Closing the knowledge gap: Helminth parasites of freshwater turtles from the Chaco-Pampa Plain, Southern South America.

Six species of freshwater turtles dominate the Chaco-Pampa Plain in southern South America and their parasites have been relatively understudied, with most records concentrated in Brazil. Particularly in Argentina, there are only scattered records of parasites for most of the turtles that inhabit the region, leaving a large knowledge gap. The purpose of the present contribution is to increase the knowledge of the internal parasites of six species of freshwater turtles from Argentina, after 15 years of fieldwork, by providing new hosts and additional geographic records for many host-parasite relationships. Some molecular sequences of the studied parasites were provided as a tool for better species identification. We processed 433 stomach and fecal samples from live individuals and visceral and soft tissue samples from 54 dissected turtles collected from a wide range and different ecoregions. We found 6230 helminths belonging to 18 taxa (one cestode, 11 digeneans and six nematodes). Fourteen new parasite-host associations are reported here, and for the first time parasites are recorded for Phrynops williamsi. This work contributes significantly to the knowledge of the parasitofauna in freshwater turtles in Argentina, providing a detailed list of parasites present in each turtle species and reporting molecular characters for future studies.

Gastrointestinal parasites of Peltocephalus dumerilianus (Testudines: Podocnemididae) from Jaú National Park, Brazilian Amazon.

The big-headed Amazon River turtle, Peltocephalus dumerilianus, is endemic to the Orinoco and Amazon River basins. It is a food source for local communities, often unsustainably. Knowledge about P. dumerilianus' parasitological fauna and host-parasite relationships is limited. Thus, ecological aspects of gastrointestinal parasitism in this species were investigated. Helminths were found in the gastrointestinal tract of 21 turtles, morphologically identified, and infection descriptors calculated. All animals harbored helminths: nematodes Ancyracanthus pinnatifidus, Paratractis hystrix, Atractis trematophila, Klossinemella conciliatus indeterminate three Klossinemella species, and digeneans Nematophila grandis, Helicotrema spirale, and Telorchis hagmanni. The highest parasite load occurred in the large intestine, followed by the small intestine and stomach. Shell length directly correlated with parasite burden of heteroxenic helminths, with males having higher burden than females. This is the first record of A. trematophila, K. conciliatus, and T. hagmanni in P. dumerilianus, and new location record for A. trematophila, P. hystrix, N. grandis, H. spirale, and T. hagmanni. Three potentially new Klossinemella species are presented.

Orientatractis bowseri n. sp. (Nematoda: Cosmocercoidea: Atractidae) parasite of Peltocephalus dumerilianus (Schweigger) (Testudines: Podocnemididae) from the Brazilian Amazon.

Peltocephalus dumerilianus (Schweigger) is a diurnal freshwater turtle widely distributed in the Orinoco and Amazon River basins in Brazil, Colombia, Peru, Ecuador, Venezuela, and French Guiana. During a helminthological survey of freshwater turtles from the Brazilian Amazon, numerous nematodes with characteristics of the genus Orientatractis Petter, 1966 were recovered from the stomach of Peltocephalus dumerilianus. These nematodes were different from all congeneric species, and we describe herein. Orientatractis bowseri n. sp. differs from all known species of the genus by having dorsal cuticular projections on the posterior region of the females, distribution of caudal papillae and by having papilla-like ornamentations on the cuticle distributed from the mid-esophagus to the mid portion of the body. This study represents the fourth specie of the genus Orientatractis reported in Brazil, the seventh species of Orientatractis in the Neotropical region, and the third found infecting freshwater turtles in Brazil.

New Recording of Toxoplasma gondii in Wild Tortoise Testudo graeca Using Nested PCR Method.

