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.

Description of B abesia coryicola sp. nov. from Florida pumas (Puma concolor coryi) from southern Florida, USA.

Previously, a high prevalence of piroplasms has been reported from Florida pumas (Puma concolor coryi) from southern Florida. In the current study, we describe the biological characteristics of a novel Babesia species in Florida pumas. Ring-stage trophozoites were morphologically similar to trophozoites of numerous small babesids of felids including B. leo, B. felis, and Cytauxzoon felis. Parasitemias in Florida pumas were very low (<1%) and hematologic values of 25 Babesia-infected Florida pumas were within normal ranges for P. concolor. Phylogenetic analysis of near full-length 18S rRNA gene, ß-tubulin, cytochrome c oxidase subunit I, cytochrome c oxidase subunit III, and cytochrome b gene sequences indicated that this Babesia species is a member of the Babesia sensu stricto clade and is related to groups of Babesia spp. from carnivores or ungulates, although the closest group varied by gene target. Internal transcribed spacer (ITS)-1 region sequences from this Babesia sp. from 19 Florida pumas were 85.7-99.5% similar to each other and ∼88% similar to B. odocoilei. Similarly, an ITS-2 sequence from one puma was 96% similar to B. bigemina and 92% similar to a Babesia sp. from a red panda (Ailurus fulgens). Infected pumas were positive for antibodies that reacted with B. odocoilei, B. canis, and B. bovis antigens with titers of 1:256, 1:128, and 1:128, respectively. No serologic reactivity was noted for Theileria equi. No molecular evidence of congenital infection was detected in 24 kittens born to 11 Babesia-infected female pumas. Pumas from other populations in the United States [Louisiana (n = 1), North Dakota (n = 5) and Texas (n = 28)], British Columbia, Canada (n = 9), and Costa Rica (n = 2) were negative for this Babesia sp. Collectively, these data provide morphologic, serologic, genetic, and natural history data for this novel Babesia sp. which we propose the name Babesia coryicola sp. nov. sp. This is the first description of a felid-associated Babesia species in North America.

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.

Diversity of Babesia spp. in skunks from selected states in the United States of America.

Babesia species are intraerythrocytic protozoan parasites that infect a variety of hosts. The goal of this study was to evaluate the piroplasm species present in skunks in various states in the United States and determine whether there was any geographic variation. Spleen, whole blood, or blood on filter paper were received from Pennsylvania, Kentucky, North Carolina, South Carolina, Georgia, Missouri, Louisiana, Texas, Kansas, and California, and were tested for Babesia sp. We tested four species of skunks including striped skunk (Mephitis mephitis, n = 72), eastern spotted skunk (Spilogale putorius, n = 28), western spotted skunk (Spilogale gracilis, n = 15), and hog-nosed skunk (Conepatus leuconotus, n = 11). A PCR assay targeting the 18S rRNA region and cox1 region were used to determine if skunks were infected with piroplasms and for phylogenetic analyses. A total of 48.4% (61/126) of skunks tested positive for a Babesia species. Both the 18S and cox1 analysis supported a skunk-specific Babesia microti-like sp. of carnivores as well as a species in the B. microti complex that is phylogenetically unique from both B. microti of humans and the B. microti-like sp. of carnivores. In the 18S analysis, there was a third species of Babesia in hog-nosed skunks in the western piroplasm group. This study shows that at least three species of piroplasms occur in skunk species in the United States and further highlights the importance of phylogenetic analyses and the use of multiple gene targets when studying piroplasms.

