Lake Michigan insights from island studies: the roles of chipmunks and coyotes in maintaining Ixodes scapularis and Borrelia burgdorferi in the absence of white-tailed deer.

Deer management (e.g., reduction) has been proposed as a tool to reduce the acarological risk of Lyme disease. There have been few opportunities to investigate Ixodes scapularis (blacklegged tick) and Borrelia burgdorferi sensu stricto dynamics in the absence of white-tailed deer (Odocoileus virginianus) in midwestern North America. A pair of islands in Lake Michigan presented a unique opportunity to study the role of alternative hosts for the adult stage of the blacklegged tick for maintaining a tick population as a deer herd exists on North Manitou Island but not on South Manitou Island, where coyotes (Canis latrans) and hares (Lepus americanus) are the dominant medium mammals. Additionally, we were able to investigate the maintenance of I. scapularis and B. burgdorferi in small mammal communities on both islands, which were dominated by eastern chipmunks (Tamias striatus). From 2011 to 2015, we surveyed both islands for blacklegged ticks by drag cloth sampling, bird mist netting, and small and medium-sized mammal trapping. We assayed questing ticks, on-host ticks, and mammal biopsies for the Lyme disease pathogen, B. burgdorferi. We detected all three life stages of the blacklegged tick on both islands. Of the medium mammals sampled, no snowshoe hares (Lepus americanus, 0/23) were parasitized by adult blacklegged ticks, but 2/2 coyotes (Canis latrans) sampled on South Manitou Island in 2014 were parasitized by adult blacklegged ticks, suggesting that coyotes played a role in maintaining the tick population in the absence of deer. We also detected I. scapularis ticks on passerine birds from both islands, providing support that birds contribute to maintaining as well as introducing blacklegged ticks and B. burgdorferi to the islands. We observed higher questing adult and nymphal tick densities, and higher B. burgdorferi infection prevalence in small mammals and in adult ticks on the island with deer as compared to the deer-free island. On the islands, we also found that 25% more chipmunks were tick-infested than mice, fed more larvae and nymphs relative to their proportional abundance compared to mice, and thus may play a larger role compared to mice in the maintenance of B. burgdorferi. Our investigation demonstrated that alternative hosts could maintain a local population of blacklegged ticks and an enzootic cycle of the Lyme disease bacterium in the absence of white-tailed deer. Thus, alternative adult blacklegged tick hosts should be considered when investigating deer-targeted management tools for reducing tick-borne disease risk, especially when the alternative host community may be abundant and diverse.

Individual and Site-Specific Variation in a Biogeographical Profile of the Coyote Gastrointestinal Microbiota.

Most knowledge of the vertebrate gut microbiota comes from fecal samples; due to difficulties involved in sample collection, the upper intestinal microbiota is poorly understood in wild animals despite its potential to inform broad interpretations about host-gut microbe relationships under natural conditions. Here, we used 16S rRNA gene sequencing to characterize the microbiota of wild coyotes (Canis latrans) along the gastrointestinal tract, including samples from the duodenum, jejunum, ileum, caecum, ascending and descending colon, and feces. We used this intestinal profile to (1) quantify how intestinal site and individual identity interact to shape the microbiota in an uncontrolled setting, and (2) evaluate whether the fecal microbiota adequately represent other intestinal sites. Microbial communities in the large intestine were distinct from those in the small intestine, with higher diversity and a greater abundance of anaerobic taxa. Within each of the small and large intestine, individual identity explained significantly more among-sample variation than specific intestinal sites, revealing the importance of individual variation in the microbiota of free-living animals. Fecal samples were not an adequate proxy for studying upper intestinal environments, as they contained only half the amplicon sequence variants (ASVs) present in the small intestine at three- to four-fold higher abundances. Our study is a unique biogeographical investigation of the microbiota using free-living mammals rather than livestock or laboratory organisms and provides a foundational understanding of the gastrointestinal microbiota in a wild canid.

An altered microbiome in urban coyotes mediates relationships between anthropogenic diet and poor health.

