Island hitchhikers: pathogen agents of Madeira and Azores ticks.

Ticks are blood-sucking arthropods that can transmit pathogens to their host. As insular ecosystems can enhance tick-host interactions, this study aimed to understand tick diversity, pathogen presence, and their respective associations in the Azores and Madeira archipelagos. Unfed or partially engorged ticks (n = 120) were collected from 58 cats and dogs in the Azores (n = 41 specimens) and Madeira (n = 79 specimens) from November 2018 to March 2019. Vector identification was based on morphology and molecular criteria. For pathogen sequencing, 18S gene fragment for Babesia/Hepatozoon and gltA for Rickettsia were performed. Sequence data was explored using BLAST and BLAST and phylogenetic inference tools. In the Azores, Ixodes hexagonus, I. ventalloi, and Rhipicephalus sanguineus (n = 6; 14.6%, n = 6; 14.6%, and n = 29; 70.7% respectively) were found and in Madeira I. ricinus and R. sanguineus (n = 78, 98.7%; and n = 1, 1.3%; respectively) were identified. Tick COI markers confirmed species highlighting confirmation of R. sanguineus s.s. and genotype A of I. ventalloi. In the Azores Islands, the detected Rickettsia massiliae was linked to R. sanguineus (dogs and cats) and I. hexagonus (dogs), and in Madeira Island, R. monacensis (dogs) and Hepatozoon silvestris (cats) were found associated with I. ricinus. Further, I. ventalloi presence in the Azores expands west its known range, and Hepatozoon silvestris in Madeira may suggest that I. ricinus could have a role as a potential vector. Finally, as R. massiliae and R. monacensis presence underlines public health risks, surveillance by health authorities is crucial as pathogen-tick interactions may drive disease spread, therefore monitoring remains pivotal for disease prevention.

Is there only one species of Hepatozoon infecting Brazilian caimans? Integrative taxonomy unveiling the parasite's diversity.

Hepatozoon spp. are the most common haemoparasites reported from reptiles around the world, however, only six species have been described infecting crocodilians. In Brazil, Hepatozoon caimani Carini, 1909 is currently the only recognized species from the caiman hosts. This study provides new data on the diversity of species of Hepatozoon infecting Caiman crocodilus (Linnaeus) using molecular data and phylogenetic analysis, with additional support of morphological data of developmental stages from host blood and tissue. Forty-four individuals were collected and screened for haemogregarines, and blood and tissue samples were analysed by light microscopy with 31 (70.45%) infected. Hepatozoon spp. blood developmental stages included immature and mature gamonts with or without cytoplasmic vacuoles and free gamonts. Additionally, merogonic developmental stages were found in the liver and spleen of infected hosts. Based on the morphological and molecular data, this study identified two possible different species of Hepatozoon, being one of them the H. caimani with intragenotypic divergence.

Genetic diversity of vector-borne pathogens in spotted and brown hyenas from Namibia and Tanzania relates to ecological conditions rather than host taxonomy.

