State University Veterinary Diagnostic Laboratory Research Articles

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.

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.

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.

PSI-4 Evaluation of chemical mitigants when soybean meal was inoculated with porcine epidemic diarrhea virus, porcine reproductive and respiratory virus, and Seneca Valley virus 1

Abstract Chemical mitigants have been found to decrease virus concentrations in feed and ingredient matrices. Continued research is needed to identify the appropriate inclusion levels and application time for different viruses in these matrices. Therefore, the objective was to evaluate different inclusion levels of two different chemical mitigants when applied either before or after inoculation of porcine epidemic diarrhea virus (PEDV), porcine reproductive and respiratory syndrome virus (PRRSV) and Seneca Valley virus 1 (SVV1) to soybean meal. The experiment was designed in a 2×2×2 factorial with two chemical mitigants, a formaldehyde-based product (Termin-8, Anitox Corp. Lawrenceville, GA) and a product containing Synergistic blend of phytochemicals and carboxylic acids (Finio, Anitox Corp. Lawrenceville, GA), two applications times, before virus inoculation (prevention) and after virus inoculation (mitigation), and either a low or high inclusion level (Termin-8: 2 or 3 kg/MT; Finio: 1.75 or 2.75 kg/MT). On d 1, samples of the respective matrices were weighed in 50 g aliquots and added to 500 mL bottles. Chemical mitigants were applied to the prevention samples at their respective inclusion levels and 50 µL each of 1×107 TCID50/mL PEDV, 1×108 TCID50/mL PRRSV, and 1×108 TCID50/mL SVV1 were added to the mitigation samples. All bottles were shaken and allowed to sit at room temperature for 24 h. On d 2, virus was added to the prevention samples and chemical mitigants were added to the mitigation bottles, samples incubated at room temperature for 1 h before 200 mL of phosphate buffer solution was added, and bottles incubated at 4°C for 18 h. After incubation, liquid from the samples was centrifuged and the supernatant analyzed via triplex PCR at Kansas State University Veterinary Diagnostic Laboratory. Cycle threshold and proportion PCR positive were analyzed using SAS GLIMMIX v 9.4 (SAS, Inc., Cary, NC) with each virus and matrix combination analyzed individually. A chemical mitigant×application time×inclusion level interaction was not observed (P > 0.05) for any virus. A chemical mitigant×application time interaction was observed (P < 0.05) in all three viruses, where less viral RNA was detected (P < 0.05) when either Termin-8 or Finio were applied after inoculation (mitigation) than when applied prior to inoculation or to the positive controls. An application time×inclusion level interaction was noticed (P < 0.05) for SVV1 and PEDV in soybean meal. Chemical mitigants applied at either the low or high inclusion level after inoculation (mitigation) had less detectable RNA (P < 0.05) than their prevention counterparts. Overall, Termin-8 and Finio can reduce detectable RNA immediately in contaminated soybean meal, with ongoing research evaluating the effect of extended contact times between the mitigants and soybean meal.

