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.

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.

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.

Dracunculus Species in Meso-mammals from Georgia, United States, and Implications for the Guinea Worm Eradication Program in Chad, Africa.

The prevalence and diversity of parasitic nematodes in wildlife have been well studied for certain species, yet for others considerable gaps in knowledge exist. The parasitic nematode Dracunculus insignis infects North American wildlife, and past research on this species has led to an increased understanding of the potential host diversity and transmission of the closely related human Guinea worm, Dracunculus medinensis (which is currently the focus of a global eradication program). Many definitive hosts have been documented for D. insignis; however, the life cycle has been studied only in laboratories, and only a single phylogenetic study has been conducted on D. insignis (from Canada). The goals of the present study were to investigate the prevalence of infections with Dracunculus species among wildlife at a single site (Di-Lane plantation) in the southeastern United States, evaluate the genetic diversity of parasites at this site, and investigate potential paratenic hosts that may be involved in transmission. Over 3 yr, we sampled 228 meso-mammals, reporting an overall prevalence of infection with Dracunculus insignis of 20% (46/228). Amphibians and fish were sampled in the same geographic area as infected meso-mammals. Dracunculus insignis third-stage larvae were recovered from 2 different species of amphibians, but all fish sampled were negative. Phylogenetic analysis of the partial cytochrome c oxidase I (COI) gene showed very little diversity of Dracunculus at Di-Lane; however, we did recover a single nematode from a Virginia opossum (Didelphis virginiana) that falls outside of the D. insignis clade, more closely aligns with Dracunculus lutrae, and may represent an undescribed species. This work documents the occurrence of D. insignis in frogs, a potential transmission pathway for D. insignis at a single geographic site in nature. When applied to the global Guinea Worm Eradication Program, and Chad, Africa, in particular, this work increases our knowledge of the potential role of aquatic animals in the transmission of Dracunculus species and informs on potential intervention strategies that may be applied to the eradication of Guinea worm in Africa.

Surveillance for Borrelia spp. in Upland Game Birds in Pennsylvania, USA.

The Borrelia genus contains two major clades, the Lyme borreliosis group, which includes the causative agents of Lyme disease/borreliosis (B. burgdorferi sensu stricto and other related B. burgdorferi sensu lato genospecies), and the relapsing fever borreliosis group (B. hermsii, B. turicatae, and B. parkeri). Other unclassified reptile- and echidna-associated Borrelia spp. (i.e., B. turcica and 'Candidatus Borrelia tachyglossi', respectively) do not belong in either of these two groups. In North America, Borrelia spp. from both of the major clades are important pathogens of veterinary and public health concern. Lyme disease is of particular interest because the incidence in the northeastern United States continues to increase in both dogs and humans. Birds have a potentially important role in the ecology of Borrelia species because they are hosts for numerous tick vectors and competent hosts for various Borrelia spp. Our goal was to investigate the prevalence of Borrelia spp. in four free-living species of upland game birds in Pennsylvania, USA including wild turkey (Meleagris gallopavo), ruffed grouse (Bonasa umbellus), ring-necked pheasants (Phasianus colchicus), and American woodcock (Scolopax minor). We tested 205 tissue samples (bone marrow and/or spleen samples) from 169 individuals for Borrelia using a flagellin gene (flab) nested PCR, which amplifies all Borrelia species. We detected Borrelia DNA in 12% (24/205) of samples, the highest prevalence was in wild turkeys (16%; 5/31), followed by ruffed grouse (13%; 16/126) and American woodcock (3%; 1/35). All pheasants (n = 13) were negative. We sequenced amplicons from all positive game birds and all were B. burgdorferi sensu stricto. Our results support previous work indicating that certain species of upland game birds are commonly infected with Borrelia species, but unlike previous studies, we did not find any relapsing fever borreliae.

Lungworm (Crenosoma goblei) Infection in Unweaned Free-Ranging Raccoons (Procyon lotor) in Washington State, USA.

Larval parasites morphologically identified as Crenosoma goblei were found in orphaned raccoons (Procyon lotor) admitted to a wildlife rehabilitation center in Washington State, US. Partial 18S rRNA and cytochrome oxidase I gene sequences were most similar to other Crenosoma spp. Although commonly reported in free-ranging raccoons throughout much of the central and eastern US, this finding extends the range of C. goblei into the Pacific Northwest of North America. Fecal samples taken within 48 h of admission were positive for larvae in 70% (21/30) of individuals and in all five group samples taken from an additional 14 raccoons. Typically, Crenosoma spp. use an intermediate host, but the unweaned status of all the raccoons in this report suggests a potential direct, possibly vertical, route of transmission. Serial fecal analysis of pooled samples collected over several subsequent months demonstrated spontaneous resolution of larval shedding. Adult worms and gross evidence of pneumonia were detected in the lungs of several juvenile raccoons at necropsy, and verminous pneumonia was confirmed histologically in the single case submitted. Clinical signs directly attributable to verminous pneumonia were not seen in most actively shedding raccoons, suggesting that subclinical disease among juveniles may be common.