Generalist species able to exploit anthropogenic food sources are becoming increasingly common in urban environments. Coyotes (Canis latrans) are one such urban generalist that now resides in cities across North America, where diseased or unhealthy coyotes are frequently reported in cases of human-wildlife conflict. Coyote health and fitness may be related to habitat use and diet via the gut microbiome, which has far-reaching effects on animal nutrition and physiology. In this study, we used stomach contents, stable isotope analysis, 16S rRNA gene amplicon sequencing, and measures of body condition to identify relationships among habitat use, diet, fecal microbiome composition, and health in urban and rural coyotes. Three distinct relationships emerged: (1) Urban coyotes consumed more anthropogenic food, which was associated with increased microbiome diversity, higher abundances of Streptococcus and Enterococcus, and poorer average body condition. (2) Conversely, rural coyotes harbored microbiomes rich in Fusobacteria, Sutterella, and Anaerobiospirillum, which were associated with protein-rich diets and improved body condition. (3) Diets rich in anthropogenic food were associated with increased abundances of Erysipelotrichiaceae, Lachnospiraceae, and Coriobacteriaceae, which correlated with larger spleens in urban coyotes. Urban coyotes also had an increased prevalence of the zoonotic parasite Echinococcus multilocularis, but there were no detectable connections between parasite infection and microbiome composition. Our results demonstrate how the consumption of carbohydrate-rich anthropogenic food by urban coyotes alters the microbiome to negatively affect body condition, with potential relationships to parasite susceptibility and conflict-prone behavior.

Serologic Evidence for the Exposure of Eastern Coyotes (Canis latrans) in Pennsylvania to the Tick-Borne Pathogens Borreliella burgdorferi and Anaplasma phagocytophilum.

Lyme disease and anaplasmosis are tick-borne bacterial diseases caused by Borreliella and Anaplasma species, respectively. A comprehensive analysis of the exposure of eastern coyotes (Canis latrans) in the northeastern United States to tick-borne pathogens has not been conducted. In this report, we assess the serological status of 128 eastern coyotes harvested in Pennsylvania in 2015 and 2017 for antibodies to Borreliella burgdorferi and Anaplasma phagocytophilum Immunoblot and dot blot approaches were employed to test each plasma sample by using cell lysates and recombinant proteins as detection antigens. The results demonstrate high seropositivity incidences of 64.8% and 72.7% for B. burgdorferi and A. phagocytophilum, respectively. Antibodies to both pathogens were detected in 51.5% of the plasma samples, indicating high potential for coinfection. Antibodies to the B. burgdorferi proteins DbpB, VlsE, DbpA, BBA36, and OspF (BBO39) were detected in 67.2, 63.3, 56.2, 51.6, and 48.4% of the plasma samples, respectively. Antibodies to the A. phagocytophilum P44 and P130 proteins were detected in 72.7 and 60.9% of the plasma samples, respectively.IMPORTANCE The incidence of Lyme disease (Borreliella burgdorferi) and anaplasmosis (Anaplasma phagocytophilum) are increasing in North America and Europe. The causative agents of these debilitating tick-transmitted infections are maintained in nature in an enzootic cycle involving Ixodes ticks and diverse mammals and birds. It has been postulated that predators directly or indirectly influence the dynamics of the enzootic cycle and disease incidence. Here, we demonstrate high seropositivity of eastern coyotes for B. burgdorferi and A. phagocytophilum As coyotes become established in urban and suburban environments, interactions with humans, companion animals, and urban/suburban wildlife will increase. Knowledge of the pathogens that these highly adaptable predators are exposed to or carry, and their potential to influence or participate in enzootic cycles, is central to efforts to reduce the risk of tick-borne diseases in humans and companion animals.

COYOTES (CANIS LATRANS) IN ARIZONA, USA, EXHIBIT IMMUNE AND GENETIC EVIDENCE OF RICKETTSIAL INFECTIONS.