Title: Diversité des Babesia spp. chez des mouffettes provenant d'États sélectionnés des États-Unis. Abstract: Les espèces de Babesia sont des protozoaires parasites intraérythrocytaires qui infectent divers hôtes. Le but de cette étude était d'évaluer les espèces de piroplasmes présentes chez les mouffettes dans divers états des États-Unis et de déterminer s'il existait une variation géographique. Des rates, du sang total ou du sang sur papier filtre ont été reçus de Pennsylvanie, du Kentucky, de Caroline du Nord, de Caroline du Sud, de Géorgie, du Missouri, de Louisiane, du Texas, du Kansas et de Californie, et ont été testés pour Babesia sp. Nous avons testé quatre espèces de mouffettes, dont la mouffette rayée (Mephitis mephitis, n = 72), la mouffette tachetée de l'Est (Spilogale putorius, n = 28), la mouffette tachetée de l'Ouest (Spilogale gracilis, n = 15) et la mouffette à nez plat (Conepatus leuconotus, n = 11). Un test PCR ciblant la région de l'ARNr 18S et la région cox1 a été utilisé pour déterminer si les mouffettes étaient infectées par des piroplasmes et pour des analyses phylogénétiques. Au total, 48,4 % (61/126) des mouffettes ont été testées positives pour une espèce de Babesia. Les analyses du 18S et du cox1 ont toutes deux confirmé une espèce de type Babesia microti de carnivores spécifique aux mouffettes ainsi qu'une espèce du complexe B. microti qui est phylogénétiquement unique à la fois par rapport à B. microti de l'homme et à l'espèce des carnivores. Dans l'analyse 18S, il y avait une troisième espèce de Babesia chez les mouffettes à nez plat du groupe des piroplasmes de l'ouest. Cette étude montre qu'au moins trois espèces de piroplasmes sont présentes chez les espèces de mouffettes aux États-Unis et souligne en outre l'importance des analyses phylogénétiques et de l'utilisation de plusieurs cibles génétiques lors de l'étude des piroplasmes.

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.

Systemic toxoplasmosis in 2 domestic rabbits in Georgia, United States.

Toxoplasma gondii infections are common in a range of mammalian and avian species, but clinical disease has been reported only rarely in domestic rabbits. Two cases of toxoplasmosis in domestic rabbits from the same premises were submitted to a diagnostic pathology facility in Athens, GA, USA. Both rabbits died after exhibiting clinical signs of gastrointestinal stasis. The gross findings observed in both rabbits comprised miliary, random, white-to-tan, necrotic foci throughout the spleen, liver, and lungs. Histologically, tachyzoites were observed within necrotizing inflammatory foci in the spleens of both rabbits, and in various other organs (tracheobronchial lymph node, lung, heart, and cecal appendix) of one rabbit. In both cases, the tachyzoites were immunoreactive with anti-Toxoplasma gondii antibodies. In addition, T. gondii DNA was detected via PCR and sequencing from a fresh lung sample from one rabbit and formalin-fixed, paraffin-embedded spleen, liver, femoral bone marrow, and haired skin from the second rabbit. Given that T. gondii can cause disease in domestic rabbits and is also a concern for other potential intermediate hosts (e.g., humans, other domestic animals), this parasite warrants consideration in the diagnostic evaluation of lagomorph tissues with compatible lesions.

Health Assessment of Adult Male Eastern Wild Turkeys (Meleagris gallopavo silvestris) from Western Kentucky, USA.

Wild turkeys (Meleagris gallopavo) are an important game species throughout the geographic range. Populations throughout multiple regions of the US have been declining, including in Kentucky, US, raising concerns among managers and resource users. To better understand the overall population health, we performed postmortem examinations and targeted pathogen, mineral, and toxicant testing on 36 adult male, apparently healthy, wild turkeys that were hunter harvested in western Kentucky during April 2018. We found that birds were in fair to good nutritional condition with no significant gross or microscopic lesions. Ticks (Amblyomma spp.) and lice (three species) were present on 94 and 31% of birds, respectively. We commonly detected intestinal nematodes and cestodes and found coccidian oocysts in 39% and capillarid eggs in 6% of birds. The prevalences of lymphoproliferative disease virus and reticuloendotheliosis virus were 39 and 11%, respectively. Spleen samples tested with PCR were positive for Borrelia burgdorferi, Haemoproteus sp., and Leucocytozoon sp. in 11, 83, and 3%, respectively. Based on a subjective histologic assessment of testis tissues, most birds had widespread and abundant sperm present. Mineral analysis and broad toxicant screening on liver samples from 32 turkeys were unremarkable. Further work is needed to assess potential population risk factors and to determine individual- and population-level impacts of pathogens on adults and poults.

