Elizabeth VanWormer

Molecules to modeling: Toxoplasma gondii oocysts at the human-animal-environment interface

Environmental transmission of extremely resistant Toxoplasma gondii oocysts has resulted in infection of diverse species around the world, leading to severe disease and deaths in human and animal populations. This review explores T. gondii oocyst shedding, survival, and transmission, emphasizing the importance of linking laboratory and landscape from molecular characterization of oocysts to watershed-level models of oocyst loading and transport in terrestrial and aquatic systems. Building on discipline-specific studies, a One Health approach incorporating tools and perspectives from diverse fields and stakeholders has contributed to an advanced understanding of T. gondii and is addressing transmission at the rapidly changing human-animal-environment interface.

Educating pastoralists and extension officers on diverse livestock diseases in a changing environment in Tanzania

East African pastoralists and their livestock are vulnerable to alterations in resource availability and disease transmission and frequently face poor access to livestock health services. Government veterinarians tasked with guiding health services must prioritize livestock health risks and allocate limited resources across disparate ecosystems with different disease threats. To identify livestock diseases of concern and strategies for improving herd health and resilience, we conducted community focus groups with pastoralists and interviewed pastoralist household leaders, village extension officers, and government veterinary officials in south-central Tanzania, an area experiencing rapid population growth and environmental change. All participants discussed pastoralist access to livestock health services, livestock disease priorities, and means to improve livestock health.Perceptions of diseases of importance differed among pastoralists, extension officers, and government veterinarians. Spatial differences in diseases of concern among study area pastoralists emphasized the need for locally adaptable livestock health service delivery. Although pastoralist strategies to improve livestock health differed by ethnic group, many pastoralists as well as extension officers and government veterinarians identified livestock health education and training for pastoralists and extension officers as a critical need.Policies designed at the regional, rather than the local, level may not reflect the disease concerns of the entire area. To effectively address veterinary health problems and make livestock herds more resilient to environmental change, conditions at the local level must be considered. Education targeted to pastoralist households and extension officers could achieve greater flexibility in the livestock health system and provide more reliable information about local conditions for governmental policymakers.

Congenital Transmission of Toxoplasma gondii in Deer Mice (Peromyscus maniculatus) After Oral Oocyst Infection

Abstract To investigate how different routes of Toxoplasma gondii transmission influence the antibody response and infection status of deer mice (Peromyscus maniculatus), 80 mice were orally infected with 1, 5, 10, or 100 T. gondii oocysts. Ten weeks postinfection, 15 T. gondii–seropositive female mice were bred and allowed to produce 2 litters. Evidence of persistent T .gondii infection in orally infected mice was detected by serology and DNA amplification in mice from all 4 oocyst treatment groups, including those that received only a single T. gondii oocyst. Congenital transmission of T. gondii was detected by polymerase chain reaction (PCR) in 7/8 first and 4/7 second litters. Toxoplasma gondii was also detected by PCR in 9/30 congenitally infected offspring 16 wk after birth, despite the fact that detectable serological titers had waned. These findings raise questions about the applicability of serological testing to assess the prevalence of T. gondii infection in deer mice and other rodents in the wild. Additionally, the detection of frequent congenital transmission suggests that deer mice could help maintain T. gondii in the environment even in the absence of definitive feline hosts.

Using molecular epidemiology to track Toxoplasma gondii from terrestrial carnivores to marine hosts: implications for public health and conservation.

Background Environmental transmission of the zoonotic parasite Toxoplasma gondii, which is shed only by felids, poses risks to human and animal health in temperate and tropical ecosystems. Atypical T. gondii genotypes have been linked to severe disease in people and the threatened population of California sea otters. To investigate land-to-sea parasite transmission, we screened 373 carnivores (feral domestic cats, mountain lions, bobcats, foxes, and coyotes) for T. gondii infection and examined the distribution of genotypes in 85 infected animals sampled near the sea otter range. Methodology/Principal Findings Nested PCR-RFLP analyses and direct DNA sequencing at six independent polymorphic genetic loci (B1, SAG1, SAG3, GRA6, L358, and Apico) were used to characterize T. gondii strains in infected animals. Strains consistent with Type X, a novel genotype previously identified in over 70% of infected sea otters and four terrestrial wild carnivores along the California coast, were detected in all sampled species, including domestic cats. However, odds of Type X infection were 14 times higher (95% CI: 1.3–148.6) for wild felids than feral domestic cats. Type X infection was also linked to undeveloped lands (OR = 22, 95% CI: 2.3–250.7). A spatial cluster of terrestrial Type II infection (P = 0.04) was identified in developed lands bordering an area of increased risk for sea otter Type II infection. Two spatial clusters of animals infected with strains consistent with Type X (P≤0.01) were detected in less developed landscapes. Conclusions Differences in T. gondii genotype prevalence among domestic and wild felids, as well as the spatial distribution of genotypes, suggest co-existing domestic and wild T. gondii transmission cycles that likely overlap at the interface of developed and undeveloped lands. Anthropogenic development driving contact between these cycles may increase atypical T. gondii genotypes in domestic cats and facilitate transmission of potentially more pathogenic genotypes to humans, domestic animals, and wildlife.