Toxoplasmosis is one of the most widespread zoonotic diseases, especially in warm and humid areas, and affects all mammals, including humans and many herbivores and carnivores. The present study investigated the Toxoplasma gondii (T. gondii) parasite in tortoises for the first time in Iraq using PCR technology. A total of 28 tortoises/Testudo graeca (T. graeca) were collected between October 2018 and March 2019 from the study stations and then sent to the Animal House, which belongs to the Department of Biology, Faculty of Education, University of AL-Qadisiyah, Iraq, to perform the dissection. The body cavity was opened, and all organs were removed. The tortoises' liver, heart, and brain were removed and kept at -20ºC until use. Afterward, the samples were subjected to DNA extraction. The Nested-PCR technique was implemented using two pairs of primers, and then the PCR products were analyzed using 1.5% agarose gel electrophoresis. The amplification of the gene during the first cycle indicated that 10 samples gave positive results with a total percentage of (11.9%), including five liver samples, three heart samples, and two brain samples (17.85%, 10.71%, and 7.14%, respectively). On the other hand, during the second cycle of the reaction, the amplification of the gene was obtained in seven samples (8.33%). The highest percentage of the presence of the gene was recorded in the tortoises' liver (14.28%) and the lowest in their brain (3.57%). This study is among the first to investigate the molecular detection of T. gondii in wild tortoises (T. graeca) in Iraq. The findings imply that tortoises have a role in transmitting T. gondii and are believed to acquire infection by feeding on small invertebrate animals or plants contaminated with the oocysts of the parasite.

Hemoparasite occurrence and hematological/serum chemistry variations in Podocnemis vogli turtles: A comparative analysis between wild-residing infected specimens and captive non-infected counterparts.

Reptiles show a high occurrence of hemoparasites in the wild; however, little is known about the impact of such infections on their hosts' physiology and health status. Podocnemis vogli is an ancient turtle distributed in South America, frequently infected by blood parasites. Specifically, we analyzed the hematological and serum chemistry parameters of 78 wild turtles. We compared these values with those obtained from non-infected turtles of the same species in ex-situ conditions, considering factors such as sex and coinfections. Two orders of hemoparasites were detected under microscopic analyses: Adelorina, represented by Haemogregarina sp. (98.72 ± 0.28%), and Haemosporida, represented by Haemocystidium sp. (30.77 ± 1.16%), the latter genus being always in coinfection with Haemogregarina. Significant differences were observed in 20 parameters between infected (free living) and uninfected (living in captivity) turtles. The ALP, PCV, Hb, and MCV were significantly different by sex; albumin, cholesterol, creatinine, percentage of heterophils, lymphocytes, and basophils differed in coinfection. This is the first report of reference intervals of P. vogli in the wild and the first study comparing hematological and blood biochemistry values between hemoparasite-infected and uninfected P. vogli turtles. However, the limited knowledge of these parameters in wild reptiles and the wide range of interval values makes it difficult to determine any significant impact on turtle health. Nevertheless, this study highlights the need for more in-depth research to explore the potential effects of blood parasite infections on turtles, including immune response and coevolution studies.

Detection of Hemopathogens in Chelonoidis carbonaria: Microscopic, Molecular, Hematological, and Clinical Biochemistry Aspects.

Background: The growing contact between men and wild animals, caused by the increase in the population in urban centers and the destruction of the habitat of these animals, has been leading to a greater circulation of pathogens between humans and wildlife. Chelonoidis carbonaria, a tortoise found throughout South America, is one of the animals most rescued from animal trafficking and illegal breeding. Considering this situation, this study aimed to verify the occurrence of hemoparasites in C. carbonaria. Materials and Methods: Blood samples from 73 C. carbonaria were collected from animals located in (1) a rural commercial breeding unit, (2) an urban zoo, and (3) a center of rescued animal screening. Genomic DNA was extracted from these animals and used in PCRs to detect specific genomic fragments of haemogregarines (i.e., Hepatozoon and Hemolivia), and members of the Anaplasmataceae Family (i.e., Ehrlichia sp. and Anaplasma sp.). Blood samples were screened for hemopathogens by direct microscopy and were used for hematological assays, and serum samples were analyzed to determine the concentration of serum components. Results: It was found that 34.2% of the tortoises presented Sauroplasma sp. in their blood samples; these animals showed clinical biochemistry changes that indicate altered liver function. Two zoo animals were positive for Ehrlichia sp. in PCR, and also presented clinical biochemistry and hematological changes. Conclusion: The present project is pioneer in the detection of Ehrlichia sp. in C. carbonaria, and was able to identify changes in clinical biochemistry that can be a result of the infection by hemopathogens in this species.

New species of Polystomoides (Monogenoidea: Polystomatidae) parasitizing the urinary bladder of a freshwater turtle in Brazil.

