Veterinary Diagnostic Laboratory Research Articles

Updating the epidemiology of canine leishmaniosis in the United Kingdom through the use of electronic health data.

Dogs infected with Leishmania infantum have been increasingly reported in the UK mostly related to imported/travelled dogs. Up-to-date epidemiologic data are essential for a better control of this zoonotic disease in such emerging areas. This study aimed for the first time, to estimate the percentage and temporal variation of dog and cat samples testing positive for L. infantum infection at commercial diagnostic laboratories, and to describe the travel history of positive dogs and cats positive to leishmaniosis in a network of UK veterinary practices. Leishmania infantum serology and PCR data were collected by the Small Animal Veterinary Surveillance Network (SAVSNET) from five UK national veterinary diagnostic laboratories between 2010 and 2022 and were analysed. In addition, electronic health records (EHRs) were collected from 251 veterinary practices across the UK between March 2014 and September 2022. Text mining tools were used to identify cases compatible with clinical leishmaniosis as recorded in the clinical narratives; these were subsequently manually validated. Data from a total of 25,327 diagnostic samples (25,201 from dogs and 126 from cats) were analysed including 20,517 sera tested by either quantitative ELISA or IFAT, and 4,810 by PCR. Leishmania infantum antibodies were detected in 39.7% of tested dog samples and 1.07% of cat samples. In dogs, seropositivity increased from 2013 to 2022. Leishmania DNA was only detected by PCR in samples from dogs (11.8%). A total of 368 dogs with canine leishmaniosis (CanL) were identified from clinical narratives. Of these, 189 had either visited, or were rescued/imported from, Spain, Greece, Cyprus and other southern European countries. Among factors associated with CanL in the UK canine population, dogs between 3-6 years of age were 4.71 times more likely to have CanL than those two years or younger. In addition, there was an increased risk of having recorded CanL clinical cases from 2017 to 2022, compared to 2014. Southeast of England was the UK region that accounted for the highest number of CanL cases (34.51%). This study provides recent trends in Leishmania infection in dogs in the UK, identifies risk factors and countries likely associated with imported cases, and provides important insights to help plan and monitor national intervention strategies.

Evaluation of Fecal Sample Pooling for Real-Time RT-PCR Testing SARS-CoV-2 in Animals

During the COVID-19 pandemic, veterinary diagnostic laboratories tested both human and animal samples and needed to ensure that they could accurately perform large numbers of diagnostic tests in a timely manner. Sample pooling, a methodology used effectively for over 80 years as a surveillance tool for screening large numbers of potentially infected individuals, was employed. Given its sensitivity, real-time polymerase chain reaction (PCR) is more suitable for employing this strategy, as compared to other less sensitive testing methods. In this study, we evaluated the capability of detecting SARS-CoV-2 in both 5-sample and 10-sample pools of feces using real-time reverse transcriptase polymerase chain reaction (rRT-PCR) as well as determined the level of sensitivity. A blinded method test (BMT) by an independent laboratory was conducted to assess the five-sample fecal pool. To complement detection capability, the stability of the genome within a PBS fecal suspension was measured under various time and temperature conditions across a 28-day period. Our results showed that the limit of detection for 5-sample and 10-sample fecal pools is 12.8 and 6.4 genome copies in a 25 µL PCR, respectively. The 5-sample and 10-sample pooling resulted in a cycle threshold (Ct) value loss of 2.35 and 3.45, as compared to Ct values of known positive individual samples, but consistent detection was still achieved in pools containing positive samples with an original Ct below 36 and 34, respectively. The simulation of clinical five-sample pooling showed that all positive samples could be detected regardless of the number (1–3) of positive samples in each pool. The BMT results demonstrated excellent sensitivity (100 copies/reaction) in five-sample pools for the detection of SARS-CoV-2 RNA even though a fecal matrix effect was observed. Finally, our results show that the SARS-CoV-2 genome remains stable over a wide range of time and temperature variations. Overall, our findings provide solid data to scale up SARS-CoV-2 testing capacity in veterinary diagnostic laboratories.

