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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.

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.

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., Graphical abstract Image 1, Highlights • We examined grey-headed albatross chicks found dead at a Subantarctic Island. • Diomedenema diomedeae (Aproctoidea: Desmidocercidae) was found in one albatross. • D. diomedeae infection caused severe air sacculitis. • The first 18S rRNA and COI sequences for a species of Aproctoidea are provided. • Phylogenetic analysis suggests Aproctoidea and Diplotriaenoidea are closely related.

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.

Larval parasites morphologically identified as

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.

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.

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.

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.

Text. Individual history of the studied African penguin. (PDF 25 kb)

Figure S1. Cytoplasmic inclusions in a lymphocyte of an African penguin (Spheniscus demersus) infected by “Candidatus Anaplasma sphenisci”. The lymphocyte with cytoplasmic inclusions (upper right) and a normal lymphocyte (lower left) are shown. Cytoplasmic inclusions had a width of 1.98 ± 0.54 μm (range = 0.86–3.10 μm). Modified Wright-Giemsa stain. Scale-bar: 5 μm. (PDF 8025 kb)

Table S1. GenBank accession codes for the sequences analyzed. (PDF 30 kb)

Figure S2. Phylogenetic relationships of “Candidatus Anaplasma sphenisci” as determined by different phylogenetic methods based on partial sequences of the 16S rRNA and groEL genes. Branch lengths are drawn proportionally to evolutionary distance (scale-bars are shown). Numbers adjacent to nodes indicate bootstrap values (a, b, d, e) or posterior probabilities (c, g). (PDF 110 kb)

The Magellanic Penguin ( Spheniscus magellanicus) is native to Argentina, Chile, and the Falkland/Malvinas Islands, and is a regular winter migrant in Uruguayan and Brazilian coastal waters. The species is known to be susceptible to a variety of gastrointestinal nematodes, cestodes, trematodes, and acanthocephalans, as well as renal trematodes and pulmonary nematodes. Schistosomes (Platyhelminthes, Trematoda, Schistosomatidae) and microfilariae (Nematoda, Secernentea, Onchocercidae) were histologically identified in Magellanic Penguins ( Spheniscus magellanicus) that died while under care at rehabilitation centers in southern Brazil. Phylogenetic analysis of the COI gene, ITS-1 region, 5.8S rRNA gene, ITS-2 region, and 28S rRNA gene sequences of the schistosome revealed that it is closely related to, but distinct from, a schistosome reported from the African Penguin ( Spheniscus demersus). The schistosomes from Magellanic and African Penguins were grouped with Gigantobilharzia huronensis, Gigantobilharzia melanoidis, and Dendritobilharzia pulvurenta; however, the lack of a clearly monophyletic origin precludes determining their genus. The incidental discovery of novel parasites during a study that did not specifically aim to investigate the occurrence of helminths underscores the value of histopathological examination as an exploratory diagnostic approach.