Assessment of accuracy of liver fluke diagnostic tests using the gold standard of total worm counts.

In many regions of New Zealand liver fluke is endemic, infecting most grazing ruminants, including cattle, sheep, and deer. Restricting the economic losses and welfare costs associated with liver fluke relies on accurately identifying those animals with a production limiting infection. This has proven a difficult goal and although several antemortem quantitative tests are available, including faecal egg counts (FEC), serum ELISA and copro-antigen ELISA, none can be considered a gold standard test of liver fluke infection. The accepted gold standard test for fascioliasis is the total fluke count, which is both laborious and can only be completed at post-mortem. This study aimed to compare the performance of four liver fluke diagnostic tests, against the results of a gold standard total fluke count test. Two groups of cattle were selected, 29 culled mixed age beef cows (MAC) and ten 30-month-old steers. The cattle were blood sampled and faecal sampled prior to slaughter and their whole livers recovered post slaughter at the abattoir. Liveweight was also recorded at slaughter. After collection, each liver was weighed, scored for gross pathology, then serum, faeces and livers were frozen at -20 °C for later analysis. Faecal egg counts and F. hepatica copro-antigen ELISA tests were completed on the faecal samples and total fluke counts were completed on the livers. Fasciola hepatica antibody concentration in serum samples were quantified using a commercial ELISA test. Poisson regression models were built to model the association between each diagnostic test and the total fluke count, and a linear regression model was built to examine the relationship between each diagnostic test and live weight at slaughter. The median fluke count was significantly higher in MAC than steers (p = 0.01), and F. hepatica eggs were present in 100% steers and 66% MAC. There was a significant effect of copro-antigen ELISA value on total fluke count (p < 0.0001), with a coproantigen ELISA value = 20.1 predicting 10 flukes and a value = 44.8 predicting 30 flukes. There was also a significant effect of FEC on total fluke count (p = 0.002) but the R-squared value for this model was lower. There was no association between liver fibrosis score or antibody ELISA test and total fluke count (p = 0.95, p = 0.73, respectively). There was a significant effect of total fluke count (p = 0.03) on liveweight at slaughter, with liveweight falling 20.4 kg for each unit increase in loge (total fluke count). There was no effect of FEC (p = 0.11), antibody ELISA (p = 0.55) or copro-antigen ELISA value (p = 0.16) on liveweight at slaughter. Taken together, these results show that the coproantigen ELISA test is the better test for estimating the true liver fluke burden and that the number of flukes in the liver has a negative effect on cattle live weights at slaughter.

Unveiling genetic variants: Tetra-primer ARMS-PCR diagnosis and structural insights into BLAD, BC, and DUMPS in Pakistani cattle herds.

BACKGROUND: Bovine leukocyte adhesion deficiency (BLAD), bovine citrullinemia (BC), and deficiency of Uridine monophosphate synthetase (DUMPS) are the common autosomal recessive disorders affecting the global dairy industry. BLAD leads to poor wound healing and recurrent infections. In BC, ammonia builds up leading to neurological disorders and death. DUMPS results in developmental abnormalities. METHODOLOGY: In this study, tetra-primer amplification refractory mutation system polymerase chain reaction (ARMS PCR) based diagnostic tests were optimized for BLAD, BC, and DUMPS. A total of 250 animals (58 indigenous and 192 Holstein Friesian (HF)) were screened from all across Pakistan. In addition to validation of ARMS-PCR results through Sanger sequencing, the protein modeling provided structural insights of the disease-associated reported SNPs. Pathway analysis illustrated gene functions under normal and mutated conditions. Furthermore, haplotype and phylogenetic analysis of ASS1 (Argininosuccinate synthetase) gene were performed on study samples and NCBI retrieved sequences. RESULTS: The study's focus was to screen the herds for prevalence of carriers of genetic disorders, as they are the main source of disease dissemination. One animal was found carrier for BC, whereas no carriers were found for BLAD and DUMPS. The protein models corroborated the reported amino acid change in BLAD, and protein truncation in both BC and DUMPS proteins. SNPs found in NCBI retrieved sequences were either silent or missense and had no effect on protein structure. DNA network presented graphical illustration of haplotype interactions and phylogenetic analysis conferred evolutionary landscape of ASS1 gene. The combination of these approaches produced an in-depth genetic picture of BC in Pakistani cattle. CONCLUSION: The development of diagnostic tests and identification of the heterozygous BC sample underscores the significance of constant monitoring to avoid the unwanted dissemination of mutant alleles among Pakistani cattle, thereby promoting the general well-being and sustainability of the dairy sector.