Proficiency test of SARS-CoV-2 Omicron variant detection in diagnostics samples by veterinary diagnostic laboratories

AbstractVeterinary diagnostic laboratories (VDLs) play a critical role in screening both human and animal samples for SARS-CoV-2. To evaluate the SARS-CoV-2 detection methods used by VDLs, a proficiency test was performed by the US Food and Drug Administration’s Veterinary Laboratory and Investigation and Response Network in collaboration with two other organizations. Thirty-two sets of 12 blind-coded samples were prepared by fortifying Molecular Transport Medium (MTM) or feline feces with SARS-CoV-2 Omicron variant or non-SARS-CoV-2 equine coronavirus RNA at various concentrations and shipped to 32 participants for blinded (unbiased) analysis. Results were analyzed according to the principles of International Organization for Standardization 16140-2:2016 using two approaches such as establishing the rate of detection (ROD) and the success rate by applying the analysis of binary outcome by logit approach. ROD provided the overall assessment of laboratories performance, whereas the novel logit approach provided an insight to more specific analysis based on the complexity of each sample. The ROD was 83% and 98% for MTM samples at 200 and 20000 genome copies per 100 µL, respectively. Fecal samples were classified as challenging exploratory, and results were not included in the assessment of performance but discussion purposes only. Fecal samples exhibited matrix interference impacting the performance. The ROD was 44% and 89% for fecal samples at 2000 and 20000 genome copies per 100 µL, respectively. The non-COVID coronavirus RNA, which was used to address the specificity, did not interfere with methods used. Establishing the success rate by evaluating the qualitative results (detected/not detected) applying a logit approach revealed that, out of thirty-two participants, twenty-eight had satisfactory results, one participant had unsatisfactory results, and three participants had questionable results for MTM samples. For fecal samples, three participants out of thirty-two did not meet the expectations at higher concentrations. Lower concentrations of fecal samples were excluded from this analysis. Again, the fecal samples were considered as challenge samples and the results were provided to assist participants in their continuous efforts to improve their performance and not to evaluate their performance.

Assessment of porcine reproductive and respiratory syndrome virus (PRRSV) farm surface contamination through environmental sampling

BackgroundDuring the fall of 2020, the porcine reproductive and respiratory syndrome virus (PRRSV) L1C.5 variant emerged and rapidly spread throughout southern Minnesota generating questions regarding possible transmission routes. This study aimed to investigate whether PRRSV could be detected on surfaces inside and outside pig barns housing L1C.5 variant PRRSV-positive pigs to illustrate the potential for indirect transmission of PRRSV. Seven Midwestern U.S. PPRS-positive breeding or growing pig farms and one PRRS-negative farm were conveniently selected. Internal and external barn surfaces were wiped using a PBS moistened cloth and the resulting liquid was submitted to the University of Minnesota Veterinary Diagnostic Laboratory for PRRSV RT-PCR testing and virus isolation.ResultsAll (n = 26) samples from PRRSV-negative farm tested negative. Nineteen (13%) out of 143 samples from positive farms yielded positive RT-PCR results. Positive samples originated primarily from exhaust fan cones and doorknobs, followed by anteroom floor and mortality carts/sleds. Virus isolation attempted on two samples did not yield positive results.ConclusionsPRRSV contamination can occur on surfaces inside and outside pig barns that are in frequent contact with farm personnel. Although virus isolation attempts were negative, our results illustrate the potential for PRRSV to be transmitted indirectly through contaminated materials or farm personnel. The study supports the implementation of biosecurity practices by farm personnel to prevent the introduction of PRRSV into farms and the prevention of PRRSV transmission between farms.

Bacillary hemoglobinuria in beef cattle infected with Fascioloides magna in Missouri.

Bacillary hemoglobinuria (BH) is an infectious disease, mostly affecting cattle, caused by Clostridium haemolyticum (C. novyi type D), with acute hepatic necrosis and intravascular hemolysis. Cattle are typically predisposed to BH by liver injury caused by Fasciola hepatica, although cases have been reported in cattle without evidence of this parasite. Here we describe a cluster of 14 BH cases from 7 counties in north-central to central Missouri submitted to a veterinary diagnostic laboratory between December 2020 and April 2023. Postmortem examination in all cases revealed hemoglobinuria and acute hepatic necrosis with large numbers of gram-positive bacilli with terminal-to-subterminal spores. Flukes, fluke ova, and/or fluke pigment consistent with Fascioloides magna were identified in 12 of 14 cases. Sequences of the nuclear ribosomal internal transcribed spacer 1 (ITS1) from one fluke had 100% identity to F. magna. C. novyi was detected by fluorescent antibody testing of liver impression smears (11 of 12 cases) and by immunohistochemistry of liver sections (7 of 7 cases). PCR on formalin-fixed, paraffin-embedded tissues amplified the C. haemolyticum beta toxin gene in each of the 7 cases tested. To our knowledge, a confirmed cluster of BH associated with F. magna has not been reported previously in cattle.