Rocky Mountain spotted fever (RMSF), caused by the bacterium Rickettsia rickettsii, was recognized as endemic in Arizona, US after a 2002 outbreak and has since been a public health concern. The brown dog tick (Rhipicephalus sanguineus sensu lato) is the principal vector of this pathogen in Arizona. Domesticated dogs (Canis lupus familiaris) are the tick's main host, so free-roaming dogs in peridomestic areas have been named the primary risk factor for human cases of RMSF. However, the sudden emergence and long-distance dispersal of the pathogen have not been adequately explained, and one possible mechanism could include wildlife. Coyotes (Canis latrans) are wide ranging in Arizona and closely related to dogs, so it is possible that brown dog ticks parasitize coyotes and infect them. Although R. rickettsii is the most severe spotted fever group (SFG) rickettsial pathogen in humans, others occur in Arizona, and antibodies raised against them are cross-reactive, so we more-broadly hypothesized that coyotes in Arizona are exposed to SFG rickettsiae. We collected coyote tissues in spring 2016 and 2017. We tested sera for antibodies to R. rickettsii and found 9% (8/94) of samples were antibody-positive with titers of ≥256. Subsequent quantitative PCR analyses of skin showed evidence for Rickettsia spp. in 2.9% (4/138) of samples. These data suggest that coyotes have a role in the maintenance of SFG rickettsiae in Arizona. Further investigation is warranted to reveal which specific pathogen-vector complexes act on coyotes in the region and whether they represent a risk to human health.

BARTONELLA ROCHALIMAE AND B. VINSONII SUBSP. BERKHOFFII IN WILD CARNIVORES FROM COLORADO, USA.

Spleen samples from 292 wild carnivores from Colorado, US were screened for Bartonella infection. Bartonella DNA was detected in coyotes ( Canis latrans ) (28%), striped skunks ( Mephitis mephitis ) (23%), red foxes ( Vulpes vulpes ) (27%), and raccoons ( Procyon lotor ) (8%) but not in black bears ( Ursus americanus ), gray foxes ( Urocyon cinereoargenteus ), and mountain lions ( Puma concolor ). Two Bartonella species, B. vinsonii subsp. berkhoffii and B. rochalimae, were identified. All 10 infected striped skunks exclusively carried B. rochalimae while coyotes, red foxes, and raccoons could be infected with both Bartonella species. Five of seven infected coyotes carried B. v. berkhoffii whereas five of seven infected red foxes and 11 of 14 infected raccoons carried B. rochalimae. Further studies are needed to understand relationships between Bartonella species, wild carnivores, and their ectoparasites.

PARASITOLOGY AND SEROLOGY OF FREE-RANGING COYOTES (CANIS LATRANS) IN NORTH CAROLINA, USA.

Coyotes (Canis latrans) have expanded recently into the eastern US and can serve as a source of pathogens to domestic dogs (Canis lupus familiaris), livestock, and humans. We examined free-ranging coyotes from central North Carolina, US, for selected parasites and prevalence of antibodies against viral and bacterial agents. We detected ticks on most (81%) coyotes, with Amblyomma americanum detected on 83% of those with ticks. Fifteen (47%) coyotes were positive for heartworms (Dirofilaria immitis), with a greater detection rate in adults (75%) than juveniles (22%). Serology revealed antibodies against canine adenovirus (71%), canine coronavirus (32%), canine distemper virus (17%), canine parvovirus (96%), and Leptospira spp. (7%). We did not detect antibodies against Brucella abortus/suis or Brucella canis. Our results showed that coyotes harbor many common pathogens that present health risks to humans and domestic animals and suggest that continued monitoring of the coyote's role in pathogen transmission is warranted.

Zoonotic Bartonella species in cardiac valves of healthy coyotes, California, USA.

We investigated whether Bartonella spp. could cause endocarditis in coyotes or localize to cardiac valves before lesions develop. Bartonella DNA was amplified more often from coyote cardiac valves than spleen. Bartonella infection apparently leads to cardiac valve tropism, which could cause endocarditis, an often lethal complication in mammals, including humans.

Prevalence and characterization of Escherichia coli and Salmonella strains isolated from stray dog and coyote feces in a major leafy greens production region at the United States-Mexico border.

