Mysterious microsporidians: springtime outbreaks of disease in Daphnia communities in shallow pond ecosystems.

Parasites can play key roles in ecosystems, especially when they infect common hosts that play important ecological roles. Daphnia are critical grazers in many lentic freshwater ecosystems and typically reach peak densities in early spring. Daphnia have also become prominent model host organisms for the field of disease ecology, although most well-studied parasites infect them in summer or fall. Here, we report field patterns of virulent microsporidian parasites that consistently infect Daphnia in springtime, in a set of seven shallow ponds in Georgia, USA, sampled every 3-4 weeks for 18 months. We detected two distinct parasite taxa, closely matching sequences of Pseudoberwaldia daphniae and Conglomerata obtusa, both infecting all three resident species of Daphnia: D. ambigua, D. laevis, and D. parvula. To our knowledge, neither parasite has been previously reported in any of these host species or anywhere in North America. Infection prevalence peaked consistently in February-May, but the severity of these outbreaks differed substantially among ponds. Moreover, host species differed markedly in terms of their maximum infection prevalence (5% [D. parvula] to 72% [D. laevis]), mean reduction of fecundity when infected (70.6% [D. ambigua] to 99.8% [D. laevis]), mean spore yield (62,000 [D. parvula] to 377,000 [D. laevis] per host), and likelihood of being infected by each parasite. The timing and severity of the outbreaks suggests that these parasites could be impactful members of these shallow freshwater ecosystems, and that the strength of their effects is likely to hinge on the composition of ponds' zooplankton communities.

Surveillance of tick-borne pathogens in domestic dogs from Chad, Africa.

BACKGROUND: Tick-borne pathogens are understudied among domestic animals in sub-Saharan Africa but represent significant threats to the health of domestic animals and humans. Specifically, additional data are needed on tick-borne pathogens in Chad, Africa. Surveillance was conducted among domestic dogs in Chad for selected tick-borne pathogens to measure (1) the prevalence of antibodies against Anaplasma spp., Borrelia burgdorferi, and Ehrlichia spp.; (2) the prevalence of infections caused by Hepatozoon spp., Ehrlichia canis, Anaplasma platys, and Babesia spp.; and (3) associations of pathogens with demographic, spatial, and temporal factors. Blood samples were collected from domestic dogs at three time points (May 2019, November 2019, June 2020) across 23 villages in southern Chad. RESULTS: Of the 428 dogs tested with the IDEXX SNAP 4Dx test in May 2019, 86% (n = 370, 95% CI = 83-90%) were positive for antibodies to Ehrlichia spp., 21% (n = 88, 95% CI = 17-25%) were positive for antibodies to Anaplasma spp., and 0.7% (n = 3, 95% CI = 0.1-2%) were positive for antibodies to Borrelia burgdorferi. Four different pathogens were detected via PCR. Hepatozoon spp. were most commonly detected (67.2-93.4%, depending on the time point of sampling), followed by E. canis (7.0-27.8%), A. platys (10.1-22.0%), and Babesia vogeli (0.4-1.9%). Dogs were coinfected with up to three pathogens at a single time point, and coinfections were most common in May 2019 compared to November 2019 and May 2020. CONCLUSIONS: Overall, this study provides new data about the epidemiology of tick-borne pathogens in domestic dogs in Chad, with potential implications for dog and human health.

Vector communities under global change may exacerbate and redistribute infectious disease risk.