Trachemys dorbigni is the most abundant freshwater turtle species in Rio Grande do Sul, southern Brazil. Chelonians are known to host a wide variety of pathogens, including viruses, bacteria, hemoparasites and helminths. Among these, nine genera of polystomatid flatworms (Monogenoidea; Polystomatidae) infect freshwater turtles: Apaloneotrema, Aussietrema, Fornixtrema, Manotrema, Pleurodirotrema, Polystomoidella, Polystomoides, Uropolystomoides and Uteropolystomoides. However, little is known about the biology of these parasites in the Neotropical Realm. Through investigative cystoscopy, specimens of Polystomatidae were located inside the urinary bladder of the host T. dorbigni. Retrieved specimens were fixed and stained whole mounts prepared for taxonomic identification. In the present paper, a new species of Polystomoides (Monogenoidea: Polystomatidae) parasitizing the urinary bladder of a freshwater turtle of the species T. dorbigni in Brazil is described. Polystomoides santamariensis n. sp. differs from the congeneric species on the length of the genital spines, which are longer. Given the enormous diversity of freshwater turtles around the world, it is likely that a large number of chelonian polystomatids are still unknown.

A quantum leap in the evolution of platyhelminths: Host-switching from turtles to hippopotamuses illustrated from a phylogenetic meta-analysis of polystomes (Monogenea, Polystomatidae).

While monogenean worms are mainly parasites of the gills and skin of fish, and to a lesser extent parasites of the oral cavity, urinary bladder, and/or conjunctival sacs of amphibians and freshwater turtles, Oculotrema hippopotamiStunkard, 1924 is the single monogenean polystome reported from a mammal, the common hippopotamus (Hippopotamus amphibius Linnaeus). Several hypotheses have been suggested in the last decade to explain the origin of this enigmatic parasite which infects the conjunctival sacs of H. amphibius. Based on a molecular phylogeny inferred from nuclear (28S and 18S) and mitochondrial (12S and COI) sequences of O. hippopotami and chelonian polystomes, we found a sister group relationship between O. hippopotami and Apaloneotrema moleri (Du Preez & Morrison, 2012). This result suggests lateral parasite transfer between freshwater turtles and hippopotamuses, thus likely reflecting one of the most exceptional known examples of host-switching in the course of vertebrate evolution. It also demonstrates that the proximity in the ecological habitat of parasites within host species is an important feature for their speciation and diversification. Because A. moleri and its host, the Florida softshell turtle (Apalone ferox (Schneider)), are restricted to the USA, we suggest that an ancestral stock of parasites may have been isolated on primitive African trionychids after they diverged from their American relatives, and then switched to hippopotamuses or anthracotheres in Africa.

Epistylis spp. Infestation in Two Species of Mud Turtles (Kinosternon spp.) in the American Southwest.

The protistan genus Epistylis contains freshwater colonial species that attach to aquatic organisms in an epibiotic or parasitic relationship. They are known to attach to the epidermis and shells of aquatic turtles, but have not been reported to cause heavy infestations or morbidity in turtles. We documented heavy infestations of Epistylis spp. in several populations of Sonoran mud turtles (Kinosternon sonoriense) inhabiting livestock ponds in Arizona, USA, and rough-footed mud turtles (Kinosternon hirtipes) from livestock ponds in Texas, USA, over the course of several years. Severe Epistylis spp. infestations on mud turtles appeared to alter diving and swimming behavior when compared to uninfested conspecifics. Infestations were cleared in captivity using tap water or a 10% salt solution, and the turtles had no permanent damage to their shell or epidermis upon clearing. While several of the mud turtles we observed had poor body condition, it is possible that the severe infestations we observed were caused by a comorbidity associated with a pathogen, parasite, or poor habitat quality that made the turtles more susceptible to the Epistylis spp. infestation. Further research on causes for these severe infestations are warranted because they contribute to changes in behavior of the heavily infested turtles and may contribute to morbidity in Kinosternon spp. when mud turtles inhabit extremely warm, shallow, eutrophic aquatic habitats, such as livestock ponds.

Revision of the systematics of the Polystomoidinae (Platyhelminthes, Monogenea, Polystomatidae) with redefinition of Polystomoides Ward, 1917 and Uteropolystomoides Tinsley, 2017.