In 2010, Romaine lettuce grown in southern Arizona was implicated in a multi-state outbreak of Escherichia coli O145:H28 infections. This was the first known Shiga toxin-producing E. coli (STEC) outbreak traced to the southwest desert leafy green vegetable production region along the United States-Mexico border. Limited information exists on sources of STEC and other enteric zoonotic pathogens in domestic and wild animals in this region. According to local vegetable growers, unleashed or stray domestic dogs and free-roaming coyotes are a significant problem due to intrusions into their crop fields. During the 2010-2011 leafy greens growing season, we conducted a prevalence survey of STEC and Salmonella presence in stray dog and coyote feces. Fresh fecal samples from impounded dogs and coyotes from lands near produce fields were collected and cultured using extended enrichment and serogroup-specific immunomagnetic separation (IMS) followed by serotyping, pulsed-field gel electrophoresis (PFGE), and antimicrobial susceptibility testing. A total of 461 fecal samples were analyzed including 358 domestic dog and 103 coyote fecals. STEC was not detected, but atypical enteropathogenic E. coli (aEPEC) strains comprising 14 different serotypes were isolated from 13 (3.6%) dog and 5 (4.9%) coyote samples. Salmonella was cultured from 33 (9.2%) dog and 33 (32%) coyote samples comprising 29 serovars with 58% from dogs belonging to Senftenberg or Typhimurium. PFGE analysis revealed 17 aEPEC and 27 Salmonella distinct pulsotypes. Four (22.2%) of 18 aEPEC and 4 (6.1%) of 66 Salmonella isolates were resistant to two or more antibiotic classes. Our findings suggest that stray dogs and coyotes in the desert southwest may not be significant sources of STEC, but are potential reservoirs of other pathogenic E. coli and Salmonella. These results underscore the importance of good agriculture practices relating to mitigation of microbial risks from animal fecal deposits in the produce production area.

Prevalence of antibodies to spotted fever group Rickettsia spp. and Ehrlichia spp. in coyotes (Canis latrans) in Oklahoma and Texas, USA.

Coyotes (Canis latrans) are commonly infested with ticks, including Amblyomma americanum, the predominant vector of Ehrlichia chaffeensis and Ehrlichia ewingii; Dermacentor variabilis, an important vector of Rickettsia rickettsii; and Amblyomma maculatum, a major vector of Rickettsia parkeri, a spotted fever group (SFG) Rickettsia. To determine the degree to which coyotes are infected with or exposed to tick-borne bacterial disease agents, serum samples collected from coyotes in Oklahoma and Texas were tested for antibodies reactive to R. rickettsii, Ehrlichia canis, E. chaffeensis, E. ewingii, Borrelia burgdorferi, and Anaplasma phagocytophilum by indirect fluorescent antibody (IFA) testing or enzyme-linked immunosorbent assay (ELISA). Of the coyotes tested, 60% (46/77) and 64% (47/74) had antibodies reactive to R. rickettsii and E. chaffeensis, respectively, on IFA. Additionally, 5% (4/77) had antibodies reactive to E. canis, but not B. burgdorferi or A. phagocytophilum, on SNAP(®) 4Dx(®) ELISA; subsequent serologic analysis by plate ELISA using species-specific peptides revealed antibodies to E. ewingii, E. canis, and E. chaffeensis in 46% (23/50), 18% (9/50), and 4% (2/50) of serum samples, respectively. Taken together, these data indicate that coyotes in this region are commonly exposed to SFG Rickettsia and E. ewingii and that further consideration of coyotes as a component of the maintenance cycle for these pathogens may be warranted.

Further investigation of exposure to Lawsonia intracellularis in wild and feral animals captured on horse properties with equine proliferative enteropathy.

This study investigated the exposure to Lawsonia intracellularis in wild birds, mice, rabbits, raccoons, coyotes and squirrels, and feral cats and pigs on 10 farms with confirmed equine proliferative enteropathy (EPE). Serum samples from all resident foals (417 samples) as well as fecal (461) and serum (106) samples from wild and feral animals were collected for serological and molecular detection of L. intracellularis following the diagnosis of EPE in index cases. A total of three cats from two farms, three mice from two farms and eight cottontail rabbits from one farm had evidence of prior exposure to L. intracellularis. These animals may be an indicator of environmental exposure or may be actively involved in the transmission of L. intracellularis to foals by acting as a potential reservoir/amplifier host.