Diagnostic utility of acute phase proteins and their ability to guide antibiotic usage in pigs, horses, and cattle: a mapping review.

To mitigate the use of antibiotics for many of the multifactorial diseases seen in pigs, horses and cattle, new diagnostic tools are needed. Acute phase protein (APP) measurements can, in humans, be used to guide antibiotic treatment initiation, evaluate treatment efficacy, and make a prognosis. The aim of this review is to collect evidence on the clinical functionality of APP measurements as a tool to guide antibiotic treatment in pigs, horses, and cattle. Literature was retrieved using Medline, CAB Abstracts and Google Scholar. The acute phase response has been investigated for a plethora of diseases and clinical signs and the major acute phase proteins are elevated in diseased compared to healthy animals. Few studies correlated acute phase response with aetiology, antibiotic treatment efficacy, prognosis, or severity of disease. The existing research does not support that APP can be used to guide antibiotic treatment, but the reported studies indicate that C-reactive protein (CRP) might be able to differentiate between bacterial and non-bacterial causes of disease in pigs. Serum amyloid A (SAA) might reflect underlying aetiology in horses and infectious or non-infectious cases of mastitis in cows.

Imaging of a subcutaneous abscessation in the back of a calf with hindlimb paralysis.

Background: Ultrasonography is not chosen as the common imaging modality to diagnose spinal cord diseases. The present report indicates good diagnostic efficacy of ultrasonography for identifying spinal cord compressed by subcutaneous mass when scanning through the defected vertebral laminae and spinous process. Case Description: A five-month-old female Holstein calf presented with progressive hindlimb paralysis following a surgical resection of a back mass conducted at 21 days of age. The mass was subsequently histopathologically diagnosed as a pulmonary choristoma. Alongside hindlimb paralysis, the calf developed a swollen back at the lumbar region where the mass was removed. This suggested regrowth of the resected mass, causing injury to the underlying spinal cord. Ultrasonography identified the subcutaneous involvement of the capsular mass, which had three anechoic cavities separated by the echogenic septal structures. The spinal cord could be ultrasonographically demonstrated as adjacent to the mass through the defected vertebral laminae and spinous process in the second and third lumbar vertebras. Ultrasound-guided centesis allowed the collection of purulent exudates in which Escherichia coli was isolated. Myelography and subsequent computed tomography (CT) revealed a partial blockage of the intradural flow of contrast media at the levels of the second and third lumbar vertebras, diagnosed as spina bifida on the CT images. When applying ultrasonography to the spinal cord within a saline pool soon after the subcutaneous abscess was successfully resected, the spinal cord was characterized by the interrupted and partly extended hyperechogenic line of the central canal within the echogenic parenchyma. The echotexture of the spinal cord showed damage due to compression from the subcutaneous abscess. The animal had a sub-optimal postoperative outcome, including limited improvement of the neurological signs. Conclusion: In the present case, combining ultrasonography, radiography (myelography), and CT was very effective for diagnosing spina bifida, with the subcutaneous abscess inducing spinal cord compression. Additionally, using intraoperative ultrasonographic scanning to evaluate the degree of spinal cord damage can contribute to predicting the postoperative outcome.

A colorimetric assay for Neospora caninum utilizing the loop-mediated isothermal amplification technique.

Neospora caninum (N. caninum) is a protozoan parasite that poses a serious risk to livestock by infecting various domestic and wild animals. Loop-Mediated Isothermal Amplification (LAMP) offers a cost-effective, highly sensitive, and specific method for detecting protozoan parasites. This study aims to develop a precise, rapid, and visually assessable colorimetric LAMP method, improving on traditional techniques. We employed a rigorous screening process to identify the optimal primer set for this experiment. Subsequently, we fine-tuned the LAMP reaction at 65 °C for 40 min with 270 µmol/L neutral red. We then confirmed the specificity of primers for N. caninum through experimental validation. The LAMP method demonstrated a lower detection limit compared to traditional Polymerase Chain Reaction (PCR) techniques. While LAMP offers clear advantages, the prevalence of DNA detected in 89 sheep serum and 59 bovine serum samples using the nested PCR method was 3.37 % (3/89) and 1.69 % (1/59), respectively. In contrast, when the LAMP method was employed, the prevalence of detected DNA rose to 5.61 % (5/89) for sheep and 3.38 % (2 /59) for bovine. A comparison of two molecular assays using the intragroup correlation coefficient (ICC) resulted in a value of 0.999 (95 % CI: 0.993-0.996, p < 0.001), indicating the LAMP method is in the "better" range according to James Lee's categorization. The LAMP technique, optimized with specific primers of N. caninum and neutral red dye, not only exhibited higher sensitivity but also provided convenience over conventional PCR methods, highlighting its potential for on-site applications and cost-effective field detection.