212 Growth and behavior of Yorkshire gilts exhibiting physiological resilience to weaning stress

Abstract Weaning is an acute early-life stressor and therefore, an excellent model for examining lasting impacts of stress resilience. Our objective was to identify gilts exhibiting physiological resilience or vulnerability to weaning stress and characterize long-term impact on growth and behavior. Purebred Yorkshire litters born to multiparous sows from a single farrowing group were used. At weaning (26 ± 1 d of age), blood samples were collected from all gilt piglets of 13 litters (n = 101) at -1 d, 0 d, and +4 d pre- and post-weaning. Serum cortisol concentrations were quantified by the Michigan State University Veterinary Diagnostic Laboratory. For each litter, serum cortisol was used to identify the gilt most capable of returning to baseline concentrations by +4 d as stress resilient (SR, n = 13) and the gilt least capable as stress vulnerable (SV, n = 13). Skin lesions were recorded at -1 d, +1 d, and +4 d at weaning and when gilts were mixed into grow-finish cohorts. Body weights were recorded at birth, 4 wk, 8 wk, and 12 wk and used to calculate average daily gain (ADG) for the farrowing, nursery, and early grow-finish stages. To assess behavior, focal gilts underwent a social dyad test in a novel arena at 7 wk, a handling test at 8 wk, and novel object test at 12 wk. We observed no differences in skin lesions at weaning, but at 8 wk SR gilts had significantly more rear lesions -1 d pre-mixing (P = 0.031) and middle lesions 4 d post-mixing (P = 0.049). No differences in body weight or ADG were observed at any stage. We did not observe differences between SR and SV pigs during the social dyad or handling tests. During the novel object test, SR gilts exhibited a tendency towards increased activity as demonstrated by a greater number of total zone crosses, 1 m line crosses, and 0.5 m line crosses (P = 0.062, 0.091, and 0.103, respectively). In conclusion, we found gilts resilient to weaning stress were involved in more agonistic behavior at mixing and exhibited increased activity levels in a novel situation. The absence of growth differences supports the potential to select for resilience without negatively impacting growth performance, and the observed behavioral differences could suggest implications for ease of handling and adaptability if females are retained for breeding.

198 Evaluation of zinc source and concentration on growth performance of nursery swine

Abstract Trace minerals (TM) supplementation in swine diets can negatively interact with other nutrients. Zinc (Zn) and copper (Cu) mutually inhibit their intestinal absorption; thus, limiting their bioavailability. Recent peer-reviewed work has suggested that nursery pigs fed a diet supplemented with increased Zn experienced impaired Cu metabolism, negatively impacting growth performance. However, it has been demonstrated that mitigation of antagonisms might be possible with the use of organic TM supplemented at decreased concentrations. To validate these findings, a total of 2,629 pigs [initial body weight (BW) = 6.2 kg] were used in a 37-d experiment. Pens of 36 pigs were randomly allotted to 1 of 8 dietary treatments with 9 replications per treatment. Dietary treatments were formulated to contain 1 of 4 Zn sources: Zn Sulfate, Zn Hydroxychloride A (Intellibond Zn [Micronutrients Inc., Indianapolis, IN), Zn Hydroxychloride B [Zn Hydroxychloride (Alltech, Inc., Nicholasville, KY)] or Organic Zn [Bioplex Zn (Alltech, Inc)] at 1 of 2 concentrations: 125 or 250 ppm. The concentrations utilized for Organic Zn treatment were 62.5 or 125 ppm per published recommendations. Experimental diets were fed in two phases: phase 1 from d 0 to 21 and phase 2 from d 22 to 37. Added Cu in both dietary phases was 10 ppm. At d 37, serum and liver samples (data not showed) were collected from 1 barrow per pen and submitted to Michigan State University Veterinary Diagnostic Laboratory to determine Zn and Cu concentration. No significant source × concentration interactions were observed for growth performance in this study. From d 0 to 37, pigs fed the greatest Zn concentration had less (P = 0.040) average daily feed intake (ADFI) and marginally decreased (P = 0.060) average daily gain (ADG), despite Zn source (Table 1). In addition, a marginal improvement (P = 0.070) in gain to feed ratio (G:F) was observed in pigs fed diets containing Organic Zn, regardless of concentration. For mineral concentrations in serum, there was a marginally significant source × concentration interaction (P = 0.078), where pigs fed diets with 125 ppm of Zn Hydroxychloride B had one of the greatest Zn concentrations, yet it was drastically reduced when pigs were fed diets with 250 ppm. Furthermore, Cu concentration in serum was decreased (P = 0.045) in pigs fed the greatest concentration of Zn, despite source. In conclusion, these results suggest that greater dietary Zn concentration impaired Cu absorption, reduced feed intake, and marginally decreased growth. Organic Zn, included at 50% reduced concentrations, marginally improved feed efficiency.

West Nile Virus Infection in a Pekin Duck ( Anas platyrhynchos domesticus ) Developer Flock.