Detection of Dirofilaria immitis and Ehrlichia species in coyotes (Canis latrans), from rural Oklahoma and Texas.

There is a lack of knowledge regarding the prevalence of Dirofilaria immitis and Ehrlichia spp. in coyotes in Oklahoma and Texas. Documenting the prevalence of these vector-borne disease agents in coyotes from Oklahoma and Texas underscores the importance of wild canids as reservoir hosts that infect companion animals and humans. To learn more about the sylvatic cycle of D. immitis and Ehrlichia spp. in coyotes from Oklahoma and Texas, we tested for infection with and exposure to, respectively, these disease agents. Coyote carcasses were collected opportunistically from animal control experts and hunters in seven counties in Oklahoma and Texas from January to March, 2010. Serum samples from 77 coyotes were tested with a commercial ELISA test. Five (6.5%) coyotes had D. immitis antigens, and four (5.2%) had antibodies to Ehrlichia spp. The overall prevalence of D. immitis was low relative to studies from the eastern United States. Little is known about the prevalence of Ehrlichia spp. throughout the United States, but coyotes from rural Oklahoma in the current study had a higher exposure rate than those reported from California, and a lower rate than data from an earlier study from Oklahoma.

Annual seroprevalence of Yersinia pestis in coyotes as predictors of interannual variation in reports of human plague cases in Arizona, United States.

Although several health departments collect coyote blood samples for plague surveillance, the association between reported human cases and coyote seroprevalence rates remains anecdotal. Using data from an endemic region of the United States, we sought to quantify this association. From 1974 to 1998, about 2,276 coyote blood samples from four Arizona counties were tested for serological evidence of exposure to Yersinia pestis, the causative agent of plague. Using a titer threshold presumed to be indicative of recent infection (serum titers of ≥1:256), we found a statistically significant relationship between years with >17% sero-positive coyotes and years with two or more human cases reported. Moreover, when the annual coyote seroprevalence rates were dichotomized at 17%, 84% of the years were correctly classified using four biologically relevant meteorological variables in a linear regression. This is the first time a statistically significant temporal association between human plague cases and coyote seroprevalence rates has been shown. However, issues with data resolution and surveillance effort that potentially limit the public health utility of using coyote seroprevalence rates are discussed.

Active use of coyotes (Canis latrans) to detect Bovine Tuberculosis in northeastern Michigan, USA.

Bovine tuberculosis (bTB) is endemic in white-tailed deer (Odocoileus virginianus) in northeastern Michigan, USA, and research suggests transmission to cattle. Prevalence of the disease in deer is estimated at 1.8%, but as prevalence decreases the difficulty of detection increases. Research suggests coyotes (Canis latrans) have a higher prevalence of bTB in Michigan than deer and sampling coyotes may be a more efficient surveillance tool to detect presence or spread of the disease. Coyotes possess suitable ecological characteristics to serve as a sentinel species, assuming transmission between coyotes is not significant. The question of whether free-ranging coyotes shed Mycobacterium bovis, the causative agent of bTB, has not been previously addressed. We actively used coyotes as a sentinel to detect bTB in infected and uninfected counties in Michigan's Northeastern Lower Peninsula. We determined whether bTB infection was present through bacteriologic culture of lymph nodes and tissues containing lesions and cultured oral/nasal swabs and feces to establish shedding. Seventeen of 171 coyotes were M. bovis culture positive, one of which was from a previously uninfected county. All oral, nasal secretions and feces were culture negative suggesting minimal, if any, shedding of M. bovis. Thus, infection of coyotes is likely to occur through ingestion of infected deer carcasses and not from interaction with conspecifics. These findings support previous research suggesting that coyotes are useful sentinels for bTB. The use of coyotes as a sentinel, may allow wildlife managers to detect the spread of bTB into naïve counties. With earlier detection managers may be able to take proactive surveillance measures to detect the disease in deer and reduce the potential risk to domestic livestock and captive deer herds.

Antigenic profiling of yersinia pestis infection in the Wyoming coyote (Canis latrans).

