Fecal contamination of urban parks by domestic dogs and tragedy of the commons.

Contamination of public parks by dogs is a potential source of conflict among park users, causing "tragedy of the commons" problems. Besides the social conflict, feces can pose serious health risks to both dogs and humans. In this study we analyzed the extent and patterns of the distribution of dog feces in the urban parks of the City of Calgary. We collected dog feces from randomly selected locations in the urban parks. The average density of dog feces by the different dog leash policies of the parks and the distribution pattern of the fecal density within the parks were assessed, and the total contamination of the public parks for the entire city was estimated. We found off-leash parks to be significantly more contaminated than other types of parks. We estimated 127.23 g/ha of dog feces are left unpicked in city parks in total every week. Dog feces were found more often and in greater amount in off-leash parks, and near park entrances and parking lots, than in on-leash parks and away from the park entrances. These results suggest that public park visitors, especially those visiting off-leash parks, are likely to be exposed to large amounts of dog feces. Designation of parks as on-leash and educating dog-owners may be an effective approach for reducing the fecal contamination.

Echinococcus multilocularis in urban coyotes, Alberta, Canada.

Echinococcus multilocularis is a zoonotic parasite in wild canids. We determined its frequency in urban coyotes (Canis latrans) in Alberta, Canada. We detected E. multilocularis in 23 of 91 coyotes in this region. This parasite is a public health concern throughout the Northern Hemisphere, partly because of increased urbanization of wild canids.

Genetic variation of the mitochondrial DNA control region across plains bison herds in USA and Canada.

Once numbering in the tens of millions, bison (Bison bison) have faced a broad range of challenges over the past century, including genetic impacts from the population bottleneck in the 1800s, and significant loss and fragmentation of habitat resulting in little opportunity for population growth on remaining small, geographically isolated reserves. To identify best practices for bison stewardship against this backdrop, managers must understand the genetic composition of existing conservation herds. This study characterized 14 plains bison (Bison bison bison) herds managed by the U.S. Department of Interior and Parks Canada Agency based on complete mtDNA control region sequences. Among 209 bison, we detected 11 major mtDNA control region haplotypes based on nucleotide substitutions and 23 sub-haplotypes where indels are considered. We determined matrilineal relationships between the herds and compared our genetic findings to historic records. The recent common ancestry of modern bison deriving from small, scattered groups combined with gene flow through foundation and translocation events between herds during the last 100 years, is reflected in Fst value (0.21), haplotype (0.48 ± 0.04) and nucleotide (0.004 ± 0.002) diversities, and mean number of pairwise differences (3.38 ± 1.74). Genetic diversity was distributed unevenly among herds, with 21.41% of genetic variation observed between herds. Median joining network, together with trends in the Tajima's D and Fs tests, revealed two patterns in the recent evolution of mtDNA sequences in bison: mutational process has generated diversity with a Hap 1 haplotype epicenter, and missed mtDNA haplotypes exist in the network due to bottleneck, loss through management practices, or incomplete sampling of specimens across conservation herds. This work significantly expands characterization of the genetic diversity among bison conservation herds, providing additional decision support for managers considering restoring gene flow to achieve long-term species viability.

Droplet digital PCR as a sensitive tool to assess exposure pressure from Echinococcus multilocularis in intermediate hosts.

A key element to understanding parasite epidemiology is assessing their prevalence in the respective wild reservoir hosts. The tapeworm Echinococcus multilocularis circulates between canid species (definite hosts) and small mammals (mostly rodents; intermediate hosts). Prevalence rates of Echinococcus multilocularis in the intermediate host are most exclusively determined through macroscopic examination of the liver generally followed by molecular or histological diagnostic for parasite species confirmation. The overall objective of the study was to investigate the suitability of Real-Time PCR and Droplet Digital PCR (ddPCR) analysis as tool to detect exposure pressure (frequency of infection events) from E. multilocularis in intermediate hosts even in the absence of macroscopic lesions in the liver. One hundred six small mammals (meadow voles and deer mice) were trapped followed by post-mortem examination including macroscopic evaluation of the liver to detect lesions indicative of infection with Echinococcus multilocularis but also by sampling a piece of liver in absence of lesion to submit it to molecular assay. Macroscopic lesions were present in the livers of two samples. Including the latter two samples, five samples yielded a positive result following Real-Time PCR, whereas 16 samples displayed three or more positive droplets upon ddPCR and were considered positive. Whether these additional cases without macroscopic lesions would have become infectious during the lifespan of the rodent or were abortive or early infections is unclear, but these data suggest levels of exposure of intermediate hosts to the parasite is much higher than assumed.