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.

Surveillance for Selected Pathogens and Parasites of Northern Bobwhite (Colinus virginianus) from Western Oklahoma, USA, 2018-20.

The Northern Bobwhite (Colinus virginianus) has been undergoing a range-wide population decline. Potential causes for declines across its historic range have been investigated for decades and include habitat loss and fragmentation and a variety of parasitic and infectious diseases. Although there have been studies on bobwhite ecology in Oklahoma, USA, relatively little is known about parasites and pathogens in the region. We evaluated the health of free-ranging bobwhites from nine sites in western Oklahoma. From 2018 to 2020, 206 bobwhites were evaluated for gross and microscopic lesions and tested for selected pathogens. In general, bobwhites were in good nutritional condition with ample muscle mass and fat stores. No significant gross lesions were observed in any bobwhite and no significant histologic lesions were detected in a subset. There was no evidence of infection with or exposure to reticuloendotheliosis virus, West Nile virus, respiratory Mycoplasmataceae species, Pasteurella multocida, intestinal Eimeria spp., or oral Trichomonas spp. Several pathogens of potential concern were detected, including avian adenovirus (8.6%), Toxoplasma gondii (2.3%), and haemosporidians (a Haemoproteus sp. (1.5%), Leucocytozoon schoutedeni (1.5%), and Plasmodium homopolare haplotype 2 [lineage LAIRI01; 3.6%]). Physaloptera sp. (12%) and Sarcocystis sp. (1%) were detected in the breast muscle. Low intraspecific genetic diversity was noted for Physaloptera sp., and sequences were most similar to Physaloptera sequences from bobwhites and grasshoppers (Orthoptera) in Texas. Low intensities of chewing lice, chiggers, and ticks were observed. A subset of bobwhites had evidence of exposure to selected toxicants and heavy metals; a small number had low levels of iron, manganese, zinc, molybdenum, and copper, which were not considered diagnostically relevant. In general, bobwhites from western Oklahoma appeared to be in good health with a low diversity of pathogens detected, but future work is needed to understand potentially changing disease risks for this population.

A single Haemoproteus plataleae haplotype is widespread in white ibis (Eudocimus albus) from urban and rural sites in southern Florida.

The American white ibis (Eudocimus albus), a common bird species in Florida, has become increasingly urban, with many populations relying heavily on urban and suburban habitats, which may alter parasite transmission. Parasites of ibis, especially haemosporidians, are understudied. Avian haemosporidia can have a wide range of impacts on birds, including decreased reproductive success or increased mortality. Because southern Florida is subtropical and has a high diversity of potential vectors for haemosporidia, we hypothesized that there will be a high prevalence and genetic diversity of haemosporidia in white ibis. A total of 636 ibis from South Florida were sampled from 2010 to 2022, and blood samples were tested for haemosporidia by examination of Giemsa-stained thin blood smears and/or nested PCRs targeting the cytochrome b gene. A total of 400 (62.9%, 95% CI 59-66.7%) ibis were positive for parasites that were morphologically identified as Haemoproteus plataleae. Sequences of 302 positives revealed a single haplotype of Haemoproteus (EUDRUB01), which was previously reported from white ibis in South Florida and captive scarlet ibis (E. ruber) in Brazil. No Plasmodium or Leucocytozoon infections were detected. Parasitemias of the 400 positive birds were very low (average 0.084%, range 0.001%-2.16% [although only 2 birds had parasitemias >1%]). Prevalence and parasitemias were similar for males and females (68% vs. 61.6% and 0.081% vs. 0.071%, respectively). Prevalence in juveniles was lower compared with adults (52% vs. 67.4%) but parasitemias were higher in juveniles (0.117% vs. 0.065%). This data shows that H. plataleae is common in ibis in South Florida. Although parasitemias were generally low, additional research is needed to determine if this parasite has subclinical effects on ibis, if additional haplotypes or parasite species infect ibis in other regions of their range, or if H. plataleae is pathogenic for other sympatric avian species.