BACKGROUND: Improved knowledge on vector-borne pathogens in wildlife will help determine their effect on host species at the population and individual level and whether these are affected by anthropogenic factors such as global climate change and landscape changes. Here, samples from brown hyenas (Parahyaena brunnea) from Namibia (BHNA) and spotted hyenas (Crocuta crocuta) from Namibia (SHNA) and Tanzania (SHTZ) were screened for vector-borne pathogens to assess the frequency and genetic diversity of pathogens and the effect of ecological conditions and host taxonomy on this diversity. METHODS: Tissue samples from BHNA (n = 17), SHNA (n = 19) and SHTZ (n = 25) were analysed by PCRs targeting Anaplasmataceae, Rickettsia spp., piroplasms, specifically Babesia lengau-like piroplasms, Hepatozoidae and filarioids. After sequencing, maximum-likelihood phylogenetic analyses were conducted. RESULTS: The relative frequency of Anaplasmataceae was significantly higher in BHNA (82.4%) and SHNA (100.0%) than in SHTZ (32.0%). Only Anaplasma phagocytophilum/platys-like and Anaplasma bovis-like sequences were detected. Rickettsia raoultii was found in one BHNA and three SHTZ. This is the first report of R. raoultii from sub-Saharan Africa. Babesia lengau-like piroplasms were found in 70.6% of BHNA, 88.9% of SHNA and 32.0% of SHTZ, showing higher sequence diversity than B. lengau from South African cheetahs (Acinonyx jubatus). In one SHTZ, a Babesia vogeli-like sequence was identified. Hepatozoon felis-like parasites were identified in 64.7% of BHNA, 36.8% of SHNA and 44.0% of SHTZ. Phylogenetic analysis placed the sequences outside the major H. felis cluster originating from wild and domestic felids. Filarioids were detected in 47.1% of BHNA, 47.4% of SHNA and 36.0% of SHTZ. Phylogenetic analysis revealed high genetic diversity and suggested the presence of several undescribed species. Co-infections were frequently detected in SHNA and BHNA (BHNA median 3 pathogens, range 1-4; SHNA median 3 pathogens, range 2-4) and significantly rarer in SHTZ (median 1, range 0-4, 9 individuals uninfected). CONCLUSIONS: The frequencies of all pathogens groups were high, and except for Rickettsia, multiple species and genotypes were identified for each pathogen group. Ecological conditions explained pathogen identity and diversity better than host taxonomy.

Investigating the Prevalence of Intestinal Parasites in Immunocompromised Patients in Bushehr Province, Southwest Iran: A Conventional and Molecular Study

Objective: The aim of this study was to determine the status of intestinal parasitic infections in immunocompromised patients in Bushehr province, southwest Iran by conventional and molecular methods. Methods: A total of 201 stool samples were collected from kidney transplant recipients, AIDS patients and patients under chemotherapy. Samples were collected from healthy people as the control group. The specimens were tested using various conventional methods. Polymerase chain reaction (PCR) testing was performed on samples identified as positive for Coccidia by direct microscopic examination. Results: Approximately 32.45% were infected with at least one type of intestinal parasite. The highest (46.8%) and lowest rates of infection (24%) were observed in AIDS and chemotherapy patients, respectively, while the infection rate of the control group was 16%. Isospora spp. and Cryptosporidium spp. were observed in all patient groups, and Sarcocystis spp. sporocysts were detected in one of the transplant recipients. All identified coccidia were confirmed by PCR. There was a significant relationship between the rate of intestinal parasite infection and certain variables. Conclusion: Given the potential risk of certain intestinal parasites in people with immune deficiency, it is recommended that diagnosis of parasitic infections in such patients be based on specific parasitological methods. Thus, it is advisable that physicians refer them to a parasitology laboratory prior to drug administration.

Phenological and intrinsic predictors of mite and haemacoccidian infection dynamics in a Mediterranean community of lizards.

Ectotherms are vulnerable to environmental changes and their parasites are biological health indicators. Thus, parasite load in ectotherms is expected to show a marked phenology. This study investigates temporal host­parasite dynamics in a lizard community in Eastern Spain during an entire annual activity period. The hosts investigated were Acanthodactylus erythrurus, Psammodromus algirus and Psammodromus edwardsianus, three lizard species coexisting in a mixed habitat of forests and dunes, providing a range of body sizes, ecological requirements and life history traits. Habitat and climate were considered as potential environmental predictors of parasite abundance, while size, body condition and sex as intrinsic predictors. Linear models based on robust estimates were fitted to analyse parasite abundance and prevalence. Ectoparasitic mites and blood parasites from two haemococcidian genera were found: Lankesterella spp. and Schellackia spp. Habitat type was the only predictor explaining the abundance of all parasites, being mostly higher in the forest than in the dunes. The results suggest that particularities in each host­parasite relationship should be accounted even when parasites infect close-related hosts under the same environmental pressures. They also support that lizard parasites can be biomarkers of environmental perturbation, but the relationships need to be carefully interpreted for each host­parasite assemblage.

Prevalence of coccidia in domestic pigs in China between 1980 and 2019: a systematic review and meta-analysis.