Molecular characterization of Giardia spp. and Cryptosporidium spp. from dogs and coyotes in an urban landscape suggests infrequent occurrence of zoonotic genotypes.

Giardia spp. and Cryptosporidium spp. are common gastrointestinal parasites with the potential for zoonotic transmission. This study aimed to (1) determine the genotypes occurring in dogs and coyotes occupying a similar urban area; (2) determine if these hosts were infected with potentially zoonotic genotypes; (3) provide baseline molecular data. In August and September 2012, 860 dog owners living in neighborhoods bordering six urban parks in Calgary, Alberta, Canada, provided faecal samples from their dogs. From March 2012 through July 2013, 193 coyote faeces were also collected from five of six of the same parks. Direct immunofluorescence microscopy (DFA) indicated that Giardia spp. and Cryptosporidium spp. infected a total of 64 (7.4%) and 21 (2.4%) dogs, as well as 15 (7.8%) and three (1.6%) coyotes, respectively. Semi-nested, polymerase chain reactions targeting the 16S small-subunit ribosomal ribonucleic acid (SSU rRNA) and 18S SSU rRNA genes of Giardia spp. and Cryptosporidium spp., respectively, were conducted on samples that screened positive by DFA, and products were sequenced and genotyped. Dogs were infected with Giardia intestinalis canid-associated assemblages C (n = 14), D (n = 13), and Cryptosporidium canis (n = 3). Similarly, G. intestinalis assemblages C (n = 1), D (n = 1) and C. canis (n = 1), were detected in coyotes, as well as G. intestinalis assemblage A (n = 1) and Cryptosporidium vole genotype (n = 1). Dogs and coyotes were predominantly infected with host-specific genotypes and few potentially zoonotic genotypes, suggesting that they may not represent a significant risk for zoonotic transmission of these parasites in urban areas where these hosts are sympatric.

A community analysis approach to parasite transmission in multi-host systems: Assemblages of small mammal prey and Echinococcus multilocularis in an urban area in North America.

BACKGROUND: Echinococcus multilocularis (Em) is a parasite with a complex life cycle whose transmission involves a predator-prey interaction. Accidental ingestion of Em eggs by humans may cause alveolar echinococcosis, a potentially fatal disease. Although previous research suggested that the composition of the assemblage of prey species may play a key role in the transmission, the relation between Em presence and the prey assemblages has never been analyzed. Herein, we propose a community analysis approach, based on assemblage similarity statistics, clustering, non-metric dimensional scaling and GLM modelling to analyze the relationships between small mammal assemblages, environmental variables, and the prevalence of Em in intermediate and definitive hosts in an urban area. RESULTS: In our study areas within the City of Calgary, Alberta (Canada), we identified three main small mammal assemblages associated with different prevalence of Em, characterized by a different proportion of species known to be good intermediate hosts for Em. As expected, assemblages with higher proportion of species susceptible to Em were observed with higher prevalence of parasite, whereas the total abundance per se of small mammals was not a predictor of transmission likely due to dilution effect. Furthermore, these assemblages were also predicted by simple environmental proxies such as land cover and terrain. CONCLUSIONS: Our results indicated that the use of a community analysis approach allows for robust characterization of these complex and multivariate relationships, and may offer a promising tool for further understanding of parasite epidemiology in complex multi-host systems. In addition, this analysis indicates that it is possible to predict potential foci of disease risk within urban areas using environmental data commonly available to city planners and land managers.

Neospora caninum DNA in Coyote Fecal Samples Collected in an Urban Environment.

