Inhibition of sexual stage-specific proteins results in reduced numbers of sexual stages and oocysts of Cystoisospora suis (Apicomplexa: Coccidia) in vitro.

Parasites of the order Coccidia (phylum: Alveolata, subphylum: Apicomplexa) have sophisticated life cycles that include a switch from asexual to sexual development, characterised by distinct cell types. During the development of gametes (gamogony), substantial changes occur at the cellular and subcellular levels, leading to cell fusion of micro- and microgametes, and the development of a zygote that forms a protective outer layer for environmental survival as an oocyst, the transmissible stage. Studies on the porcine coccidian Cystoisospora suis already identified changes in transcription profiles during different time points in the parasite's development and identified proteins with potential roles in the sexual development of this parasite. Here, we focus on three proteins that are possibly involved in the sexual development of C. suis. Enkurin and hapless protein 2 (HAP2) play important roles in signal transduction and gamete fusion during the fertilisation process, and oocyst wall forming protein 1 (OWP1) is a homologue of oocyst wall forming proteins of related parasites. We evaluated their locations in the different life cycle stages of C. suis and their inhibition by specific antibodies in vitro. Immunolocalization detected enkurin in merozoites and sporulated oocysts, HAP2 in merozoites and microgamonts, and OWP2 in merozoites, macrogamonts, oocysts and sporozoites. Up to 100% inhibition of the development of sexual stages and oocyst formation with purified chicken immunoglobulin IgY sera against recombinant enkurin, HAP2, and especially OWP1, were demonstrated. We conclude that the three investigated sexual stage-specific proteins constitute targets for in vivo intervention strategies to interrupt parasite development and transmission to susceptible hosts.

Changes in serum biomarkers of inflammation in bovine besnoitiosis.

BACKGROUND: Acute and chronic besnoitiosis in extensive natural-service herds can have relevant effects in the health of bulls and negative consequences in their productive performance. Recent progress has been made in order to elucidate the pathogenesis of this disease. In this context, the study of biomarkers of inflammation in serum would contribute to gaining knowledge about the physiopathology of bovine besnoitiosis. Serological biomarkers could help in early diagnosis and prognosis, as seropositive bulls may have mild or severe testicular lesions. METHODS: Herein, we have investigated the diagnostic and/or prognostic value of a panel of serum (serological) biomarkers related to inflammation, including total protein, globulin and albumin, haptoglobin (Hp), adenosine deaminase (ADA) paraoxonase-1 (PON-1) and acetylcholinesterase (AChE) in naturally and experimentally B. besnoiti-infected males classified according to different clinical phases of the disease (acute, chronic and subclinical besnoitiosis). RESULTS: Results showed a similar response pattern in these biomarkers for naturally and experimentally infected cattle, with a few relevant variations. Most significant changes occurred during the acute phase of infection, although significant changes in a few biomarkers were also observed during the chronic infection. Haptoglobin, albumin, PON-1 and ADA were identified as the biomarkers that showed changes of higher magnitude in the acute phase of the infection, whereas high total protein and globulin values were found in chronically infected cattle. We have described the changes of a panel of inflammatory biomarkers of acute and chronic bovine besnoitiosis. CONCLUSIONS: In summary, several biomarkers with promising diagnostic value have been identified. The biomarkers associated with acute infection are related to previously reported molecular biomarkers in testicular parenchyma of infected bulls and could help in the diagnosis of early infections and complement results from specific immunoglobulin M (IgM) detection.

A case report and statewide surveillance of "weak meat" condition of Alaska weathervane scallops, Patinopecten caurinus, linked to a recently identified pathogenic parasite, Merocystis kathae (Apicomplexa: Aggregatidae).