BACKGROUND: Swine coccidiosis, a protozoan disease caused by coccidia, can result in diarrhoea and weight loss in piglets and even economic losses in the pig industry. Here, we report the first systematic review and meta-analysis of the prevalence of coccidia (including Eimeria spp. and Cystoisospora suis) in pigs in China. METHODS: Five databases (PubMed, ScienceDirect, Chinese Web of Knowledge, Wanfang, and Chongqing VIP) were searched and 50 studies (46,926 domestic pigs, 22 provinces) ultimately identified pertaining to the prevalence of coccidia infection from 1980 to 2019. We incorporated the effect size using the random-effects model in the "meta" package in R software and conducted univariate and multivariate meta-regression analyses using a mixed-effects model. RESULTS: The pooled prevalence rate of coccidia in pigs was 21.9%, including the C. suis infection rate of 9.1%. The highest prevalence of coccidia (39.6%) was found in northwest China, and this region also presented the lowest prevalence of C. suis (4.7%). In the subgroup analysis based on sampling year, the highest prevalence of coccidia was detected in 2001 or earlier (32.6%), whereas the lowest rate was found in 2012 or later (14.3%). An opposite trend was observed for C. suis (5.5% in 2000 or earlier vs 14.4% in 2000 or later). The prevalence of coccidia in extensive farming systems (29.5%) was higher than that in intensive farming systems (17.3%). In contrast, the point estimate of C. suis prevalence was lower in the extensive farming systems (5.1%) than in the intensive farming systems (10.0%), but the difference was not significant (P > 0.05). Among the four age categories, the highest total coccidia prevalence (26.2%) was found in finishing pigs, followed by suckling piglets (19.9%), whereas the highest prevalence of C. suis (14.9%) was observed in suckling piglets. CONCLUSIONS: Our findings suggest that coccidia infection in Chinese pigs is common, although the prevalence of C. suis in pigs does not receive sufficient attention. We recommend the rational use of anticoccidial drugs to avoid drug resistance and the development of preventive and control measures for C. suis to reduce the incidence of swine coccidiosis.

Cytochrome c oxydase I phylogenetic analysis of Haemogregarina parasites (Apicomplexa, Coccidia, Eucoccidiorida, Haemogregarinidae) confirms the presence of three distinct species within the freshwater turtles of Tunisia.

Species of Haemogregarina are apicomplexan blood parasites that use vertebrates as intermediate hosts. Due to limited interspecific morphological characters within the genus during the last decade, 18S rRNA gene sequences were widely used for species identification. As coinfection patterns were recently reported from nuclear molecular data for two sympatric freshwater turtles Mauremys leprosa and Emys orbicularis from Tunisia, our objectives were to design COI specific primers to confirm the presence of three distinct species in both host species. Blood samples were collected from 22 turtles, from which DNAs were extracted and used as templates for amplification. Following different rounds of PCR and nested PCR, we designed specific Haemogregarina COI primers that allowed the sequencing of nine distinct haplotypes. Phylogenetic Bayesian analysis revealed the occurrence of three well-differentiated sublineages that clustered together into a single clade. Based on pairwise genetic distances (p-distance), we confirmed the occurrence of three distinct but phylogenetically closely related species coinfecting M. leprosa and E. orbicularis in the same aquatic environments. Our results demonstrate that the use of fast evolving genes within Haemogregarina will help to investigate the parasite diversity within both intermediate vertebrate and definitive invertebrate hosts, and to assess the evolution, historical biogeography and specificity of haemogregarines.

Molecular screening indicates high prevalence and mixed infections of Hepatozoon parasites in wild felines from South Africa.

Genetic diversity within partial 18S rRNA sequences from Hepatozoon protozoan parasites from wild felines in South Africa was assessed and compared with data from domestic cats to assess patterns of host specificity. Lions, leopards, servals, a caracal and an African wildcat were all positive for parasites of the Hepatozoon felis-complex. However, haplotypes were not species-specific, and potential mixed infections were widespread. Additional genetic markers are needed to untangle the extremely complex situation of these parasites in both domestic cats and wild felines in South Africa.

A new species, Dactylosoma piperis n. sp. (Apicomplexa, Dactylosomatidae), from the pepper frog Leptodactylus labyrinthicus (Anura, Leptodactylidae) from Mato Grosso State, Brazil.