Definitive hosts of Neospora caninum are species of canids, such as domestic dogs ( Canis lupus familiaris), coyotes ( Canis latrans) and foxes ( Vulpes vulpes), whereas ruminants, such as cattle ( Bos taurus), sheep ( Ovis aries), and deer (Cervidae) serve as intermediate hosts. We investigated the presence of N. caninum in feces of coyotes in Calgary, Alberta, Canada. One hundred coyote fecal samples were collected from five city parks. Following DNA extraction, the presence of N. caninum DNA was determined by using real-time PCR analysis with a primer and probe pair targeting the Nc5 gene. Ten of the 100 samples contained detectable amounts of N. caninum DNA. White-tailed deer ( Odocoileus virginianus) and mule deer ( Odocoileus hemionus) are common in Calgary's city parks, and although we did not examine them for the presence of N. caninum bradyzoites, the presence of N. caninum DNA in coyote fecal samples is indicative that a sylvatic host of N. caninum exists within this urban environment.

High species diversity of trichostrongyle parasite communities within and between Western Canadian commercial and conservation bison herds revealed by nemabiome metabarcoding.

BACKGROUND: Many trichostrongylid nematode species are reported to infect bison, some of which are major causes of disase and production loss in North American bison herds. However, there is little information on the species distribution and relative abundance of these parasites in either commercial or conservation herds. This is largely because trichostrongylid nematode species cannot be distinguished by visual microscopic examination of eggs present in feces. Consequently, we have applied ITS2 rDNA nemabiome metabarcoding to describe the trichostrongyle parasite species diversity in 58 bison production groups derived from 38 commercial North American plains bison (Bison bison bison) herds from across western Canada, and two bison conservation herds located in Elk Island National Park (EINP) [plains bison and wood bison (Bison bison athabascae)] and one in Grasslands National Park (GNP) (plains bison). RESULTS: We report much higher infection intensities and parasite species diversity in commercial bison herds than previously reported in beef cattle herds grazing similar latitudes. Predominant trichostrongyle parasite species in western Canadian commercial bison herds are those commonly associated with Canadian cattle, with Ostertagia ostertagi being the most abundant followed by Cooperia oncophora. Combined with high fecal egg counts in many herds, this is consistent with significant clinical and production-limiting gastrointestinal parasitism in western Canadian bison herds. However, Haemonchus placei was the most abundant species in five of the production groups. This is both surprising and important, as this highly pathogenic blood-feeding parasite has not been reported at such abundance, in any livestock species, at such northerly latitudes. The presence of Trichostrongylus axei as the most abundant parasite in four herds is also unusual, relative to cattle. There were striking differences in parasite communities between the EINP and commercial bison herds. Most notably, Orloffia bisonis was the predominant species in the wood bison herd despite being found at only low levels in all other herds surveyed. CONCLUSIONS: This study represents the most comprehensive description of parasite communities in North American bison to date and illustrates the power of deep amplicon sequencing as a tool to study species diversity in gastrointestinal nematode communities.

Wilderness in the 'city' revisited: different urbes shape transmission of Echinococcus multilocularis by altering predator and prey communities.

The urbanization of Echinococcus multilocularis, the agent of alveolar echinococcosis (AE), is a public health concern worldwide. Here we propose to consider 'urban' habitats under a broad ecological perspective and discuss the effects of human settlements (urbes) on host communities and the process of parasite urbanization. We argue that interactions between landscape features (i.e., landscape composition and configuration) and host communities can shape the heterogeneity of transmission gradients observed within and across different types of human settlement. Due to unique ecological characteristics and public health management priorities, we envisage urban landscapes as a model system to further increase our understanding of host-parasite interactions shaping the circulation of E. multilocularis.

Feeding ecology informs parasite epidemiology: prey selection modulates encounter rate with Echinococcus multilocularis in urban coyotes.