PSIX-26 Effectiveness of mineral supplementation during range grazing on mineral status in ewes

Abstract Mineral supplementation in the intermountain west region of the US may be necessary for maintenance of healthy flocks since important trace minerals are deficient in this region. However, rough terrain can make mineral supplementation challenging. The objective of this study was to evaluate the trace mineral status of ewes on range. We hypothesized that ewes with access to a mineral supplement would have decreased risk of mineral deficiency. In this study, ewes (n = 59) grazed four summer pastures on Cedar Mountain near Cedar City, Utah. Ewes between the ages of 1 and 6 yr of age were stratified by initial body weight (BW) = 87 kg ± 13 kg and breed [black face (BLF; Suffolk) and white face breeds (WF; Targhee, Rambouillet, and their crosses)] into two treatment groups 1) trace mineral supplement ad libitum (TRT; n = 7 BLF and 23 WF) or 2) no mineral supplement (CON; n = 15 BLF and 14 WF). All ewes received a diet of mixed alfalfa and triticale hay without mineral supplementation 30 d before the trial began. A SmartFeed (C-lock Inc.) was used to track individual animal mineral intake. Plasma samples were collected via jugular venipuncture at d 0, 34, 77, and 116. A clinical veterinarian from Utah State University performed liver biopsies at d 0 and 116 on a subset of ewes (n = 10). After the grazing season, plasma and liver samples were sent to the Iowa State University Veterinary Diagnostic Laboratory for analysis of Cu, Mn, Se, and Zn. The mixed procedure of SAS was used to analyze plasma mineral content with day as a repeated measure and individual ID as a random variable. The Glimmix procedure of SAS was used to analyze liver mineral content. Plasma Cu and Se increased (P ≤ 0.05) by day in all ewes throughout the grazing season. No differences related to treatment or breed (P > 0.10) were observed for plasma Cu or Se concentrations. Plasma Mn and Zn decreased (P < 0.0001) by 1.5 ppb and 0.4 ppm, respectively, between d 0 and 116 with no treatment effect (P > 0.10). BLF ewes had 0.3 ppm (P = 0.01) greater plasma Mn concentrations than WF ewes, with no difference (P = 0.6) observed due to treatment. Hepatic Cu and Se increased (P ≤ 0.05) by 114 ppm and 1 ppm, respectively, between d 0 and 116. Hepatic Mn concentrations did not differ (P > 0.10) by day or treatment. Hepatic Zn concentrations tended to increase by day (P = 0.07), 23 ppm between d 0 and 116. CON ewes had 28 ppm greater (P = 0.04) hepatic Zn concentrations than TRT ewes. Additional research is warranted to understand why CON had greater hepatic Zn concentrations than TRT, which may indicate that diverse rangeland forages may provide sufficient concentrations for sheep production in summer months.

A retrospective study of lingual lesions in 793 dogs and 406 cats at the Athens Veterinary Diagnostic Laboratory, 2010-2020.

