The postmortem diagnosis of an Armillifer infection case in a cynomolgus macaque (Macaca fascicularis).

Armillifer moniliformis belongs to the order Porocephalida and family Porocephalidae, and it can cause zoonotic pentastomiasis. A suspected parasitic infection was incidentally discovered in the abdominal cavity of a cynomolgus macaque that died of persistent diarrhea. 18S rDNA amplification and sequencing revealed a high similarity (99.83%) to the Armillifer moniliformis Guangxi isolate. The isolated parasite was named the Armillifer moniliformis Yunnan isolate (GenBank accession no. HM048870). Our report presents a case of Armillifer moniliformis infection in macaques. The results indicated that early quarantine and diagnosis should be employed for animal health.

Prevalence and risk factors of Linguatula spp. in slaughter animals in Tabriz, Iran, and methods for nymphal stage inactivation.

Linguatula is a food born zoonotic parasite in which carnivores and herbivores serve as final and intermediate hosts, respectively. Human infection with Linguatula spp. occurs following consumption of raw or undercooked infected internal organs of the intermediate host and/or consumption of water and/or vegetables contaminated with eggs released from final hosts. The aim of this study was to determine prevalence and risk factors of Linguatula spp. in sheep, cattle, buffalo, goats and camels slaughtered at the Tabriz abattoir, Iran. In addition, effect of temperature and sodium chloride (NaCl) on survival time of Linguatula spp. nymphs was assessed. For this purpose, 25,520 mesenteric lymph nodes from 2552 animals and the livers and lungs from 656 animals were collected randomly and examined. To evaluate the effect of temperature and NaCl on the survival of Linguatula spp. nymphs in infected livers and lungs, 30 g of each liver and lung with dimensions of 2 × 3 × 4 cm, were exposed to temperatures of -20, 10, 50, 60 and 72 °C and NaCl concentrations of 5 %, 10 %, 15 % and 20 % for 3, 6, 12, 24, 48 and 72 h, in triplicate. Based on the mesenteric lymph nodes, 25.7 % (656 of 2552 animals) were infected with Linguatula spp. Of the 656 liver and lungs assessed, 141 (21.5 %) and 62 (9.5 %) were infected with Linguatula spp., respectively. The rate of infection of mesenteric lymph nodes in all animals was significant with age (P < 0.05), with more older animals infected. In regards to sex, except for camels, more female animals were infected than male animals (P < 0.05). There was a significant difference in survival of nymphs based on temperature and/or NaCl and time (P < 0.0001). At 72 h, all temperatures assessed except 10 °C, resulted in all nymphs being inactivated. Sodium chloride was more effective against Linguatula spp. nymphs in livers than in lungs with 100 % efficacy only achieved against nymphs in livers at 20 % concentration after 48 h and at 10 and 15 % concentration after 72 h. Based on these results, heating and application of common salt as a food preservative in meat products reduces the survival time of Linguatula spp. nymphs and their use could decrease the risk of food-born microorganisms.

Unusual parasitoses in urban dogs: Urinary capillariosis and linguatulosis, cases report.

Parasitic diseases caused by uncommonly diagnosed parasites may pose a threat to companion animals' health in urban environments where they are least expected. The pentastomid Linguatula serrata (tongue worm) and the capillarid Pearsonema plica (syn. Capillaria plica) are parasites with an indirect life cycle, infecting both domestic and wild carnivores. The present report describes two cases: the first one of urinary capillariosis and the other of linguatulosis, in two dogs living in the urban environment of Athens, Greece. In the case of capillariosis, the dog never lived out of the city, so it was presumably infected in the urban environment. On the contrary, in the case of linguatulosis, the dog was adopted at a young age from a rural environment but remained asymptomatic for several months while living in the city. Both dogs had mild symptoms, compatible with these infections. Urinary capillariosis and linguatulosis are uncommon in owned, pet dogs, living in cities due to epizootiological characteristics, i.e. need for wildlife reservoir for P. plica and consumption of raw infected viscera for L. serrata. Different factors contribute to the fact that such infections may occur in scenarios where they are least expected. Recent studies show a progressive worldwide increase in the number of uncommon parasitoses in pet animals, that in some cases, such as linguatulosis, are of zoonotic relevance. Regular parasitological examinations and preventive antiparasitic schemes are necessary in order to treat and prevent infections in pet animals and safeguard the health of both animals and humans under the concept of One Health.