Lameness prevalence in lactating and dry cows - expert's, student's and farmers' assessments.

OBJECTIVE: The aim of this study was to evaluate differences in lameness prevalence among Austrian dairy farms for both lactating and dry cows as assessed by locomotion scoring (LCS 1-5) by an expert and a student who had received brief training in locomotion scoring and by farmers' estimates and by farmers' lameness assessments. MATERIALS AND METHODS: In total, 632 cows from 11 farms were evaluated, including 553 lactating and 79 dry cows. Lameness prevalence was calculated for all farms collectively as well as for individual farms. The ratio of lameness prevalence determined by the student/expert (Student's Assessment Index: SAI), the ratio between farmers' assessments/expert assessments (Farmers' Assessment Index: FAI) and the ratio between farmers' estimates/expert assessments (Farmers' Detection Index: FDI) were calculated. RESULTS: For the expert, the mean lameness prevalence (LCS≥2) among all 632 cows was 63.1%, while for the student it was 60.2%, and the farmers' assessment was 37.3%, resulting in a mean difference of 25.8% between the expert and the farmers. In cows with LCS 2, the SAI was 99.6%, in cows with LCS 3, it was 84.4%, and in cows with LCS≥4, it was 88.6%. The mean FDI and FAI for all cows on the 11 farms were 35.1% and 58.2%, respectively, with wide variation across farms (6.8-79.1% and 17.8-94.7%, respectively). Overall, lactating and dry cows exhibited a high mean lameness prevalence (63.9% vs. 59.6%), which differed by only 4.3%. CONCLUSIONS AND CLINICAL RELEVANCE: Farmers should actively assess their cattle for lameness on a regular basis. Moreover, they should be trained to identify lame cows (lactating and dry cows), especially those showing mild lameness (LCS 2). Since the recently trained student achieved similar high recognition rates as the expert, it is assumed that a training in locomotion scoring can contribute to higher recognition rates.

Determining lactate concentrations in Korean indigenous calves and evaluating its role as a predictor for acidemia in calf diarrhea.

BACKGROUND: Calf diarrhea leads to high mortality rates and decreases in growth and productivity, causing negative effects on the livestock industry. Lactate is closely associated with metabolic acidosis in diarrheic calves. However, there have been no reports on lactate concentrations in Korean indigenous (Hanwoo) calves, especially those with diarrhea. This study aimed to determine the reference range of L-lactate and D-lactate concentrations in Hanwoo calves and to better understand the utility of lactate as predictive factors for acidemia in diarrheic calves. RESULTS: L-lactate and D-lactate concentrations were measured in healthy (n = 44) and diarrheic (n = 93) calves, and blood gas analysis was performed on diarrheic calves. The reference range in healthy calves was 0.2-2.25 mmol/L for L-lactate and 0.42-1.38 mmol/L for D-lactate. Diarrheic calves had higher concentrations of L-lactate and D-lactate than healthy calves. In diarrheic calves, L-lactate and D-lactate each had weak negative correlation with pH (r = - 0.31 and r = - 0.35). In diarrheic calves with hyper-L-lactatemia, the combined concentrations of L-lactate and D-lactate had moderate correlation with pH (r = - 0.51) and anion gap (r = 0.55). Receiver operating characteristic analysis showed D-lactate had fair predictive performance (AUC = 0.74) for severe acidemia, with an optimal cut-off value of > 1.43 mmol/L. The combined concentrations of L-lactate and D-lactate showed fair predictive performance for predicting acidemia (AUC = 0.74) and severe acidemia (AUC = 0.72), with cut-off values of > 6.05 mmol/L and > 5.95 mmol/L. CONCLUSIONS: The determined reference ranges for L-lactate and D-lactate in Hanwoo calves enable the identification of hyper-L-lactatemia and hyper-D-lactatemia. Diarrheic calves exhibited increased lactate concentrations correlated with acid-base parameters. While the concentrations of L-lactate and D-lactate have limitations as single diagnostic biomarkers for predicting acidemia or severe acidemia, their measurement remains important, and L-lactate has the advantage of being measurable at the point-of-care. Assessing lactate concentrations should be considered by clinicians, especially when used alongside other clinical indicators and diagnostic tests. This approach can improve calf diarrhea management, contributing positively to animal welfare and providing economic benefits to farms.