Vector-borne parasites may be transmitted by multiple vector species, resulting in an increased risk of transmission, potentially at larger spatial scales compared to any single vector species. Additionally, the different abilities of patchily distributed vector species to acquire and transmit parasites will lead to varying degrees of transmission risk. Investigation of how vector community composition and parasite transmission change over space due to variation in environmental conditions may help to explain current patterns in diseases but also informs our understanding of how patterns will change under climate and land-use change. We developed a novel statistical approach using a multi-year, spatially extensive case study involving a vector-borne virus affecting white-tailed deer transmitted by Culicoides midges. We characterized the structure of vector communities, established the ecological gradient controlling change in structure, and related the ecology and structure to the amount of disease reporting observed in host populations. We found that vector species largely occur and replace each other as groups, rather than individual species. Moreover, community structure is primarily controlled by temperature ranges, with certain communities being consistently associated with high levels of disease reporting. These communities are essentially composed of species previously undocumented as potential vectors, whereas communities containing putative vector species were largely associated with low levels, or even absence, of disease reporting. We contend that the application of metacommunity ecology to vector-borne infectious disease ecology can greatly aid the identification of transmission hotspots and an understanding of the ecological drivers of parasite transmission risk both now and in the future.

Parasitaemia data and molecular characterization of Haemoproteus catharti from New World vultures (Cathartidae) reveals a novel clade of Haemosporida.

BACKGROUND: New World vultures (Cathartiformes: Cathartidae) are obligate scavengers comprised of seven species in five genera throughout the Americas. Of these, turkey vultures (Cathartes aura) and black vultures (Coragyps atratus) are the most widespread and, although ecologically similar, have evolved differences in morphology, physiology, and behaviour. Three species of haemosporidians have been reported in New World vultures to date: Haemoproteus catharti, Leucocytozoon toddi and Plasmodium elongatum, although few studies have investigated haemosporidian parasites in this important group of species. In this study, morphological and molecular methods were used to investigate the epidemiology and molecular biology of haemosporidian parasites of New World vultures in North America. METHODS: Blood and/or tissue samples were obtained from 162 turkey vultures and 95 black vultures in six states of the USA. Parasites were identified based on their morphology in blood smears, and sequences of the mitochondrial cytochrome b and nuclear adenylosuccinate lyase genes were obtained for molecular characterization. RESULTS: No parasites were detected in black vultures, whereas 24% of turkey vultures across all sampling locations were positive for H. catharti by blood smear analysis and/or PCR testing. The phylogenetic analysis of cytochrome b gene sequences revealed that H. catharti is closely related to MYCAMH1, a yet unidentified haemosporidian from wood storks (Mycteria americana) in southeastern USA and northern Brazil. Haemoproteus catharti and MYCAMH1 represent a clade that is unmistakably separate from all other Haemoproteus spp., being most closely related to Haemocystidium spp. from reptiles and to Plasmodium spp. from birds and reptiles. CONCLUSIONS: Haemoproteus catharti is a widely-distributed parasite of turkey vultures in North America that is evolutionarily distinct from other haemosporidian parasites. These results reveal that the genetic diversity and evolutionary relationships of avian haemosporidians are still being uncovered, and future studies combining a comprehensive evaluation of morphological and life cycle characteristics with the analysis of multiple nuclear and mitochondrial genes will be useful to redefine the genus boundaries of these parasites and to re-evaluate the relationships amongst haemosporidians of birds, reptiles and mammals.

Possible Role of Fish as Transport Hosts for Dracunculus spp. Larvae.

To inform Dracunculus medinensis (Guinea worm) eradication efforts, we evaluated the role of fish as transport hosts for Dracunculus worms. Ferrets fed fish that had ingested infected copepods became infected, highlighting the importance of recommendations to cook fish, bury entrails, and prevent dogs from consuming raw fish and entrails.

Guinea Worm (Dracunculus medinensis) Infection in a Wild-Caught Frog, Chad.

A third-stage (infective) larva of Dracunculus medinensis, the causative agent of Guinea worm disease, was recovered from a wild-caught Phrynobatrachus francisci frog in Chad. Although green frogs (Lithobates clamitans) have been experimentally infected with D. medinensis worms, our findings prove that frogs can serve as natural paratenic hosts.

Possible Role of Fish and Frogs as Paratenic Hosts of Dracunculus medinensis, Chad.