First record of Sebekia oxycephala (Pentastomida: Sebekidae) infecting Helicops angulatus (Reptilia: Colubridae) in Northeast Brazil.

In this study, we investigated the infection by pulmonary parasites in the water snake Helicops angulatus. We found four nymphs of the pentastomide parasite Sebekia oxycephala, parasitizing two adult females H. angulatus collected in the municipality of Rio Tinto, Paraíba, Brazil. This occurrence is likely due to the nymphs of this parasite directly infecting aquatic intermediate hosts, such as fish. The result of this study represents the first record of pentastomid infecting H. angulatus.

Systematic assessment of nymphs of Diesingiinae (Pentastomida: Sebekidae) parasitizing Palloceros harpagos (Cyprinodontiformes: Poeciliidae) from Parque Nacional Iguazú, Argentina: filling gaps in an incomplete phylogenetic framework.

During a search for parasites in fishes from Iguazu River, Argentina, a nymph of pentastomid was found encysted on mesenteries of Phalloceros harpagos. The 28S rDNA and COI mt-DNA were used to compare with the sequences deposited in the GenBank. Pentastomid nymphs belong to the subfamily Diesingiinae (Sebekidae) for having chloride cell pores distributed in a single row per annulus; also, the hooks and rows of spines of our material differ to other genera. Present specimens are most likely Diesingia sp., having 63-74 annuli, a U shaped oral cadre with fibers closing anteriorly and a peg like extension of the oral cadre. The 28S rDNA analysis places our samples into a sister group of Alofia sp., but the COI mt-DNA situate them on the base of the clade. In conclusion, our pentastomid positively belongs to Diesingia sp., however, indisputable assignation requires a reliable description of the nymph, or the availability of sequences linking nymphs and adults, which even could provide evidence of a new hitherto undescribed genus. Aditionally, the systematic position of Sebekia oxycephala previously described by P. harpagos is discussed on the basis of our results, allowing us to suggest a re-assignation of such specimens to the genus Diesingia.

Linguatula serrata (Pentastomida: Linguatulidae) infection in a paucisymptomatic greyhound imported from Romania to Italy: A case report and literature overview.

Background: Linguatula serrata is a pentastomid zoonotic parasite with worldwide distribution. Although some cases of L. serrata infection have been reported in dogs, the epidemiology of this parasite remains largely unknown in developed countries. In recent years, canine linguatulosis has been repeatedly linked to cases of imported infections. This study aims to focus attention on this uncommon parasite through the presentation of a case report and an overview of the literature. Case Description: A 1-year-old intact female Borzoi imported from Romania to Italy sneezed spontaneously a worm-like parasite specimen. Morphological and molecular diagnosis identified the parasite as a female of the zoonotic pentastomid L. serrata (so-called European tongue worm) that lives in the nasopharyngeal tract of canids. Eggs of Linguatula were detected in the feces. Molecular identification (99%-100% homology) was based on DNA extraction, polymerase chain reaction of a 700-bp fragment of the mitochondrial cytochrome c oxidase subunit I gene, and alignment with BLAST analysis. Due to the possibility of other adult or juvenile specimens of the parasite still occurring in the dog, a treatment attempt with a combination of febantel/pyrantel/praziquantel was made. No parasite eggs were detected in fecal samples after the drug was administered. Endoscopy confirmed the absence of adult parasites and slight pathological changes. A follow-up examination conducted 3 months after the treatment did not reveal any clinical and laboratory abnormalities. Conclusion: Linguatula serrata appears to be currently prevalent in some European countries, but there are no recent extensive studies on the prevalence of canine linguatulosis, so the parasite frequently remains undetected and unreported in dogs as the diagnosis is often overlooked. Parasites not commonly found such as L. serrata can become increasingly prevalent and may be detected in imported dogs. Therefore, veterinarians must be aware of the possible presence of uncommon and exotic pathogens in these dogs, be able to recognize the relevant clinical signs, and diagnose the infection quickly. This will improve the prognosis in individual dogs, reduces the risk of possible public health implications, and reduces the risk of uncommon and exotic pathogens establishing new endemic foci.