Polystomatids are platyhelminth parasites that infect mainly amphibians and freshwater turtles. For more than seven decades, chelonian polystomes were classified into three genera according to the number of hamuli, i.e. absent for Neopolystoma, one pair for Polystomoidella and two pairs for Polystomoides. Following re-examination of morphological characters, seven new genera were erected the past six years, namely Apaloneotrema, Aussietrema, Fornixtrema, Manotrema, Pleurodirotrema, Uropolystomoides and Uteropolystomoides. However, the polyphyly of Neopolystoma and Polystomoides on the one hand, and the nested position of Uteropolystomoides within a clade encompassing all Neopolystoma and Polystomoides spp. on the other, still raised questions about the validity of these genera. We therefore re-examined several types, paratypes and voucher specimens, and investigated the molecular phylogeny of polystomes sampled from the oral cavity of North American turtles to re-evaluate their systematic status. We show that all Polystomoides Ward, 1917, sensu Du Preez et al., 2022, Neopolystoma Price, 1939, sensu Du Preez et al., 2022 and Uteropolystomoides Tinsley, 2017 species, display vaginae that are peripheral and extend well beyond the intestine. We thus reassign all species of the clade to Polystomoides and propose nine new combinations; however, although Uteropolystomoides is nested within this clade, based on its unique morphological features, we propose to keep it as a valid taxon. Polystomoides as redefined herein groups all polystome species infecting either the oral cavity or the urinary bladder of cryptodires, with peripheral vaginae and with or without two pairs of small hamuli. Uteropolystomoides nelsoni (Du Preez & Van Rooyen 2015), originally described from Pseudemys nelsoni Carr is now regarded as Uteropolystomoides multifalx (Stunkard, 1924) n. comb. infecting three distinct Pseudemys species of North America.

Title: Révision de la systématique des Polystomoidinae (Plathelminthes, Monogenea, Polystomatidae) avec redéfinition des genres Polystomoides Ward, 1917 et Uteropolystomoides Tinsley, 2017. Abstract: Les Polystomatidés sont des plathelminthes parasites qui infestent principalement les amphibiens et les tortues d'eau douce. Pendant plus de sept décennies, les polystomes de chéloniens ont été classés en trois genres selon le nombre d'hamuli, absents pour Neopolystoma, une paire pour Polystomoidella et deux paires pour Polystomoides. Suite au réexamen des caractères morphologiques, sept nouveaux genres ont été érigés ces six dernières années, à savoir Apaloneotrema, Aussietrema, Fornixtrema, Manotrema, Pleurodirotrema, Uropolystomoides et Uteropolystomoides. Cependant, la polyphylie de Neopolystoma et Polystomoides d'une part, et la position imbriquée d'Uteropolystomoides au sein d'un clade englobant toutes les espèces de Neopolystoma et Polystomoides d'autre part, soulèvent encore des questions sur la validité de ces trois genres. Nous avons donc réexaminé plusieurs types, paratypes et vouchers et étudié la phylogénie moléculaire de polystomes prélevés dans la cavité buccale de tortues d'Amérique du Nord pour réévaluer leur statut systématique. Nous montrons que toutes les espèces de Polystomoides Ward, 1917, sensu Du Preez et al., 2022, Neopolystoma Price, 1939, sensu Du Preez et al., 2022 et Uteropolystomoides Tinsley, 2017, présentent des vagins périphériques qui s'étendent bien au-delà de l'intestin. Nous réattribuons ainsi toutes les espèces du clade à Polystomoides et proposons neuf nouvelles combinaisons; cependant, nous proposons de conserver Uteropolystomoides sur la base de ses caractéristiques morphologiques exceptionnelles, bien que son espèce soit imbriquée au sein de ce clade. Polystomoides tel que redéfini ici regroupe toutes les espèces de polystomes infectant soit la cavité buccale, soit la vessie des cryptodires, avec des vagins périphériques, et deux paires de petits hamuli ou sans hamuli. Uteropolystomoides nelsoni (Du Preez & Van Rooyen 2015), l'unique espèce décrite à l'origine à partir de Pseudemys nelsoni Carr est maintenant considérée comme Uteropolystomoides multifalx (Stunkard, 1924) n. comb., qui infecte trois espèces distinctes de Pseudemys d'Amérique du Nord.

Dolphins and sea turtles may host zoonotic parasites and pathogenic bacteria as indicators of anthropic pressure in the Gulf of Taranto (Northern Ionian Sea, Central-Eastern Mediterranean Sea).