Weathervane scallop, Patinopecten caurinus, the largest scallop species in the world, is distributed from northern California, U.S.A., to the Bering Sea, and is only commercially harvested in Alaska. The fishery is considered well managed by the State of Alaska (U.S.A) Department of Fish and Game (ADF&G) and federal government, with many precautionary measures in place to avoid overharvest. There have been episodic declines in some management areas due to unknown causes. Fishermen also encounter scallops with abnormal adductor muscles, a condition colloquially termed "weak meat", characterized by the retention of muscle when shucked, an obvious darkened discoloration, and/or an abnormal texture making the product unacceptable for marketing. A similar syndrome in Atlantic sea scallops, Placopecten magellanicus, described as "gray meat", occurs in the eastern U.S. and Canada, and proposed causes include senescence, loss of bioenergetics due to chronic infestations, or a synergism of these factors. Recently a severe apicomplexan infection was found to cause a gray meat condition in Iceland scallops, Chlamys islandica, and the collapse of that stock. This parasite was subsequently detected in Atlantic sea scallops with the gray meat condition off the U.S. East Coast. Studies that followed identified the parasite as Merocystis kathae, previously described from the common whelk, Buccinum undatum, more than 100 years ago. In 2015 Bering Sea fishermen reported weak meat in their catch, so samples were submitted to ADF&G for diagnosis. Adductor muscles from all affected scallops had many large foci of an apicomplexan associated with necrosis, fibrosis, and muscular atrophy. Given the reduced quality, marketability, and possibly fitness of affected scallops, we performed a survey to estimate prevalence, intensity, and geographic distribution of this apicomplexan in Alaskan weathervane scallops. We sampled 180 scallops, from individual beds within each of the three major geographically broad scallop areas in Alaska. Overall prevalence was about 82%, ranging from 69 to 100% by district. Overall mean infection intensity, based on the number of parasite foci/section, was about 9 (range of 5-29, by location), with scallops from the Bering Sea and Southwest Kodiak being most severely infected. Molecular analyses confirmed that the Alaskan parasite is M. kathae, i.e., the same apicomplexan that caused the collapse of Icelandic scallops and a suspected cause for gray meat and mass mortality of Atlantic sea scallops in northeast North America.

Effects of Vaccination Against Coccidiosis on Gut Microbiota and Immunity in Broiler Fed Bacitracin and Berry Pomace.

Feeding practices have been found to influence gut microbiota which play a major role in immunity of poultry. In the present study, changes in cecal microbiota and humoral responses resulting in the 55 ppm bacitracin (BACI), 1% each of cranberry (CP1) and wild blueberry (BP1) pomace alone or in combination (CP+BP) feeding in broiler Cobb 500 vaccinated or not against coccidiosis were investigated. In the non-vaccinated group, no significant treatment effects were observed on performance parameters. Vaccination significantly affected bird's performance parameters particularly during the growing phase from 10 to 20 days of age. In general, the prevalence of coccidiosis and necrotic enteritis (NE) was reduced by vaccination (P < 0.05). BACI-treated birds showed low intestinal lesion scores, and both CP1 and BP1 feed supplementations reduced Eimeria acervulina and Clostridium perfringens incidences similar to BACI. Vaccination induced change in serum enzymes, minerals, and lipid levels in 21-day old birds while, levels of triglyceride (TRIG) and non-esterified fatty acids (NEFA) were higher (P < 0.05) in CP1 treated non-vaccinated group than in the control. The levels of NEFA were lower in BACI- and CP1-fed birds than in the control in non-vaccinated day 28 old birds. The highest levels of all estimated three immunoglobulins (IgY, IgM, and IgA) were found in the vaccinated birds. Metagenomics analysis of the cecal bacterial community in 21-day old birds showed the presence of Firmicutes (90%), Proteobacteria (5%), Actinobacteria (2%), and Bacteroidetes (2%). In the vaccinated group, an effect of BACI was noted on Proteobacteria (P = 0.03). Vaccination and/or dietary treatments influenced the population of Lactobacillaceae, Enterobacteriaceae, Clostridiaceae, and Streptococcaceae which were among the most abundant families. Overall, this study revealed that besides their beneficial effects on performance, alike bacitracin, berry pomaces in poultry feed have profound impacts on the chicken cecal microbiota and blood metabolites that could be influenced by vaccination against coccidiosis.

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.

Recent achievements and doors opened for coccidian parasite research and development through transcriptomics of enteric sexual stages.

