375; P = 0.055). Moreover, quantitative correlation between the qPCR intensity and the numbers of growing colonies was observed in half of the 60 samples with qualitative matching results. Conclusions: Both methods give identical qualitative and quantitative results with approximately a half and a quarter of the mono- and bi-bacterial samples respectively. Several reasons can explain the differences. The multiplex qPCR assay only targets the most important mammary gland pathogens and can detect DNA of bacteria both alive and dead. Conversely, bacteria only grow when alive and the MALDI-TOF MS databases do not include all bovine milk-associated bacterial species yet. Significance and Impact of the Study This study further highlights the limitations and complementarity of the genetic and phenotypic tests for the identification of bacteria present in milk samples.

Gastrointestinal nematode (GIN) infections are a common constraint in pasture-based herds and cause a decrease in animal health, productivity and farm profitability. Current control practices to prevent production losses of GIN infections in livestock depend largely on the use of anthelmintic drugs. However, due to the continued use of these drugs over more than three decades, the industry is now increasingly confronted with nematode populations resistant to the available anthelmintics. This emerging anthelmintic resistance (AR) in cattle nematodes emphasizes the need for a change toward more sustainable control approaches that limit, prevent or reverse the development of AR. The uptake of diagnostic methods for sustainable control could enable more informed treatment decisions and reduce excessive anthelmintic use. Different diagnostic and targeted or targeted selective anthelmintic control approaches that slow down the selection pressure for anthelmintic resistance have been developed and evaluated recently. Now it is time to transform these insights into guidelines for sustainable control and communicate them across the farmer community. This article reviews the current uptake of such sustainable practices with a focus on farmer's socio-psychological factors affecting this uptake. We investigate communication as a possible tool to change current behavior and successfully implement more sustainable anthelmintic treatment strategies.

The aim of this study was to determine the intramammary dose of benzylpenicillin required to maintain a concentration in the milk above the MIC for the Gram-positive bacteria that cause mastitis. The product used in this study was a commercially available procaine benzylpenicillin in an oily suspension with micronized particles. Three dose levels were used: 200,000, 300,000, and 600,000IU. Concentrations of benzylpenicillin in cow milk and plasma were determined after a single intramammary dose was administered into one quarter of each of the five cows in each treatment group. Samples were analyzed using an HPLC-MS/MS method, which was validated during the study. Concentrations in the milk were well above the MIC for the target pathogens for all doses tested. There was a linear dose-dependent increase in the mean AUCs of benzylpenicillin concentrations in plasma and milk. At the first milking, 12hr after dosing, there was a significant difference between the mean milk benzylpenicillin concentrations in cows treated with a dose of 600,000IU, and those treated with 200,000 or 300,000IU. Although this study shows a linear relationship between the dose of procaine benzylpenicillin administered and the concentration in the milk in the healthy udder, it would be useful to conduct studies on cows with mastitis to define the optimum dose and duration of intramammary treatment with benzylpenicillin.

Matrix-assisted laser desorption/ionization timeof-flight mass spectrometry (MALDI-TOF MS) is a fast and reliable method to identify the most common pathogenic bacteria in humans and animals. The goals of this study were to amend a commercial database with additional species, evaluate the amended database for identification of bacterial genera and species causing bovine mastitis, and describe the plethora of species involved. In total, 500 udder pathogenic isolates were subjected to MALDI-TOF MS using bacterial or fungal colony material; 93.5% could be identified to the species level, and 6.5% were identified only to the genus level. Isolates identified to the genus level required further identification to the species level by conventional methods or 16S rDNA sequencing. Mass spectra from verified species were used to expand the MALDI-TOF MS database to improve future identification ability. A total of 24 genera and 61 species were identified in this study. Identified isolates were mainly staphylococci, streptococci, Enterobacteriaceae, and coryneforme bacteria. In conclusion, MALDI-TOF MS is a powerful, rapid, and reliable technique to identify the most common microorganisms causing bovine mastitis, and the database can be continuously expanded and improved with additional species.