Canine Distemper in an Isolated Population of Fishers (Martes pennanti) from California

Four fishers (Martes pennanti) from an insular population in the southern Sierra Nevada Mountains, California, USA died as a consequence of an infection with canine distemper virus (CDV) in 2009. Three fishers were found in close temporal and spatial relationship; the fourth fisher died 4 mo later at a 70 km distance from the initial group. Gross lesions were restricted to hyperkeratosis of periocular skin and ulcera-tion of footpads. All animals had necrotizing bronchitis and bronchiolitis with syncytia and intracytoplasmic inclusion bodies. Inclusion bodies were abundant in the epithelia of urinary bladder and epididymis but were infrequent in the renal pelvis and the female genital epithelia. No histopathologic or immu-nohistochemical evidence for virus spread to the central nervous system was found. One fisher had encephalitis caused by Sarcocystis neurona and another had severe head trauma as a consequence of predation. The H gene nucleo-tide sequence of the virus isolates from the first three fishers was identical and was 99.6% identical to the isolate from the fourth fisher. Phylogenetically, the isolates clustered with other North American isolates separate from classical European wildlife lineage strains. These data suggest that the European wildlife lineage might consist of two separate subgroups that are genetically distinct and endemic in different geographic regions. The source of infection as well as pertinent transmission routes remained unclear. This is the first report of CDV in fishers and underscores the significance of CDV as a pathogen of management concern.

Prevalence and risk factors associated with Sarcocystis neurona infections in opossums (Didelphis virginiana) from central California.

Sarcocystis neurona, a protozoal parasite shed by opossums (Didelphis virginiana), has been shown to cause significant morbidity and mortality in horses, sea otters, and other marine mammals. Over the course of 3 years (fall 2005-summer 2008), opossums from central California were tested for infection with S. neurona. Of 288 opossums sampled, 17 (5.9%) were infected with S. neurona based on the molecular characterization of sporocysts from intestinal scrapings or feces. Risk factors evaluated for association with S. neurona infection in opossums included: age, sex, location, season, presence of pouch young in females, concomitant infection, and sampling method (live-trapped or traffic-killed). Multivariate logistic regression analysis identified that opossums in the Central Valley were 9 times more likely to be infected than those near the coast (p=0.009). Similarly, opossum infection was 5 times more likely to be detected during the reproductive season (March-July; p=0.013). This first investigation of S. neurona infection prevalence and associated risk factors in opossums in the western United States can be used to develop management strategies aimed at reducing the incidence of S. neurona infections in susceptible hosts, including horses and threatened California sea otters (Enhydra lutris neries).

Surveillance for Toxoplasma gondii in California mussels (Mytilus californianus) reveals transmission of atypical genotypes from land to sea.

Coastal habitat contamination with Toxoplasma gondii is a health risk to humans and marine wildlife, with infections documented in both nearshore and pelagic marine mammals. Due to lack of sensitive methods for detection of T. gondii in water, this study utilized an alternative surveillance approach for evaluating marine habitat contamination using wild mussels. The objectives of this study were to (i) validate sensitive molecular tools for T. gondii detection in mussels and (ii) apply optimized methods in a surveillance study to determine the prevalence and genotype(s) of T. gondii in mussels. Simplex polymerase chain reaction screening and multiplex genotyping assays were validated and then applied on 959 wild-caught mussels collected from central California. Thirteen mussels (1.4%) had detectable T. gondii DNA and the presence of T. gondii in mussels was significantly associated with proximity to freshwater run-off and collection during the wet season. Molecular characterization revealed alleles from T. gondii types I, II/III, X at the B1 locus, and a novel atypical B1 allele that was recently documented in T. gondii-infected carnivores from California. Findings demonstrate higher than previously reported T. gondii contamination of California coastlines, and describe novel strains of the parasite that further link terrestrial sources with marine contamination.

Attempted Detection of Toxoplasma gondii Oocysts in Environmental Waters Using a Simple Approach to Evaluate the Potential for Waterborne Transmission in the Galápagos Islands, Ecuador

Toxoplasmosis is a health concern for wildlife and humans, particularly in island ecosystems. In the Galápagos Islands, exposure to Toxoplasma gondii has been found in marine avifauna on islands with and without domestic cats. To evaluate potential waterborne transmission of T. gondii, we attempted to use filtration and epifluorescent microscopy to detect autofluorescent T. gondii oocysts in fresh and estuarine surface water samples. T. gondii oocyst-like structures were microscopically visualized but were not confirmed by polymerase chain reaction and sequence analyses. Further research is needed to refine environmental pathogen screening techniques and to evaluate disease risk of waterborne zoonoses such as T. gondii for wildlife and humans, particularly in the Galápagos and other naive island ecosystems.

Coastal development and precipitation drive pathogen flow from land to sea: evidence from a Toxoplasma gondii and felid host system

Rapidly developing coastal regions face consequences of land use and climate change including flooding and increased sediment, nutrient, and chemical runoff, but these forces may also enhance pathogen runoff, which threatens human, animal, and ecosystem health. Using the zoonotic parasite Toxoplasma gondii in California, USA as a model for coastal pathogen pollution, we examine the spatial distribution of parasite runoff and the impacts of precipitation and development on projected pathogen delivery to the ocean. Oocysts, the extremely hardy free-living environmental stage of T. gondii shed in faeces of domestic and wild felids, are carried to the ocean by freshwater runoff. Linking spatial pathogen loading and transport models, we show that watersheds with the highest levels of oocyst runoff align closely with regions of increased sentinel marine mammal T. gondii infection. These watersheds are characterized by higher levels of coastal development and larger domestic cat populations. Increases in coastal development and precipitation independently raised oocyst delivery to the ocean (average increases of 44% and 79%, respectively), but dramatically increased parasite runoff when combined (175% average increase). Anthropogenic changes in landscapes and climate can accelerate runoff of diverse pathogens from terrestrial to aquatic environments, influencing transmission to people, domestic animals, and wildlife.

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.