The Dactylosomatidae Jakowska and Negrelli, 1955 are one of four families belonging to adeleorinid coccidia and comprise the genera Babesiosoma Jakowska and Nigrelli, 1956 and Dactylosoma Labbé, 1894. These blood protozoa occur in peripheral blood of lower vertebrates, and are commonly reported parasitising amphibians. The present study describes Dactylosoma piperis n. sp. from the pepper frog Leptodactylus labyrinthicus (Spix, 1824) (Anura: Leptodactylidae), collected in 2018 at the municipality of Araguaiana, Mato Grosso State, Brazil, based on morphology of intra-erythrocytic trophozoite, primary and secondary merogonic stages and a molecular analysis (partial 18S rDNA). Dactylosoma piperis n. sp. forms a well-supported clade with other Dactylosomatidae. This is the first molecular characterization of a species of Dactylosoma from a Brazilian anuran.

TITLE: Une nouvelle espèce, Dactylosoma piperis n. sp. (Apicomplexa, Dactylosomatidae), parasite de la grenouille Leptodactylus labyrinthicus (Anura, Leptodactylidae) de l'état du Mato Grosso, Brésil. ABSTRACT: Les Dactylosomatidae Jakowska et Negrelli, 1955 sont l'une des quatre familles appartenant aux coccidies Adeleorina et comprennent les genres Babesiosoma Jakowska et Nigrelli, 1956 et Dactylosoma Labbé, 1894. Ces protozoaires sanguins se trouvent dans le sang périphérique des vertébrés inférieurs et sont fréquemment signalés comme parasitant des amphibiens. Ce travail décrit Dactylosoma piperis n. sp. de la grenouille Leptodactylus labyrinthicus (Spix, 1824) (Anura : Leptodactylidae), collectée en 2018 dans la municipalité d'Araguaiana, État du Mato Grosso, Brésil, d'après la morphologie du trophozoïte intra-érythrocytaire, des stades mérogoniques primaires et secondaires et une analyse moléculaire (ADNr 18S partiel). Dactylosoma piperis n. sp. forme un clade bien soutenu avec d'autres Dactylosomatidae. Il s'agit de la première caractérisation moléculaire d'une espèce de Dactylosoma à partir d'un anoure brésilien.

Molecular detection of Apicomplexan hemoparasites in anurans from Brazil.

Amphibians are among the most threatened vertebrate groups in the world, and the main causes include climate change, habitat destruction, and emerging diseases. Herein, we investigated the occurrence and characterized molecularly Apicomplexa in anurans from southeastern Brazil. Forty individuals from seven anuran species were sampled in São Paulo state. In the molecular analyses, one Leptodactylus latrans and one Rhinella diptycha were positive in PCR assays for species of Hepatozoon. Two L. latrans were also positive for coccidian infections (Lankesterella sp. and an unidentified coccidian species). Phylogenetic analysis based on 18S rDNA clustered the sequences detected in anurans from the present study with Hepatozoon spp. detected in reptiles and other anurans from Brazil, albeit they were separate from Hepatozoon haplotypes detected in frogs from Africa and North America. Our study showed, for the first time, the molecular detection of Lankesterella sp. and another coccidian in L. latrans. Additionally, co-infection by different species of Hepatozoon haplotypes and an unidentified coccidian in anurans from Brazil was documented.

Molecular detection of tick-borne pathogens in caracals (Caracal caracal) living in human-modified landscapes of South Africa.

BACKGROUND: Wild carnivores living alongside humans and domestic animals are vulnerable to changes in the infectious disease dynamics in their populations. The aims of this study were to determine the prevalence and diversity of selected tick-borne pathogens (TBPs) of veterinary and/or zoonotic concern in wild populations of caracals (Caracal caracal) occurring in human-modified landscapes in South Africa. Using molecular techniques, we screened 57 caracal blood samples for infection by rickettsial bacteria and piroplasms in three regions of South Africa: rangeland in the Central Karoo (n = 27) and Namaqualand (n = 14) as well as the urban edge of the Cape Peninsula (n = 16) of South Africa. To characterise pathogen identity, we sequenced the 18S rRNA and 16S rRNA genes from positive samples and analysed sequences within a phylogenetic framework. We also examine the diversity of potential tick vectors. RESULTS: All individuals tested were infected with at least one tick-borne pathogen. Pathogens included Hepatozoon felis, Babesia felis, Babesia leo and a potentially novel Babesia species. An Anaplasma species previously described in South African domestic dogs was also found in 88% of urban edge caracals. Higher rates of co-infection characterised urban edge caracals (81% vs 15% and 0% in the two rangeland populations), as well as a greater incidence of mixed infections. Host attached tick species include Haemaphysalis elliptica, an important pathogen vector among carnivore hosts. CONCLUSIONS: This study confirms the occurrence of previously undocumented tick-borne pathogens infecting free-ranging caracals in human-modified landscapes. We identify clear differences in the pathogen profiles among our study populations and discuss the likely health costs to caracals living adjacent to urban areas.