Copepods infected with Dracunculus medinensis larvae collected from infected dogs in Chad were fed to 2 species of fish and tadpoles. Although they readily ingested copepods, neither species of fish, Nile tilapia (Oreochromis niloticus) nor fathead minnow (Pimephalis promelas), were found to harbor Dracunculus larvae when examined 2-3 weeks later. Tadpoles ingested copepods much more slowly; however, upon examination at the same time interval, tadpoles of green frogs (Lithobates [Rana] clamitans) were found to harbor small numbers of Dracunculus larvae. Two ferrets (Mustela putorius furo) were fed fish or tadpoles that had been exposed to infected copepods. Only the ferret fed tadpoles harbored developing Dracunculus larvae at necropsy 70-80 days postexposure. These observations confirm that D. medinensis, like other species in the genus Dracunculus, can readily survive and remain infective in potential paratenic hosts, especially tadpoles.

Vector species richness increases haemorrhagic disease prevalence through functional diversity modulating the duration of seasonal transmission.

Although many parasites are transmitted between hosts by a suite of arthropod vectors, the impact of vector biodiversity on parasite transmission is poorly understood. Positive relationships between host infection prevalence and vector species richness (SR) may operate through multiple mechanisms, including (i) increased vector abundance, (ii) a sampling effect in which species of high vectorial capacity are more likely to occur in species-rich communities, and (iii) functional diversity whereby communities comprised species with distinct phenologies may extend the duration of seasonal transmission. Teasing such mechanisms apart is impeded by a lack of appropriate data, yet could highlight a neglected role for functional diversity in parasite transmission. We used statistical modelling of extensive host, vector and microparasite data to test the hypothesis that functional diversity leading to longer seasonal transmission explained variable levels of disease in a wildlife population. We additionally developed a simple transmission model to guide our expectation of how an increased transmission season translates to infection prevalence. Our study demonstrates that vector SR is associated with increased levels of disease reporting, but not via increases in vector abundance or via a sampling effect. Rather, the relationship operates by extending the length of seasonal transmission, in line with theoretical predictions.

Prevalence of Baylisascaris procyonis in wild rodents in central Georgia, USA.

Raccoon roundworm, Baylisascaris procyonis, is a zoonotic parasite of raccoons (Procyon lotor) that needs a One Health approach to better inform risks to human and animal health. The few studies on B. procyonis in wild rodents have primarily focused on white-footed mice (Peromyscus leucopus). This study aimed to determine the prevalence and rodent host range of B. procyonis in Georgia (USA) and investigate differences in prevalence at urban/fragmented sites and rural/agriculture sites. We sampled 99 rodents of five species. Larvae were recovered from seven of 78 (9.0%) white-footed mice with a mean of 4.4 larvae (range 1-12). One mouse had a single larva in the brain. Prevalence was not different between urban and rural sites. This report extends the geographic range of this parasite and confirms that rodents serve as paratenic hosts in the southern range. Therefore, baylisascariasis should be considered a differential for neurologic domestic animals, wildlife, or people in this region.

Distribution and prevalence of antibodies to Trichinella spp. and Toxoplasma gondii in wild pigs (Sus scrofa) in the United States.

Invasive wild pigs (Sus scrofa) are a reservoir for over 100 viral, bacterial, and parasitic pathogens that are transmissible to humans, livestock, domestic animals, and wildlife in North America. Numerous historical local surveys and results from a nation-wide survey (2006-2010) indicated that wild pigs in the United States act as reservoirs for Trichinella spp. and Toxoplasma gondii, two zoonotic pathogens of importance for human and animal health. Since that time, wild pig populations have expanded and increased in density in many areas. Population expansion of wild pigs creates opportunities for the introduction of pathogens to new areas of the country, increasing health risks. The goal of this study was to investigate the current geographic distribution and prevalence of Trichinella spp. and T. gondii antibodies in wild pigs using serum samples collected from 2014 to 2020. Serum samples from 36 states were tested for antibodies to Trichinella spp. (n = 7467) and T. gondii (n = 5984) using commercially available enzyme-linked immunosorbent assays. Seroprevalence for Trichinella spp. (12.4%, 927/7467) and T. gondii (40.8%, 2444/5984) are significantly higher compared to a previous 2006-2010 study across all regions. Results from this study also showed a lower seroprevalence (4.8%) for Trichinella spp. in the West region compared to the other regions (South: 13.4%; Midwest: 18.4%; Northeast: 19.1%). There were new detection records for antibodies to Trichinella spp. in 11 states, mostly in the West, Midwest, and Northeast regions compared to a previous study in 2014. Males and juveniles were less likely to be positive for Trichinella spp. antibodies, compared to females and older animals, respectively. Seroprevalence was similar for T. gondii across the regions (31.8-56%) with some states having particularly high seroprevalence (e.g., Hawaii 79.4% and Pennsylvania 68%). There were new T. gondii antibody detection records for 12 states, mostly in the West, Midwest, and Northeast regions. Adults were more likely than juveniles and subadults to be seropositive. These data confirm that the distribution and prevalence of antibodies for Trichinella spp. and T. gondii are increasing in the United States, likely driven by wild pig population growth and range expansion.