Duodenal ileus caused by a cage magnet in a Fleckvieh cow.

The present report aims to describe the case of a duodenal obstruction ileus in a dairy cow that was caused by a cage magnet. The 4.7-year-old German Fleckvieh cow was hospitalized because of symptoms of intestinal obstruction such as anorexia, noticeable drop in milk yield, reduced defecation, dehydration and positive percussion and swinging auscultation in a circumscribed area cranial of the right flank over the last 2 ribs. Six months as well as 3 days prior to hospitalization the cow had already been treated for signs of hardware disease, which included administration of a cage magnet.After the initial clinical diagnostic procedure on hospital admission, a diagnostic laparotomy in the right paralumbar fossa was performed to identify the cause of the ileus. The cranial part of the duodenum was markedly dilated, and a solid foreign body was found obstructing the intestine immediately aboral to the duodenal sigmoid flexure. This was identified as a cage magnet, which was massaged in retrograde direction into the pyloric antrum and removed via abomasotomy. The cow recovered from surgical intervention and was discharged from the hospital 6 days later.The present report describes an unusual complication of cage magnet administration, which is a standard veterinary procedure and generally considered a safe treatment option in cows with clinical signs of acute traumatic reticuloperitonitis.

Benchmarking analysis of computer vision algorithms on edge devices for the real-time detection of digital dermatitis in dairy cows.

Digital dermatitis (DD) is a bovine claw disease responsible for ulcerative lesions on the coronary band of the foot. It causes significant animal welfare and economic losses to the cattle industry. Early detection of DD can lead to prompt treatment and decrease lameness. Current detection and staging methods require a trained individual to evaluate the interdigital space on each foot for clinical signs of DD. Computer vision (CV), a type of artificial intelligence for image analysis, has demonstrated promising results on object detection tasks. However, farms require robust solutions that can be deployed in harsh conditions including dust, debris, humidity, precipitation, other equipment issues. The study aims to train, deploy, and benchmark DD detection models on edge devices. Images were collected from commercial dairy farms with the camera facing the interdigital space on the plantar surface of the foot. Images were scored for M-stages of DD by a trained investigator using the M-stage DD classification system. Models were trained to detect and score DD lesions and embedded on an edge device. The Tiny YOLOv4 model deployed on a CV specific integrated camera module connected to a single board computer achieved a mean average precision (mAP) of 0.895, an overall prediction accuracy of 0.873, and a Cohen's kappa of 0.830 for agreement between the computer vision model and the trained investigator. The model reached a final inference speed of 40 frames per second (FPS) and ran stably without any interruptions. The CV model was able to detect DD lesions on an edge device with high performance and speed. The CV tool can be used for early detection and prompt treatment of DD in dairy cows. Real-time detection of DD on edge device will improve health outcomes, while simultaneously decreasing labor costs. We demonstrate that the deployed model can be a low-power and portable solution for real-time detection of DD on dairy farms. This result is a step towards applying CV algorithms to veterinary medicine and implementing real-time detection of health outcomes in precision farming.

Development of polymerase chain reaction-lateral flow dipstick assay for detection of Mycoplasma bovis in cattle.

Mycoplasma bovis (M. bovis) is capable of causing a range of diseases in cattle, encompassing calf pneumonia, arthritis, conjunctivitis, meningitis, and mastitis. It is widely recognized as one of the predominant pathogens posing a significant threat to the global cattle industry. Therefore, accurate and sensitive methods are urgently needed to detect M. bovis. This study aims to detect M. bovis by combining colloidal gold with biotin-labeled oligonucleotides to improve detection sensitivity and form a chromogenic detection probe based on signal amplification technology. Here, we developed a sensitive and specific polymerase chain reaction-lateral flow dipstick assay (PCR-LFD) strip for efficient nucleic acid detection of M. bovis. A pair of specific primers with 5' ends labeled with biotin and digoxigenin probes was designed for PCR experiments. Colloidal gold particles-labeled anti-digoxigenin IgG coated gold-labeled test strip was prepared, streptavidin was used as the detection probe, and nitrocellulose membrane coated goat anti-mouse IgG was used as the control line. Our results showed that the detection limit of the PCR-LFD was 89 fg/µL for the M. bovis DNA. The results from the test strip were highly consistent with those from real-time qPCR. This assay were highly specific for M. bovis, as there were no cross-reactions with other microorganisms tested and the detection sensitivity of the test was also relatively high (97.67%). The novel strips present a promising tool for the cost-effective and sensitive diagnosis of M. bovis.

