Accurate identification of mastitis-causing bacteria supports effective management and can be used to implement selective use of antimicrobials for treatment. The objectives of this study were to compare the results from a culture-based mastitis pathogen detection test kit ('VetoRapid', Vétoquinol) with standard laboratory culture and to evaluate the potential suitability of the test kit to inform a selective treatment programme. Overall 231 quarter milk samples from five UK dairy farms were collected. The sensitivity and specificity of the test kit for the identification of Escherichia coli, Staphylococcus aureus, coagulase-negative staphylococci, Streptococcus uberis and Enterococcus spp. ranged from 17 per cent to 84 per cent and 92 per cent to 98 per cent, respectively. In total, 23 of 68 clinical samples were assigned as meeting the requirement for antimicrobial treatment (Gram-positive organism cultured) according to standard culture results, with the test kit results having sensitivity and specificity of 91 per cent and 78 per cent, respectively. Several occurrences of misidentification are reported, including S. aureus being misidentified as coagulase-negative staphylococci and vice versa. The test kit provides rapid preliminary identification of five common causes of bovine mastitis under UK field conditions and is likely to be suitable for informing selective treatment of clinical mastitis caused by Gram-positive organisms.

Periodontitis is an infectious polymicrobial, immuno-inflammatory disease of multifactorial aetiology that has an impact on the health, production and welfare of ruminants. The objective of the present study was to determine the microbial profiles present in the gingival sulcus of cattle considered periodontally healthy and in the periodontal pocket of animals with periodontitis lesions using high-throughput bacterial 16S rRNA gene sequencing. Subgingival biofilm samples were collected from 40 cattle with periodontitis and 38 periodontally healthy animals. In total, 1923 OTUs were identified and classified into 395 genera or higher taxa. Microbial profiles in health differed significantly from periodontitis in their composition (p < 0.0001, F = 5.30; PERMANOVA) but no statistically significant differences were observed in the diversity of healthy and periodontitis microbiomes. The most prevalent taxa in health were Pseudomonas, Burkholderia and Actinobacteria, whereas in disease these were Prevotella, Fusobacterium and Porphyromonas. The most discriminative taxa in health were Gastranaerophilales, Planifilum and Burkholderia, and in disease these were Elusimicrobia, Synergistes and Propionivibrio. In conclusion, statistically significant difference exists between the microbiome in bovine oral health and periodontitis, with populations showing 72.6% dissimilarity. The diversity of the bacteria found in health and periodontitis were similar and bacteria recognised as periodontal pathogens showed increased abundance in disease. In this context, the main components of bacterial homeostasis in the biofilm of healthy sites and of dysbiosis in periodontal lesions provide unprecedented indicators for the evolution of knowledge about bovine periodontitis.

Periodontitis is a polymicrobial infectious disease that causes occlusion change, tooth loss, difficulty in rumination, and premature culling of animals. This study aimed to detect species of the genera Porphyromonas and Prevotella present in the periodontal pocket of sheep with lesions deeper than 5mm (n=14) and in the gingival sulcus of animals considered periodontally healthy (n=20). The presence of microorganisms was evaluated by polymerase chain reaction (PCR) using specific primers for Porphyromonas asaccharolytica, Porphyromonas endodontalis, Porphyromonas gingivalis, Porphyromonas gulae, Prevotella buccae, Prevotella intermedia, Prevotella loescheii, Prevotella melaninogenica, Prevotella nigrescens, Prevotella oralis, and Prevotella tannerae. Prevalence and risk analysis were performed using Student's t-test and Spearman's correlation. Among the Prevotella and Porphyromonas species detected in the periodontal lesions of sheep, P. melaninogenica (85.7%), P. buccae (64.3%), P. gingivalis (50%), and P. endodontalis (50%) were most prevalent. P. gingivalis (15%) and P. oralis (10%) prevailed in the gingival sulcus. P. gulae and P. tannerae were not detected in the 34 samples studied. Data evaluation by t-test verified that occurrence of P. asaccharolytica, P. endodontalis, P. gingivalis, P. buccae, P. intermedia, P. melalinogenica, and P. nigrescens correlated with sheep periodontitis. The findings of this study will be an important contribution to research on pathogenesis of sheep periodontitis and development of its control measures.

Mastitis, inflammation of the bovine mammary gland, is generally caused by intramammary infection with bacteria, and antimicrobials have long been a corner stone of mastitis control. As societal concern about antimicrobial use in animal agriculture grows, there is pressure to reduce antimicrobial use in dairy farming. Point-of-care tests for on-farm use are increasingly available as tools to support this. In this Research Reflection, we consider available culture-dependent and culture-independent tests in the context of ASSURED criteria for low-resource settings, including convenience criteria, scientific criteria and societal criteria that can be used to evaluate test performance. As tests become more sophisticated and sensitive, we may be generating more data than we need. Special attention is given to the relationship between test outcomes and treatment decisions, including issues of diagnostic refinement, antimicrobial susceptibility testing, and detection of viable organisms. In addition, we explore the role of technology, big data and people in improved performance and uptake of point-of-care tests, recognising that societal barriers may limit uptake of available or future tests. Finally, we propose that the 3Rs of reduction, refinement and replacement, which have been used in an animal welfare context for many years, could be applied to antimicrobial use for mastitis control on dairy farms.