First report of Davaneidae and Strongylida parasitizing Ramphocelus carbo (Aves: Passeriformes: Thraupidae) in the southwest of the Brazilian Amazon region.

In Brazil, species of the genus Ramphocelus with the presence of various endoparasites have already been reported. Coccidia have been the parasites most frequently found. All species of this genus have similar habitats and ecological niches, which makes parasite transmission easy. The aim of this study was to diagnose the presence of endoparasites in fecal samples from specimens of Ramphocelus carbo that were caught in the Cazumbá-Iracema Extractive Reserve (Cazumbá Resex), in the State of Acre. The specimens (n = 30) were caught in mist nets arranged in different ecosystems of the Reserve. After identification, the bird specimens were placed in cloth bags for a maximum of 30 minutes to collect feces. Among the 22 samples collected, 63.63% (n = 14) were positive for endoparasites. The coccidia were the parasites most frequently. Helminths belonging to Ascaridiidae (Nematoda), Strongylida (Nematoda) and Davaneidae (Cestoda) were recorded for the first time in R. carbo in the State of Acre. Parasites belonging to Strongylida and Davaneidae were recorded for the first time in a species of Ramphocelus. These findings add information on the parasitic fauna of wild birds, since in the Amazon region there are few studies on this subject.

The Coccidia (Apicomplexa) of the Archosauria (Crocodylia: Eusuchia) of the World.

The order Crocodylia (suborder Eusuchia) includes 27 species of alligators, caimans, crocodiles, and gharials that are cosmopolitan in distribution, inhabiting subtropical and tropical locations. Numerous surveys (many of them trivial, with small sample sizes) have reported a wide variety of blood and intestinal apicomplexans from 17/27 (63%) crocodilian hosts, but neither a summation for the group nor a revisionary systematic approach to species evaluation has ever been provided. Herein, we summarize information on the 16 species of apicomplexans that we consider to be valid, including 8 Eimeria, 1 Haemogregarina (that eventually may be transferred to Hepatozoon), 4 Hepatozoon, 2 Isospora, and 1 Progarnia species. In addition, there are 46 apicomplexan forms that we have relegated to species inquirendae and/or only partially identified forms that await further study. We hope this review provides a foundation for future research between parasitologists and herpetologists on parasitism of all reptiles and their relatives. Since many apicomplexans seem to be reasonably host-specific, knowledge of shared species and/or genera may assist herpetologists to better understand the phylogenetic relationships among the New World crocodilians and the direction of their dispersal in the New World.

Coccidian parasites from birds at rehabilitation centers in Portugal, with notes on Avispora bubonis in Old World.

Portugal has some rehabilitation centers for wild animals, which are responsible for the rehabilitation and reintroduction of birds, among other animals, into the wild. Coccidian parasites of these wild birds in rehabilitation centers are especially important because these centers can introduce coccidian species into new environments through the reintroduction of their respective hosts. In this context, the current study aimed to identify intestinal coccidia from wild birds at two rehabilitation centers for wild animals located in two municipalities of Portugal. Eighty-nine wild birds of 9 orders and 11 families were sampled, of which 22 (25%) were positive for Coccidia. Avispora spp. were found in raptors. Sporocysts of Sarcocystinae subfamily were recovered from owls. An Isospora sp. was found in Turdus merula Linnaeus, 1758, and an Eimeria sp. was found in Fulica atra Linnaeus, 1758. Among the coccidian species, Avispora bubonis (Cawthorn, Stockdale, 1981) can be highlighted. The finding of this species indicates that transmission of coccidians from the New World to the Old World may be occurring, potentially through dispersion by Bubo scandiacus (Linnaeus, 1758) through Arctic regions or by means of anthropic activities, and/or through other unknown mechanisms.