Microscopic, Molecular and Antigen Detection and Isolation of Cryptosporidium parvumParasites in Diarrheal Disease of Calves in Iran.

Cryptosporidiosis is a parasitic disease caused by the protozoan Cryptosporidium in vertebrates. In livestock, especially ruminants, infants develop diarrheal syndromes. The infection is common worldwide , including Iran, where it is reported in several species. Morphological diagnosis of Cryptosporidium species is associated with many limitations and has no taxonomic value on its own, so the use of molecular methods can overcome these limitations to some extent. The present aims at microscopic, molecular and antigen detection and isolation of Cryptosporidium parvum parasites. Firstly, 300 samples were collected from different parts of Iran. Subsequently oocysts from feces were purified by the method of Casemore et al. using the flotation technique and stained by the modified Ziehl-Neelsen method (Henriksen method) and identified by diagnostic keys. ELISA test was also performed on the samples with results ranging from 1 to 4 positive. The results of our study show that, of the 300 cases tested for Cryptosporidium, 48 cases (16%) and 54 (18%) were positive in ELISA and PCR, respectively. Microscopic evaluation also mainly confirmed the ELISA results. These cases were collected in summer, autumn, and winter, with, more than 50% of the positive cases found among the samples collected in autumn. In addition, 54 positive cases were found by PCR test, which is 6 cases more than ELISA results. Finally, the results of PCR detection and ELISA were subjected to chi-square analysis, where no significant difference was found between the collected data (p=0.0587).

A systematic review of predictive, diagnostic, and prognostic biomarkers for detecting reproductive diseases in cattle using traditional and omics approaches.

Reproductive diseases and illnesses pose significant challenges in cattle farming, affecting fertility, milk production, and overall herd health. In recent years, the integration of various omics approaches, including transcriptomics, proteomics, metagenomics, miRNAomics, and metabolomics, has revolutionized the study of these conditions. This systematic review summarised the findings from studies that investigated reproductive disease biomarkers in both male and female cattle. After extracting 6137 studies according to exclusion and inclusion criteria, a total of 60 studies were included in this review. All studies identified were associated with female cattle and none were related to reproductive diseases in bulls. The analysis highlights specific biomarkers, metabolic pathways, and microbial compositions associated with bovine reproductive disease conditions, providing valuable insights into the underlying molecular mechanisms of disease. Pro-inflammatory cytokines such as IL-1ß, IL-8, IL-4, IL-6, TNFα and acute-phase response proteins such as SAA and HP have been identified as promising biomarkers for bovine reproductive diseases. However, further research is needed to validate these markers clinically and to explore potential strategies for improving cow reproductive health. The role of bulls as carriers of venereal diseases has been underestimated in the current literature and therefore needs more attention to understand their impact on infectious reproductive diseases of female cattle.

Molecular identification of haemoparasites in animals using blood lysate PCR: a quick and inexpensive alternative to purified whole genomic DNA.

Haemoparasitic diseases constitute a significant constraint to economic livestock farming. Diagnostic techniques that are inexpensive, rapid, reliable, and precise are crucial for the management of diseases. In this context, PCR assays are very valuable yet expensive since the samples must be processed before being included in the PCR reaction. Accordingly, the goal of the current study was to lower the PCR costs without jeopardizing the assay's sensitivity and specificity. For that purpose, the alkaline solution was optimized for low cost and quick DNA extraction (blood lysate), and PCR reagents were modified for optimum reaction. In comparison to purified whole blood genomic DNA, the currently developed and optimized blood lysate method was found to be 95.5% less expensive, as well as being equally sensitive and specific for the molecular detection (PCR) of haemoparasites like Babesia, Theileria, Trypanosoma and rickettsiales in cattle, buffaloes, horses, and dogs. The assay was also demonstrated to be quick, less likely to cross-contaminate, and appropriate for use in laboratories with limited resources. Therefore, the currently developed and optimized blood lysate method could serve as a viable alternative to purified whole blood genomic DNA for molecular detection (PCR) of haemoparasites in animals particularly in resource-limited settings.

An evaluation form to aid dairy producers to systematically assess cows prior to culling.

