Annette Roug

Prevalence, Environmental Loading, and Molecular Characterization of Cryptosporidium and Giardia Isolates from Domestic and Wild Animals along the Central California Coast

ABSTRACT The risk of disease transmission from waterborne protozoa is often dependent on the origin (e.g., domestic animals versus wildlife), overall parasite load in contaminated waterways, and parasite genotype, with infections being linked to runoff or direct deposition of domestic animal and wildlife feces. Fecal samples collected from domestic animals and wildlife along the central California coast were screened to (i) compare the prevalence and associated risk factors for fecal shedding of Cryptosporidium and Giardia species parasites, (ii) evaluate the relative importance of animal host groups that contribute to pathogen loading in coastal ecosystems, and (iii) characterize zoonotic and host-specific genotypes. Overall, 6% of fecal samples tested during 2007 to 2010 were positive for Cryptosporidium oocysts and 15% were positive for Giardia cysts. Animal host group and age class were significantly associated with detection of Cryptosporidium and Giardia parasites in animal feces. Fecal loading analysis revealed that infected beef cattle potentially contribute the greatest parasite load relative to other host groups, followed by wild canids. Beef cattle, however, shed host-specific, minimally zoonotic Cryptosporidium and Giardia duodenalis genotypes, whereas wild canids shed potentially zoonotic genotypes, including G. duodenalis assemblages A and B. Given that the parasite genotypes detected in cattle were not zoonotic, the public health risk posed by protozoan parasite shedding in cattle feces may be lower than that posed by other animals, such as wild canids, that routinely shed zoonotic genotypes.

Serosurveillance for Livestock Pathogens in Free-Ranging Mule Deer (Odocoileus hemionus)

Routine disease surveillance has been conducted for decades in mule deer (Odocoileus hemionus) in California for pathogens shared between wildlife and domestic ruminants that may have implications for the animal production industry and wildlife health. Deer sampled from 1990 to 2007 (n = 2,619) were tested for exposure to six pathogens: bluetongue virus (BTV), epizootic hemorrhagic disease virus (EHDV), bovine viral diarrhea virus (BVDV), Leptospira spp., Anaplasma spp. and Brucella spp. We evaluated the relationship between exposure to these pathogens and demographic risk factors to identify broad patterns in seroprevalence across a large temporal and spatial scale. The overall seroprevalence for the entire study period was 13.4% for BTV, 16.8% for EHDV, 17.1% for BVDV, 6.5% for Leptospira spp., 0.2% for Brucella spp., and 17% for Anaplasma spp. Antibodies against BTV and EHDV were most prevalent in the deer populations of southern California. Antibodies against Leptospira spp. and Anaplasma spp. were most prevalent in coastal and central northern California whereas antibodies against BVDV were most prevalent in central-eastern and northeastern California. The overall seroprevalence for Anaplasma spp. was slightly lower than detected in previous studies. North and central eastern California contains large tracts of federal land grazed by livestock; therefore, possible contact between deer and livestock could explain the high BVDV seroprevalence found in these areas. Findings from this study will help to establish baseline values for future comparisons of pathogen exposure in deer, inform on long-term trends in deer population health and provide relevant information on the distribution of diseases that are shared between wildlife and livestock.

EPIDEMIOLOGY AND POTENTIAL LAND-SEA TRANSFER OF ENTERIC BACTERIA FROM TERRESTRIAL TO MARINE SPECIES IN THE MONTEREY BAY REGION OF CALIFORNIA

