The order Testudines comprises some of the most endangered groups of vertebrates. Under specific circumstances, infectious and parasitic diseases may affect the survival and fecundity of these animals, potentially threatening Testudines populations. In Sardinia, besides the three species of tortoises present in the wild (Testudo hermanni, Testudo graeca and Testudo marginata), many others are kept as pets. However, epidemiological studies on these animals have not been conducted so far. Thus, the aim of the work was to investigate the presence of gastrointestinal parasites in captive and wild tortoises of Sardinia, Italy, with particular regard to those of zoonotic importance. For the 215 animals examined (n = 36 wild caught and n = 179 private-owned), fecal samples were collected and processed by flotation and modified Ziehl-Neelsen technique. An overall prevalence of 81.4% for endoparasites was detected, with oxyurids being the most prevalent (74.4%), followed by Nyctotherus spp. (18.6%), Cryptosporidium spp. (12.6%), Angusticaecum spp. (2.8%), strongyles (0.9%), Balantidium spp. (0.9%), coccidia (0.9%), cestodes (0.5%), and Giardia spp. (0.5%). Data suggest that tortoises are affected by a great variety of endoparasites, and further molecular analysis are required to assess the impact of Cryptosporidium and Giardia species in these hosts. Therefore, regular health screenings are of importance for the management of these animals and for preventing emerging infectious diseases.

BACKGROUND: European wildcats (Felis silvestris) are widely distributed in Europe and a strictly protected species in Germany. Lately, anthropogenic protective efforts lead to increasing numbers of wildcats in southwestern Germany. Moreover, in recent years the numbers of domestic cats are increasing. Thus, the contact between domestic and wildcats may lead to the spread of zoonotic pathogens in both animal species. As data on vector-borne pathogens (VBPs) in wildcats from Germany are limited to date, the objective of this study was to investigate the presence and current distribution of VBPs in wildcats from southwestern Germany.
METHODS: Skin and spleen samples from 117 European wildcats, originating from a regional carcass-monitoring program in southwestern Germany, were examined by real-time and conventional polymerase chain reaction (PCR) for the presence of Anaplasma phagocytophilum, Neoehrlichia mikurensis, Rickettsia spp., Bartonella spp., and Piroplasmida.
RESULTS: In total, 6.8% (n = 8) of the wildcats were Rickettsia-positive, specified as R. helvetica. Three wildcats were positive for A. phagocytophilum (2.6%), one for Bartonella spp., namely B. taylorii (0.8%), and 84 for Cytauxzoon spp. (71.8%). Out of these 84 samples, 23 were further sequenced revealing very high identity levels (99.84-100%) to C. europaeus, which is considered to be pathogenic for domestic cats. All wildcats were negative for the presence of N. mikurensis DNA.
CONCLUSIONS: European wildcats in southwestern Germany are hosting several VBPs. With the exception of Cytauxzoon spp., low prevalence rates of most examined pathogens suggest that wildcats are primarily incidental hosts for sylvatic pathogens associated with rodents, in contrast to domestic cats. However, the high prevalence of the cat-associated pathogen C. europaeus suggests that wildcats in southwestern Germany may serve as reservoirs for this pathogen.

Males and females often differ in ecology, behaviour and lifestyle, and these differences are expected to lead to sex differences in parasite susceptibility. However, neither the sex differences in parasite prevalence, nor their ecological and evolutionary drivers have been investigated across a broad range of taxa using phylogenetically corrected analyses. Using the most extensive dataset yet that includes 755 prevalence estimates from 151 wild bird species in a meta-analytic framework, here we compare sex differences in blood and gastrointestinal parasites. We show that despite sex differences in parasite infection being frequently reported in the literature, only Haemoproteus infections were more prevalent in females than in males. Notably, only seasonality was strongly associated with the sex-specific parasite prevalence of both Leucocytozoon and Haemoproteus, where birds showed greater female bias in prevalence during breeding periods compared to the non-breeding period. No other ecological or sexual selection variables were associated with sex-specific prevalence of parasite prevalence. We suggest that much of the variation in sex-biased prevalence could be idiosyncratic, and driven by local ecology and behavioural differences of the parasite and the host. Therefore, breeding ecology and sexual selection may only have a modest influence on sex-different parasite prevalence across wild birds.