Developer ducks are ducks being reared for breeding. Like breeder candidate chickens, they are raised with appropriate light and feed programs. A commercial Pekin duck (Anas platyrhynchos domesticus) developer flock experienced an extraordinary, elevated mortality event at 6 wk of age. Weekly mortality rate that week was 162 ducklings out of a flock of 6420 (2.5%). Mortality jumped to 988 (15.4%) ducklings the next week. On first elevated mortality, six dead ducks from that flock were submitted for diagnostic investigation at Michigan State University Veterinary Diagnostic Laboratory. Myocarditis, pale striping or diffuse pallor of the epicardium, was grossly evident in five of the six submitted ducklings. All of the ducklings had hydropericardium, three had ascites, and three had congested meninges. Histology confirmed myocarditis with myocardial necrosis. Cerebrum and brainstem had lymphocytic vasculitis with rare neuronal necrosis in affected areas, as well as Purkinje cells in the cerebellum. West Nile virus was confirmed by PCR the day after submittal and by immunohistochemistry soon thereafter.

Detection and disease diagnosis trends (2017–2022) for Streptococcus suis, Glaesserella parasuis, Mycoplasma hyorhinis, Actinobacillus suis and Mycoplasma hyosynoviae at Iowa State University Veterinary Diagnostic Laboratory

BackgroundAccurate measurement of disease associated with endemic bacterial agents in pig populations is challenging due to their commensal ecology, the lack of disease-specific antemortem diagnostic tests, and the polymicrobial nature of swine diagnostic cases. The main objective of this retrospective study was to estimate temporal patterns of agent detection and disease diagnosis for five endemic bacteria that can cause systemic disease in porcine tissue specimens submitted to the Iowa State University Veterinary Diagnostic Laboratory (ISU VDL) from 2017 to 2022. The study also explored the diagnostic value of specific tissue specimens for disease diagnosis, estimated the frequency of polymicrobial diagnosis, and evaluated the association between phase of pig production and disease diagnosis.ResultsS. suis and G. parasuis bronchopneumonia increased on average 6 and 4.3%, while S. suis endocarditis increased by 23% per year, respectively. M. hyorhinis and A. suis associated serositis increased yearly by 4.2 and 12.8%, respectively. A significant upward trend in M. hyorhinis arthritis cases was also observed. In contrast, M. hyosynoviae arthritis cases decreased by 33% average/year. Investigation into the diagnostic value of tissues showed that lungs were the most frequently submitted sample, However, the use of lung for systemic disease diagnosis requires caution due to the commensal nature of these agents in the respiratory system, compared to systemic sites that diagnosticians typically target. This study also explored associations between phase of production and specific diseases caused by each agent, showcasing the role of S. suis arthritis in suckling pigs, meningitis in early nursery and endocarditis in growing pigs, and the role of G. parasuis, A. suis, M. hyorhinis and M. hyosynoviae disease mainly in post-weaning phases. Finally, this study highlighted the high frequency of co-detection and -disease diagnosis with other infectious etiologies, such as PRRSV and IAV, demonstrating that to minimize the health impact of these endemic bacterial agents it is imperative to establish effective viral control programs.ConclusionsResults from this retrospective study demonstrated significant increases in disease diagnosis for S. suis, G. parasuis, M. hyorhinis, and A. suis, and a significant decrease in detection and disease diagnosis of M. hyosynoviae. High frequencies of interactions between these endemic agents and with viral pathogens was also demonstrated. Consequently, improved control programs are needed to mitigate the adverse effect of these endemic bacterial agents on swine health and wellbeing. This includes improving diagnostic procedures, developing more effective vaccine products, fine-tuning antimicrobial approaches, and managing viral co-infections.

Nocardia farcinica abortion in a goat.