Rickettsia species in ticks collected from wild pigs (Sus scrofa) and Philippine deer (Rusa marianna) on Guam, Marianna Islands, USA.

The prevalence and diversity of ticks on wildlife species on Guam is understudied, as to date no work has been conducted on the infection of these ticks with Rickettsia (obligate intracellular pathogens that use a variety of ectoparasites as vectors and can cause disease in humans, domestic animals, and wildlife species). The goal of our study was to investigate the presence of Rickettsia species on the island of Guam by testing ticks found on Philippine deer (Rusa marianna) and wild pigs (Sus scrofa). Increasing numbers of these species have led to increased interactions with humans, including hunting, highlighting the importance of studies on vector prevalence and associated zoonotic pathogens. In this study, ticks were removed from Philippine deer and wild pigs in March and April of 2015 and tested for Rickettsia spp. using nested PCR. Overall, a low prevalence of Rickettsia spp. was detected (5.4% (6/112 ticks)). Ticks removed from wild pigs were identified as Amblyomma breviscutatum, one of which was positive for Rickettsia ambylommatis. Ticks recovered from Philippine deer were identified as Rhipicephalus microplus, and five were positive for Rickettsia; two with R. amblyommatis and one with 'Candidatus Rickettsia senegalensis', a recently proposed species in the R. felis cluster. The remaining two sequences were short and species classification was not possible. Rickettsia felis is a known zoonotic pathogen in the spotted fever group of Rickettsia and there is evidence that 'C. R. senegalensis' can also cause illness in people. This study confirms the occurrence of Rickettsia in ticks on Guam and highlights the presence of potential human pathogenic species in the R. felis cluster.

A search for tiny dragons (Dracunculus medinensis third-stage larvae) in aquatic animals in Chad, Africa.

Dracunculus medinensis, or human Guinea worm (GW), causes a painful and debilitating infection. The global Guinea Worm Eradication Program (GWEP) has successfully reduced human GW cases from 3.5 million in 21 countries in 1986 to only 30 cases in three remaining countries in 2017. Since 2012, an increase in GW infections in domestic dogs, cats and baboons has been reported. Because these infections have not followed classical GW epidemiological patterns resulting from water-borne transmission, it has been hypothesized that transmission occurs via a paratenic host. Thus, we investigated the potential of aquatic animals to serve as paratenic hosts for D. medinensis in Chad, Africa. During three rainy and two dry season trips we detected no GW larvae in 234 fish, two reptiles and two turtles; however, seven GW larvae were recovered from 4 (1.4%) of 276 adult frogs. These data suggest GW infections may occur from ingestion of frogs but the importance of this route is unknown. Additional studies are needed, especially for other possible routes (e.g., ingestion of fish intestines that were recently shown to be a risk). Significantly, 150 years after the life cycle of D. medinensis was described, our data highlights important gaps in the knowledge of GW ecology.

A novel candidate species of Anaplasma that infects avian erythrocytes.

BACKGROUND: Anaplasma spp. are Gram-negative obligate intracellular bacteria transmitted by ticks. Even though numerous studies have detected DNA from Anaplasma spp. in the blood of birds, thus far mammals were the only vertebrates demonstrated to serve as competent hosts to these organisms. We report a novel candidate species of Anasplasma that was associated with cytoplasmic inclusions in the erythrocytes of an African penguin (Spheniscus demersus) in South Africa. METHODS: Cytoplasmic inclusions were morphologically characterized from freshly-produced blood smears, and phylogenetic analysis of 16S rRNA and groEL genes were used to evaluate the evolutionary relationships of the organism to other Anaplasmataceae. RESULTS: Dark-purple round or oval inclusions consistent with Anaplasmataceae morulae were observed in the cytoplasm of erythrocytes. Phylogenetic trees produced using different methods agreed that the organism detected in this study belongs to the genus Anaplasma, and suggested that it is most closely related to the cluster comprising A. centrale, A. capra, A. marginale and A. ovis. We propose provisionally naming the strain detected in this study as "Candidatus Anaplasma sphenisci". CONCLUSIONS: This is the first species of Anaplasma shown to produce cytoplasmic inclusions in avian cells, opening the possibility that cytoplasmic inclusions in avian erythrocytes that had previously been attributed to Aegyptianella sp. might in fact correspond to Anaplasma. Further studies on the molecular biology of avian-infecting Anaplasmataceae will be valuable to provide insight into the evolution and epidemiology of these organisms.

Molecular characterization and lesions associated with Diomedenema diomedeae (Aproctoidea: Desmidocercidae) from grey-headed albatrosses (Thalassarche chrysostoma) on Subantarctic Marion Island.