The Coccidia is the largest group of parasites within the Apicomplexa, a phylum of unicellular, obligate parasites characterized by the possession of an apical complex of organelles and structures in the asexual stages of their life cycles, as well as by a sexual reproductive phase that occurs enterically in host animals. Coccidian sexual reproduction involves morphologically distinct microgametes and macrogametes that combine to form a diploid zygote and, ultimately, following meiosis and mitosis, haploid, infectious sporozoites, inside sporocysts within an oocyst. Recent transcriptomic analyses have identified genes involved in coccidian sexual stage development and reproduction, including genes encoding for microgamete- and macrogamete-specific proteins with roles in gamete motility, fusion and fertilization, and in the formation of the resilient oocyst wall that allows coccidians to persist for long periods in the environment. Transcriptomics has also provided important clues about the regulation of gene expression in the transformation of parasites from one developmental stage to the next, a complex sequence of events that may involve transcription factors such as the apicomplexan Apetala2 (ApiAP2) family, alternative splicing, regulatory RNAs and MORC (a microrchida homologue and regulator of sexual stage development in Toxoplasma gondii). The molecular dissection of coccidian sexual development and reproduction by transcriptomic analyses may lead to the development of novel transmission-blocking strategies.

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.

Polyphyletic origin, intracellular invasion, and meiotic genes in the putatively asexual agamococcidians (Apicomplexa incertae sedis).

Agamococcidians are enigmatic and poorly studied parasites of marine invertebrates with unexplored diversity and unclear relationships to other sporozoans such as the human pathogens Plasmodium and Toxoplasma. It is believed that agamococcidians are not capable of sexual reproduction, which is essential for life cycle completion in all well studied parasitic apicomplexans. Here, we describe three new species of agamococcidians belonging to the genus Rhytidocystis. We examined their cell morphology and ultrastructure, resolved their phylogenetic position by using near-complete rRNA operon sequences, and searched for genes associated with meiosis and oocyst wall formation in two rhytidocystid transcriptomes. Phylogenetic analyses consistently recovered rhytidocystids as basal coccidiomorphs and away from the corallicolids, demonstrating that the order Agamococcidiorida Levine, 1979 is polyphyletic. Light and transmission electron microscopy revealed that the development of rhytidocystids begins inside the gut epithelial cells, a characteristic which links them specifically with other coccidiomorphs to the exclusion of gregarines and suggests that intracellular invasion evolved early in the coccidiomorphs. We propose a new superorder Eococcidia for early coccidiomorphs. Transcriptomic analysis demonstrated that both the meiotic machinery and oocyst wall proteins are preserved in rhytidocystids. The conservation of meiotic genes and ultrastructural similarity of rhytidocystid trophozoites to macrogamonts of true coccidians point to an undescribed, cryptic sexual process in the group.

Head-to-head comparisons of Toxoplasma gondii and its near relative Hammondia hammondi reveal dramatic differences in the host response and effectors with species-specific functions.

Toxoplasma gondii and Hammondia hammondi are closely-related coccidian intracellular parasites that differ in their ability to cause disease in animal and (likely) humans. The role of the host response in these phenotypic differences is not known and to address this we performed a transcriptomic analysis of a monocyte cell line (THP-1) infected with these two parasite species. The pathways altered by infection were shared between species ~95% the time, but the magnitude of the host response to H. hammondi was significantly higher compared to T. gondii. Accompanying this divergent host response was an equally divergent impact on the cell cycle of the host cell. In contrast to T. gondii, H. hammondi infection induces cell cycle arrest via pathways linked to DNA-damage responses and cellular senescence and robust secretion of multiple chemokines that are known to be a part of the senescence associated secretory phenotype (SASP). Remarkably, prior T. gondii infection or treatment with T. gondii-conditioned media suppressed responses to H. hammondi infection, and promoted the replication of H. hammondi in recipient cells. Suppression of inflammatory responses to H. hammondi was found to be mediated by the T. gondii effector IST, and this finding was consistent with reduced functionality of the H. hammondi IST ortholog compared to its T. gondii counterpart. Taken together our data suggest that T. gondii manipulation of the host cell is capable of suppressing previously unknown stress and/or DNA-damage induced responses that occur during infection with H. hammondi, and that one important impact of this T. gondii mediated suppression is to promote parasite replication.