Coccidian parasites from birds at rehabilitation centers in Portugal, with notes on Avispora bubonis in Old World / Coccídios parasitas de aves em centros de reabilitação em Portugal, com notas sobre Avispora bubonis no Velho Mundo

Abstract Portugal has some rehabilitation centers for wild animals, which are responsible for the rehabilitation and reintroduction of birds, among other animals, into the wild. Coccidian parasites of these wild birds in rehabilitation centers are especially important because these centers can introduce coccidian species into new environments through the reintroduction of their respective hosts. In this context, the current study aimed to identify intestinal coccidia from wild birds at two rehabilitation centers for wild animals located in two municipalities of Portugal. Eighty-nine wild birds of 9 orders and 11 families were sampled, of which 22 (25%) were positive for Coccidia. Avispora spp. were found in raptors. Sporocysts of Sarcocystinae subfamily were recovered from owls. An Isospora sp. was found in Turdus merula Linnaeus, 1758, and an Eimeria sp. was found in Fulica atra Linnaeus, 1758. Among the coccidian species, Avispora bubonis (Cawthorn, Stockdale, 1981) can be highlighted. The finding of this species indicates that transmission of coccidians from the New World to the Old World may be occurring, potentially through dispersion by Bubo scandiacus (Linnaeus, 1758) through Arctic regions or by means of anthropic activities, and/or through other unknown mechanisms.

Resumo Portugal possui alguns centros de reabilitação de animais silvestres, responsáveis ​​pela reabilitação e reintrodução de aves, entre outros animais, na natureza. Os coccídios parasitas dessas aves silvestres em centros de reabilitação são especialmente importantes porque esses centros podem introduzir espécies de coccídios em novos ambientes através da reintrodução de seus respectivos hospedeiros. Neste contexto, o presente estudo visou identificar coccídios intestinais de aves silvestres em dois centros de reabilitação de animais silvestres localizados em dois municípios de Portugal. Oitenta e nove aves silvestres de 9 ordens e 11 famílias foram amostradas, das quais 22 (25%) foram positivas para coccídios. Avispora spp. foram encontradas em aves de rapina. Esporocistos de coccídios da subfamilia Sarcocystinae foram encontrados em corujas. Uma Isospora sp. foi encontrada em Turdus merula Linnaeus, 1758 e uma Eimeria sp. foi encontrada em Fulica atra Linnaeus, 1758. Entre as espécies de coccídios, Avispora bubonis (Cawthorn, Stockdale, 1981) pode ser destacada. O encontro dessa espécie indica que a transmissão de coccídios do Novo Mundo para o Velho Mundo pode estar ocorrendo, potencialmente através da dispersão por Bubo scandiacus (Linnaeus, 1758) pelas regiões árticas ou por meio de atividades antrópicas, e/ou através de outros mecanismos desconhecidos.

Mysteries of host switching: Diversification and host specificity in rodent-coccidia associations.

Recent studies show that host switching is much more frequent than originally believed and constitutes an important driver in evolution of host-parasite associations. However, its frequency and ecological mechanisms at the population level have been rarely investigated. We address this issue by analyzing phylogeny and population genetics of an extensive sample, from a broad geographic area, for commonly occurring parasites of the genus Eimeria within the abundant rodent genera Apodemus, Microtus and Myodes, using two molecular markers. At the most basal level, we demonstrate polyphyletic arrangement, i.e. multiple origin, of the rodent-specific clusters within the Eimeria phylogeny, and strong genetic/phylogenetic structure within these lineages determined at least partially by specificities to different host groups. However, a novel and the most important observation is a repeated occurrence of host switches among closely related genetic lineages which may become rapidly fixed. Within the studied model, this phenomenon applies particularly to the switches between the eimerians from Apodemus flavicollis/Apodemus sylvaticus and Apodemus agrarius groups. We show that genetic differentiation and isolation between A. flavicollis/A. sylvaticus and A. agrarius faunas is a secondary recent event and does not reflect host-parasite coevolutionary history. Rather, it provides an example of rapid ecology-based differentiation in the parasite population.