Lingual biopsies are a common type of sample submission at the Athens Veterinary Diagnostic Laboratory (AVDL). Here we describe the pathology diagnoses of 793 canine and 406 feline lingual biopsies submitted to the AVDL in a 10-y period. Non-neoplastic lesions accounted for 450 diagnoses (57%) in dogs and 239 diagnoses (59%) in cats. Canine non-neoplastic lesions consisted of inflammatory lesions (286 cases; 64% of non-neoplastic lesions) and tumor-like proliferative lesions (164 cases; 36% of non-neoplastic lesions). Feline non-neoplastic lesions consisted of inflammatory lesions (228 cases; 95% of non-neoplastic lesions) and tumor-like proliferative lesions (11 cases; 5% of non-neoplastic lesions). The most common canine neoplasms were melanocytic neoplasms (103 cases; 30% of neoplasms) and epithelial neoplasms (102 cases; 30% of neoplasms), followed by mesenchymal neoplasms (90 cases; 26% of neoplasms) and round cell neoplasms (48 cases; 14% of neoplasms). Approximately 43% of melanocytic neoplasms affected Chow Chows and Labrador Retrievers, and 20% of epithelial neoplasms affected Labrador Retrievers. In cats, most tumors were epithelial (158 cases; 94% of neoplasms), followed by mesenchymal (8 cases; 5% of neoplasms) and round cell neoplasms (1 case; 1% of neoplasms). Over 50% of neoplasms of cats affected domestic shorthair cats. Although the percentage of lingual biopsies that had a neoplastic diagnosis was roughly the same between species, the diversity of neoplasms was much greater in dogs than in cats.

A New Variant of Avian Encephalomyelitis Virus Associated with Neurologic Signs in Turkey Poults.

Avian encephalomyelitis (AE) is a disease caused by the avian encephalomyelitis virus (AEV) of the genus Tremovirus in the family Picornaviridae. Recently, cases of turkey poults showing neurological signs were submitted to the veterinary diagnostic laboratories at South Dakota State University and the University of Minnesota. The affected birds were showing nervous neurological signs such as tremors, inability to stand, torticollis, and wing drop. Clinical signs were observed by 3 weeks of age. Necropsy of birds revealed no significant gross lesions in the internal organs, including the brain. There was no significant bacterial growth in the brains. Microscopic examination of various sections of the brain revealed multifocal lymphocplasmacytic perivascular cuffs in the cerebellum and cerebral cortex. The brain samples were processed for detection and whole genome sequencing by next-generation sequencing. Three full-length polyprotein sequences (6405 nt) of AEV were assembled. All three sequences shared 99.9-100% nucleotide and 100% amino acid identities with each other. Only 77.7-78.5% of nucleotide and 90.3-92.5% of amino acid identities with AEV field strains and vaccine sequences were available in GenBank. This indicates that a new divergent variant of AEV is circulating in the field and causing AE outbreaks in the Midwest region.

Reverse zoonosis of the 2022–2023 human seasonal H3N2 detected in swine

The Iowa State University Veterinary Diagnostic Laboratory detected nineteen human-to-swine reverse zoonoses of the 2022–2023 human seasonal H3N2 between November 2022 and November 2023. Cases from seven U.S. locations were detected: 3 Colorado, 1 Illinois, 1 Indiana, 2 Missouri, 7 North Carolina, 1 Ohio, and 1 Pennsylvania. One additional case was detected in Mexico and two cases were identified from Chile. Case samples were comprised of 4 nasal swabs and 15 oral fluids. Virus was successfully isolated from two of four nasal swab samples, but isolation from oral fluids was unsuccessful. The swine detections of H3 human viruses were classified to one of two human-seasonal H3 clades, 3C.2a1b.2a.2b and 3C.2a1b.2a.2a.1. Phylogenetic inference indicated at minimum 7 reverse zoonotic events occurred, with possible swine-to-swine transmission following the initial spillover. Twelve neuraminidase genes were sequenced, and nine were classified as human-seasonal H3N2 lineage: the remaining were endemic swine IAV NA genes from the N2.2002B, N2.1998, or the N1.Classical lineage, suggesting reassortment. The two viral isolates obtained from nasal swab samples were sequenced and were entirely human-lineage viruses. Seven swine samples with human seasonal H3 were sequenced and revealed co-detections with H1 1A.3.3.3 (gamma), with internal gene segments from both the triple reassortant internal gene (TRIG) and pandemic 2009 lineages. Serologic investigation of samples from swine production systems provided evidence for infection with human seasonal H3N2. One farm in the United States and four farms in Mexico had concurrent virologic evidence. The swine-isolated 3C.2a1b.2a.2b H3N2 was antigenically distinct from endemic 1990.4.A, 2010.1, and 2010.2 swine H3N2 lineages, but retained antigenic similarity to a recent human seasonal H3N2 (A/Darwin/6/2021). Pigs experimentally inoculated with a representative isolate demonstrated replication in the nose and lungs and minimal to mild macroscopic and microscopic lung lesions, but primary pigs did not transmit the virus to indirect contacts. If sustained in the pig population, this human seasonal H3 would represent the first new lineage detected in pigs the 2020 decade and present an emerging threat to swine health and production.