Characterization of the Tongue Worm, Linguatula serrata (Pentastomida), Identified from Hares (Lepus europaeus) in Romania.

Linguatula serrata (Frölich, 1789) is a widespread parasite known as the tongue worm belonging to the family Linguatulidae. The adult form of the parasite is usually located in the upper respiratory tract of domestic and wild carnivores while the larval forms are located in the visceral organs of intermediate hosts (various herbivorous mammals). Twenty-four European brown hares (Lepus europaeus) were examined in this study, of which two were positive with L. serrata nymphs. The collected nymphs were examined morphologically using electron-microscopic analysis and molecularly by amplification of 18S rRNA and COX1 genes. Lung tissue samples were also collected and histopathological examination was performed. Histopathological examination revealed the following lesions: generalized inflammatory oedema, granulomas with necrosis, calcification and fibrosis in the bronchial tree. The results of molecular sequencing for L. serrata specimens collected from the European brown hares are deposited in GenBank. This study presents the first report on Linguatula serrata nymphs collected from L. europaeus in Romania, using molecular and morphological characterization simultaneously.

Abdominal Calcifications in a Patient From the Congo Basin with History of Snake Meat Consumption: Radiographic Evidence of Prior Armillifer Infection.

We report a case of a 60-year-old asymptomatic male with history of consumption of uncooked snake meat while living in the Congo basin and prior imaging showing multiple abdominal calcifications. Patient had multiple subepithelial colonic lesions identified during screening colonoscopy and microscopic examination of the lesions demonstrated a calcified nodule in the submucosa with overlying normal mucosa. However, no parasite was identified within the calcified nodule. Given the history of consumption of uncooked snake meat and the typical radiographic feature of multiple abdominal calcifications, it is very likely that the patient's radiographic abnormalities are due to prior Armillifer armillatus infection, a parasitic infection acquired from consumption of uncooked snake meat. Patient was asymptomatic at the time of evaluation and was not given anti-parasitic treatment.

Natural history of the parasite Waddycephalus in the Townsville region of north-east Australia.

Waddycephalus is an understudied genus of pentastomids native to Australia and south-east Asia. The genus was recognized in 1922 but there has been little research on these pentastomid tongue worms over the last century. A few observations suggest a complex life cycle through 3 trophic levels. We aimed to add knowledge to the Waddycephalus life cycle in woodland habitats in the Townsville region of north-east Australia. We used camera trapping to identify the most likely first-intermediate hosts (coprophagous insects), we conducted gecko surveys to identify multiple new gecko intermediate host species and we dissected road-killed snakes to identify additional definitive hosts. Our study paves the way for further research into the intriguing life cycle of Waddycephalus, investigation of spatial variation in prevalence and impacts of the parasite on host species.

The invasive pentastome Raillietiella orientalis in a banded water snake from the pet trade.

Raillietiella orientalis is established in Florida and rapidly spreading both geographically and in known host species. A banded water snake (Nerodia fasciata), purchased in Michigan at a regional reptile show, expectorated a pentastome whose morphology and DNA sequence indicated that it was R. orientalis. This event indicates that, through the pet trade, R. orientalis has been spread 1,500 km from its previously known distribution limit. Fecal sample analyses indicated that the snake was shedding large numbers of embryonated eggs for at least several months. The diversity of reptile species that are both known hosts of R. orientalis in Florida and are commonly sold in the pet trade indicates that this invasive pentastome may become a widespread health concern for pet owners and veterinarians.

TONGUE WORM (PENTASTOMIDA) PARASITES OF NORTH AMERICAN HERPETOFAUNA: CHECKLIST OF SPECIES, IDENTIFICATION KEY, AND NEW STATE AND HOST RECORDS FROM MEXICO.

