key: cord-0887785-ziwp8t09 authors: Smith, Fauna Leah; Heller, Meera C.; Crossley, Beate M.; Clothier, Kristin A.; Anderson, Mark L.; Barnum, Samantha S.; Pusterla, Nicola; Rowe, Joan D. title: Diarrhea outbreak associated with coronavirus infection in adult dairy goats date: 2022-02-15 journal: J Vet Intern Med DOI: 10.1111/jvim.16354 sha: 099c889c267756fa6594ffc305d1701554726eec doc_id: 887785 cord_uid: ziwp8t09 BACKGROUND: Infection by coronaviruses cause gastrointestinal disease in many species. Little is known about its prevalence and importance in goats. OBJECTIVE: Identify the etiology, demographics, and clinical features of an outbreak of diarrhea in adult goats. HYPOTHESIS: Bovine coronavirus (BCoV) PCR would detect viral material in feces of goats in the herds involved in the diarrhea outbreak. ANIMALS: Twelve herds with 4 to 230 adult goats were affected. Goats sampled for fecal PCR were ≥1‐year‐old: 25 from affected herds and 6 from a control herd. METHODS: This is a cross‐sectional descriptive study of an outbreak of diarrheal disease in adult goats. BCoV PCR primers for the spike (S) or nucleocapsid (N) proteins were used to test fecal material from affected goats. The N protein sequencing and phylogenetic analysis was performed. Herd records and owner surveys were used to characterize morbidity, clinical signs, and treatment. RESULTS: In 2 affected herds 18/25 of animals had at least 1 positive BCoV PCR test. Goats from affected herds were significantly more likely to be PCR positive than the control herd (OR 8.75, 95% CI 1.11‐104, P = .05). The most common clinical signs were change in fecal consistency (19/20) and decreased milk production (14/15). Phylogenetic analysis of the N protein showed this virus was closely related to a bovine‐like coronavirus isolated from a giraffe. CONCLUSIONS AND CLINICAL IMPORTANCE: Bovine coronavirus primers detected nucleic acids of the N and S proteins in feces of goats in affected herds. Coronavirus shedding frequency was temporally associated with the outbreak. Coronaviruses (CoVs) are enveloped, positive-stranded RNA viruses with a tropism for the gastrointestinal and respiratory tracts causing diseases in a wide range of hosts. 1 In cattle, CoV is associated with neonatal calf diarrhea, 2 winter dysentery in adult cattle 2 and outbreaks of respiratory disease. 3, 4 Bovine-like CoVs are associated with diarrhea outbreaks in adult captive wild ruminants 5 and CoVs are identified in diarrhetic sheep. [6] [7] [8] Little is reported about CoV in goats. It is not an important pathogen in neonatal diarrhea in goats. [8] [9] [10] There is variable seropositivity rates (1%-43.1%) to bovine coronavirus (BCoV) in adult goats. [11] [12] [13] Molecular detection of BCoV occurs in 4.5% goats comingled with cattle but is not associated with disease. 14 PCR detection of BCoV is reported in captive wild ruminants with diarrhea. 5 Animal exhibitions present a risk for disease transmission, because of comingling different herds and species. Biosecurity practices by individual herds vary with many having poor protocols, increasing the risk of disease transmission. 15 Disease outbreaks associated with livestock exhibitions are reported, 16, 17 as well as frequent anecdotal accounts. Dispersal of animals after an event can make outbreak investigations difficult. Our objectives were to identify common clinical signs in adult dairy goats in an outbreak of diarrheal disease and to determine the relevance of BCoV positive PCR in affected herds. We hypothesized that BCoV PCR could detect viral antigen in goat feces and a greater frequency of positive results would occur in the affected herds. The outbreak involved dairy goat herds that attended 2 exhibitions between July 24th and August 3rd, 2017 in Northern California. Twelve herds were identified as having adult goats with diarrhea, by contacting exhibitors using the fairs' entry lists. Five herds were affected after the first exhibition, 7 additional herds were affected after the second (Figure 1 ). Exhibition 1 had >30 goat herds in attendance and shared wash racks, barn airspace and movement corridors with dairy cattle (Figure 1 ). Exhibition 2 had 13 herds in attendance, 4 were affected herds from the first fair, and the facility only housed goats ( Figure 1 ). Exhibition 2 required an entry veterinary inspection, but animals developing diarrhea during the fair remained on the premises. A survey was sent to the 12 herds reporting diarrheal cases regarding clinical signs, animals affected, treatments used and outcomes of affected animals. Two of the 12 affected herds (herd A and herd B), both affected after exhibition 2 (Figure 1 ), submitted fecal samples to Laboratory A (California Animal Health and Food Safety Laboratory, University of California, Davis, California) for fecal culture, Salmonella spp. PCR, clostridial toxin testing, fecal egg count for trichostrongyles, Coccidia, and Cryptosporidium. Two samples were submitted for electron F I G U R E 1 Facility configuration and location of affected herds at 2 consecutive exhibitions associated with outbreaks of adult goat diarrhea. Diagram illustrates proximity of affected herds as well as shared facilities such as movement corridors; show rings, holding pens and wash racks at the 2 exhibitions. The 2 herds (herds A and B) that were followed extensively were affected after exhibition 2 microscopy (EM). Feces were tested for CoV using PCR with BCoV primers. These samples were submitted within 3 days of the start of the outbreak from clinically affected animals (3 from herd A and 2 from herd B). Herd health records from both herds were analyzed for clinical signs, duration of illness and recurrence of disease. Fecal sample/rectal swabs were collected from 6 adult (>2-years-old) dairy breed goats from a herd that had no animal movement in >6 months. The samples were collected at the same time as the initial sampling of herds A and B. BCoV PCR testing at the Laboratory A was performed as described. 18 In brief, RNA extraction was performed according to the manufac- Twelve herds reported animals with alterations in fecal consistency, ranging from soft-formed to watery. Herd size ranged from 4 to 230 adult animals. All 12 herds had affected adult females of dairy breeds: Nubian (3/12), Toggenburg (2/12), Alpine (2/12), Mixed including Alpine, La Mancha, Nubian, Saanen, Nigerian Dwarf, and crossbred (5/12). The survey had a response rate of 75% (9/12). Some herds reported affected animals that did not attend exhibitions, including adult males (2/9) and weaned kids (>4 months of age; 4/9). The reported morbidity ranged from 15% to 90%. One death was reported. Other clinical signs reported included: acute drop in milk production Note: Changes in fecal consistency was the most common clinical finding, followed by decreased milk production in lactating animals. (9/9), anorexia (9/9), fever (5/9), and cough ( Sick animals were no more likely than healthy animals to be BCoV PCR positive (OR 0.58, 95% CI 0.048-5.65, P > .99). There was no correlation between the PCR Cq value, as a viral load proxy, and being a sick or healthy animal in an affected herd (P = .74; Figure 4 ). Coronavirus has not been reported in association with diarrheal disease in adult goats. During this outbreak, 12 affected herds were identified with more than 300 affected animals, with morbidity ranging from 15% to 90%. Affected herds attended 1 of 2 exhibitions over a 10-day period. Exhibitions result in the comingling of animals from different sources and often different species providing an opportunity for infectious pathogens to transmit between herds and possibly species. 16, 17 A survey of exhibitors attending exhibition 1 indicates poor biosecurity practices are common. 15 Practices at exhibitions result in multiple points for possible disease transmission between herds. At exhibition 2 animal movement required the use of narrow aisleways flanked by affected herds and both exhibitions had common-use wash racks and holding pens. Similar to winter dysentery this outbreak occurred when animals were comingled in a high-density environment. 3, 4, 19 Change in fecal consistency and decreased milk production were the most common clinical signs, however, cough was noted in 3 herds. BCoV is associated with coincident respiratory and gastrointestinal disease. 3 The present study provides evidence that this outbreak was associated with CoV, however, the longitudinal testing occurred after the from the affected herds were significantly more likely to be BCoV positive than the control herd. Additionally, the shedding frequency was much higher than is reported in surveilled clinically normal goats. 14 Our results also indicate viral shedding may occur in animals that never exhibited clinical signs. Subclinical shedders present challenges to control of transmission, as a potential source of exposure. There was a strong temporal association between the outbreak and the frequency of CoV shedding in the 2 affected herd, but further work is needed to determine the definitive cause. Given the wide host range and ability of CoVes to be zoonotic, 24, 25 good hygiene and biosecurity is important when dealing with animals shedding CoV. 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Heller https://orcid.org/0000-0002-0080-5736