Infection of parthenogenetic lizards by blood parasites does not support the "Red Queen hypothesis" but reveals the costs of sex.

Sexual organisms should be better suited than asexual ones in a context of continuous evolution in response to opposite organisms in changing environments ("Red Queen" hypothesis of sex). However, sex also carries costs associated with the maintenance of males and mating (sex cost hypothesis). Here, both non-mutually excluding hypotheses are tested by analysing the infestation by haemogregarines of mixed communities of Darevskia rock lizards composed of parthenogens generated by hybridisation and their bisexual relatives. Prevalence and intensity were recorded from 339 adult lizards belonging to six species from five syntopic localities and analysed using Generalized Mixed-Models (GLMM). Both infestation parameters depended on host-size (like due to longer exposure with age), sex and, for intensity, species. Once accounting for locality and species, males were more parasitized than conspecific females with bisexual species, but no signal of reproductive mode itself on parasitization was recovered. Essentially, male-male interactions increased haemogregarine intensity while females either sexual or asexual had similar reproductive costs when in the same conditions. These findings deviate from the predictions from "Red Queen" dynamics while asymmetric gender costs are here confirmed. Thus, increased parasitization pressure on males adds to other costs, such as higher social interactions and lower fecundity, to explain why parthenogenetic lizards apparently prevail in the short-term evolutionary scale. How this is translated in the long-term requires further phylogenetic analysis.

Social dominance, but not parasite load, affects sperm quality and sperm redox status in house sparrows.

Sperm performance is an important component of male reproductive success. However, sperm production is costly and males need to optimize their investment in sperm quality versus the somatic traits involved in mating success, e.g. their social status. As oxidative stress affects both sperm performance and somatic functions, it has been hypothesized to mediate such a trade-off. According to the oxidation-based soma/germline trade-off hypothesis, dominant males should favour the antioxidant protection of their somatic tissues, and subordinate males should favour the antioxidant protection of their sperm. We tested this hypothesis by experimentally infecting wild-caught house sparrows Passer domesticus with Coccidia Isopora sp., an internal parasite known to deplete antioxidant resources. We predicted that (i) increased parasite load affects sperm oxidative status and sperm performance and that (ii) males with experimentally high parasite load adjust the antioxidant protection of their soma versus their sperm according to their social status. Despite a 5400% increase in parasite load, sperm performance and somatic and spermatic oxidative status remained unaffected, irrespective of male social status. Nevertheless, males increased their sperm performance over time, a pattern mirrored by an increase in the antioxidant protection of their sperm. Moreover, males at the lower end of the hierarchy always produced sperm of lower velocity, suggesting that they were constrained and privileged their soma over their germline. To conclude, high parasite loads do not necessarily affect sperm performance and oxidative status. In contrast, social hierarchy and the relative investment in soma versus sperm antioxidant protection are determinants of sperm performance.

Both candidate gene and neutral genetic diversity correlate with parasite resistance in female Mediterranean mouflon.

BACKGROUND: Parasite infections can have substantial impacts on population dynamics and are accordingly a key challenge for wild population management. Here we studied genetic mechanisms driving parasite resistance in a large herbivore through a comprehensive approach combining measurements of neutral (16 microsatellites) and adaptive (MHC DRB1 exon 2) genetic diversity and two types of gastrointestinal parasites (nematodes and coccidia). RESULTS: While accounting for other extrinsic and intrinsic predictors known to impact parasite load, we show that both neutral genetic diversity and DRB1 are associated with resistance to gastrointestinal nematodes. Intermediate levels of multi-locus heterozygosity maximized nematodes resistance, suggesting that both in- and outbreeding depression might occur in the population. DRB1 heterozygosity and specific alleles effects were detected, suggesting the occurrence of heterozygote advantage, rare-allele effects and/or fluctuating selection. On the contrary, no association was detected between genetic diversity and resistance to coccidia, indicating that different parasite classes are impacted by different genetic drivers. CONCLUSIONS: This study provides important insights for large herbivores and wild sheep pathogen management, and in particular suggests that factors likely to impact genetic diversity and allelic frequencies, including global changes, are also expected to impact parasite resistance.

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.