An updated checklist of tongue worms (Pentastomida) which parasitize wild North American amphibians and reptiles is presented: a total of 14 species grouped in 6 genera, 4 families, and 2 orders are registered; these infect a total of 58 species of reptiles and 3 amphibians in the region. An illustrated identification key for the taxa listed is proposed. Kiricephalus coarctatus (Diesing, 1860) (Porocephalidae) is recorded for the first time in the snakes Arizona elegans Kennicott, 1859 (Colubridae); Micrurus tener Baird and Girard, 1853 (Elapidae); and Bothrops asper (Garman, 1884) (Viperidae); the exotic Raillietiella taegueselfiRiley, McAllister, and Freed, 1988 (Raillietiellidae) is recorded in the exotic lizard Hemidactylus frenatus Duméril and Bibron, 1836 (Gekkonidae) for the first time, and in Hemidactylus turcicus (Linnaeus, 1758) (Gekkonidae) for the first time in Mexico, this is also the first time it is recorded in a species of lizard native of the Americas, Sceloporus cyanogenys Cope, 1885 (Phrynosomatidae), this latter record represents a parasite spillover event; finally, Porocephalus stilesiSambon, 1910 (Porocephalidae) is formally recorded for the first time in Mexico in the snake Bothrops asper.

Sebekia sp. (Eupentastomida, Sebekidae) in Pygocentrus piraya (Actinopterygii: Serrasalmidae) from marginal lagoons of the middle São Francisco River basin, Brazil.

Pygocentrus piraya is an endemic species of the São Francisco River basin. In this study, 342 carnivorous fish were examined, nymphs specimens of Sebekia sp. and its ecological indexes are recorded only in piranhas, of the total of 53 collected from three marginal lagoons of the middle São Francisco River basin: 17 specimens from Curral de Varas lagoon; 15 from Maris lagoon, both of state of Minas Gerais; and 21 specimens from Mocambo lagoon, state of Bahia. The prevalence (P) and mean abundance (MA) of Sebekia sp. nymphs in intermediate hosts were as follows: in Curral de Varas lagoon, P = 11.8%, MA = 0.18; in Maris lagoon, P = 46.6%, MA = 0.47; and in Mocambo lagoon, P = 14.3%, MA = 1.05. Despite the lower number of fish examined from Maris lagoon, this lagoon had the highest number of piranhas (seven) parasitized by Sebekia sp. Pentastomids have zoonotic importance and evaluation of the indirect life cycle of sebekids in lagoons is necessary for defining the intermediate and final hosts involved. This record is novel and stems from collection of piranhas in the marginal lagoons of the São Francisco River, in the states of Minas Gerais and Bahia, Brazil.

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.

Tongue worm (subclass: Pentastomida) infection and treatment in two domesticated reptiles - A case report.

Tongue worms (Subclass: Pentastomida) are endoparasites found in carnivorous reptiles, fish, amphibians, birds, and mammals. Several pentastomids cause pentastomiasis, a zoonotic disease. We encountered tongue worm infection in two reptiles imported into Japan: a Wahlberg's velvet gecko (Homopholis wahlbergii) bred in Germany and a wild-caught green tree python (Morelia viridis) from the Republic of Indonesia. The infecting tongue worms were identified as Raillietiella sp. and Armillifer sp. in the gecko and python, respectively, based on morphological and molecular analyses. Gecko is a newly recorded definitive host harboring Raillietiella tongue worm. Live tongue worms were first detected in domestic reptiles in Japan, and this indicates a risk that these tongue worms could be spread to other animals, as well as humans. Therefore, the diagnosis and treatment of infected pets are of importance. In the case of the two infected reptiles, the availability of fecal egg examination for diagnosis was shown. In addition, the effectiveness of treatment with 200 µg/kg/day ivermectin orally once weekly was demonstrated, without no side-effects and no re-detection of eggs after the cessation of treatment.

A Preliminary Study on the Antigenic Proteins of Linguatula serrata Nymphal Stage in Sheep with Visceral Linguatulosis.