A female goat fetus was received by the Colorado State University-Veterinary Diagnostic Laboratory following an isolated abortion of twins by a reportedly healthy doe. Postmortem examination did not reveal any gross abnormalities. Histologic evaluation revealed pyogranulomatous and necrotizing bronchopneumonia with intracellular and extracellular gram-positive and non-acid-fast filamentous bacilli. Aerobic culture of the stomach contents and pooled lung and liver tissue yielded light growth of Nocardia sp., which was identified by MALDI-TOF MS and 16s rDNA sequencing as Nocardia farcinica.

PSII-16 Comparison of Microencapsulated Zinc Oxide and Conventional Zinc Oxide on Weanling Pig Small Intestinal Morphology

Abstract A total of 300 pigs (DNA 200 × 400; initially 6.04 ± 0.08 kg BW) were weaned at an average of 21 d of age and used in a 42-d experiment with 12 replications per treatment. At weaning (experiment d 0), pigs were randomly allotted to one of five dietary treatments: 1) negative control (standard nursery diet containing 110 ppm Zn from trace mineral premix); 2) control diet with 3,000 ppm added Zn in the form of ZnO in phase 1 and 2,000 ppm added Zn in the form of ZnO in phase 2 (High-ZnO); 3) control diet with 400 ppm added Zn in the form of ZnO in phases 1 and 2 (Low-ZnO); 3) 3,000 ppm added Zn in the form of microencapsulated ZnO in phase 1 and 2,000 ppm added Zn in the form of microencapsulated ZnO in phase 2 (High-MZnO); 5) 400 ppm added Zn in the form of microencapsulated ZnO in phases 1 and 2 (Low-MZnO). Pigs were fed dietary treatments from d 0 to 28. On d 28, a subset of pigs with a BW closest to the treatment mean (n = 30 total; 6 per treatment) were euthanized via captive bolt and necropsied for collection of small intestinal tissue to evaluate morphology at the Kansas State University Veterinary Diagnostic Laboratory. Briefly, after pigs were slaughtered, they were opened viscerally, and the small intestine was identified. Two samples (approximately 2 cm long) were cut from the proximal and distal ends of the ileum and fixed in formalin. Samples were then embedded in paraffin and 4 µm cross-sections were cut and stained with hematoxylin and eosin. Slides were analyzed using a microscope at 10× magnification. Measurements [villus height (VH), crypt depth (CD), and villus height:crypt depth ration (VH:CD)] were evaluated from 10 unbroken villi and their corresponding crypts. Data were analyzed using the GLIMMIX procedure of SAS (version 9.4) with individual pig as the experimental unit. No evidence of differences in proximal ileal VH (P = 0.34), proximal ileal CD (P = 0.52), and proximal ileal VH:CD (P = 0.49) were observed between dietary treatments (Table 1). Likewise, there was no evidence of differences in distal ileal VH (P = 0.63), CD (P = 0.53), or VH:CD (P = 0.80) between dietary treatments. In summary, feeding a microencapsulated form of ZnO did not appear to alter small intestinal morphology in weanling pigs compared with conventional, unprotected ZnO.

Case report: Diagnostic findings of bovine neurological cases at the Iowa State University Veterinary Diagnostic Laboratory between January 2017 and May 2021