Detection and differentiation of low virulence and virulent Orthoavulavirus javaense using a molecular beacon with RT-LAMP

Newcastle disease (ND), an economically important disease in poultry, is caused by virulent strains of the genetically diverse Orthoavulavirus javaense (OAVJ). Laboratories rely on quantitative real-time reverse transcription PCR (qRT-PCR) to detect OAVJ and differentiate between OAVJ pathotypes. This study demonstrates that a fusion cleavage site based molecular beacon with reverse transcription loop mediated isothermal amplification (MB-RT-LAMP) assay can detect and differentiate OAVJ pathotypes in a single assay. Data show that the assay can rapidly identify diverse OAVJ genotypes with sensitivity only one log-fold lower than the current fusion qRT-PCR assay (104 copies), exhibits a high degree of specificity for OAVJ, and the molecular beacon can differentiate mesogenic/velogenic sequences from lentogenic sequences. Further, data show that a two-minute rapid lysis protocol preceding MB-RT-LAMP can detect and differentiate OAVJ RNA from both spiked samples and oropharyngeal swabs without the need for RNA isolation. As the MB-RT-LAMP assay can rapidly detect and discriminate between lentogenic and mesogenic/velogenic sequences of OAVJ within one assay, without the need for RNA isolation, and is adaptable to existing veterinary diagnostic laboratory workflow without additional equipment, this assay could be a rapid primary screening tool before qRT-PCR based validation in resource limited settings.

Evaluation of the diagnostic sensitivity and specificity of two pen-side tests for detecting African swine fever virus in experimentally infected pigs

African swine fever virus (ASFV) has spread through many countries and regions worldwide, causing significant losses. Timely detection of ASFV-infected pigs is crucial for disease control. In this study, we assessed the performance of two pen-side tests: a portable real-time PCR (qPCR) test for detecting viral genomic DNA and a lateral flow immunoassay (LFIA) for detecting viral antigens. To determine the time from infection to the earliest detection, 10 ASFV-seronegative pigs were inoculated intramuscularly with 104.0 hemadsorption dose 50 of a highly virulent ASFV strain. Whole blood and oral swab samples were alternately collected from each group of five pigs daily until all succumbed to the infection. Samples were promptly subjected to the two pen-side tests upon collection, and a subset was transported to a veterinary diagnostic laboratory for analysis using a reference qPCR assay. Viral genomic DNA was consistently detected by the reference qPCR assay in all blood samples from 2 days postinfection (dpi), preceding the onset of clinical signs, and in oral swabs from 4 dpi onwards. The portable qPCR test demonstrated comparable performance to the reference qPCR assay for both whole blood and oral swab samples. The LFIA exhibited 100% specificity when testing with whole blood samples but showed reduced sensitivity, particularly for blood samples collected early or late after infection. The antigen test did not perform well with oral swabs.

Neoplastic and non-neoplastic lesions in biopsy samples from pet rabbits in Hong Kong: a retrospective analysis, 2019-2022.

Rabbits are popular pets in the urban environment of Hong Kong, ranking third behind cats and dogs. Here we describe the frequency of neoplastic and non-neoplastic lesions in biopsies from pet rabbits submitted to the CityU Veterinary Diagnostic Laboratory between 2019 and 2022, comprising 247 tissue samples from 243 rabbits collected by veterinarians in 19 veterinary clinics. Among the 243 rabbits, there were 128 females (65 spayed), 114 males (54 castrated); sex information was not provided for 1 rabbit. The rabbit breeds included 45 Lionhead, 35 Dwarf, 14 Lop, 11 Dwarf Lop, 5 French Lop, 3 Angora, 2 Dutch, 2 Holland Lop, and 1 each of Netherland Dwarf, Velveteen, Mini Lop, and New Zealand White. The mean ages of rabbits with neoplastic and non-neoplastic lesions were 7.1 and 5.7 y, respectively. The most common neoplastic lesions were adenocarcinoma (26.4%), trichoblastoma (21.4%), sarcoma (9.4%), and thymoma (8.2%). The most common non-neoplastic lesion was uterine cystic endometrial hyperplasia (14.8%), followed by dermal abscess formation in the ventral abdomen or skin of the head (12.5%). Although a broad spectrum of other lesions was described, our findings in biopsies from pet rabbits in Hong Kong are consistent with those in other jurisdictions.

