Coxiella burnetii in domestic doe goats in the United States, 2019-2020.

Coxiella burnetii is a bacterial pathogen capable of causing serious disease in humans and abortions in goats. Infected goats can shed C. burnetii through urine, feces, and parturient byproducts, which can lead to infections in humans when the bacteria are inhaled. Goats are important C. burnetii reservoirs as evidenced by goat-related outbreaks across the world. To better understand the current landscape of C. burnetii infection in the domestic goat population, 4,121 vaginal swabs from 388 operations across the United States were analyzed for the presence of C. burnetii by IS1111 PCR as part of the United States Department of Agriculture, Animal Plant Health Inspection Service, Veterinary Services' National Animal Health Monitoring System Goats 2019 Study. In total, 1.5% (61/4121) of swabs representing 10.3% (40/388) (weighted estimate of 7.8, 95% CI 4.4-13.5) of operations were positive for C. burnetii DNA. The quantity of C. burnetii on positive swabs was low with an average Ct of 37.9. Factors associated with greater odds of testing positive included suspected Q fever in the herd in the previous 3 years, the presence of wild deer or elk on the operation, and the utilization of hormones for estrus synchronization. Factors associated with reduced odds of testing positive include the presence of kittens and treatment of herds with high tannin concentrate plants, diatomaceous earth, and tetrahydropyrimidines. In vitro analysis demonstrated an inhibitory effect of the tetrahydropyrimidine, pyrantel pamoate, on the growth of C. burnetii in axenic media as low as 1 µg per mL. The final multivariable logistic regression modeling identified the presence of wild predators on the operation or adjacent property (OR = 9.0, 95% CI 1.3-61.6, p value = 0.0248) as a risk factor for C. burnetii infection.

Antimicrobial Resistance Characteristics of Fecal Escherichia coli and Enterococcus Species in U.S. Goats: 2019 National Animal Health Monitoring System Enteric Study.

Escherichia coli and Enterococcus species are normal bacteria of the gastrointestinal tract and serve as indicator organisms for the epidemiology and emergence of antimicrobial resistance in their hosts and the environment. Some E. coli serovars, including E. coli O157:H7, are important human pathogens, although reservoir species such as goats remain asymptomatic. We describe the prevalence and antimicrobial resistance of generic E. coli, E. coli O157:H7, and Enterococcus species collected from a national surveillance study of goat feces as part of the National Animal Health Monitoring System (NAHMS) Goat 2019 study. Fecal samples were collected from 4918 goats on 332 operations across the United States. Expectedly, a high prevalence of E. coli (98.7%, 4850/4915) and Enterococcus species (94.8%, 4662/4918) was found. E. coli O157:H7 prevalence was low (0.2%; 10/4918). E. coli isolates, up to three per operation, were evaluated for antimicrobial susceptibility and 84.7% (571/674) were pansusceptible. Multidrug resistance (MDR; ≥3 classes) was uncommon among E. coli, occurring in 8.2% of isolates (55/674). Resistance toward seven antimicrobial classes was observed in a single isolate. Resistance to tetracycline alone (13.6%, 92/674) or to tetracycline, streptomycin, and sulfisoxazole (7.0% 47/674) was the most common pattern. All E. coli O157:H7 isolates were pansusceptible. Enterococcus isolates, up to four per operation, were prioritized by public health importance, including Enterococcus faecium and Enterococcus faecalis and evaluated. Resistance to lincomycin (93.8%, 1232/1313) was most common, with MDR detected in 29.5% (388/1313) of isolates. The combination of ciprofloxacin, lincomycin, and quinupristin resistance (27.1%, 105/388) was the most common pattern detected. Distribution and characteristics of antimicrobial resistance in E. coli and Enterococcus in the U.S. goat population from this study can inform stewardship considerations and public health efforts surrounding goats and their products.

Detection and molecular characterization of Salmonella species on U.S. goat operations.

Salmonella species are an important cause of gastrointestinal disease in animals, including goats. Additionally, Salmonella species are among the top five U.S. foodborne pathogens causing illness to humans. The goat industry is rapidly expanding in the U.S. yet estimates of Salmonella prevalence within these populations is lacking. The aim of this study was to investigate the fecal prevalence, antimicrobial resistance (AMR), biofilm potential, and virulence profile of Salmonella species isolated from goat feces as part of the United States Department of Agriculture (USDA) National Animal Health Monitoring System (NAHMS) Goat 2019 study, enteric microbe component. A total of 4917 fecal samples were collected from 332 operations, from September 2019-March 2020. Salmonella were isolated using standard enrichment and culture methods; antimicrobial susceptibility was determined by broth microdilution. Biofilm production was assessed using a crystal violet assay and normalized to a positive control strain, and PCR was used to detect virulence genes. Overall, we detected a low prevalence (0.7%, n = 35/4917) of Salmonella in goat feces and identified a broad range of serotypes including S. Bareilly (35%) and a single rare S. Sharon. All isolates were pansusceptible to 14 antimicrobials except one, which was resistant to only tetracycline (MIC ≥ 32 µg/mL). All strains were found to possess the majority of virulence determinants screened, and 40% (14 of 35) formed weak, moderate, or strong biofilm. We found a low prevalence of Salmonella, and characteristics of Salmonella in the U.S. goat population informs ongoing public health efforts to manage risk of animal food products and animal interactions.