Although Yersinia pestis is classified as a "high-virulence" pathogen, some host species are variably susceptible to disease. Coyotes (Canis latrans) exhibit mild, if any, symptoms during infection, but antibody production occurs postinfection. This immune response has been reported to be against the F1 capsule, although little subsequent characterization has been conducted. To further define the nature of coyote humoral immunity to plague, qualitative serology was conducted to assess the antiplague antibody repertoire. Humoral responses to six plasmid-encoded Y. pestis virulence factors were first examined. Of 20 individual immune coyotes, 90% were reactive to at least one other antigen in the panel other than F1. The frequency of reactivity to low calcium response plasmid (pLcr)-encoded Yersinia protein kinase A (YpkA) and Yersinia outer protein D (YopD) was significantly greater than that previously observed in a murine model for plague. Additionally, both V antigen and plasminogen activator were reactive with over half of the serum samples tested. Reactivity to F1 was markedly less frequent in coyotes (35%). Twenty previously tested antibody-negative samples were also examined. While the majority were negative across the panel, 15% were positive for 1-3 non-F1 antigens. In vivo-induced antigen technology employed to identify novel chromosomal genes of Y. pestis that are up-regulated during infection resulted in the identification of five proteins, including a flagellar component (FliP) that was uniquely reactive with the coyote serum compared with immune serum from two other host species. Collectively, these data suggest that humoral immunity to pLcr-encoded antigens and the pesticin plasmid (pPst)-encoded Pla antigen may be relevant to plague resistance in coyotes. The serologic profile of Y. pestis chromosomal antigens up-regulated in vivo specific to C. latrans may provide insight into the differences in the pathogen-host responses during Y. pestis infection.

Epizootiologic survey of Mycobacterium bovis in wildlife and farm environments in northern Michigan.

Bovine tuberculosis (bovine TB), caused by Mycobacterium bovis, has reemerged in northern Michigan, USA, with detections in white-tailed deer (Odocoileus virginianus) in 1994 and in cattle in 1998. Since then, significant efforts have been directed toward reducing deer densities in the area in the hopes of reducing the bovine TB prevalence rate in deer and eliminating spillover of the disease into cattle. Despite the success of the efforts to reduce deer densities, additional cattle herds have become infected. Other mammals can be infected with M. bovis, and some carnivores and omnivores had been found to be infected with the disease in northern Michigan, USA. We conducted a multiyear surveillance effort to detect bovine TB in wild species of mammals in the Michigan, USA, outbreak area. From 2002 to 2004, tissue samples from 1,031 individual animals of 32 species were collected, processed, and cultured for M. bovis. Only 10 (1.0%) were culture-positive for M. bovis (five raccoons [Procyon lotor], four opossums [Didelphis virginiana], and one grey fox [Urocyon cinereoargenteus]). We also found two raccoons and four opossums to be positive for Mycobacterium avium. We collected 503 environmental samples from cattle farms recently identified as bovine TB positive; none yielded positive M. bovis culture results. Finally, we used infrared cameras to document wildlife use of four barns in the area. Many avian and mammalian species of wildlife were observed, with raccoons being the most commonly observed species. This surveillance study identified no new wildlife species that should be considered significant reservoirs of bovine TB in the outbreak area in northern Michigan, USA. However, the relatively high, apparent bovine TB prevalence rates in some carnivorous and omnivorous species, their relatively long life spans, and their frequent use of barns, suggests that removal of raccoons, opossums, foxes, and coyotes (Canis latrans) should be considered when a newly infected farm is depopulated of cattle.

Bartonella rochalimae in raccoons, coyotes, and red foxes.

To determine additional reservoirs for Bartonella rochalimae, we examined samples from several wildlife species. We isolated B. rochalimae from 1 red fox near Paris, France, and from 11 raccoons and 2 coyotes from California, USA. Co-infection with B. vinsonii subsp. berkhoffii was documented in 1 of the coyotes.

Hematology, parasitology, and serology of free-ranging coyotes (Canis latrans) from South Carolina.