Cytauxzoon spp. have been detected in Brazil infecting mainly asymptomatic domestic cats and wild felids. However, the supposed genetic similarity with the pathogenic Cytauxzoon felis is questionable because it is based on analysis of short sequences of the 18S rRNA gene. Herein, we describe a novel Cytauxzoon species infecting an asymptomatic little-spotted-cat (Leopardus tigrinus) based on morphological, histopathological, and molecular analyses. The animal was attended presenting a history of a run-over with multiple traumas. Although the little-spotted-cat was stabilized, he died a few days later. Ring-shaped merozoites within erythrocytes were found on blood smears and in the abdominal effusion. In addition, schizonts were observed in histiocytes in the liver. Phylogenetic analyses based on both near-complete 18S rRNA and cytb genes positioned the obtained sequences in a unique clade, albeit closely related to Cytauxzoon felis from the USA. Genetic divergences ranging from 0.004 and 0.067-0.068 were found between the near-complete 18S rRNA and cytb sequences of Cytauxzoon sp. detected in the little-spotted-cat and C. felis, respectively. This study evidenced the circulation of a novel Cytauxzoon species, herein named Cytauxzoon brasiliensis sp. nov., in an asymptomatic wild felid species from Brazil. Further studies are necessary to identify Cytauxzoon species from domestic and wild felids in the country.

Tritrichomonas foetus is a flagellated and anaerobic parasite able to infect cattle and felines. Despite its prevalence, there is no effective standardized or legal treatment for T. foetus-infected cattle; the vaccination still has limited success in mitigating infections and reducing abortion risk; and nowadays, the diagnosis of T. foetus presents important limitations in terms of sensitivity and specificity in bovines. Here, we characterize the plasma membrane proteome of T. foetus and identify proteins that are represented in different isolates of this protozoan. Additionally, we performed a bioinformatic analysis that revealed the antigenicity potential of some of those proteins. This analysis is the first study to identify common proteins at the plasma membrane of different T. foetus isolates that could be targets for alternative diagnostic or vaccine techniques in the future.

Histomoniasis, caused by the protozoan, Histomonas meleagridis, is an economically important disease of turkeys, and it also affects several other species of domesticated and wild Galliformes, including chickens. Under natural conditions, the parasite is transmitted through eggs of a nematode, Heterakis gallinarum, that shares its hosts with Hi. meleagridis. The protozoan infects tissues of both male and female He. gallinarum and eventually is carried within the worm egg. Histomonas meleagridis more readily infects and develops in chickens, and the proximity of chicken farms is a major risk factor for outbreaks in turkeys. Chemoprophylaxis had controlled Hi. meleagridis in turkeys very successfully, but histomoniasis has recently reemerged in turkeys because anti-histomonal drugs are no longer permitted by the United States Food and Drug Administration because of the concerns for residual toxins in poultry meat. Horizontal transmission of the protozoan in the absence of worm eggs remains a mystery because the flagellate trophozoite excreted in the feces of turkeys is not viable for any length of time. A proposed resistant stage of the protozoan has not yet been conclusively demonstrated. Here we review the discovery of the protozoan and the current status of the disease and its control.

BACKGROUND: Trichomonosis is a common infection in small animals, mostly manifesting in gastrointestinal symptoms such as diarrhea. Although oral trichomonads are also known, the species found colonizing the large intestine are more frequently detected protozoa.
METHODS: In the present study, four wildcats, 94 domestic cats, and 25 dogs, originating from 18 different locations in Hungary, were investigated for the presence of oral and large intestinal trichomonads based on the 18S rRNA gene and ITS2.
RESULTS: All oral swabs were negative by polymerase chain reaction (PCR). However, Tritrichomonas foetus was detected in a high proportion among tested domestic cats (13.8%) and dogs (16%), and Pentatrichomonas hominis only in two domestic cats. In addition, a novel Tritrichomonas genotype was identified in one cat, probably representing a new species that was shown to be phylogenetically most closely related to Tritrichomonas casperi described recently from mice. All positive dogs and half of the positive cats showed symptoms, and among cats, the most frequent breed was the Ragdoll.
CONCLUSIONS: With molecular methods, this study evaluated the prevalence of oral and intestinal trichomonads in clinical samples of dogs and cats from Hungary, providing the first evidence of T. foetus in dogs of this region. In contrast to literature data, P. hominis was more prevalent in cats than in dogs. Finally, a hitherto unknown large intestinal Tritrichomonas species (closely related to T. casperi) was shown to be present in a cat, raising two possibilities. First, this novel genotype might have been a rodent-associated pseudoparasite in the relevant cat. Otherwise, the cat was actually infected, thus suggesting the role of a predator-prey link in the evolution of this trichomonad.