The objective of this study was to survey drying-off practices and use of dry cow therapy (DCT) in Finland through an online questionnaire. The questionnaire was accessible to all dairy farmers of the Finnish dairy herd recording system in 2016 (approximately 5,400 farms). In total, 715 dairy producers across the country, representative of the Finnish dairy industry, participated in the survey. Cows were dried off gradually in most of the farms. Most farms (78%) reported using selective DCT, whereas 9% of farms did not use any DCT, and 13% of farms applied blanket DCT. A significant trend was observed with increasing herd size and proportion of farms using blanket DCT. Percentage of farms using blanket DCT was also higher in farms with automatic milking system. Farmer's own experience was the most commonly reported reason for choosing a particular approach to DCT. Microbiological testing of milk samples at dry-off was the preferred method of selecting cows for DCT; 82 and 64% of farms using selective and blanket DCT approach, respectively, reported testing milk samples before treatment. The second most common criteria for using antibiotic DCT were clinical mastitis history and high somatic cell count. A high number of farms using selective DCT reported treating only up to one-fourth of their cows at dry-off. Information acquired on drying-off practices in Finland allows for future monitoring of prudent antimicrobial usage at dry-off.

Staphylococcus aureus is one of the most common pathogens that cause mastitis in dairy cows. Various subtypes, virulence genes and mobile genetic elements have been associated with isolates from bulk tank milk and clinical mastitis. So far, no Danish cattle associated S. aureus isolates have been whole-genome sequenced and further analyzed. Thus, the main objective was to investigate the population structure and genomic content of isolates from bulk tank milk and clinical mastitis, using whole-genome sequencing. This may reveal the origin of strains that cause clinical mastitis. S. aureus isolates from bulk tank milk (n = 94) and clinical mastitis (n = 63) were collected from 91 and 24 different farms, respectively and whole-genome sequenced. The genomic content was analyzed and a phylogenetic tree based on single nucleotide polymorphisms was constructed. In general, the isolates from both bulk tank milk and clinical mastitis were of similar genetic background. This suggests that dairy cows are natural carriers of the S. aureus subtypes that cause clinical mastitis if the right conditions are present and that a broad range of subtypes cause mastitis. A phylogenetic cluster that mostly consisted of ST151 isolates carried three mobile genetic elements that were primarily found in this group. The prevalence of resistance genes was generally low. However, the first ST398 methicillin resistant S. aureus isolate from a Danish dairy cow with clinical mastitis was detected.

Escherichia coliis a major udder pathogen causing clinical mastitis in dairy cattle and its heat stable endotoxin in powdered infant formula milk is a potential risk factor in neonatal infections. Cephalosporins are frequently used for treatment of mastitis caused by mastitis; however, use of these antimicrobials may induce antimicrobial resistance inE. coli. The objective of this study was to explore thein vitroeffect of subminimum inhibitory concentrations (sub-MIC) of cefalotin (CF) and ceftazidime (CAZ) on the morphology, antimicrobial resistance, and endotoxin releasing characteristics of 3E. coliisolates recovered from bovine clinical mastitis. The parentE. coliisolates, which were susceptible to CF and CAZ, were exposed to CF or CAZ separately at sub-MIC levels to produce 9 generations of induced isolates. Colonies of the CAZ-induced isolates from all 3 parentE. coliwere smaller on blood agar and the bacteria became filamentous, whereas the CF-induced isolates did not demonstrate prominent morphological changes. After induction by CF or CAZ, many induced isolates showed resistance to cefoxitin, CAZ, CF, kanamycin, ampicillin, and amoxicillin/clavulanic acid while their parent isolates were susceptible to these antimicrobials. Notably, 5 CAZ-induced isolates from the same parent isolate were found to produce extended-spectrum beta-lactamase (ESBL) though none of the tested ESBL related genes could be detected. All CAZ-induced isolates released more endotoxin with a higher release rate, whereas endotoxin release of CF-inducedE. coliisolates was not different from parent isolates. The exposure of cephalosporins at sub-MIC levels induced resistantEscherichia coli.We inferred that cephalosporins, especially CAZ, should be used prudently for treatment of clinicalE. colimastitis.

A total of 90 cows from three commercial farms were used to evaluate the relationship between subclinical mastitis and clinical mastitis and thermal nociceptive threshold. Milk strips from all udder quarters were tested for clinical mastitis with visual inspection of milk and udder alterations and for subclinical mastitis using California Mastitis Test. Milk yield was recorded, milk was sampled and further analyzed for somatic cells count (SCC). Cows were considered healthy when SCC200 000 cells/ml and no visual alterations in milk and/or udder, with moderate subclinical mastitis when SCC>500 000 cells/ml and no visual alterations in milk and/or udder and with clinical mastitis when visual alterations in milk and/or udder were detected. Nociceptive threshold was evaluated with the thermal threshold meter apparatus applied to the rear legs. Thermal threshold (TT) decreased when we compared healthy cows with cows presenting clinical mastitis and tended to decrease when we compare healthy cows with those with moderate subclinical mastitis. TT was lower at the ipsilateral rear leg compared with the contralateral leg to the infected mammary gland. TT linearly decreases as log10SCC increased and it showed sharp decrease as log10SCC exceed the value of 6.4. Increase in one unit of log10SCC increased the odds of low thermal threshold (lower than 55.8°C). Subclinical mastitis might be a welfare issue as it tended to decrease nociceptive thermal threshold.