A Parasitologic and Molecular Survey of Hepatozoon canis Infection In Stray Dogs In Northeastern Iran.

Canine hepatozoonosis, caused by Hepatozoon canis, is a tick-borne disease in domestic and wild dogs that is transmitted by ingestion of Rhipicephalus sanguineus ticks. The aim of the study was to detect H. canis in stray dogs in Iran with the use of blood smear examination and molecular techniques. From October 2014 to September 2015, 150 EDTA blood samples were collected from stray dogs in northeastern Iran. Blood smears were microscopically examined for the presence of Hepatozoon gamonts; whole blood was evaluated by PCR with subsequent sequencing and phylogenetic analysis. Hepatozoon spp. gamonts were observed in the neutrophils of 5/150 (3.3%) blood smears, whereas Hepatozoon spp. 18S rDNA was detected in 12/150 (8.0%) blood samples from stray dogs. There was a good correlation between microscopy and PCR methods (κ = 0.756). The highest seasonal rate of infection was detected in the summer ( P < 0.05). The frequency of Hepatozoon spp. infection did not differ significantly by gender or age ( P > 0.05). The alignment analysis of the sequenced samples showed ≥99% similarity with other nucleotide sequences of Hepatozoon spp. in GenBank. The phylogenetic tree also revealed that the nucleotide sequences in this study were clustered in the H. canis clade and different from the Hepatozoon felis and Hepatozoon americanum clades. Accordingly, we conclude that H. canis infection is present among dogs in northeastern Iran.

The Evolutionary Biology, Ecology and Epidemiology of Coccidia of Passerine Birds.

Coccidia are intracellular parasites of the phylum Apicomplexa that cause a range of pathologies collectively termed coccidiosis. Species of coccidia of commercial importance have been well studied, with the effect of other species on passerine birds receiving increasing attention. In this chapter, we review the literature on coccidia in passerines, with a particular focus on wild populations. The taxonomy and life cycle of passerine coccidia are covered, as is their impact on the health of passerines, their epidemiology and their role in parasite-mediated natural and sexual selection. Coccidia can pose a significant threat to the health of wild passerine populations, and high rates of mortality have been observed in some studies. We examine some of the genetic factors that influence host resistance to coccidia and discuss how these parasites may be important in relation to sexually selected traits. General patterns are beginning to emerge with regard to the epidemiology of the parasites, and the influence of different aspects of the host's ecology on the prevalence and intensity of coccidia is being revealed. We examine these, as well exceptions, in addition to the phenomenon of diurnal oocyst shedding that can bias studies if not accounted for. Finally, we discuss potential future directions for research on coccidia in passerines and the importance of understanding parasite ecology in the management of threatened species.

An optimized DNA extraction method for molecular identification of coccidian species.

Molecular identification of Eimeria parasites infecting poultry and livestock has been commonly used for more than 20 years. An important step of the molecular identification technique is the rupturing of the oocyst wall for DNA extraction. Previously, DNA extraction methods included pre-treatment with sodium hypochlorite and osmotic shock with saturated salt solution. Here, we present a modification of this technique for a more sensitive and efficient identification of Eimeria spp. in field samples. The disruption extent of the oocyst walls, yield of DNA extraction, and identification of species-specific DNA sequences by PCR were used to evaluate this optimized method. Incubation of oocysts in sodium hypochlorite for 1.5 h at 4 °C followed by treatment with a saturated salt solution for 1 h at 55 °C broke up the walls of most Eimeria tenella oocysts, as well as other coccidian species of chicken and rabbit, such as Eimeria intestinalis and even Cryptosporidium cuniculus. Notably, polymerase chain reaction (PCR) amplification of the intervening transcribed sequence 1 (ITS-1) was successfully performed with genomic DNA extracted from just 50 oocysts using this optimized method. Our findings will greatly promote the development of molecular diagnosis methods of coccidiosis and simplify coccidian species identification and categorization as well as infection prevalence, providing a significant advancement in the development of techniques for coccidiosis control and prevention.