Enhanced detection of Giardia duodenalis mixed assemblage infections in pre-weaned dairy calves using next generation sequencing.

Giardia duodenalis is one of the most common parasitic causes of gastrointestinal illness in humans worldwide with widespread infections in mammalian hosts. It frequently infects cattle, producing a high number of cysts. Cattle can harbor both host-adapted assemblage E and human pathogenic assemblages A and B. Previous studies have demonstrated that conventional molecular methods lack the sensitivity required for detecting mixed infections and that the occurrence of mixed infections in cattle are likely underestimated. To investigate the presence of mixed assemblage infections in cattle, 2539 pre-weaned dairy calves from the United Stated were screened for the presence of G. duodenalis using molecular tools. Next generation amplicon sequencing (NGS) was then performed for a subset of around 30% of positive samples (n = 314) and the ability of NGS and Sanger sequencing to detect mixed assemblage infections was compared. The overall prevalence of G. duodenalis in pre-weaned dairy calves in the sample using PCR was high (1013/2539; 39.9%). Molecular genotyping identified only assemblage A and E, with assemblage E as the predominant assemblage. Out of the 314 samples examined by both Sanger and NGS, 9 samples (2.9%) were identified as mixed A/E infections by Sanger while NGS identified 56 samples (17.8%), which was six-times more mixed infections compared with Sanger sequencing. NGS demonstrated superior sensitivity to Sanger in detecting assemblages present in low abundances. The percentage of mixed A/E infections found in the sampled dairy calves was higher than was hypothesized using values from the literature. This underestimation could be present in the wider cattle population as well, though further exploration would be needed to verify that claim. These findings highlight the advantages of NGS application in molecular epidemiological studies of Giardia. To better understand Giardia epidemiology, establish routes of transmission, and assess the potential role of cattle and other animals as a source of environmental contamination with zoonotic assemblages it is necessary to uncover mixed assemblage infections.

Mycoplasma ovis infection in domestic sheep (Ovis aries) in the United States: Prevalence, distribution, associated risk factors, and associated outcomes.

Mycoplasma ovis is a hemotropic bacterium reported to infect sheep, goats, and deer species. Infection in these species can result in anemia, jaundice, and ill-thrift. Although of worldwide distribution, only rare reports of this bacterium in the United States exist. The objectives of this retrospective study were to identify the prevalence and distribution of M. ovis, and identify associated demographic and management risk factors, and reproductive and production outcomes associated with infection on domestic sheep (Ovis aries) operations in the United States. As part of the United States Department of Agriculture (USDA), Animal Plant Health Inspection Service, Veterinary Services' National Animal Health Monitoring System (NAHMS) Sheep 2001 and 2011 studies, blood was collected and sera banked from 21,369 ewes in 2001 and 13,128 ewes in 2011. Participating premises were located in 22 states across the United States for each sample year. In 2015 the USDA, Agricultural Research Service, Animal Disease Research Unit received aliquots of these sera, and DNA was extracted and analyzed by PCR for the presence of M. ovis genomic DNA. Flock presence and mean within-flock prevalence of M. ovis were 73.3% and 23.2%, respectively. Model selection using Mallow's Cp Criterion was used to determine which variables significantly affected flock presence and within-flock prevalence. The final flock presence model included flock size, year of blood collection, region, and vaccine administration. The final within-flock prevalence model included year of blood collection, interaction between flock size and region, and interaction between reported abortions and grazing with sheep from other operations. Medium and large operations had a higher flock presence and within-flock prevalence. Flock presence was higher in operations that administered any vaccines. Operations that reported any abortions and grazed with sheep from other operations had a higher within-flock prevalence.

Zoonotic and genetically diverse subtypes of Blastocystis in US pre-weaned dairy heifer calves.

Blastocystis is an emerging zoonotic pathogen with global distribution. However, limited data exist on the prevalence and genetic diversity of Blastocystis in the USA and in food animals. We conducted the first large-scale molecular investigation of Blastocystis in the USA by testing 2539 fecal samples from dairy heifer calves from 13 states. Blastocystis was detected in 73 (2.9%) fecal samples and in 10 of the 13 tested states. Molecular characterization showed a wide diversity of subtypes. Eleven subtypes were identified, seven previously reported (ST-3, ST-4, ST-5, ST-10, ST-14, ST-17, and ST-21) and four potentially novel subtypes (named ST-23 to ST-26). Zoonotic subtypes 3, 4, and 5 were found in 67% (49) of the positive specimens in this population. Our results suggest that cattle could serve as reservoirs of infection for humans and other domestic animals highlighting the potential risk of zoonotic transmission for Blastocystis.