Some remarks on the distribution and dispersion of Coccidia from icterid birds in South America: Isospora guaxi n. sp. and Isospora bellicosa Upton, Stamper & Whitaker, 1995 (Apicomplexa: Eimeriidae) from the red-rumped cacique Cacicus haemorrhous (L.) (Passeriformes: Icteridae) in southeastern Brazil.

A new species of coccidian, Isospora guaxi n. sp., and Isospora bellicosa Upton, Stamper & Whitaker, 1995 (Protozoa: Apicomplexa: Eimeriidae) are recorded from red-rumped caciques Cacicus haemorrhous (L.) in the Parque Nacional do Itatiaia, Brazil. Isospora guaxi n. sp. has sub-spheroidal oöcysts, measuring on average 30.9 × 29.0 µm, with smooth, bi-layered wall c.1.9 µm thick. Micropyle and oöcyst residuum are absent, but a polar granule is present. Sporocysts are ellipsoidal, measuring on average 19.3 × 13.8 µm. Stieda body is knob-like and sub-Stieda body is prominent and compartmentalized. Sporocyst residuum is composed of scattered granules. Sporozoites are vermiform, with one refractile body and a nucleus. Isospora bellicosa has sub-spheroidal to ovoidal oöcysts, measuring on average 27.1 × 25.0 µm, with smooth, bi-layered wall c.1.5 µm thick. Micropyle and oöcyst residuum are absent, but one or two polar granules are present. Sporocysts are ellipsoidal, measuring on average 18.1 × 10.9 µm. Stieda body is knob-like and sub-Stieda body is rounded to rectangular. Sporocyst residuum is composed of a cluster of compact or diffuse granules. Sporozoites are vermiform, with one refractile body and a nucleus. Isospora bellicosa was originally described from the Peruvian meadowlark Sturnella bellicosa deFilippi, a trans-Andean icterid which is allopatric with the cis-Andean C. haemorrhous. Therefore, in conclusion, this current study reveals the dispersion of coccidia from Icteridae across the Andes Mountains, besides describing the sixth isosporoid coccidium infecting an icterid bird.

Diversity and host specificity of coccidia (Apicomplexa: Eimeriidae) in native and introduced squirrel species.

Introduction of alien species into new areas can have detrimental effects on native ecosystems and impact the native species. The present study aims to identify coccidia infecting native and introduced squirrels in Italy, to gain insight into possible transmission patterns and role of monoxenous coccidia in mediating the competition between alien and native hosts. We collected 540 faecal samples of native red squirrels, Sciurus vulgaris, invasive alien grey squirrels, S. carolinensis, and introduced Pallas's squirrels, Callosciurus erythraeus. Total prevalence of Eimeria spp. was 95.6% in S. vulgaris, 95.7% in S. carolinensis and only 4.1% in C. erythraeus. Morphological examination revealed 3 Eimeria morphotypes. Phylogenetic analyses of Eimeria DNA based on 18S, ITS, cox I markers displayed fairly distinct monophyletic clades in the microscopically indistinguishable E2 morphotype, proving indisputable distinction between the isolates from red and grey squirrels. Grey squirrels successfully introduced E. lancasterensis from their native range, but this species does not spill over to native red squirrels. Similarly, there is no evidence for the transmission of E. sciurorum from red to grey squirrels. The possible transmission and the potential role of monoxenous coccidia in mediating the competition between native and invasive squirrels in Italy were not confirmed.

Infections by Intestinal Coccidia and Giardia duodenalis.

The coccidians Cryptosporidium spp, Cyclospora cayetanensis, and Cystoisospora belli and the flagellate Giardia duodenalis are pathogenic protozoa associated with gastrointestinal manifestations. Diagnosis relies heavily on microscopy, and although ova-and-parasite examinations can detect Giardia and Cystoisospora, Cryptosporidium and Cyclospora often require specific diagnostic requests. Approved non-microscopy methods are available for Giardia and Cryptosporidium, although negative results are frequently followed by microscopic assays. Polymerase chain reaction-based methods are not frequently used for diagnosis of Giardia and Cryptosporidium and have been used primarily for epidemiologic or outbreak investigations of Giardia and Cryptosporidium.