The Prince Edward Islands are Subantarctic islands in the southwest Indian Ocean that are of global importance as seabird nesting sites, and are breeding grounds for five species of albatrosses (Procellariiformes: Diomedeidae). In March-April 2016 numerous chicks of one of these species, the grey-headed albatross (Thalassarche chrysostoma), were found dead at colonies on Marion Island (46°57'S 37°42'E), the larger of the two Prince Edward Islands. Affected chicks were weak, prostrated, apathetic, had drooping wings, and many eventually died while sitting on the nest. Five carcasses were necropsied, and samples were obtained for pathological and parasitological analysis. Four chicks appeared to have died from starvation, and one died due to air-sac helminthiasis, with extensive hemorrhage in the air sacs and multifocal pyogranulomatous air-sacculitis. The air sac parasites were identified as Diomedenema diomedeae (Aproctoidea: Desmidocercidae). Phylogenetic analysis of the nuclear 18S rRNA gene and mitochondrial COI gene confirmed that D. diomedeae belongs to the suborder Spirurina and showed that it is most closely related to the Diplotriaenidae (superfamily Diplotriaenoidea), a family of parasites that infect the air sacs and subcutaneous tissues of a variety of bird species. To our knowledge this is the first record of the occurrence of a nematode in the respiratory tract of an albatross and the first study to provide DNA sequences for a species of the superfamily Aproctoidea.

Prevalence and genetic characterization of Dirofilaria lutrae Orihle, 1965 in North American river otters (Lontra canadensis).

Dirofilaria lutrae Orihle, 1965 is a subcutaneous filarial worm found in North American river otters (Lontra canadensis). Previous studies reported the geographical range of this worm to include Louisiana, Florida, and North Carolina, USA. Although D. lutrae does not cause disease in otters, it must be distinguished from Dirofilaria immitis (Leidy, 1856), which causes heartworm disease in otters. The goal of this study was to determine the prevalence of D. lutrae in several states and to investigate the phylogenetic relationship of D. lutrae and other Dirofilaria species. Adult D. lutrae were collected from 32 of 40 (80%) otters from North Carolina (n = 38), South Carolina (n = 1), and Kentucky (n = 1), whereas all otters from Georgia (n = 1), Pennsylvania (n = 2), Arkansas (n = 184) and California (n = 1) were negative for D. lutrae. A portion of the cytochrome c oxidase I (COI) was amplified from nine North Carolina specimens and one South Carolina specimen. Sequence analysis indicated that all but one were identical to each other and D. lutrae was phylogenetically most closely related to D. immitis; however, few Dirofilaria sequences are available for comparison. Because other subcutaneous parasites may be detected in otters, parasites should be carefully examined to confirm their identity. Finally, given the high prevalence of D. lutrae in otters in some southeastern states, microfilaria found in blood samples of otters must be correctly identified because treatment for D. lutrae is generally not warranted and some pharmaceutical treatments can cause mortality in otters.

Dracunculus infections in domestic dogs and cats in North America; an under-recognized parasite?

We reviewed 62 new cases and 18 published reports of Dracunculus infections in domestic dogs and cats to describe the epidemiology of this parasite in dogs and cats in North America. We collected host and parasite data when available, including age, sex, and breed of dog, nematode location in the host, and any clinical signs at presentation and/or description of the apparent lesion. For dogs, infections were noted in six of the AKC breed groups, but none was reported from the toy group or the miscellaneous breed class. Age of infected dogs ranged from 7 months to 19 years (median 4 years; average 5.3 years), and infection rates were similar in male and female dogs. Most nematodes were associated with the distal extremities, but worms were also found in the chest/thorax, abdomen, head, and flank. Although most infected dogs had a single worm, three dogs had two or more worms that were collected from multiple lesions. Three new cat cases, with similar lesions, presentations and seasonality, were detected in Alabama, North Carolina and Texas. Cases were reported from a wide geographic range throughout eastern North America, during every month of the year, but 72% of infections were diagnosed in the late winter to early spring (December to May). All collected worms were larvigerous females which cannot be identified to species based on morphologic characters. Thus, we attempted to amplify and sequence a portion of the cytochrome c oxidase subunit I (COI) gene for specific identification. Although 13 worms from 12 cases were available, sequences were obtained for only eight worms from seven cases. These eight worms were D. insignis, a common parasite of raccoons (Procyon lotor) and other primarily carnivorous mammals. Female worms are the most likely to be detected in dogs and cats because male worms do not emerge, parasites should be preserved in ethanol for molecular identification. Although this study used convenience sampling of available data, we found that the parasite is widespread throughout the eastern US and Canada and that Dracunculus infections in dogs are more common than is revealed in published literature. However, more research is needed to understand the epidemiology, including transmission route(s), prevalence, and distribution of this parasite.