This study reviews the Iowa State University Veterinary Diagnostic Laboratory (ISU-VDL) computer records of bovine neurological cases submitted from January 2017 to May 2021. There were 236 cases diagnosed as follows: listeriosis (83), polioencephalomalacia (44), thrombomeningoencephalitis (23), miscellaneous bacteria (22), lead toxicosis (21), nitrate toxicosis (18), urea toxicosis (9), nervous coccidiosis (5), hypomagnesemia (5), sodium toxicosis (3), rabies (1), herpesvirus type 6 (1), and epsilon toxin from Clostridium perfringens type D (1). Most histories did not provide specific descriptions of the type of neurological signs, which could better target diagnostic investigation. However, each condition had specific peculiarities relevant from a post-mortem diagnostic perspective. For listeriosis, confirmation of Listeria monocytogenes using IHC was considered superior to bacterial culture, with 16/19 (84%) positive by IHC compared to 23/59 (38.9%) of cases positive by culture. In 44/83 (53.0%) of cases, histopathological examination mentioned that cerebellum and cerebrum were unremarkable, implying that without submission of the entire brain, particularly brain stem and obex, listeriosis cannot be ruled out. In 44 cases histologically diagnosed as polioencephalomalacia, when ultraviolet light (Wood’s lamp) was performed, it revealed multifocal areas of apple-green autofluorescence in 13/21 (61.9%) of brains, indicating that false negatives can occur with this technique. A substantial number of submissions were toxicoses that required specific sample types to confirm a diagnosis. For lead, whole blood, liver or kidney was necessary. For nitrate and urea toxicosis, appropriate samples were ocular fluid or the incriminating feed/water source. Confirmation of sodium toxicosis relied on sodium analysis of brain tissue.

Diagnosis of Anaplasma marginale in cattle at the Iowa State University veterinary diagnostic laboratory 2003–2021

Anaplasma marginale is a blood-borne rickettsia-like organism that infects cattle erythrocytes and causes anaplasmosis. This study reviews diagnostic data of all A. marginale diagnostics performed from 2003 to August 2021 in the Iowa State Veterinary Diagnostic Laboratory. Typically, the referring veterinarian's initial tentative diagnosis was based on presenting clinical signs or necropsy findings. Confirmatory testing at the ISU-VDL consisted of light microscopy evaluation of stained blood smears or molecular diagnostic procedures. A total of 94 cases were submitted with tissue samples from deceased animals, of which 79 were from Iowa and 15 were from other states. The most typical gross lesions were widespread yellow adipose tissue and splenomegaly. Typical histopathological lesions included marked bile stasis and hemosiderin-laden macrophages in the liver and spleen, respectively. Starting in 2013, when PCR was implemented to confirm cases of anaplasmosis, 315/1125 (28%) were positive to A. marginale, and 810 were negative, using a cut-off of 35.0 Ct. The average (±SD) of the positive PCR Ct was 19.5 (±6.0), and the first and third quartiles were 14.9 and 23.4. Most cases occurred between August and November, peaking in September, whether from necropsies or positive blood samples by PCR. The most common tick observed in Iowa, Dermacentor variabilis, is likely the main vector for transmission. Further surveys should be conducted to estimate seroprevalence by geographical location, the density of cattle populations, distribution of known vectors according to season, and strains of A. marginale.

Abstract P3-08-02: Prospective Use of Feline Mammary Carcinomas to Study MiR-10b and Metastasis

Abstract Despite many advances in cancer treatment, metastatic disease is estimated to be responsible for 90% of all cancer-related deaths. Current treatments for metastatic disease target various aspects of carcinogenesis but not specifically the metastatic process, representing a major unmet clinical need. MicroRNA-10b, a small non-coding RNA, offers tremendous potential as a treatment target for metastatic disease. It is implicated in invasion, migration, and viability of metastatic cells across a variety of cancer types, and it is upregulated in metastases compared to their matched primary tumor, establishing miR-10b as a potential treatment target unique to metastatic niche. We have previously developed a therapeutic targeting miR-10b in metastases and tested it in murine models of metastatic breast cancer. This therapeutic, consisting of an anti-miR-10b antisense oligonucleotide conjugated to iron oxide-based nanoparticles, prevents metastasis and eradicates pre-existing metastases in murine breast cancer models. With an outlook to clinical translation of our approach, we seek to test our therapeutic strategy in larger animal models. Feline mammary carcinomas are considered by most to be the best large animal model for human breast cancer due to similarities such as relative age of onset, histopathology, metastatic patterns, and treatment response. To support the use of this model with our therapeutic, we investigated the characteristics of miR-10b expression in spontaneous metastatic breast cancer in companion cats. Archival blocks of matched primary tumors and metastatic lymph nodes from companion cats diagnosed with mammary carcinoma (n=9, 44%TNBC, 56%HER2+) were obtained from the tissue bank of the Michigan State University (MSU) Veterinary Diagnostic Laboratory (VDL). Tissues were analyzed for miR-10b and its target HOXD10 expression using qRT-PCR and in situ hybridization. qRT-PCR revealed that miR-10b expression was significantly upregulated in 55.5% of lymph node metastases compared to their matched primary tumor, mirroring findings in human metastatic cancer. This was validated by qRT-PCR for HOXD10 gene expression (a direct target of miR-10b), which was significantly downregulated in these metastases compared to their matched primary tumor. In situ hybridization demonstrated that miR-10b expression was increased at the invasive edge of tumors and in actively invading cells, suggesting miR-10b plays a similar role in invasion in feline breast cancer as it does in human breast cancer. Altogether, these findings support the use of feline mammary carcinomas as a model of human breast cancer and as an excellent candidate for treatment with our therapeutic. Citation Format: Alan Halim, N. Anna Savan, Paulo Vilar Saavedra, Vilma Yuzbasiyan-Gurkan, Matti Kiupel, Lorenzo Sempere, Anna Moore. Prospective Use of Feline Mammary Carcinomas to Study MiR-10b and Metastasis [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P3-08-02.