We investigated the role of urban coyote feeding ecology in the transmission of Echinococcus multilocularis, the causative agent of Alveolar Echinococcosis in humans. As coyotes can play a main role in the maintenance of this zoonotic parasite within North American urban settings, such study can ultimately aid disease risk management. Between June 2012 and June 2013, we collected 251 coyote feces and conducted trapping of small mammals (n = 971) in five parks in the city of Calgary, Alberta, Canada. We investigated E. multilocularis epidemiology by assessing seasonal variations of coyote diet and the selective consumption of different rodent intermediate host species. Furthermore, accounting for small mammal digestibility and coyote defecation rates we estimated the number of small mammal preys ingested by coyote and consequently, coyote encounter rates with the parasite. Dominant food items included small mammals, fruit and vegetation, although hare and deer were seasonally relevant. The lowest frequency of occurrence per scat of small mammals was recorded in winter (39.4%), when consumption of deer was highest (36.4%). However, highest encounter rates (number of infected hosts predated/season) with E. multilocularis (95% CI: 1.0-22.4), combined with the lack of predation on non-competent small mammal species, suggest that winter is the critical season for transmission and control of this parasite. Within the small mammal assemblage, voles (Microtus pennsylvanicus and Myodes gapperi) were the selected preys of urban coyotes and likely played a key role for the maintenance of the urban sylvatic life-cycle of E. multilocularis in Calgary.

Echinococcus multilocularis in North America: the great unknown.

Over the last decade, studies have begun to shed light on the distribution and genetic characterization of Echinococcus multilocularis, the causative agent of alveolar echinococcosis (AE), in North America. Recent findings indicate that the parasite is likely expanding its range in the central region of the United States and Canada and that invasions of European strains might have occurred. In our review, we present the available data on E. multilocularis infections in wild and domestic animals and humans in North America and emphasize the lack of knowledge on the distribution of the parasite in wild and domestic hosts. Furthermore, we stress the need to better understand the complexity of host communities and their roles in shaping the transmission and distribution of the parasite. We hypothesize that a lack of knowledge about AE by North American physicians might result in the misdiagnosis of cases and an underestimation of disease incidence. The endemic presence of the parasite in urban areas and a recent human case in Alberta, Canada, suggest that the scientific community may need to reconsider the local public health risks, re-assess past cases that might have been overlooked and increase surveillance efforts to identify new cases of human AE.

Spatial heterogeneity and temporal variations in Echinococcus multilocularis infections in wild hosts in a North American urban setting.

Echinococcus multilocularis, the causative agent of human alveolar echinococcosis, has the potential to circulate in urban areas where wild host populations and humans coexist. The spatial and temporal distribution of infection in wild hosts locally affects the risk of transmission to humans. We investigated the spatial and temporal patterns of E. multilocularis infection in coyotes and rodent intermediate hosts within the city of Calgary, Canada, and the association between spatial variations in coyote infection and the relative composition of small mammal assemblages. Infection by E. multilocularis was examined in small mammals and coyote faeces collected monthly in five city parks from June 2012 to June 2013. Coyote faeces were analysed using a ZnCl(2) centrifugation and sedimentation protocol. Infection in intermediate hosts was assessed through lethal trapping and post-mortem analysis. Parasite eggs and metacestodes were morphologically identified and molecularly confirmed through species-specific PCR assays. Of 982 small mammals captured, infection was detected in 2/305 (0.66%) deer mice (Peromyscus maniculatus), 2/267 (0.75%) meadow voles (Microtus pennsylvanicus), and 1/71 (1.41%) southern red backed voles (Myodes gapperi). Overall faecal prevalence in coyotes was 21.42% (n = 385) and varied across sites, ranging from 5.34% to 61.48%. Differences in coyote faecal prevalence across sites were consistent with local variations in the relative abundance of intermediate hosts within the small mammal assemblages. Infections peaked in intermediate hosts during autumn (0.68%) and winter (3.33%), and in coyotes during spring (43.47%). Peaks of infections in coyote faeces up to 83.8% in autumn were detected in a hyper-endemic area. To the best of our knowledge, our findings represent the first evidence of a sylvatic life-cycle of E. multilocularis in a North American urban setting, and provide new insights into the complexity of the parasite transmission ecology.

Unexpected diversity of the cestode Echinococcus multilocularis in wildlife in Canada.