The occurrence of protozoan parasites Giardia duodenalis and Cryptosporidium spp. such as the pathogenic bacteria Salmonella spp. and Escherichia coli was molecularly investigated in the following free ranging species of striped dolphins (Stenella coeruleoalba), Risso's dolphins (Grampus griseus) as well as loggerhead (Caretta caretta) and green (Chelonia mydas) sea turtles living in the Gulf of Taranto (Mediterranean Sea). Out of forty-one investigated individuals belonging to the 4 species, 13 (31.7%) were positive to one or more pathogens and zoonotic G. duodenalis assemblage A, Cryptosporidium parvum and S. enterica were identified in striped dolphins, loggerhead and green sea turtles. In this work, the presence of these opportunistic pathogens has been investigated in fecal samples of free ranging dolphin and sea turtle species for the first time. Moreover, this is the first record of C. parvum in loggerhead sea turtles. These results may provide baseline data for the potential role of cetaceans and sea turtles as potential sentinel species for zoonotic and terrestrial pathogens in the marine environment.

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.

Vogtnema asymmetrica n. gen., n. sp. (Nematoda: Cosmocercoidea: Atractidae) parasite of Rhinoclemmys punctularia (Daudin) (Testudines: Geoemydidae) from eastern Amazon, Brazil.

Nematodes collected from the stomach and large intestine of Rhinoclemmys punctularia (Daudin) from the eastern Amazon, Brazil, are assigned to a new genus, Vogtnema n. gen., allocated in the family Atractidae (Cosmocerdoidea). Vogtnema n. gen. differs from all other genera of Atractidae based on the combination of the following morphological characters: four lips V-shaped (two subdorsal and two subventral) without sclerotized structures associated, absence of sclerotized formations and/or pieces surrounding the oral opening, oral aperture surrounded by flange internal membranous structures; in males the larger spicule is alate. The type species Vogtnema asymmetrica n. gen., n. sp. has the five precloacal pairs of papillae situated on the right side of the body that are more prominent than those on the left side. We also provide an identification key to the genera of the family Atractidae.

Pathological changes by spirorchiid eggs in hawksbill sea turtle (Eretmochelys imbricata) stranded off brazilian coast.

Eretmochelys imbricata (hawksbill turtle) is classified as a critically endangered species at world level; in Brazil, it is listed among the threatened species. Spirorchiids are parasites of the circulatory system of chelonians which may cause serious lesions in the various tissues of the host due to deposition of eggs in the bloodstream. In this context, the aim of the present study was to describe the pathology caused by spirorchiid eggs in E. imbricata from the Brazilian over a five year period. A total of 29 animals were analyzed, of which nine (31%) presented lesions associated to spirorchiids eggs. The lesions were: granulomatous enteritis in six (66.66%), granulomatous splenitis in five (55.55%), granulomatous hepatitis in three (33.33%), granulomatous pneumonia in three (33.33%), granulomatous pancreatitis in two (22.22%), and granulomatous adrenalitis in one specimen (11.11%). Concluded the main lesion in Eretmochelys imbricata was giant-cell granulomatous inflammatory reaction to the parasite's eggs. Animals exhibiting poor physical health were more susceptible to presenting such lesions.

Blood parasite diversity (Apicomplexa: Haemogregarinidae) within the western populations of the European pond turtle Emys orbicularis.

Molecular tools have revolutionized assessments of blood parasites in freshwater turtles. In the Iberian Peninsula and North Africa, two native species of terrapins occur, Emys orbicularis (Linnaeus) and Mauremys leprosa (Schweigger). Both have been identified as hosts for the blood parasite Haemogregarina stepanowi Danilewsky, 1885, which has also been found in related species. However, recent assessments of M. leprosa have identified several distinct genetic lineages of these parasites in this host, while only three haemogregarine lineages were identified in E. orbicularis in Tunisia. Here, we screened 215 individuals of E. orbicularis from the Iberian Peninsula, Menorca Island and Morocco for haemogregarine parasites using partial 18S rRNA gene sequences to estimate relationships. Three unrelated lineages of parasites were detected, one presumed H. stepanowi and two lineages previously known from M. leprosa. A considerable undescribed diversity of parasites exists within these vertebrate host species, while mixed infection and host-sharing is also widespread. Considering that E. orbicularis is near threatened in this region, it is of great importance to identify the parasites infecting it, and to further assess the potential deleterious impact of these diverse parasites on their hosts.

Discordant phylogeographic patterns between the tortoise tick Hyalomma aegyptium and their Testudo graeca hosts.