Marine mammals are at risk for infection by fecal-associated zoonotic pathogens when they swim and feed in polluted nearshore marine waters. Because of their tendency to consume 25–30% of their body weight per day in coastal filter-feeding invertebrates, southern sea otters (Enhydra lutris nereis) can act as sentinels of marine ecosystem health in California. Feces from domestic and wildlife species were tested to determine prevalence, potential virulence, and diversity of selected opportunistic enteric bacterial pathogens in the Monterey Bay region. We hypothesized that if sea otters are sentinels of coastal health, and fecal pollution flows from land to sea, then sea otters and terrestrial animals might share the same enteric bacterial species and strains. Twenty-eight percent of fecal samples tested during 2007–2010 were positive for one or more potential pathogens. Campylobacter spp. were isolated most frequently, with an overall prevalence of 11%, followed by Vibrio cholerae (9%), Salmonella spp. (6%), V. parahaemolyticus (5%), and V. alginolyticus (3%). Sea otters were found positive for all target bacteria, exhibiting similar prevalences for Campylobacter and Salmonella spp. but greater prevalences for Vibrio spp. when compared to terrestrial animals. Fifteen Salmonella serotypes were detected, 11 of which were isolated from opossums. This is the first report of sea otter infection by S. enterica Heidelberg, a serotype also associated with human clinical disease. Similar strains of S. enterica Typhimurium were identified in otters, opossums, and gulls, suggesting the possibility of land-sea transfer of enteric bacterial pathogens from terrestrial sources to sea otters.

Zoonotic fecal pathogens and antimicrobial resistance in county fair animals

Livestock fairs present a unique opportunity for the public to experience close contact with animals, but may also expose people to zoonotic pathogens through contact with animal feces. The goal of this study was to screen cattle, sheep, goat, chicken, rabbit and horse feces from a livestock fair in California for the potentially zoonotic pathogens Escherichia coli O157:H7, Salmonella, Campylobacter, Vibrio, Cryptosporidium and Giardia spp., as well as determining the level of antimicrobial resistance in E. coli and Salmonella. Notably, E. coli O157:H7 was reported for the first time in a pig at a county fair in California. Campylobacter jejuni as well as Salmonella enterica serovars Derby and Thompson were also isolated from pigs, cattle, sheep, goats or chickens, whereas horses and rabbits were negative for all target pathogens. The prevalence of antimicrobial resistance as well as multi-drug resistance patterns were highest for E. coli and Salmonella spp. cultured from pigs and chickens, were generally widespread but at lower levels for other animal groups, and included resistance to ampicillin and streptomycin, two antimicrobial drugs of importance for human medicine. This study provides data that highlight the importance of practicing good hygiene in livestock fair settings to avoid transmission of zoonotic microbes, particularly pathogens with antimicrobial resistance, to fair visitors and among animal populations.

Comparison of intervention methods for reducing human exposure to Mycobacterium bovis through milk in pastoralist households of Tanzania

Bovine tuberculosis (bTB), caused by Mycobacterium bovis, is a disease of zoonotic concern, especially in countries with no control programs in livestock and where routine pasteurization of milk is not practiced. In Tanzania, bTB is widespread in livestock and has been diagnosed in humans; however, herd bTB testing is primarily carried out for bTB-free certification in commercial dairy herds at the expense of the dairy cattle owner. For rural livestock holders, such an expense is prohibitive, and consequently there is no control of bTB in most areas. Although effective long-term solutions to control bTB in livestock are desirable, there is a need to assess the effect of preventive measures on reducing human exposure to bTB in such settings. We utilized locally relevant cattle herd characteristics and management data from the Health for Animals and Livelihood Improvement (HALI) project in south-central Tanzania to build a Reed-Frost model that compared the efficacy of alternative methods aimed at reducing the exposure of humans to infectious milk from a typical pastoralist cattle herd. During a 10-year simulation period, the model showed that boiling milk 80% of the time is necessary to obtain a reduction in liters of infectious milk approximately equivalent to what would be obtained with a standard 2-year testing and removal regimen, and that boiling milk was more effective than animal test and removal early in the time period. In addition, even with testing and removing infected cattle, a residual risk of exposure to infectious milk remained due to imperfect sensitivity of the skin test and a continuous risk of introduction of infectious animals from other herds. The model was sensitive to changes in initial bTB prevalence but not to changes in herd size. In conclusion, continuous complimentary treatment of milk may be an effective strategy to reduce human exposure to M. bovis-infected milk in settings where bTB is endemic and a comprehensive bTB control program is yet to be implemented.