Avian haemosporidians of the genera Plasmodium and Haemoproteus are a group of widely distributed blood parasites that can negatively affect the fitness of their hosts. Colombia contains the greatest diversity of birds on the planet, but knowledge about the associations between haemosporidian and its avifauna is scarce and fragmented. We collected blood samples from 255 birds (203 residents and 52 neotropical migrants) belonging to 27 families and 108 species. The study was conducted in six localities in the inter-Andean valleys of the Cauca and Magdalena rivers. Parasites of the genera Plasmodium and Haemoproteus were identified in the samples by morphological and molecular analysis of a fragment of the mitochondrial gene cyt b. Among the samples, 9.3% (n = 24) were positive for Plasmodium or Haemoproteus. Co-infection with Plasmodium and Haemoproteus was found in Red-eyed Vireo. Seventeen haemosporidian lineages were identified, five of which were reported for the first time in resident birds (Common Ground Dove, Checker-throated Stipplethroat, Tropical Kingbird, Pale-breasted Thrush, and Ruddy-breasted Seedeater) and one in the Summer Tanager (neotropical migrant). The research results confirm the wide diversity of haemosporidian present in tropical lowlands and the possible role of neotropical migratory birds in dissemination on haemosporidian along their migratory routes.

BACKGROUND: In recent years, the trichomonosis in raccoon dogs in China had occurred frequently. Pentatrichomonas hominis had been described in raccoon dogs in China in some previous studies. PURPOSE TO REVEAL: whether raccoon dogs can be infected by other trichomonad species besides P. hominis, and clarify the prevalence and species distribution of trichomonad in raccoon dogs.
METHODS: Herein, the 389 fecal samples were collected from farm-raised raccoon dogs in Hebei Province, all the samples were detected using the microscopic examination and several fecal samples containing trichomonad-like organisms were processed, cultured, stained, and photographed. Meanwhile, all the samples were screened by the species-specific nested PCR based on the small subunit rRNA (SSU rRNA) gene of P. hominis,Tritrichomonas foetus and Tetratrichomonas buttreyi, respectively, and all positive secondary PCR amplications obtained in this study were sequenced, aligned and analysed.
RESULTS: 62 fecal samples (15.9%,62/389) were trichomonad-positive under light microscopy, and the trichomonad-like cells were clearly observed in the culture contents. The PCR results showed that 100 samples were trichomonad-positive, including 45 P. hominis-positive samples (11.6%,45/389), 32 T. foetus-positive samples (8.2%,32/389), and 33 T. buttreyi-positive samples (8.5%,33/389), respectively. Double mixed infections were observed in 10 samples. The prevalence of T. foetus and P. hominis were both significantly higher in raccoon dogs with diarrhea (13.9%, and 25.0%) than that in raccoon dogs without diarrhea (7.6%, and 9.3%) (p < 0.05).All samples confirmed as trichomonad-positive under microscopy were also found to be trichomonad-positive by PCR analysis. The sequencing and phylogenetic analysis demonstrated the sequences obtained in this study belonged to P. hominis, T. foetus and T. buttreyi SSU rRNA, respectively. Among them, the T. buttreyi SSU rRNA sequences obtained in this study harbored the new sequence polymorphisms. Based on preliminary morphological and molecular analyses, raccoon dogs are considered as the new host of T. foetus and T. buttreyi.
CONCLUSIONS: This is the first report about the identifcation and prevalence of T. foetus and T. buttreyi in raccoon dogs in China, and the results increase our knowledge about the host range and prevalence of trichomonad species.

Forest regeneration is becoming a powerful tool to combat land conversion which covers 30 % of the Neotropical territory. However, little is known about the effect of forest regeneration on vector-borne diseases. Here, we describe the haemosporidian lineage composition across a successional gradient within an Atlantic Forest bird community. We test whether forest successional stages, in addition to host life history traits affect haemosporidian infection probability. We sampled birds at 16 sampling units with different successional stages between 2017 and 2018 within a forest remnant located in Antonina, Paraná, Brazil. We captured bird individuals using mist-nets, identified them to the species level, and collected blood samples to detect and identify Plasmodium and Haemoproteus lineages based on molecular analysis. We used a Bayesian phylogenetic linear model with a Bernoulli distribution to test whether the haemosporidian infection probability is affected by nest type, foraging stratum, and forest successional stage. We captured 322 bird individuals belonging to 52 species and 21 families. We found 31 parasite lineages and an overall haemosporidian prevalence of 23.9 %, with most infections being caused by Plasmodium (21.7 % of prevalence). The Plasmodium probability of infection was associated with forest successional stage and bird foraging stratum. Birds from the secondary forest in an intermediate stage of succession are more likely to be infected by the parasites than birds from the primary forests (β = 1.21, 95 % CI = 0.11 - 2.43), birds from upper strata exhibit a lower probability of infection than birds from lower foraging strata (β = -1.81, 95 % CI = -3.80 - -0.08). Nest type did not affect the Plasmodium probability of infection. Our results highlight the relevance of forest succession on haemosporidian infection dynamics, which is particularly relevant in a world where natural regeneration is the main tool used in forest restoration.