Tortoise ticks, Hyalomma aegyptium, are considered so strongly associated with their hosts that they are even used as indirect indicators for them. In such a case, a robust pattern of congruence between host and parasite could be expected, with phylogeographic breaks within the host being reflected in their parasites. We sequenced two mitochondrial partial gene regions (12S rRNA and Cytochrome Oxidase 1) from ticks across northern Africa and Anatolia, and compared patterns of variation with those identified in its main host, Testudo graeca. Two distinct haplogroups were identified, both of which were found distributed across much of northern Africa. This pattern does not reflect the known variation within the host, which has multiple, geographically disjunct subspecies in this region, but rather the major climatic zones. This relationship can be explained by adaptive processes to environmental conditions influenced by the climate, as well as by the spatial structure of the communities of tick potential hosts in larval and nymphal stages. Extensive anthropogenic movement of tortoises may also obscure congruence patterns between H. aegyptium ticks and their hosts.

Diversity of microorganisms in Hyalomma aegyptium collected from spur-thighed tortoise (Testudo graeca) in North Africa and Anatolia.

Ticks carry a diverse community of microorganisms including non-pathogenic symbionts, commensals, and pathogens, such as viruses, bacteria, protozoans, and fungi. The assessment of tick-borne microorganisms (TBM) in tortoises and their ticks is essential to understand their eco-epidemiology, and to map and monitor potential pathogens to humans and other animals. The aim of this study was to characterize the diversity of microorganisms found in ticks collected from the spur-thighed tortoise (Testudo graeca) in North Africa and Anatolia. Ticks feeding on wild T. graeca were collected, and pathogens were screened by polymerase chain reaction using group-specific primers. In total, 131 adult Hyalomma aegyptium ticks were collected from 92 T. graeca in Morocco (n = 48), Tunisia (n = 2), Algeria (n = 70), and Turkey (n = 11). Bacteria and protozoa detected included Hemolivia mauritanica (22.9%), Midichloria mitochondrii (11.4%), relapsing-fever borreliae (8.4%), Ehrlichia spp. (7.6%), Rickettsia spp. (3.4%), Borrelia burgdorferi s.l. (0.9%), Francisella spp. (0.9%), and Wolbachia spp. (0.8%). The characterization of Rickettsia included R. sibirica mongolitimonae (Algeria), R. aeschlimannii (Turkey), and R.africae (Morocco). Hemolivia mauritanica and Ehrlichia spp. prevalence varied significantly with the sampling region/country. We did not detect significant associations in microorganism presence within ticks, nor between microorganism presence and tick mitochondrial DNA haplogroups. This is the first report of Francisella persica-like, relapsing fever borreliae, M. mitochondrii, and Wolbachia spp. in H. aegyptium ticks collected from wild hosts from the South and Eastern Mediterranean region, and of R. sibirica mongolitimonae and R. africae in H. aegyptium from Algeria and Morocco, respectively. Given that T. graeca is a common species in commercial and non-commercial pet trade, the evaluation of the role of this species and its ticks as hosts for TBM is particularly relevant for public health.

Helminth fauna and histopathology associated with parasitic infections in Phrynops geoffroanus (Schweigger, 1812) (Testudines, Chelidae) in a Brazilian river subjected to anthropogenic activities.

The objective of this study was to determine the structure of the helminth fauna and identify the macroscopic and histopathological alterations associated with parasitic infections in Phrynops geoffroanus. Freshwater turtles of both sexes were captured during the dry and rainy seasons in four municipalities along the Capibaribe River. The study included 63 animals, of which 79.37% (50/63) were parasitized by one or more helminths. In total, 933 helminths of seven taxa were recovered: Serpinema monospiculatus, Spiroxys figueiredoi, Nematophila grandis, Polystomoides brasiliensis, Cheloniodiplostomum testudinis, Telorchis birabeni, and Prionosomoides scalaris. Monogeneans and digenetic trematodes were more sensitive to environmental pressures, since the prevalences varied significantly between areas. Nematodes proved to be more resistant to environmental pressure and caused severe injuries to their hosts: nodules in the stomach and small intestine, adhesions in the liver capsule, and pulmonary emphysema. Parasitic granulomas were recorded at the infection sites and in the lungs and liver, the latter caused by migration of S. figueiredoi larvae. This is the first record of P. brasiliensis, N. grandis, C. testudinis, and T. birabeni parasitizing P. geoffroanus in the state of Pernambuco. Histopathology proved to be an important tool for studies on the impact of parasites at the individual, population, and ecosystem levels. Considering the use of the Capibaribe River for public water supply, fishing, and other activities, within the One Health perspective, this study demonstrates that the anthropogenic impact affects parasites and their hosts, in addition to the human population that uses this ecosystem.