Surveillance of Feral Swine (Sus scrofa) in the Western USA for Antibodies to Vesicular Stomatitis Virus, 2013-21.

Vesicular stomatitis virus (VSV) outbreaks periodically occur in livestock in the western US and are thought to originate from outside this country. Feral swine (Sus scrofa) have been identified as an amplifying host for vesicular stomatitis New Jersey virus (VSNJV) and have been used to better understand the epidemiology of this virus through serosurveillance. This study aimed to determine if antibodies to vesicular stomatitis Indiana virus (VSIV) and VSNJV were present in feral swine in the western US and to determine if seropositive animals were associated with areas of previously detected VSV in livestock. A total of 4,541 feral swine samples was tested using virus neutralization (VN); samples exhibiting neutralizing activity against one or more of the viruses were confirmed using competitive ELISA (cELISA). Eight sera exhibited neutralizing activity by VN assay and a single serum sample from an animal from Kinney County, Texas sampled in December 2019 tested positive for antibodies to VSIV by cELISA. This finding is supported by a local outbreak of VSIV in horses in the same county in June 2019. The low prevalence of antibodies against VSNJV and VSIV was unexpected but indicates that feral swine in the western US do not represent an endemic reservoir for either of these viruses.

Species composition of Culicoides (Diptera: Ceratopogonidae) in the Ridge and Valley region of Tennessee, USA.

Biting midges in the genus Culicoides Latreille (Diptera: Ceratopogonidae) are known to transmit many pathogens of veterinary and medical concern. Although much work has been done globally and in certain regions of North America, Culicoides spp. research in rural Appalachia is limited. To begin characterizing the distribution and community structure of Culicoides spp. in Appalachia, we surveyed 2 distinct sites in the Ridge and Valley ecoregion of northeastern Tennessee, USA, from April 2021-September 2021. Culicoides spp. were sampled using 2 methods: Centers for Disease Control ultraviolet LED light traps and potential larval habitat substrate collection (coupled with water chemistry values). Site 1 was dominated by natural features, and Site 2 was a beef cattle operation. During 96 trap nights, a total of 1,568 Culicoides were collected, representing 24 species. Site 1 yielded the highest diversity, with 24 species, while Site 2 yielded 12 species. Overall, the most abundant species in light traps were C. stellifer Coquillett (44%), C. bergi Cochrane (18%), C. haematopotus Malloch (12%), and C. debilipalpis Lutz (11%). From substrate sampling, 8 species were identified. Culicoides haematopotus was the most abundant and was collected during each sampling period. Water chemistry values taken at the time of substrate collection were not significantly related to which Culicoides spp. emerged from a given substrate. Our results indicate a diverse community of Culicoides spp. in our study area, however, further work is needed to identify Culicoides species composition across a variety of landscapes in Appalachia and inform research on vector presence and associated vector disease dynamics.

Case report: Disseminated larval trematodiasis caused by Clinostomum marginatum in a green tree frog (Hyla cinerea).