Echinococcus multilocularis is a zoonotic cestode with a distribution encompassing the northern hemisphere that causes alveolar hydatid disease in people and other aberrant hosts. E. multilocularis is not genetically uniform across its distribution, which may have implications for zoonotic transmission and pathogenicity. Recent findings of a European-type haplotype of E. multilocularis in wildlife in one location in western Canada motivated a broader survey of the diversity of this parasite in wildlife from northern and western Canada. We obtained intact adult cestodes of E. multilocularis from the intestines of 41 wild canids (wolf - Canis lupus, coyote - Canis latrans, and red fox - Vulpes vulpes), taeniid eggs from 28 fecal samples from Arctic fox (Vulpes lagopus), and alveolar hydatid cysts from 39 potential rodent intermediate hosts. Upon sequencing a 370-nucelotide region of the NADH dehydrogenase subunit 1 (nad1) mitochondrial locus, 17 new haplotypes were identified. This constitutes a much higher diversity than expected, as only two genotypes (European and an Asian/North American) had previously been identified using this locus. The European-type strain, recently introduced, may be widespread in wildlife within western Canada, possibly related to the large home ranges and wide dispersal range of wild canids. This study increased understanding of the biogeographic distribution, prevalence and genetic differences of a globally important pathogenic cestode in northern and western Canada.

A new intermediate host for Echinococcus multilocularis: the southern red-backed vole (Myodes gapperi) in urban landscape in Calgary, Canada.

Human Alveolar Echinococcosis (HAE) is a potentially fatal parasitic disease caused by Echinococcus multilocularis, a cestode characterized by a sylvatic life-cycle involving several species of rodents and lagomorphs as intermediate hosts and canids as definitive hosts. Despite the wide distribution of the parasite in North America, the number of competent intermediate host species identified to date is still relatively small, and mainly includes the northern vole (Microtus oeconomus), brown lemming (Lemmus sibiricus), northern red-backed vole (Myodes rutilus), deer mouse (Peromyscus maniculatus) and meadow vole (Microtus pennsylvanicus). By monitoring the infections in rodents in the city of Calgary (Alberta, Canada), we have detected a case of severe alveolar echinococcosis in a southern red-backed vole (Myodes gapperi), a species never reported before as an intermediate host for this parasite. Observation of protoscolices in the intra-abdominal multilocular cysts indicates that M. gapperi could act as a competent intermediate host for the transmission of E. multilocularis. Since M. gapperi can be found in close proximity to, and within metropolitan areas, this species could play a role in the establishment and maintenance of the sylvatic life-cycle of E. multilocularis in urban landscapes, where the potential for zoonotic transmission is higher. The new intermediate host reported needs to be taken into account in future surveys and transmission models for this parasite.

Sensitivity of double centrifugation sugar fecal flotation for detecting intestinal helminths in coyotes (Canis latrans).

Fecal analysis is commonly used to estimate prevalence and intensity of intestinal helminths in wild carnivores, but few studies have assessed the reliability of fecal flotation compared to analysis of intestinal tracts. We investigated sensitivity of the double centrifugation sugar fecal flotation and kappa agreement between fecal flotation and postmortem examination of intestines for helminths of coyotes (Canis latrans). We analyzed 57 coyote carcasses that were collected between October 2010 and March 2011 in the metropolitan area of Calgary and Edmonton, Alberta, Canada. Before analyses, intestines and feces were frozen at -80 C for 72 hr to inactivate Echinococcus eggs, protecting operators from potential exposure. Five species of helminths were found by postmortem examination, including Toxascaris leonina, Uncinaria stenocephala, Ancylostoma caninum, Taenia sp., and Echinococcus multilocularis. Sensitivity of fecal flotation was high (0.84) for detection of T. leonina but low for Taenia sp. (0.27), E. multilocularis (0.46), and U. stenocephala (0.00). Good kappa agreement between techniques was observed only for T. leonina (0.64), for which we detected also a significant correlation between adult female parasite intensity and fecal egg counts (R(s)=0.53, P=0.01). Differences in sensitivity may be related to parasite characteristics that affect recovery of eggs on flotation. Fecal parasitologic analyses are highly applicable to study the disease ecology of urban carnivores, and they often provide important information on environmental contamination and potential of zoonotic risks. However, fecal-based parasitologic surveys should first assess the sensitivity of the techniques to understand their biases and limitations.