The welfare impact of birth on newborn calves has rarely been studied. Dystocia in particular may have significant welfare costs for calves. While analgesia is sometimes provided to calves born to difficult parturition by veterinary surgeons in practice, it is not known if this is actually beneficial. On a commercial dairy farm, we examined the behavioural time budget of 39 Holstein heifer calves born with the aid of farmer assistance and 36 calves born without assistance; half of each group were randomly allocated to receive either a single dose of the non-steroidal anti-inflammatory drug ketoprofen or a saline placebo in a two by two factorial design. The calves were group housed and their behaviour was recorded for 48 hours postpartum and analysed using instantaneous focal sampling (every 5 minutes in alternate hours). Regardless of analgesic treatment, calves born with assistance showed behaviours consistent with experiencing a less positive welfare state (lying with their head down and in lateral recumbency, and less time playing than unassisted calves). Behavioural differences between calves treated with ketoprofen and calves treated with saline (in particular increased play) suggest that the birth experience may be painful for all calves, even if no assistance is required. Our findings suggest that a single dose of ketoprofen in the immediate postpartum period may improve calf welfare regardless of assistance status and has the potential to contribute to significant welfare gains in dairy calves.

Dystocia is considered painful and stressful for both the dam and the calf, although systematic evidence of this is limited. Few studies have investigated biochemical markers of stress and pain postpartum and whether any adverse effects are ameliorated by administration of analgesia. In this study, cow-calf pairs experiencing both mild to moderate farmer assistance and no assistance at parturition were randomly assigned to either treatment or placebo group in a two-by-two design (animals subject to veterinary intervention were excluded). The treatments were the NSAID ketoprofen or saline, administered within three hours of parturition. Blood samples taken in the immediate postpartum period, and at 24 hours, 48 hours and 7 days after parturition, were analysed for plasma concentrations of creatine kinase and cortisol (cows and calves) and plasma L-lactate and total protein concentration (calves). Stress biomarkers were highest in the immediate postpartum period and declined over time (P<0.05). Cow plasma cortisol was higher in animals experiencing assisted parturition in the immediate postpartum period (P=0.023); by 24 hours no difference was evident. Intervention with NSAID analgesia did not result in beneficial changes in stress biomarkers. Based on biomarkers alone, this suggests limited benefits of NSAID treatment in unassisted or mild to moderately assisted parturition.

To limit the use of antimicrobials in dairy cattle, farmers are increasingly encouraged to adopt targeted treatment decisions based on knowledge of the pathogens causing clinical mastitis (CM), whereby treatment of non-severe CM is generally recommended for gram-positive mastitis but not for gram-negative or culture-negative mastitis. The objectives of this study were to conduct a laboratory-based evaluation of the performance of a simplified slide test as a tool to differentiate gram-positive CM from other cases of CM, and to compare its performance against a commercially available on-farm test that is commonly used in our area (VétoRapid). Test outcomes after 24–48 h incubation were compared to results from bacteriological culture and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS). Milk samples (n = 156) were obtained from cases of severe and non-severe CM on seven farms and collected by farm personnel. After removal of small numbers of contaminated samples and organisms with unknown species identity, the simplified slide test showed high sensitivity and accuracy (>80%), similar to the comparator test. For most outcomes of interest (culture positive, Escherichia coli, or gram-positive growth), the specificity of the slide test was higher than the specificity of the comparator test. When considering non-severe cases of CM only, and interpreting detection of gram-positive organisms as indicative of the need for antimicrobial treatment, the simplified test had higher specificity (77.4% v. 60.4%) and higher positive predictive value (79.7% v. 70.0%) than the comparator test and similar sensitivity (83.9% v. 87.5%). The proportion of sampled CM cases, contaminated samples and gram-positive mastitis cases – which affects the positive and negative predictive value, the economic value of diagnostic testing and its potential to reduce antimicrobial use – differed between farms. The simplicity and accuracy of the slide test could make it an attractive tool for farmers to target antimicrobial treatment of non-severe clinical mastitis.

Neonatal calves are commonly affected by diarrhea caused by different pathogens, but not always bacteria. Yet, antibiotics are routinely used as a treatment to an unknown extent. It was our goal to survey antibiotic use for the treatment of neonatal calf diarrhea in different countries and to identify influencing factors. A total of 873 farmers and veterinarians in Austria, Belgium, Portugal, and Scotland participated in a voluntary online survey. The data were analyzed using classification and regression tree analyses and chi2 tests. Overall, 52.5% of the participants stated that they use antibiotics when treating neonatal calf diarrhea. Of those, 27% use them always, and 45% use highest priority critically important antibiotics. The most important factor differentiating antibiotic use practices was the country the participants were from, which could be due to regulatory differences between the countries. All antibiotic products stated were licensed for use in cattle, but several were not licensed for the treatment of diarrhea in calves. Our study shows that there is an urgent need for more scientific evidence to define best practices for the treatment of neonatal calf diarrhea. Furthermore, consensual criteria for antibiotic therapy must be defined, and targeted training for farmers and veterinarians must be provided.