Blood and feces were collected from 34 adult (19 males, 15 females) and seven juvenile (three males, one female, three not reported) free-ranging coyotes (Canis latrans) on the US Department of Energy's Savannah River Site (South Carolina, USA). Significant (P<0.05) hematologic differences by sex were noted for red blood cell counts, hemoglobin, and hematocrit. Biochemical differences by sex occurred only for albumen (P<0.05). Twenty-one adults were antibody positive for at least one of four viruses: canine adenovirus type 1 (CAV-1; 68%), West Nile virus (WNV; 60%), Eastern equine encephalitis virus (EEEV; 38%), and Canine distemper virus (CDV; 15%). Of the seven Leptospira serovars tested for, seven (25%) of 28 adults were positive for one or more of five serovars: Pomona, Grippotyphosa, Icterohaemorrhagiae, Bratislava, and Autumnalis. Three (43%) of seven juveniles had seropositivity for a virus, one each for CDV, CAV-1, and WNV. No juveniles were seropositive for EEEV or any of the seven Leptospira serovars. Blood smears of 12 adults were positive for Dirofilaria immitis microfilaria, but blood smears from all juveniles were negative. Parvovirus was identified by electron microscopy from the feces of one adult. Ancylostoma spp., Trichuris spp., and Isospora spp. were observed in fecal samples. These data may aid in understanding the role of coyotes in disease ecology.

Colorado animal-based plague surveillance systems: relationships between targeted animal species and prediction efficacy of areas at risk for humans.

Human plague risks (Yersinia pestis infection) are greatest when epizootics cause high mortality among this bacterium's natural rodent hosts. Therefore, health departments in plague-endemic areas commonly establish animal-based surveillance programs to monitor Y. pestis infection among plague hosts and vectors. The primary objectives of our study were to determine whether passive animal-based plague surveillance samples collected in Colorado from 1991 to 2005 were sampled from high human plague risk areas and whether these samples provided information useful for predicting human plague case locations. By comparing locations of plague-positive animal samples with a previously constructed GIS-based plague risk model, we determined that the majority of plague-positive Gunnison's prairie dogs (100%) and non-prairie dog sciurids (85.82%), and moderately high percentages of sigmodontine rodents (71.4%), domestic cats (69.3%), coyotes (62.9%), and domestic dogs (62.5%) were recovered within 1 km of the nearest area posing high peridomestic risk to humans. In contrast, the majority of white-tailed prairie dog (66.7%), leporid (cottontailed and jack rabbits) (71.4%), and black-tailed prairie dog (93.0%) samples originated more than 1 km from the nearest human risk habitat. Plague-positive animals or their fleas were rarely (one of 19 cases) collected within 2 km of a case exposure site during the 24 months preceding the dates of illness onset for these cases. Low spatial accuracy for identifying epizootic activity prior to human plague cases suggested that other mammalian species or their fleas are likely more important sources of human infection in high plague risk areas. To address this issue, epidemiological observations and multi-locus variable number tandem repeat analyses (MLVA) were used to preliminarily identify chipmunks as an under-sampled, but potentially important, species for human plague risk in Colorado.

Surveillance of coyotes to detect bovine tuberculosis, Michigan.

Bovine tuberculosis (TB) is endemic in white-tailed deer (Odocoileus virginianus) in the northeastern portion of Michigan's Lower Peninsula. Bovine TB in deer and cattle has created immense financial consequences for the livestock industry and hunting public. Surveillance identified coyotes (Canis latrans) as potential bio-accumulators of Mycobacterium bovis, a finding that generated interest in their potential to serve as sentinels for monitoring disease risk. We sampled 175 coyotes in the bovine TB-endemic area. Fifty-eight tested positive, and infection prevalence by county ranged from 19% to 52% (statistical mean 33%, SE 0.07). By contrast, prevalence in deer (n = 3,817) was lower (i.e., 1.49%; Mann-Whitney U4,4 = 14, p<0.001). By focusing on coyotes rather than deer, we sampled 97% fewer individuals and increased the likelihood of detecting M. bovis by 40%. As a result of reduced sampling intensity, sentinel coyote surveys have the potential to be practical indicators of M. bovis presence in wildlife and livestock.