Objective: The objective of this prospective observational research project was to have dairy producers use and assess the utility of a cull cow evaluation form. Animals: Cull dairy cows. Procedure: Veterinarians were recruited to enrol a purposively selected group of dairy producers into a project to evaluate a cull cow evaluation form. Producers were provided with evaluation forms and asked to complete a form for every cow they culled from their herd from January to June 2017, inclusive. Results: A total of 44 producers used the form to record information on 323 cows prior to transport. Conclusion and clinical relevance: Despite the completion of 323 forms, only ~1/3 were completed fully, with compliance highest for body condition score, lameness, and temperature recordings (> 90% of forms). A cull cow evaluation form may improve the thoroughness and consistency of dairy producer assessment of cull dairy cows for fitness for transport.

Un formulaire d'évaluation pour aider les producteurs laitiers à évaluer systématiquement les vaches avant la réforme. Objectif: L'objectif de ce projet de recherche observationnelle prospective était d'amener les producteurs laitiers à utiliser et à évaluer l'utilité d'un formulaire d'évaluation des vaches de réforme. Animaux: Vaches laitières réformées. Procédure: Des vétérinaires ont été recrutés pour inscrire un groupe de producteurs laitiers sélectionnés à dessein dans un projet visant à évaluer un formulaire d'évaluation des vaches réformées. Les producteurs ont reçu des formulaires d'évaluation et ont été invités à remplir un formulaire pour chaque vache qu'ils ont éliminée de leur troupeau de janvier à juin 2017 inclusivement. Résultats: Au total, 44 producteurs ont utilisé le formulaire pour enregistrer des informations sur 323 vaches avant le transport. Conclusion et pertinence clinique: Malgré la complétion de 323 formulaires, seulement environ 1/3 ont été entièrement remplis, avec une conformité plus élevée pour le score d'état corporel, les boiteries et les enregistrements de température (> 90 % des formulaires). Un formulaire d'évaluation des vaches laitières réformées peut améliorer la rigueur et la cohérence de l'évaluation par le producteur laitier des vaches laitières réformées quant à leur aptitude au transport.(Traduit par Dr Serge Messier).

Integrating multi-wet laboratory diagnostics to study staphylococci in animals in Uganda.

BACKGROUND: Several diagnostic environments in Uganda lack real-time, robust and high-throughput technologies for comprehensive typing of microbes, which is a setback to infectious disease surveillance. This study combined various wet laboratory diagnostics to understand the epidemiology of pathogenic staphylococci isolated from animals in Uganda and the implications for global health security priorities. METHODS: A retrospective study was conducted employing records and pathogenic staphylococci (from animals) archived at the Central Diagnostic Laboratory (CDL), Makerere University, Uganda, between January 2012 and December 2019. The bacteria were speciated by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and tested for virulence factors [beta lactamases, lecithinase, deoxyribonuclease (DNase), haemolysins] and resistance to ten antimicrobials of clinical and veterinary relevance. Tetracycline and methicillin resistance genes were also tested. RESULTS: The prevalent diseases were mastitis in cattle and skin infections in dogs. Of the 111 staphylococci tested by MALDI-TOF MS, 79 (71.2%) were Staphylococcus aureus, 27 (24.3%) were Staphylococcus pseudintermedius and 5 (4.5%) were Staphylococcus schleiferi. All these strains expressed haemolysins. The prevalence of strains with lecithinase, penicillinase, cephalosporinase and DNase was 35.9% (14/39), 89.7% (35/39), 0.0% (0/39) and 87.2% (34/39), respectively. Staphylococci were primarily resistant to early penicillins (over 80%), tetracycline (57.7%), and chloramphenicol (46.2%). Minimal resistance was noted with cloxacillin (0.0%), ciprofloxacin (9.6%), and cefoxitin (3.8%). The prevalence of multidrug resistance (MDR) was 78.8% for general staphylococci, 82.2% for S. aureus, 73.1% for S. pseudintermedius, and 60.0% for S. schleiferi. Multidrug resistant staphylococci were significantly more prevalent in the cattle isolates than in the dog isolates (P < 0.05). The prevalence of methicillin-resistant staphylococci (MRS) tested by resistance to cefoxitin and mecA carriage was 3.8%. These four strains were all isolated from dog skin infections. The tetK gene was the most predominant (35.4%), followed by tetM (25.0%). CONCLUSION: In resource-constrained settings, the approach of integrated diagnostics promises sustainable disease surveillance and the addressing of current capacity gaps. The emergence of MRS (zoonotic bacteria) in companion animals creates a likelihood of reduced treatment options for related human infections, a threat to global health.

