Lorenzo Viora

Evaluation of a culture-based pathogen identification kit for bacterial causes of bovine mastitis

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.

Black-pigmented anaerobic bacteria associated with ovine 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 Students t-test and Spearmans 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.

Periodontal lesions in slaughtered cattle in the west of Scotland

PERIODONTITIS is a multifactorial infection elicited by a complex of bacterial species that interact with host tissues and cells causing the release of a broad array of inflammatory cytokines, chemokines and mediators, some of which lead to destruction of the periodontal structures, including the tooth supporting tissues, alveolar bone and periodontal ligament (Holt and Ebersole 2005). Although cattle are of worldwide economic importance in the dairy and beef industries, their dentition has not been investigated as thoroughly as that of other species. Cattle are diphyodont and hypsodont, and their permanent dentition has 32 teeth. The permanent incisors erupt sequentially between 1.5 and 4 years of age. All permanent premolars and the second and third molars erupt between one and three years of age (Page and Schroeder 1982). The importance of bovine periodontal disease has largely been overlooked by veterinarians and scientists. In Brazil, periodontal disease affects cattle kept in areas where grazing has been recently formed or reformed in the Atlantic Forest, Cerrado, Pantanal and Amazon biomes (Dobereiner and others 2000). The disease is characterised by a purulent, progressive periodontitis, with periodontal pocket formation, development of a chronic periostitis ossificans and, in some cases, systemic illness. The tooth roots become exposed with eventual loss of teeth (Dobereiner and others 1974). Reported studies in the UK are scarce. Of 501 heads of cows examined …

Postpartum ketoprofen treatment does not alter stress biomarkers in cows and calves experiencing assisted and unassisted parturition: a randomised controlled trial

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.

Microbiomes associated with bovine periodontitis and oral health

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.

Ruminant neurological disease: a retrospective cohort study

Between January 2006 and June 2016, 96 ruminants with neurological signs were donated to the Scottish Centre for Production Animal Health and Food Safety (SCPAHFS), University of Glasgow, by veterinarians in the field representing 5.4 per cent of all submissions. Forty-seven different neurological presenting signs were reported with 79 per cent of the donated patients presenting with abnormal gait. All cases presenting with abnormalities in more than 4 out of 10 neurological categories died or were euthanased on welfare grounds. Calves were significantly more likely to present with neurological disorders than adult cattle compared with the proportion of calves: cows in the Scottish cattle population and total case population donated to SCPAHFS. Lesions were most commonly localised to the spinal cord in sheep 47 per cent (16), the peripheral nervous system in cattle 45 per cent (28) and to the brain in the overall population 41 per cent (39). The most common aetiology of neurological pathologies observed was infectious or inflammatory 28 per cent (27). Definitive diagnoses could be reached in 84 per cent (81) of patients. When postmortem reports were available, they produced a diagnosis in 70 per cent (52) of cases and contradicted clinical diagnoses in 38 per cent (26) of cases. The most frequently diagnosed conditions in ruminants over the 10 years were spastic paresis, vertebral osteomyelitis and listeriosis.

Pilot study into milk haptoglobin as an indicator of udder health in heifers after calving

Mastitis, inflammation of the mammary gland, is often caused by intramammary infection with bacterial organisms. It impacts on dairy cattle welfare, production, udder health and longevity in the herd. Current detection methods for mammary inflammation and infection all have limitations, particularly for on-farm diagnosis of non-clinical mastitis after calving. Acute phase proteins have been suggested as alternative early indicators of the disease and can potentially be used as cow-side test with results in real time. In this study, milk haptoglobin concentrations were investigated over the first week postpartum to explore haptoglobins potential as indicator of udder health in dairy heifers. Haptoglobin concentration was highest on day 3 of lactation, and was positively correlated with somatic cell count, a commonly used marker of inflammation (rs=0.68). Haptoglobin level was also associated with bacteriological culture results, a key indicator of infection status, whereby median haptoglobin concentration on days 3 and 5 was higher in quarters that were infected at calving than quarters that were non infected at calving. Sensitivity and specificity of haptoglobin concentration as indicator of infection were low, both for lenient and strict culture-based definitions of intramammary infection (57 or 60% and 61 or 63%, respectively). Although haptoglobin was a poor biomarker for intramammary infection with coagulase negative staphylococci in heifers during the first week after calving, it may have value as an indicator of major pathogen infections, particularly in large scale dairy herds where pre-partum heifers are managed off-site.

Evaluation of tissue levels of Toll-like receptors and cytokine mRNAs associated with bovine periodontitis and oral health

Bovine periodontitis is a progressive and purulent infection associated with an anaerobic subgingival biofilm, which induces irreversible damage to the dentition of affected animals. The aetiopathogenesis of the disease is unclear and treatment and control of the disease process in cattle are almost unknown. The aim of this study was to investigate the innate immune response by quantifying expression of Toll-like receptor (TLR) and cytokine genes in gingival tissue samples from cattle with and without periodontitis. Postmortem biopsies of gingival tissues were collected from 20 cattle with periodontitis and 20 cattle with no clinical signs of periodontal lesions. Tissue expression of TLR2, TLR4, TNF-α, IFN-γ, IL-1β and IL-4 genes were determined using quantitative real-time PCR. Statistically significant increases in mRNA levels encoding TLR2 (p = 0.025), TLR4 (p = 0.037), TNF-α (p = 0.025), IFN-γ (p = 0.014), IL-1β (p < 0.001) and IL-4 (p = 0.014) were observed in animals with periodontitis when compared to periodontally healthy animals. Increased levels of TLRs and inflammatory cytokines in periodontal tissue indicate an induction of the innate immune response of cattle and suggest that a substantial microbial challenge may be involved in the aetiopathogenesis of bovine periodontitis.