A survey of vitamin and trace mineral ranges for diagnostic lab reporting from conventionally raised swine

Objective: The purpose of this study was to survey the vitamin and mineral levels in various pig tissues at different phases of the life cycle. Materials and methods: Forty-eight healthy pigs of different stages of production were used for sampling of different tissues. Seven sows and a minimum of 10 animals from each phase of production (suckling, nursery, and finishing) were selected for sampling. A blood sample was collected via sterile venipuncture for serum vitamin and mineral analysis. After euthanasia, the diaphragm and liver were collected. Samples were submitted to the Iowa State University Veterinary Diagnostic Laboratory for analysis. Data were analyzed using SAS (version 9.4; SAS Institute Inc) and presented as minimum and maximum concentrations with standard error. The experimental unit was the animal. Results: Levels of vitamin A, vitamin E, copper, zinc, selenium, iron, and manganese were higher in liver tissues than in serum and diaphragm tissues. Diaphragm muscle had similar levels of phosphorus as the liver tissue. Serum had similar levels of calcium as the liver tissue. Implications: These data provide a sampling of vitamin and mineral levels present in tissues and serum of commercial pigs and suggests that vitamin and mineral levels differ between sampling sites.

Use of the ISU FLUture multisequence identity tool for rapid interpretation of swine influenza A virus sequences in the United States.

Rapid and reliable identification of the hemagglutinin (HA) and neuraminidase (NA) genetic clades of an influenza A virus (IAV) sequence from swine can inform control measures and multivalent vaccine composition. Current approaches to genetically characterize HA or NA sequences are based on nucleotide similarity or phylogenetic analyses. Public databases exist to acquire IAV genetic sequences for comparison, but personnel at the diagnostic or production level have difficulty in adequately updating and maintaining relevant sequence datasets for IAV in swine. Further, phylogenetic analyses are time intensive, and inference drawn from these methods is impacted by input sequence data and associated metadata. We describe here the use of the IAV multisequence identity tool as an integrated public webpage located on the Iowa State University Veterinary Diagnostic Laboratory (ISU-VDL) FLUture website: https://influenza.cvm.iastate.edu/. The multisequence identity tool uses sequence data derived from IAV-positive cases sequenced at the ISU-VDL, employs a BLAST algorithm that identifies sequences that are genetically similar to submitted query sequences, and presents a tabulation and visualization of the most genetically similar IAV sequence and associated metadata from the FLUture database. Our tool removes bioinformatic barriers and allows clients, veterinarians, and researchers to rapidly classify and identify IAV sequences similar to their own sequences to augment interpretation of results.

Gross and histologic description of trematodosis in fetal and neonatal beef calves in North Dakota and Minnesota.