Fatal Systemic Haemosporidiosis in a Free-Ranging Greater Sage-Grouse (Centrocercus urophasianus).

A wild adult Greater Sage-grouse (Centrocercus urophasianus) from northeastern Oregon, US, was found dead, with multiorgan necrosis and inflammation associated with protozoal schizonts and phanerozoites. Molecular characterization identified these protozoa as Plasmodium homopolare H2 and a novel Leucocytozoon sp. lineage. Fatal systemic haemosporidiosis has not been previously reported in Greater Sage-grouse.

Application of a universal parasite diagnostic test to biological specimens collected from animals.

A previously described universal parasite diagnostic (nUPDx) based on PCR amplification of the 18S rDNA and deep-amplicon sequencing, can detect human blood parasites with a sensitivity comparable to real-time PCR. To date, the efficacy of this assay has only been assessed on human blood. This study assessed the utility of nUPDx for the detection of parasitic infections in animals using blood, tissues, and other biological sample types from mammals, birds, and reptiles, known to be infected with helminth, apicomplexan, or pentastomid parasites (confirmed by microscopy or PCR), as well as negative samples. nUPDx confirmed apicomplexan and/or nematode infections in 24 of 32 parasite-positive mammals, while also identifying several undetected coinfections. nUPDx detected infections in 6 of 13 positive bird and 1 of 2 positive reptile samples. When applied to 10 whole parasite specimens (worms and arthropods), nUPDx identified all to the genus or family level, and detected one incorrect identification made by morphology. Babesia sp. infections were detected in 5 of the 13 samples that were negative by other diagnostic approaches. While nUPDx did not detect PCR/microscopy-confirmed trichomonads or amoebae in cloacal swabs/tissue from 8 birds and 2 reptiles due to primer template mismatches, 4 previously undetected apicomplexans were detected in these samples. Future efforts to improve the utility of the assay should focus on validation against a larger panel of tissue types and animal species. Overall, nUPDx shows promise for use in both veterinary diagnostics and wildlife surveillance, especially because species-specific PCRs can miss unknown or unexpected pathogens.

Infection with a novel pentastome (Raillietiella sp.) in a juvenile endangered Galápagos tortoise (Chelonoidis niger).

Pentastomes are crustacean endoparasites in the lungs of herpetofauna and rarely mammals and birds. To date, the only pentastomes reported in chelonians, all aquatic turtles, are Diesingia megastomum from Brazil and Pelonia africana from South Africa. In March 2021, 185 juvenile tortoises (Chelonoidis niger) were confiscated after an attempted illegal exportation from the Galápagos. It is believed the tortoises were removed from nests on Santa Cruz Island. The young tortoises were individually wrapped in plastic and at seizure ten were dead and another 25 tortoises subsequently died. One of the tortoises, estimated to be 3 months old, had 11 pentastomes in the lungs. The pentastomes were identified as a Raillietiella sp. based on morphology. The specimens had a claviform body that tapers into a bifid tail, a 165.3 µm × 92.1 µm buccal cadre, and sharp tipped anterior and posterior hooks with the posterior hooks being larger than the anterior. The males have smooth copulatory spicules with a rounded, smooth base. All females were gravid. Molecular analysis confirmed that the parasites were a Raillietiella sp. Based on the COI gene, it was most similar (82.7% identical) to an undescribed Raillietiella species from a Caribbean anole (Anolis cristatellus) found in Florida followed by R. hebitihamata (81%), R. indica (80.7%), and R. orientalis (78.8-80.7%). Based on the 18S rRNA gene sequence (1799 bp), it was most similar (99.3% identical) to two undescribed Raillietiella species followed by R. aegypti from a berber skink (Eumeces schneideri) from Saudia Arabia. Phylogenetically, with both molecular targets, the Raillietiella sp. from the Galápagos tortoise grouped with other Raillietiella spp. and was basal within the group. Currently, the origin of this parasite (native to Galápagos or introduced) and the life cycle are unknown. Because some pentastome species, especially when in aberrant hosts, can be pathogenic, additional studies of parasites in native and introduced reptile and amphibian species in the Galápagos are needed to better understand the risk this parasite poses to Galápagos tortoises.