Comparative Analysis of Methicillin-Resistant Staphylococcus pseudintermedius Prevalence and Resistance Patterns in Canine and Feline Clinical Samples: Insights from a Three-Year Study in Germany.

The emergence of methicillin-resistant Staphylococcus pseudintermedius (MRSP) presents a significant public health concern globally, particularly within veterinary medicine. MRSP's resistance to multiple antibiotics is limiting treatment options and potentially leading to severe infections in companion animals. This study aimed to understand antimicrobial resistance in dogs and cats, focusing on MRSP resistance patterns and its prevalence in Germany. We analyzed results of bacterial diagnostic samples from canines and felines, sourced from a German veterinary diagnostic microbiology laboratory between 2019 and 2021. This dataset included samples from 3491 veterinary practices, covering 33.1% of veterinary practices and clinics in Germany. MRSP rates were detailed by host species, sample types and co-resistance patterns. Analysis of 175,171 bacterial examination results revealed S. pseudintermedius in 44,880 samples, yielding a 25.6% isolation rate. S. pseudintermedius was more prevalent in dogs (35.0%) than cats (3.6%). Methicillin resistance was found in 7.5% of all S. pseudintermedius isolates. MRSP prevalence was higher in feline samples (16.1%, 95% CI 14.4-17.8) compared to canine samples (7.1%, 95% CI 6.8-7.0). S. pseudintermedius showed high resistance rates to ampicillin (cats: 48.6%, dogs: 67.6%) and clindamycin (cats: 37.2%, dogs: 32.7%), while MRSP exhibited high co-resistance to clindamycin (cats: 82.8%, dogs: 85.4%) and sulfamethoxazole + trimethoprim (cats: 66.4%, dogs: 66.2%). Our study revealed distinct resistance patterns of MRSP in cats compared to dogs, highlighting the need for tailored treatment approaches and the importance of antimicrobial resistance surveillance.

Prevalence, risk factors, and hematologic changes in dogs from Baja California with presence of Ehrlichia spp., and coinfection with Anaplasma spp.

Ehrlichia and Anaplasma are obligate intracellular, gram-negative bacteria with tropism for hematopoietic cells, especially leukocytes and platelets. There are several Ehrlichia species that infect dogs. Ehrlichia canis is transmitted by Rhipicephalus sanguineus and replicates within monocytes and macrophages, leading to canine monocytic ehrlichiosis, a disease of worldwide distribution. The clinical signs are varied and non-specific. Anaplasma has two species that infect dogs, Anaplasma phagocytophilum and Anaplasma platys, the second also transmitted by Rhipicephalus sanguineus. This study aimed to evaluate the epidemiology and hematologic changes associated with the presence of Ehrlichia spp. and Anaplasma spp. coinfection in dogs from Baja California. Complete hematological analysis, examination of buffy coat smears, and ELISA tests were performed on blood samples from three veterinary diagnostic laboratories from Mexicali and Tijuana cities in Baja California, Mexico. A total of 5,469 dog samples were analyzed. The overall prevalence of Ehrlichia spp., was 4.79%, with a distribution of 6.3% in Mexicali (OR: 2.39 CI: 1.69-3.17) and 2.5% in Tijuana. The peak of infection was found in September. Contact with other dogs and tick infestation were the risk factors associated with infection (P < 0.05). There was 96% co-infection only in Tijuana and 0% in Mexicali. Anemia, thrombocytopenia, and hyperproteinemia are associated with Ehrlichia spp., and Ehrlichia/Anaplasma infection. In view of the foregoing, we have to maintain epidemiologic vigilance, as well as look further into the ticks present in the state and the possibility of transmission of unusual pathogens.

The role of veterinary diagnostic laboratories during COVID-19 response in the United States.