Economics of reducing response time to foreign-animal disease in the United States with point-of-care diagnostic tests.

BACKGROUND: As low probability events, United States producers, value chain actors, and veterinary services (VS) have limited experience with identifying foreign animal disease (FAD), which can allow FADs to spread undetected. Point-of-care (POC) diagnostic testing may help reduce the time from detecting an initial suspect case to implementing actionable interventions compared to the current approach of only using laboratory diagnostic testing for disease diagnosis and confirmation. To evaluate the value of the reduced response time, we compare the associated costs between the two diagnostic approaches while accounting for the uncertainty surrounding the size of a FAD event. METHODS: We apply a state-contingent approach (SCA) to model the uncertainty surrounding a FAD through alternative events, where the event defines the scale of outbreak size and its duration. We apply this approach within a cost-benefit framework (CBA) to determine the economic value from the two testing investment strategies to help explain the policymaker's response (and costs) to alternative FAD events while also considering the cost impacts on the producers from each event. RESULTS: Compared to the current laboratory strategy, a POC strategy that reduces response time by 0.5-days (swine, cattle scenarios) and 1.5-days (poultry scenario) may provide cost-saving to both producers and public response efforts. The benefit-cost analysis further suggests that despite the higher fixed costs to adopt the POC strategy, the swine and cattle sectors may benefit while the benefits may not be as pronounced in the poultry sector. DISCUSSION: POC testing that can reduce the time between detection and response during a FAD event may be a sound strategy for public expenditure and provide cost-savings for producers, especially when minimal fixed costs are incurred. However, to fully determine the value of POC testing, the consequences (costs) associated with potential actions if something goes wrong, (e.g. false positive results), should be considered in future studies.

The first isolation of Clostridium massiliodielmoense from a dead beef cow in Japan and pitfalls to be aware of in identifying the Clostridium species.

Clostridium sp. was detected in the organs of a cow with black watery diarrhea in Japan. Results identifying this species were inconsistent; Clostridium novyi type A infection was suggested by PCR assay targeting Clostridium fliC region (fliC-multiplex PCR), while 16S rRNA gene sequencing identified the isolated bacteria as Clostridium massiliodielmoense. Sequencing of fliC-multiplex PCR products from the isolates revealed the presence of fliC region in C. massiliodielmoense, which had 92.7% nucleotide similarity to that of C. novyi type A JCM 1406T, leading to the false positive detection of C. novyi by the PCR. This is the first C. massiliodielmoense isolation from clinical specimens, suggesting the need for further research on its pathogenicity and improvement in fliC-multiplex PCR.

Epidemiology and diagnostic accuracy of Clostridium perfringens toxins in the intestinal contents of camels, sheep, and cattle: a cross-sectional study in Dakahlia governorate, Egypt.

This study aimed to establish an accurate epidemiological surveillance tool for the detection of different C. perfringens types from 76 diseased and 34 healthy animals in Dakhalia Governorate, Egypt. A total of 110 intestinal content samples were randomly collected from camels, sheep, and cattle. C. perfringens was isolated and biochemically identified by the VITEK2 system. Toxinotyping and genotyping of C. perfringens isolates were specified by a multiscreen ELISA and real-time qPCR (rt-qPCR). The occurrence of C. perfringens was highest among camels (20% in healthy and 25% in diseased) and was lowest in cattle (23.1% and 14.7%). The cpa toxin was detected in all isolates by rt-qPCR and in 7 isolates by ELISA, ext toxin was detected in 7 isolates by rt-qPCR and in 6 isolates by ELISA, and cpb toxin was detected in 2 isolates by both rt-qPCR and ELISA. Four types of C. perfringens were identified by rt-qPCR, type A (65.2%), B (4.3%), C (4.3%), and D (26.1%), and three types by ELISA, type D (17.4%), A (8.7%) and C (4.3%). Our study indicated the prevalence of infection in Dakahlia by C. perfringens type A and D, particularly camels, and recommends adopting an appropriate vaccination strategy among the studied animals.

Establishing a surveillance programme for Salmonella Dublin in Austrian dairy herds by comparing herd-level vs. individual animal detection methods.