Cerebral Larva Migrans Caused by Baylisascaris spp. in a Free-ranging North American Porcupine (Erethizon dorsatum)

Abstract A free-ranging North American porcupine (Erethizon dorsatum) from Utah, US, exhibited neurologic symptoms and was submitted for necropsy. Histologic examination of the brain revealed severe encephalitis with an intralesional nematode larva consistent with Baylisascaris spp. Neurologic larva migrans had not been reported in free-ranging porcupines, or from wildlife in Utah.

Comparison of trace mineral concentrations in tail hair, body hair, blood, and liver of mule deer (Odocoileus hemionus) in California

Measuring trace mineral concentrations can be an important component of assessing the health of free-ranging deer. Trace mineral concentrations in liver most accurately reflect the trace mineral status of an individual, but, in live animals, whole blood or serum are the most commonly used sample types. Trace minerals measured in serum, such as copper, zinc, and iron, do not always accurately correlate to liver concentrations, and supplementary samples for evaluating the trace mineral status in live deer would be useful. We evaluated the utility of body and tail hair for measuring selenium, copper, zinc, iron, and manganese in free-ranging mule deer (Odocoileus hemionus) by using Spearman rank correlations and linear regression. Correlations were strongest at the time of or shortly after growth of the winter coat and in resident deer. In live deer, strong correlations and moderate linear associations (R2 = 0.57) were detected between body and tail hair and whole blood selenium in December. In postmortem-sampled deer, a strong correlation and linear association (R2 = 0.80) were found between liver and body hair selenium in August–November. Results indicate that body hair, if collected during or shortly after growth of the winter coat, can be used as a supplementary sample for measuring selenium concentrations in deer. None of the other correlations and linear associations were found to be sufficiently strong to conclude that hair can reliably be utilized as a complementary sample for measuring these trace mineral concentrations.

Spatial predictors of bovine tuberculosis infection and Brucella spp. exposure in pastoralist and agropastoralist livestock herds in the Ruaha ecosystem of Tanzania.

While many studies investigate animal-related risk factors for disease, few have considered environmental or spatial risk factors in the dynamics of bovine tuberculosis (bTB) and brucellosis. In the Ruaha ecosystem of Tanzania, we investigated the role of household location as a predictor for infection with Mycobacterium bovis and exposure to Brucella in pastoralist and agropastoralist cattle herds in a typical African wildlife-livestock-human interface. ArcGIS was utilized to calculate Euclidian distances between households and the nearest river, village center, protected area, and other infected households, followed by multivariate logistic regression to assess the association between risk factors and herd-level bTB and Brucella outcomes. Global and local spatial clustering of bTB-infected and Brucella-exposed herds was explored using the Cuzick-Edward’s test and SaTScan spatial scan statistics. Households located farther from rivers and closer to village centers and herds belonging to agropastoralists were more likely to have bTB-positive cattle. Risk of Brucella exposure increased with proximity to protected areas. One spatial cluster of households with Brucella spp. seropositive cattle was identified. Spatial factors may be useful for assessing disease risk and for formulating intervention and control strategies for households that manage cattle in ecosystems characterized by seasonally limited resources and intense wildlife-livestock interfaces.

Comparison of Two Bacterial Transport Media for Culture of Tonsilar Swabs from Bighorn Sheep (Ovis canadensis) and Mountain Goats (Oreamnos americanus)

Abstract Duplicate tonsilar swabs were collected from 77 bighorn sheep (Ovis canadensis) and 19 mountain goats (Oreamnos americanus) in Utah. Swabs were refrigerated in bacterial transport medium or frozen in cryopreservation medium prior to bacteriologic culture. The cryopreservation medium yielded comparable or superior bacterial growth while permitting more flexibility in specimen shipment to the laboratory.

Utility of a Fecal Real-time PCR Protocol for Detection of Mycobacterium bovis Infection in African Buffalo (Syncerus caffer)

Abstract A real-time PCR protocol for detecting Mycobacterium bovis in feces was evaluated in bovine tuberculosis–infected African buffalo (Syncerus caffer). Fecal samples spiked with 1.42×103 cells of M. bovis culture/g and Bacille Calmette–Guérin standards with 1.58×101 genome copies/well were positive by real-time PCR but all field samples were negative.