Characterization of the genetics and epidemiology of Brugia sp. in domestic dogs in Chad, Africa.

Neglected tropical diseases pose a threat to domestic animal health, as domestic animals can serve as reservoirs for certain zoonotic parasitic infections, including Guinea worm (Dracunculus medinensis) and lymphatic filariasis. Surveillance for these parasites in domestic animals is needed to understand infection prevalence and transmission cycles, with the goal of instituting appropriate interventions. The goal of this research was to report our finding of Brugia sp. infection in dogs from Chad, Africa, and to characterize the genetics and epidemiology of the parasite. During a recent Chadian canine pathogen surveillance project, we identified Brugia sp. infections in a total of 46 out of 428 dogs (10.7%) sampled at three time points in 2019-2020. We found high levels of sequence similarity to B. malayi and B. pahangi based on amplification of 18S rRNA, 5.8S rRNA, and ITS-2 regions. Phylogenetic analysis of 18S rRNA gene sequences placed the Chadian Brugia sp. in a clade with other Brugia spp. but grouped it separately from both B. malayi and B. pahangi. Analysis of Hha I sequences showed the greatest similarity with B. patei, a parasite previously reported from dogs, cats, and wildlife hosts in Kenya. Epidemiologic analysis using generalized linear regression modeling found significantly higher odds of Brugia sp. detection among dogs in villages in southern Chad compared to those in the northern region. Further, within the northern region, there were higher odds of detection in the dry season, compared to the wet season, which is consistent with the ecology of a presumably mosquito-borne parasite. The same 428 dogs were tested for Dirofilaria immitis antigen using a commercial assay (IDEXX SNAP 4Dx) at the earliest time point of the study, with 119 dogs testing positive. However, no association was noted between Brugia infection and a dog being positive for Di. immitis antigen, with only seven of the 119 Di. immitis antigen-positive dogs being Brugia-positive. This is the first report of Brugia sp. in domestic dogs in Chad and additional research is needed to definitively identify the species present, elucidate transmission, and understand potential risks to canine and human health.

A survey of piroplasms in white-tailed deer (Odocoileus virginianus) in the southeastern United States to determine their possible role as Theileria orientalis hosts.

In 2017, clinical disease and mortality in cattle associated with Theileria orientalis Ikeda was reported in Virginia, U.S. The exotic tick, Haemaphysalis longicornis, is a competent vector for this species. White-tailed deer (Odocoileus virginianus) are commonly infested with H. longicornis in the eastern U.S. and are also infected with several genotypes of piroplasms such as a Theileria sp. (often called Theileria cervi-like), Babesia odocoilei, and Babesia sp. H10. However, it is currently unknown if deer are susceptible to T. orientalis and can act as potential hosts. In this study, we tested 552 white-tailed deer samples from the southeastern U.S. to determine the presence of T. orientalis. We used a PCR-RFLP to test 293 (53%) of these samples to distinguish between piroplasm genera. A total of 189 white-tailed deer were positive with Theileria, 47 were positive with Babesia, and 57 did not amplify. Because this assay does not determine species, we sequenced 30 random samples targeting a fragment of the 18S rRNA gene. Although a high diversity of Theileria and Babesia spp. were detected, none were T. orientalis. All 552 samples were then screened with a T. orientalis specific real-time PCR protocol, but none were positive for T. orientalis. Our data suggests that white-tailed deer are commonly infected with piroplasm species but not T. orientalis.

Investigating Flubendazole as an Anthelmintic Treatment for Guinea Worm (Dracunculus medinensis): Clinical Trials in Laboratory-Reared Ferrets and Domestic Dogs in Chad.