Putative intranuclear coccidium in Mauremys leprosa (Schweigger) from Morocco.

Although intranuclear coccidiosis was first identified in chelonians less than 30 years ago, it is now considered an important emerging disease. Symptoms include anorexia, weakness and weight loss, potentially leading to death of the infected animal. The use of molecular tools has led to improved diagnosis and has also led to an increase in known host species. Here we report a putative intranuclear coccidium in Mauremys leprosa (Schweigger), from Morocco, based on 18S rDNA sequence analysis. This is, to the best of our knowledge, the first report of this parasite from a freshwater terrapin species.

MOLECULAR IDENTIFICATION OF JUVENILE NEOECHINORHYNCHUS SPP. (PHYLUM: ACANTHOCEPHALA) INFECTING OSTRACOD AND SNAIL HOSTS PROVIDES INSIGHT INTO ACANTHOCEPHALAN HOST USE.

The role of invertebrates in some acanthocephalan life cycles is unclear because juvenile acanthocephalans are difficult to identify to species using morphology. Most reports suggest acanthocephalans from turtle definitive hosts use ostracods as intermediate hosts and snails as paratenic hosts. However, laboratory studies of the life cycle suggest that ostracods and snails are both required hosts in the life cycle. To elucidate the role of ostracods and snails in acanthocephalan life cycles better, we collected 558 freshwater snails of 2 species, including Planorbella cf. Planorbella trivolvis and Physa acuta, from 23 wetlands in Oklahoma, U.S.A., and examined them for acanthocephalan infections. Additionally, we examined 37,208 ostracods of 4 species, Physocypria sp. (morphotype 1), Cypridopsis sp., Stenocypris sp., and Physocypria sp. (morphotype 2) for juvenile acanthocephalans from 2 wetlands in Oklahoma. Juvenile acanthocephalans were morphologically characterized, and the complete internal transcribed spacer (ITS) region of nuclear rDNA was sequenced from acanthocephalans infecting 11 ostracod and 13 snail hosts. We also sampled 10 red-eared slider turtles, Trachemys scripta elegans, and 1 common map turtle, Graptemys geographica, collected from Oklahoma, Arkansas, and Texas and recovered 1,854 adult acanthocephalans of 4 species. The ITS of 17 adult acanthocephalans of 4 species from turtle hosts were sequenced and compared to juvenile acanthocephalan sequences from ostracod and snail hosts from this study and GenBank to determine conspecificity. Of the 23 locations sampled for snails, 7 (30%) were positive for juvenile acanthocephalans in the genus Neoechinorhynchus. The overall prevalence and mean intensity of acanthocephalans in Planorbella cf. P. trivolvis and P. acuta were 20% and 2 (1-6) and 2% and 1 (1), respectively. In contrast, only 1 of 4 species of ostracods, Physocypria sp. (morphotype 1), was infected with larval/juvenile Neoechinorhynchus spp. with an overall prevalence of 0.1% and a mean intensity of 1 (1-2). Although 4 species of acanthocephalans infected turtle definitive hosts, including Neoechinorhynchus chrysemydis, Neoechinorhynchus emydis, Neoechinorhynchus emyditoides, and Neoechinorhynchus pseudemydis, all the ITS sequences from cystacanths infecting snail hosts were conspecific with N. emydis. In contrast, the ITS sequences from larval/juvenile acanthocephalans from ostracods were conspecific with 2 species of acanthocephalans from turtles (N. emydis and N. pseudemydis) and 1 species of acanthocephalan from fish (Neoechinorhynchus cylindratus). These results indicate that N. emydis infects freshwater snails, whereas other species of Neoechinorhynchus appear not to infect snail hosts. We document new ostracod and snail hosts for Neoechinorhynchus species, including the first report of an ostracod host for N. pseudemydis, and we provide novel molecular barcodes that can be used to determine larva, juvenile, and adult conspecificity of Neoechinorhynchus species.