Clinostomum spp. are common parasites of piscivorous birds. Metacercaria are typically observed in the muscles or just under the skin of fish and rarely amphibians. We describe an unusually severe case of Clinostomum marginatum infection in an adult female green tree frog (Hyla cinerea) from Georgia (USA). The frog was found in November 2015 with a high number (>250) of widely disseminated, raised, subcutaneous nodules. The frog died in December. At necropsy, it was emaciated, and the skin was covered in raised uniform, tan-green, subcutaneous, ∼2-3 mm diameter nodules. Each nodule contained 1-3 C. marginatum metacercariae. Microscopically, high numbers of trematodes were within subcutaneous tissues and in coelomic and oral cavities, lung, liver, kidney, ovary, orbit and calvarium. Small to large numbers of lymphocytes and melanomacrophages were in connective tissues and epidermis. A 732 bp region of COI was 98.8-99.8% similar to numerous sequences of C. marginatum and, phylogenetically it grouped with these C. marginatum sequences. The ITS-1 region was 100% similar to a C. marginatum sample from a great egret (Ardea alba) from Mississippi. This report represents a novel finding of severe trematodiasis in a free-ranging amphibian with C. marginatum infection.

Patterns of Hemorrhagic Disease in White-Tailed Deer (Odocoileus virginianus) in the Great Plains of the USA, 1982-2020.

Hemorrhagic disease (HD) of deer is caused by epizootic hemorrhagic disease virus (EHDV) or bluetongue virus (BTV) and is considered one of the most important viral diseases of white-tailed deer (Odocoileus virginianus). Despite evidence of changing patterns of HD in the northeastern and upper midwestern US, the historical and current patterns of HD in the Great Plains remain poorly described. We used results from an annual survey documenting HD mortality to characterize historic and current patterns of HD in the northern and central Great Plains (North Dakota, South Dakota, Nebraska, Kansas, and Oklahoma), US, between 1982 and 2020. Further, we assessed temporal change using linear regression to determine change in annual reporting intensity (percentage of counties in a state with reported HD) and change in reporting frequency (the number of years a county or state reported HD) during each decade between 1982 and 2020. Across the 38-yr study period, HD reports expanded northeast across latitude and longitude. Intensity of HD reports significantly increased during this period for three (North Dakota, South Dakota, Kansas) of five states examined. Frequency of reports also increased for all five states. Such changes in northern latitudes might lead to increased deer mortality in regions where HD epizootics have been historically less frequent. Understanding how patterns of HD are changing on the landscape is important when considering future deer management in the face of other mortality factors.

Prevalence and diversity of spotted fever group Rickettsia species in ixodid ticks from domestic dogs in Chad, Africa.

Tick-borne pathogens in the genus Rickettsia are the causative agents of severe and potentially fatal spotted fever group (SFG) and typhus group diseases in dogs and humans. Climate, habitat, and land-use changes are impacting vector ranges, with expansions potentially resulting in novel pathogens being introduced into naïve locations. Despite the public health importance of SFG Rickettsia, there are relatively few data on the prevalence and diversity of rickettsial pathogens in sub-Saharan Africa. The aim of this study was to characterize the SFG Rickettsia prevalence and diversity in ixodid ticks (104 Amblyomma spp., 160 Rhipicephalus spp., and one Hyalomma truncatum) collected from domestic dogs in Chad, Africa. Ticks were screened for Rickettsia spp. using a nested PCR targeting the 17-kDa gene. Species identification was through bidirectional Sanger sequencing of the 17-kDa, ompA, ompB, and/or gltA gene targets. A total of 43.3 % (115/265) ticks were positive for Rickettsia spp. and six Rickettsia species were identified: R. africae, R. massiliae, R. conorii, R. felis, R. monacensis and Candidatus Rickettsia muridii. Seven additional samples were positive for Rickettsia of undetermined species. Rickettsia africae, an important zoonotic pathogen, was found in 81 % (79/97) of A. variegatum and 29 % (2/7) of an A. marmoreum complex species, a group that infests a wide range of birds and mammals, including humans. Finally, we detected a high diversity of Rickettsia spp., most of which were zoonotic, in Rh. muhsamae. Collectively these data indicate there is a risk of rickettsiosis in Chad and further studies on ticks and rickettsial pathogens in this region are warranted.