Endemic diseases can be counted among the most serious sources of losses for livestock production. In dairy farms in particular, one of the most common diseases is Johne's disease, caused by Mycobacterium avium ssp. paratuberculosis (MAP). Infection with MAP causes direct costs because it affects milk production, but it has also been suspected to increase the risk of clinical mastitis (CM) among infected animals. This might contribute to further costs for farmers. We asked whether MAP infection represents a risk factor for CM and, in particular, whether CM occurrences were more common in MAP-infected animals. Our results, obtained by survival analysis, suggest that MAP-infected cows had an increased probability of experiencing CM during lactation. These results highlight the need to account for the interplay of infectious diseases and other health conditions in economic and epidemiological modeling. In this case, accounting for MAP-infected cows having an increased CM occurrence might have nonnegligible effects on the estimated benefit of MAP control.

Background: The Finnish dairy herd recording system maintains production and health records of cows and herds. Veterinarians and farmers register veterinary treatments in the system. Milk samples for microbiological analysis are routinely taken from mastitic cows. The laboratory of the largest dairy company in Finland, Valio Ltd., analyzes most samples using real-time PCR. This study addressed pathogen-specific microbiological data and treatment and culling records, in combination with cow and herd characteristics, from the Finnish dairy herd recording system during 2010-2012. Results: The data derived from 240,067 quarter milk samples from 93,529 dairy cows with mastitis; 238,235 cows from the same herds served as the control group. No target pathogen DNA was detected in 12% of the samples. In 49% of the positive samples, only one target species and in 19%, two species with one dominant species were present. The most common species in the samples with a single species only were coagulase-negative staphylococci (CNS) (43%), followed by Staphylococcus aureus (21%), Streptococcus uberis (9%), Streptococcus dysgalactiae (8%), Corynebacterium bovis (7%), and Escherichia coli (5%). On average, 36% of the study cows and 6% of the control cows had recorded mastitis treatments during lactation. The corresponding proportions were 16 and 6% at drying-off. For more than 75% of the treatments during lactation, diagnosis was acute clinical mastitis. In the milk samples from cows with a recorded mastitis treatment during lactation, CNS and S. aureus were most common, followed by streptococci. Altogether, 48% of the cows were culled during the study. Mastitis was reported as the most common reason to cull; 49% of study cows and 18% of control cows were culled because of mastitis. Culling was most likely if S. aureus was detected in the milk sample submitted during the culling year. Conclusions: The PCR test has proven to be an applicable method also for large-scale use in bacterial diagnostics. In the present study, microbiological diagnosis was unequivocal in the great majority of samples where a single species or two species with one dominating were detected. Coagulase-negative staphylococci and S. aureus were the most common species. S. aureus was also the most common pathogen among the culled cows, which emphasizes the importance of preventive measures.

International audience; Explorations of how the complex microbial communities that inhabit different body sites might contribute to health and disease have prompted research on the ways the harmonious relationship between a host and its microbiota could be used to keep animals healthy in their production conditions. In particular, there is a growing interest in the bacterial signatures that can be found in the milk of healthy or mastitic dairy cows. The concept of sterility of the healthy mammary gland of dairy ruminants has been challenged by the results of studies using bacterial DNA-based methodology. The newly obtained data have led to the concept of the intramammary microbiota composed of a complex community of diverse bacteria. Accordingly, mammary gland infections are not mere infections by a bacterial pathogen, but the consequence of mammary dysbiosis. This article develops the logical implications of this paradigm shift and shows how this concept is incompatible with current knowledge concerning the innate and adaptive immune system of the mammary gland of dairy ruminants. It also highlights how the concept of mammary microbiota clashes with results of experimental infections induced under controlled conditions or large field experiments that demonstrated the efficacy of the current mastitis control measures.