Robust testing capacity was necessary for public health agencies to respond to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the coronavirus disease 19 (COVID-19) pandemic. As the nation faced the need for robust testing capacity, it became necessary to use all possible resources. In many cases, veterinary diagnostic laboratories rose to meet this demand because these facilities routinely perform high throughput diagnostic testing of large animal populations and are typically familiar with pathogens of high pandemic concern. In this study, we evaluated the impact of veterinary diagnostic laboratories in the United States on SARS-CoV-2 testing. Results of surveys, semi-structured interviews, and analysis of publicly available information showed that veterinary diagnostic laboratories had a substantial impact on human health through population-level testing in the COVID-19 response, supporting timely and informed public health interventions. This success was not without significant hurdles, as many participating veterinary diagnostic laboratories experienced restriction in their response due to difficulties obtaining the Clinical Laboratory Improvement Amendments (CLIA) certification required to conduct human diagnostic testing. Our results point out the importance of reducing hurdles before the next major public health emergency to enhance access to testing resources overall and to ultimately improve population health.

Prevalence and molecular epidemiology of carbapenemase-producing Enterobacterales isolated from dog and cat faeces submitted to veterinary laboratories in the USA.

To estimate the prevalence of carbapenemase-producing Enterobacterales (CPE) carriage among pets using faecal specimens submitted to veterinary diagnostic laboratories throughout the US. A secondary aim was to employ whole-genome sequencing (WGS) to characterize isolates of CPE from companion animals and compare them to publicly available CPE genomes. To estimate the prevalence of CPE in companion animals in the USA, a multicenter surveillance study including 8 different veterinary diagnostic laboratories from across the USA was conducted. Briefly, remnant faecal specimens from dogs and cats were screened using two selective agar plates (CHROMID Carba and MacConkey with 1 mg/L cefotaxime and 0.125 mg/L meropenem) and presumptive CPE isolates screened by the modified carbapenemase inactivation method for carbapenemase production. A total of 2393 specimens were screened and yielded 196 isolates for carbapenemase screening. A total of 5 isolates from 4 dogs and 1 cat at 3 different veterinary diagnostic laboratories were confirmed to produce a carbapenemase (0.21%). Whole-genome sequencing (WGS) revealed two E. coli (ST167) isolates that both produced an NDM-5 carbapenemase, two Enterobacter hormaechei (ST171) isolates that produced an NDM-5 carbapenemase and a KPC-4 carbapenemase respectively and one Klebsiella oxytoca (ST199) that produced an Oxa-48-type carbapenemase. Both E. coli isolates were found to be within at least 22 SNPs of previously characterized canine and human CPE isolates. This study demonstrates that the prevalence of CPE among companion animals is relatively low (0.21%) but that given the genetic relatedness of animal isolates to human isolates, additional surveillance is needed.

Retrospective study of laboratory-based surveillance of rabies in wild and domestic animals in the southern United States, 2010-2021.

We performed a retrospective study of all case submissions for the rabies virus (RABV) direct fluorescent antibody test (DFAT) requested of the Tifton Veterinary Diagnostic and Investigational Laboratory (Tifton, GA, USA) between July 2010 and June 2021. Submitted were 792 samples from 23 animal species from 89 counties in Georgia, and 4 neighboring counties in Florida, 1 in South Carolina, and 1 in Alabama. In 13 (1.6%) cases, the DFAT result was inconclusive; 779 (98.4%) cases had a conclusive (positive or negative) test result. Of these 779 cases, 79 (10.1%) tested positive across 10 species. The remaining 700 (89.9%) cases were negative. The main reason for submission for RABV testing was human exposure to a potentially rabid animal in 414 (52.3%) cases. Among the 79 positive cases, 74 (93.7%) involved wildlife; raccoons (51 cases; 68.9%) were the primary host confirmed with RABV infection, followed by skunk and fox (8 cases each; 10.8%), bobcat (5 cases; 6.8%), and bats (2 cases; 2.7%). Only 5 domestic animals (6.3% of the positive cases) tested positive during our study period; one from each of the bovine, canine, caprine, equine, and feline species. Hence, the sylvatic cycle plays the predominant role in circulating RABV infection in our study area.

The use of telepathology in veterinary medicine: a scoping review.