Dracunculus medinensis (Guinea worm [GW]), a zoonotic nematode targeted for eradication, has been managed using interventions aimed at humans; however, increases in domestic dog GW infections highlight the need for novel approaches. We conducted two clinical trials evaluating the efficacy of subcutaneously injected flubendazole (FBZ) as a treatment of GW infection. The first trial was conducted administering FBZ to experimentally infected ferrets; the second trial involved administering FBZ or a placebo to domestic dogs in the Republic of Tchad (Chad). We found contrasting results between the two trials. When adult gravid female GW were recovered from ferrets treated with FBZ, larvae presented in poor condition, with low to no motility, and an inability to infect copepods. Histopathology results indicated a disruption to morulae development within uteri of worms from treated animals. Results from the trial in Chadian dogs failed to indicate significant treatment of or prevention against GW infection. However, the difference in treatment intervals (1 month for ferrets and 6 months for dogs) or the timing of treatment (ferrets were treated later in the GW life-cycle than dogs) could explain different responses to the subcutaneous FBZ injections. Both trials provided valuable data guiding the use of FBZ in future trials (such as decreasing treatment intervals or increasing the dose of FBZ in dogs to increase exposure), and highlighted important lessons learned during the implementation of a field-based, double-blinded randomized control trial in Chadian dogs.

Surveillance for diseases, pathogens, and toxicants of muskrat (Ondatra zibethicus) in Pennsylvania and surrounding regions.

Using diagnostic data and contemporary sampling efforts, we conducted surveillance for a diversity of pathogens, toxicants, and diseases of muskrats (Ondatra zibethicus). Between 1977 and 2019, 26 diagnostic cases were examined from Kansas and throughout the Southeast and Mid-Atlantic, USA. We identified multiple causes of mortality in muskrats, but trauma (8/26), Tyzzer's disease (5/6), and cysticercosis (5/26) were the most common. We also conducted necropsies, during November 2018-January 2019 Pennsylvania muskrat trapping season, on 380 trapper-harvested muskrat carcasses after the pelt was removed. Tissue samples and exudate were tested for presence of or exposure to a suite of pathogens and contaminants. Gastrointestinal tracts were examined for helminths. Intestinal helminths were present in 39.2% of necropsied muskrats, with Hymenolepis spp. (62%) and echinostome spp. (44%) being the most common Molecular testing identified a low prevalence of infection with Clostridium piliforme in the feces and Sarcocystis spp. in the heart. We detected a low seroprevalence to Toxoplasma gondii (1/380). No muskrats were positive for Francisella tularensis or Babesia spp. Cysticercosis was detected in 20% (5/26) of diagnostic cases and 15% (57/380) of our trapper-harvested muskrats. Toxic concentrations of arsenic, cadmium, lead, or mercury were not detected in tested liver samples. Copper, molybdenum, and zinc concentrations were detected at acceptable levels comparative to previous studies. Parasite intensity and abundance were typical of historic reports; however, younger muskrats had higher intensity of infection than older muskrats which is contradictory to what has been previously reported. A diversity of pathogens and contaminants have been reported from muskrats, but the associated disease impacts are poorly understood. Our data are consistent with historic reports and highlight the wide range of parasites, pathogens and contaminants harbored by muskrats in Pennsylvania. The data collected are a critical component in assessing overall muskrat health and serve as a basis for understanding the impacts of disease on recent muskrat population declines.

A multi-seasonal study investigating the phenology, host and habitat associations, and pathogens of Haemaphysalis longicornis in Virginia, U.S.A.