Hepatic trematodes, such as Fasciola hepatica and Fascioloides magna, have variable distribution throughout the United States. F. magna is endemic in the upper midwestern United States, and F. magna infections are diagnosed frequently in weaned calves and adult beef cattle at the North Dakota State University Veterinary Diagnostic Laboratory (NDSU-VDL). Rarely, liver fluke infestation has also been observed in much younger calves, including aborted fetuses. We describe here, in 2 fetal and 7 neonatal beef calves submitted to the NDSU-VDL between 2011 and 2020, parasitic migration tracts in livers, consisting of regionally extensive, random, linear tracts of fibrosis admixed with black porphyrin pigment, along with foci of necrosis and hemorrhage, and mixed inflammatory cells, which were caused presumptively by F. magna infection. Samples were not available from our 9 cases for PCR assay and sequencing, but we did confirm F. magna within liver samples collected from regional cattle in 2020 and 2021. Fetal and neonatal trematodosis was often concurrent with other common causes of fetal abortion and neonatal calf loss in our cases; however, based on the prepatent period of F. magna, fetal and neonatal beef calf trematode infestations occurred in utero.

Salmonella enterica serovar Brandenburg abortions in dairy cattle.

Two separate late-term abortion outbreaks in Jersey heifers in July 2020 and December 2020 were investigated by the Iowa State University Veterinary Diagnostic Laboratory. We evaluated 3 whole fetuses and 11 sets of fresh and formalin-fixed fetal tissues during the course of the outbreaks. The late-term abortions were first identified at a heifer development site and subsequently observed at the dairy farm. Aborted fetuses had moderate-to-marked postmortem autolysis with no gross lesions identified. Observed clinical signs in cows at the dairy farm ranged from intermittent loose stools to acute post-abortion pyrexia and reduced feed intake. Routine histopathology and reproductive bacterial culture revealed acute, suppurative placentitis with moderate-to-heavy growth of Salmonella spp. group B from stomach contents, liver, placenta, and heifer fecal contents. Serotyping identified Salmonella enterica subsp. enterica serovar Brandenburg in all 14 fresh tissue cases, as well as individual and pooled heifer feces. Whole-genome sequencing analysis revealed that all isolates belonged to ST type 873 and possessed typhoid toxin genes, several fimbrial gene clusters, type III secretion system genes, and several pathogenicity islands. Abortions caused by Salmonella Brandenburg have not been reported previously in dairy cattle in the United States, to our knowledge.

Prevalence, Mechanism, Genetic Diversity, and Cross-Resistance Patterns of Methicillin-Resistant Staphylococcus Isolated from Companion Animal Clinical Samples Submitted to a Veterinary Diagnostic Laboratory in the Midwestern United States.

Methicillin-resistant Staphylococcus (MRS) is a leading cause of skin and soft tissue infections in companion animals, with limited treatment options available due to the frequent cross-resistance of MRS to other antibiotics. In this study, we report the prevalence, species distribution, genetic diversity, resistance mechanism and cross-resistance patterns of MRS isolated from companion animal (mostly dog and cat) clinical cases submitted to Iowa State University Veterinary Diagnostic Laboratory (ISU VDL) between 2012 and 2019. The majority of isolates were identified as Staphylococcus pseudintermedius (68.3%; 2379/3482) and coagulase-negative Staphylococcus (CoNS) (24.6%; 857/3482), of which 23.9% and 40.5% were phenotypically resistant to methicillin, respectively. Cross resistance to other β-lactams (and to a lesser extent to non-β-lactams) was common in both methicillin-resistant S. pseudintermedius (MRSP) and CoNS (MRCoNS), especially when oxacillin MIC was ≥4 μg/mL (vs. ≥0.5–<4 μg/mL). The PBP2a protein was detected by agglutination in 94.6% (521/551) MRSP and 64.3% (146/227) MRCoNS. A further analysis of 31 PBP2a-negative MRS isolates (all but one MRCoNS) indicated that 11 were mecA gene-positive while 20 were negative for mecA and other mec genes by PCR. The resistance to last-resort anti-staphylococcal human drugs (e.g., tigecycline, linezolid, vancomycin) among the MRS tested was none to very low. Even though genotyping indicated an overall high level of genetic diversity (87 unique PFGE patterns and 20 MLST types) among a subset of MRSP isolates tested (n = 106), certain genotypes were detected from epidemiologically connected cases at the same or different time points, suggesting persistence and/or nosocomial transmission. These results indicate a relatively high prevalence of MRS from companion animals in the Midwestern US; therefore, it is important to perform routine susceptibility testing of Staphylococcus in veterinary clinical settings for the selection of appropriate antimicrobial therapy.