Telepathology, as a subset of teleconsulting, is pathology interpretation performed at a distance. Telepathology is not a new phenomenon, but since ~2015, significant advances in information technology and telecommunications coupled with the pandemic have led to unprecedented sophistication, accessibility, and use of telepathology in human and veterinary medicine. Furthermore, telepathology can connect veterinary practices to distant laboratories and provide support for underserved animals and communities. Through our scoping review, we provide an overview of how telepathology is being used in veterinary medicine, identify gaps in the literature, and highlight future areas of research and service development. We searched MEDLINE, CAB Abstracts, and the gray literature, and included all relevant literature. Despite the widespread use of digital microscopy in large veterinary diagnostic laboratories, we identified a paucity of literature describing the use of telepathology in veterinary medicine, with a significant gap in studies addressing the validation of whole-slide imaging for primary diagnosis. Underutilization of telepathology to support postmortem examinations conducted in the field was also identified, which indicates a potential area for service development. The use of telepathology is increasing in veterinary medicine, and pathologists must keep pace with the changing technology, ensure the validation of innovative technologies, and identify novel uses to advance the profession.

Multi-drug resistant Salmonella Dublin cultured from cryopreserved Holstein semen

Salmonella enterica subspecies enterica serotype Dublin, com­monly referred to as Salmonella Dublin, has emerged as one of the most isolated serotypes of Salmonella in U.S. cattle. A highly multi-drug resistant isolate of Salmonella Dublin was cultured at the Wisconsin Veterinary Diagnostic Laboratory (WVDL) from both the raw and cryopreserved, extended semen of a 17-month-old clinically healthy Holstein bull. The objective of this study was to use antemortem and postmortem diagnos­tic testing to evaluate the systemic health of the bull and deter­mine where he was harboring the Salmonella Dublin infection.

100 Evaluating the effect of liver copper concentration on vaccine response in lightweight dairy-beef steers

Abstract Dairy-beef crossbred calves often face greater copper (Cu) exposure than native beef calves, which may impact their response to vaccines. To investigate this, 28 weaned dairy-beef steers (90.25 ± 1.28 kg; ~8 wk old) were enrolled in a study to determine the effect of Cu status on the antibody response to vaccine. Steers were stratified by body weight (BW) and assigned to one of two dietary treatments: low Cu (no supplemental Cu) and high Cu (supplementing 20 mg Cu/kg DM). Steers were housed in pens of six to eight animals for 82 d before d 0 of the vaccination period to create distinct liver Cu groupings. Seven days before initial vaccine dose, liver biopsies were collected, and steers were grouped into low (n = 13) and high (n = 15) Cu treatments. Liver Cu for low Cu averaged 291 ± 24 mg/kg DM (range 240 to 376 mg/kg DM) and for high Cu averaged 665 ± 23 mg Cu/kg liver DM (range 519 to 893 mg/kg DM). All calves were vaccinated on d 0 and boostered 21 d later with Bovilis Vista 5 SQ (Merck Animal Health, Rahway, NJ) and an ovalbumin vaccine. Blood and BW were collected on these days and again on d 49 to end the study. Serum antibody titers for BRSV, BHV1, BVDV1, and BVDV2 were analyzed by the Iowa State University Veterinary Diagnostic Laboratory (Ames, IA). Response to ovalbumin vaccination was determined using an ELISA. Plasma Cu concentrations were determined on d 0, 21, and 49 via ICP-OES. Ovalbumin antibodies were determined by calculating the ratio of each sample to a positive control. Titers, ovalbumin response, and plasma Cu data from d 0, 21, and 49 were analyzed using the Mixed procedure of SAS 9.4 (Cary, NC) with the fixed effect of Cu treatment and day as repeated. Plasma Cu decreased across days (P = 0.0008). Given both treatments had adequate liver Cu, it is unsurprising plasma Cu did not differ by treatment (P = 0.2; 1.02 and 1.07 mg Cu/L for low and high Cu, respectively). Antibodies for BRSV, BHV1, and BVDV2 increased across days (P < 0.0001), but there were no differences by treatment (P ≥ 0.12). Ovalbumin antibodies were affected by treatment by day (P = 0.04), where high Cu had greater response to ovalbumin than low Cu on d 21, but by d 49 ovalbumin antibodies were similar between Cu treatments. These results suggest excessive liver Cu concentration may not affect response to vaccination in dairy-beef steers; however, more research is needed to determine the effects of increased liver Cu concentration on oxidative stress and immune system function.