Due to its increasing occurrence in cattle farms in various countries, leading to significant economic losses in affected livestock, Salmonella enterica subspecies enterica serovar Dublin (S. Dublin) has become a highly investigated pathogen in cattle production. In Austria, there have been occasional human cases of S. Dublin as well as an increase in laboratory-confirmed cases in cattle, indicating the need for a screening programme to determine the current status in Austria. The aims of this study were, firstly, to determine the seroprevalence of S. Dublin in dairy herds through bulk milk screenings in two federal states (Salzburg, Tyrol) of Austria. Secondly, the study aimed to identify the infection status of the herds through individual animal and herd level detection, comparing microbiological, molecular and serological detection methods. The results of the study will allow the development of a sampling strategy for a surveillance programme in Austria. A total of 6973 dairy farms were tested through serological bulk milk screening. The seroprevalence for the federal state of Tyrol was 14.8 % and for Salzburg it was 18.2 %, resulting in an average seroprevalence of 16.5 %. At an individual animal level, 205 (11.3 %) animals tested positive for shedding of S. Dublin in the faeces through microbiological detection, and 268 (17.0 %) animals had positive values (ct value ≤ 38) by qPCR. The association between microbiological and molecular detection was statistically significant (p < 0.001), with a calculated kappa value of 0.65 ± 0.27 (p ≤ 0.001), assuming a substantial level of agreement. In 17 herds, where an individual animal tested positive for shedding of S. Dublin, environmental sampling and testing were carried out. At a herd level 16 (94.1 %) out of the 17 participating herds, tested positive for S. Dublin either microbiologically or by molecular assay in boot swab samples. Bulk milk samples from 14 out of the 17 participating herds were analysed for antibodies to S. Dublin and 12 samples (85.7 %) were positive. In total 111 (18.9 %) out of 587 blood samples tested positive for S. Dublin antibodies, demonstrating a statistically significant correlation (p < 0.001) both with microbiological (κ = 0.32 ± 0.49; p ≤ 0.001) and molecular (κ=0.23 ± 0.06; p ≤ 0.001) findings. It was possible to identify S. Dublin by culture from boot swabs in 14 (82.4 %) out of 17 herds and by molecular assay using qPCR in 15 (88.2 %) out of 17 herds, indicating a suitable sample type for screening on a herd level-basis for acute infections, but not for identifying chronic infections or asymptomatic carriers. Other environmental samples, such as sponge-sticks, are only suitable to a limited extent for the detection of S. Dublin. The results of this study demonstrate a moderate S. Dublin prevalence in dairy herds in the selected Austrian regions, signalling further screening and management programmes for the future.

Evaluating machine learning algorithms to predict lameness in dairy cattle.

Dairy cattle lameness represents one of the common concerns in intensive and commercial dairy farms. Lameness is characterized by gait-related behavioral changes in cows and multiple approaches are being utilized to associate these changes with lameness conditions including data from accelerometers, and other precision technologies. The objective was to evaluate the use of machine learning algorithms for the identification of lameness conditions in dairy cattle. In this study, 310 multiparous Holstein dairy cows from a herd in Northern Colorado were affixed with a leg-based accelerometer (Icerobotics® Inc, Edinburg, Scotland) to obtain the lying time (min/d), daily steps count (n/d), and daily change (n/d). Subsequently, study cows were monitored for 4 months and cows submitted for claw trimming (CT) were differentiated as receiving corrective claw trimming (CCT) or as being diagnosed with a lameness disorder and consequent therapeutic claw trimming (TCT) by a certified hoof trimmer. Cows not submitted to CT were considered healthy controls. A median filter was applied to smoothen the data by reducing inherent variability. Three different machine learning (ML) models were defined to fit each algorithm which included the conventional features (containing daily lying, daily steps, and daily change derived from the accelerometer), slope features (containing features extracted from each variable in Conventional feature), or all features (3 simple features and 3 slope features). Random forest (RF), Naive Bayes (NB), Logistic Regression (LR), and Time series (ROCKET) were used as ML predictive approaches. For the classification of cows requiring CCT and TCT, ROCKET classifier performed better with accuracy (> 90%), ROC-AUC (> 74%), and F1 score (> 0.61) as compared to other algorithms. Slope features derived in this study increased the efficiency of algorithms as the better-performing models included All features explored. However, further classification of diseases into infectious and non-infectious events was not effective because none of the algorithms presented satisfactory model accuracy parameters. For the classification of observed cow locomotion scores into severely lame and moderately lame conditions, the ROCKET classifier demonstrated satisfactory accuracy (> 0.85), ROC-AUC (> 0.68), and F1 scores (> 0.44). We conclude that ML models using accelerometer data are helpful in the identification of lameness in cows but need further research to increase the granularity and accuracy of classification.