Scant information is available on the immunological aspect of Linguatula serrata causing linguatulosis in humans and animals. The present study aimed to analyze the content of crude somatic extracts and excretory-secretory products of L. serrata nymphs to detect the immune response of sheep and immunogenic proteins of the parasite. After collecting the nymphs, somatic extracts were prepared by sonication. Excretory secretory products were prepared by the incubation of nymphs in RPMI medium at 37°C with 5% CO2. Somatic and excretory-secretory proteins were isolated using SDS-PAGE. The immunogenic properties of the resulting proteins were determined using immunoblotting and positive sera from sheep infected with visceral linguatulosis. The total content of somatic extracts and excretory-secretory products of L. serrata nymphs analyzed by SDS-PAGE (12% gel) revealed two protein patterns with more than 18 and 9 strong bands, respectively. Immunoblots using sera samples of sheep infected with the parasite, somatic extracts and excretory-secretory products demonstrated 12 and 3 antigenic proteins with molecular weights mostly in the range of 24-100 kDa and an antigen more than 180 kDa. Three common immunodominant antigenic proteins with molecular weights of 38 and 57, as well as an antigen of more than 180 kDa, were detected in the somatic extracts and excretory-secretory products of L. serrata nymphs in sheep with visceral linguatulosis. These antigens can be considetered prime candidates for future serodiagnosis and immunoprotective studies of the parasite.

Characterization of tongue worms, Linguatula spp. (Pentastomida) in Romania, with the first record of an unknown adult Linguatula from roe deer (Capreolus capreolus Linnaeus).

Specimens of the pentastomid parasite, Linguatula serrata, have been reported from several animals in Romania, including some domestic dogs translocated to other parts of Europe. In this study, gray wolves (Canis lupus, n = 80), golden jackals (C. aureus, n = 115), red foxes (Vulpes vulpes, n = 236), and roe deer (Capreolus capreolus, n = 1) were examined for pentastomes. Overall, 17.5% of wolves were found to be infected with specimens of Linguatula, with a range of infections of one to five individuals per animal. Golden jackals and foxes had much lower infection levels, with 1.73% of golden jackals and 1.69% of foxes infected; both host species were found to be infected with one or two individual pentastomes per animal. The single deer specimen was infected with three individual pentastomes. The pentastomes collected from the wolves and golden jackals were determined to be immature and mature adult specimens of L. serrata based on morphological examination and molecular analysis using the 18S rRNA gene. No pentastomes collected from the red foxes were available for identification. The pentastomes collected from the roe deer were expected to be L. arctica but determined to be mature adult male specimens of an unknown Linguatula, herein, referred to as Linguatula sp. based on its morphology; the results of molecular sequencing for the Linguatula specimen collected from the deer were inconclusive, preventing a final species identification. This study presents the first report of L. serrata in any hosts from Romania through both morphological and molecular characterization, and also presents the first report of a Linguatula sp. in Ca. capreolus, utilizing morphological characterization. Issues of morphological variability are discussed, including the presence of spines in the hook pit of specimens of Linguatula. This study highlights the need to examine all specimens of Linguatula to confirm the stage of development. Despite the inconclusive molecular result for some specimens, the authors still urge future researchers to incorporate a combined molecular and morphological approach in identifying specimens of Linguatula.

28S rRNA sequences for Linguatula spp.

Identification of specimens belonging to the genus Linguatula (Pentastomida) is relatively easy due to their unique morphology. However, differentiation between species of Linguatula can be challenging for several reasons, including considerable differences between different developmental stages of the parasite within and between species. Currently, 18S rRNA and Cox1 sequences are the only available comparable sequences in GenBank, but recent research has discussed the utility of 28S rRNA for pentastomid phylogenetics. This study presents 28S rRNA gene sequences for two members of the genus Linguatula. Sequences of 28S rRNA were successfully obtained from well-identified samples of L. serrata (collected in Australia) and L. nuttalli (collected in South Africa), with voucher specimens. Phylogenetic analysis of the 28S rRNA region showed 6% difference between L. serrata and L. nuttalli, with low levels of intraspecific variation. In comparison, 18S rRNA and Cox1 sequences from the same specimens showed 0.23% and 13% interspecific differences, respectively. The results of this study show that 28S rRNA has greater genetic diversity to allow for improved differentiation between species of Linguatula than 18S rRNA but is on par with Cox1. Records that do not provide adequate morphological or molecular data to justify independent specific diagnoses must be regarded cautiously, and the need for continued research on species of Linguatula, using a combined morphological and molecular analysis, across a number of different hosts, development stages, geographical regions and molecular markers is highlighted.