Understanding the abiotic and biotic variables affecting tick populations is essential for studying the biology and health risks associated with vector species. We conducted a study on the phenology of exotic Haemaphysalis longicornis (Asian longhorned tick) at a site in Albemarle County, Virginia, United States. We also assessed the importance of wildlife hosts, habitats, and microclimate variables such as temperature, relative humidity, and wind speed on this exotic tick's presence and abundance. In addition, we determined the prevalence of infection with selected tick-borne pathogens in host-seeking H. longicornis. We determined that the seasonal activity of H. longicornis in Virginia was slightly different from previous studies in the northeastern United States. We observed nymphal ticks persist year-round but were most active in the spring, followed by a peak in adult activity in the summer and larval activity in the fall. We also observed a lower probability of collecting host-seeking H. longicornis in field habitats and the summer months. In addition, we detected H. longicornis on several wildlife hosts, including coyote (Canis latrans), eastern cottontail (Sylvilagus floridanus), raccoon (Procyon lotor), Virginia opossum (Didelphis virginiana), white-tailed deer (Odocoileus virginianus), woodchuck (Marmota monax), and a Peromyscus sp. mouse. This latter record is the first detection of a larval H. longicornis on a North American rodent host important to the enzootic maintenance of tick-borne pathogens of humans and animals. Finally, we continued to detect the exotic piroplasm parasite, Theileria orientalis Ikeda, in H. longicornis as well as other pathogens, including Rickettsia felis, Anaplasma phagocytophilum (AP-1), and a Hepatozoon sp. previously characterized in Amblyomma americanum. These represent some of the first detections of arthropod-borne pathogens native to the United States in host-seeking H. longicornis. These data increase our understanding of H. longicornis biology in the United States and provide valuable information into the future health risks associated with this tick and pathogens.

Copepod consumption by amphibians and fish with implications for transmission of Dracunculus species.

Parasitic nematodes in the genus Dracunculus have a complex life cycle that requires more than one host species in both aquatic and terrestrial habitats. The most well-studied species, Dracunculus medinensis, is the causative agent of human Guinea worm disease (dracunculiasis). There are several other Dracunculus species that infect non-human animals, primarily wildlife (reptiles and mammals). The classic route of D. medinensis transmission to humans is through the ingestion of water containing the intermediate host, a cyclopoid copepod, infected with third-stage larvae (L3s). However, many animal hosts (e.g., terrestrial snakes, dogs) of other Dracunculus sp. appear unlikely to ingest a large number of copepods while drinking. Therefore, alternative routes of infection (e.g., paratenic or transport hosts) may facilitate Dracunculus transmission to these species. To better understand the role of paratenic and transport hosts in Dracunculus transmission to animal definitive hosts, we compared copepod ingestion rates for aquatic species (fish, frogs [tadpoles and adults], and newts) which may serve as paratenic or transport hosts. We hypothesized that fish would consume more copepods than amphibians. Our findings confirm that African clawed frogs (Xenopus laevis) and fish consume copepods, but that fish ingest, on average, significantly higher numbers (68% [34/50]) than adult African clawed frogs (36% [18/50]) during a 24-h time period. Our results suggest that amphibians and fish may play a role in the transmission of Dracunculus to definitive hosts. Still, additional research is required to determine whether, in the wild, fish or frogs are serving as paratenic or transport hosts. If so, they may facilitate Dracunculus transmission. However, if these animals simply act as dead-end hosts or as means of copepod population control, they may decrease Dracunculus transmission.

Susceptibility of anurans, lizards, and fish to infection with Dracunculus species larvae and implications for their roles as paratenic hosts.

Dracunculus spp. are parasitic nematodes that infect numerous species of mammals and reptiles. The life cycles of Dracunculus species are complex, and unknowns remain regarding the role of paratenic and transport hosts in transmission to definitive hosts. We had two primary objectives: to assess the susceptibility of several species of anurans, lizards, and fish as paratenic hosts for Dracunculus species, and to determine the long-term persistence of Dracunculus infections in African clawed frogs (Xenopus laevis). Animals were orally exposed to copepods infected with infectious third-stage larvae (L3s) of either Dracunculus insignis or D. medinensis. Dracunculus L3s were recovered from four anuran species, two lizard species, and one fish species, demonstrating that Dracunculus can infect tissues of a diversity of species. In long-term persistence trials, D. medinensis L3s were recovered from African clawed frogs tissues up to 58 days post-infection, and D. insignis L3s were recovered up to 244 days post-infection. Our findings regarding the susceptibility of novel species of frogs, lizards, and fish to infection with Dracunculus nematodes, and long-term persistence of L3s in paratenic hosts, address pressing knowledge gaps regarding Dracunculus infection in paratenic hosts and may guide future research regarding the transmission of Dracunculus to definitive mammalian hosts.