High Prevalence and Broad Distribution of Trichomonas gypaetinii in Bald Eagles (Haliaeetus leucocephalus) in the USA.

Trichomonas gypaetinii was detected in 117 (88%) of 133 Bald Eagles (Haliaeetus leucocephalus) and in 0/7 Golden Eagles (Aquila chrysaetos) in the USA, with no sex or age prevalence difference. All eagles lacked associated lesions. This study indicated that T. gypaetinii is common and widespread in Bald Eagles, but rarely associated with disease.

Otterly diverse - A high diversity of Dracunculus species (Spirurida: Dracunculoidea) in North American river otters (Lontra canadensis).

The genus Dracunculus contains numerous species of subcutaneous parasites of mammals and reptiles. In North America, there are at least three mammal-infecting species of Dracunculus. Reports of Dracunculus infections have been reported from river otters (Lontra canadensis) since the early 1900s; however, little is known about the species infecting otters or their ecology. Most reports of Dracunculus do not have a definitive species identified because females, the most common sex found due to their larger size and location in the extremities of the host, lack distinguishing morphological characteristics, and few studies have used molecular methods to confirm identifications. Thus, outside of Ontario, Canada, where both D. insignis and D. lutrae have been confirmed in otters, the species of Dracunculus in river otters is unknown. In the current study, molecular characterization of nematodes from river otters revealed a high diversity of Dracunculus species. In addition to confirming D. insignis infections, two new clades were detected. One clade was a novel species in any host and the other was a clade previously detected in Virginia opossums (Didelphis virginiana) from the USA and a domestic dog from Spain. No infections with D. lutrae were detected and neither new lineage was genetically similar to D. jaguape, which was recently described from a neotropical otter (Lontra longicaudis) from Argentina. These data also indicate that Dracunculus spp. infections in otters are widespread throughout Eastern North America. Currently the life cycles for most of the Dracunculus spp. infecting otters are unknown. Studies on the diversity, life cycle, and natural history of Dracunculidae parasites in wildlife are important because the related parasite, D. medinensis (human Guinea worm) is the subject of an international eradication campaign and there are increasing reports of these parasites in new geographic locations and new hosts, including new species in humans and domestic dogs.

Hepatozoon spp. infection in wild canids in the eastern United States.

BACKGROUND: Hepatozoon spp. are apicomplexan parasites known to cause musculoskeletal disease in a variety of animals. Two species are known to infect wild and domestic canids in the US: Hepatozoon canis and H. americanum. METHODS: In this study, blood, heart, and/or spleen samples were collected from 278 wild canids (180 coyotes, 93 red foxes, and 5 gray foxes) in the eastern US and tested via PCR for Hepatozoon. Histology slides of heart and skeletal muscle were assessed for Hepatozoon cysts and associated inflammation when fresh tissue was available (n = 96). RESULTS: Hepatozoon spp. were found in 24.2% (59/278) of individuals, with Hepatozoon canis in 14.0% (34/278) and H. americanum in 10.7% (26/278). One coyote was positive for both H. canis and H. americanum. Foxes were more likely to be positive for H. canis than coyotes (23% and 7% respectively, P = 0.0008), while only coyotes were positive for H. americanum. Of the eight sampled states, H. canis was present in six (Louisiana, North Carolina, Pennsylvania, South Carolina, Tennessee, and Virginia) while H. americanum was found in two southern states (South Carolina and Louisiana). Infection status was positively correlated with myositis and myocarditis, and heart or muscle cysts were found in 83% (5/6) of H. americanum-positive coyotes. CONCLUSION: This survey showed a moderate prevalence of H. canis and H. americanum in states where the parasite was previously unrecorded including South Carolina and Pennsylvania.