ATP7A, ATP7B, and RETN genotypes in Labrador Retrievers with and without copper-associated hepatopathy.

To determine the frequency of previously reported coding variants in the ATP7A, ATP7B, and RETN genes in a US population of Labrador Retrievers and to explore potential associations of these genotypes with pathologic hepatic copper accumulation. Archived hepatic specimens from 90 Labrador Retrievers collected between 2013 and 2021. The Michigan State University Veterinary Diagnostic Laboratory database was searched to identify archived tissues from Labrador Retrievers that had undergone hepatic histopathologic assessment. Cases were classified into control, copper-associated hepatopathy (CAH), and intermediate populations on the basis of histopathologic features and hepatic copper accumulation. The DNA was extracted from archived tissues and genotyped for reported variants in the 3 genes. Allele frequencies were determined, and associations of genotypes with CAH status were evaluated. 29 control dogs, 45 CAH dogs, and 16 intermediate dogs were included in the study. The overall ATP7A and RETN variant allele frequencies were 30% and 13%, respectively, and were not significantly different among control, CAH, and intermediate populations. The ATP7B variant allele frequency was significantly higher in the CAH population (30%) as compared to the control population (13%). However, 21 of 45 (47%) CAH dogs did not have an ATP7B variant allele whereas 7 of 28 (25%) control dogs did have an ATP7B variant allele. Study results supported a contributory role for the ATP7B variant in CAH pathogenesis in Labrador Retrievers. However, the application of genetic testing in a clinical setting is complicated by genotypic variability within healthy and diseased dogs.

Assessment of porcine Rotavirus-associated virome variations in pigs with enteric disease

Enteric disease is the predominant cause of morbidity and mortality in young mammals including pigs. Viral species involved in porcine enteric disease complex (PEDC) include rotaviruses, coronaviruses, picornaviruses, astroviruses and pestiviruses among others. The virome of three groups of swine samples submitted to the Kansas State University Veterinary Diagnostic Laboratory for routine testing were assessed, namely, a Rotavirus A positive (RVA) group, a Rotavirus co-infection (RV) group and a Rotavirus Negative (RV Neg) group. All groups were designated by qRT-PCR test results for Porcine Rotavirus A, B, C and H such that samples positive for RVA only went in the RVA group, samples positive for > 1 rotavirus went in the RV group and samples negative for all were grouped in the RVNeg group. All of the animals had clinical enteric disease resulting in scours and swollen joints/lameness, enlarged heart and/or a cough. All samples were metagenomic sequenced and analyzed for viral species composition that identified 14 viral species and eight bacterial viruses/phages. Sapovirus and Escherichia coli phages were found at a high prevalence in RVA and RV samples but were found at low or no prevalence in the RVNeg samples. Picobirnavirus was identified at a high proportion and prevalence in RVNeg and RV samples but at a low prevalence in the RVA group. Non-rotaviral diversity was highest in RVA samples followed by RV then RV Neg samples. A sequence analysis of the possible host of Picobirnaviruses revealed fungi as the most likely host. Various sequences were extracted from the sample reads and a phylogenetic update was provided showing a high prevalence of G9 and P[23] RVA genotypes. These data are important for pathogen surveillance and control measures.