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Microtubules and specialized microtubule-containing structures are assembled from tubulins, an ancient superfamily of essential eukaryotic proteins. Here, we use bioinformatic approaches to analyze features of tubulins in organisms from the phylum Apicomplexa. Apicomplexans are protozoan parasites that cause a variety of human and animal infectious diseases. Individual species harbor one to four genes each for α- and β-tubulin isotypes. These may specify highly similar proteins, suggesting functional redundancy, or exhibit key differences, consistent with specialized roles. Some, but not all apicomplexans harbor genes for δ- and ε-tubulins, which are found in organisms that construct appendage-containing basal bodies. Critical roles for apicomplexan δ- and ε-tubulin are likely to be limited to microgametes, consistent with a restricted requirement for flagella in a single developmental stage. Sequence divergence or the loss of δ- and ε-tubulin genes in other apicomplexans appears to be associated with diminished requirements for centrioles, basal bodies, and axonemes. Finally, because spindle microtubules and flagellar structures have been proposed as targets for anti-parasitic therapies and transmission-blocking strategies, we discuss these ideas in the context of tubulin-based structures and tubulin superfamily properties.

Considering the original nature of carnivorism among dogs and cats, nowadays these pets are increasingly fed BARF (Biologically Appropriate Raw Food/Bone And Raw Food) diet to improve their health conditions. However, this diet may also carry health risks, such as vitamin or mineral deficiencies, and infection with pathogens including parasites. In our study, fecal samples of 89 pet animals kept on BARF diet were subjected to coprological examination followed by molecular analyses. Six of them shed eggs of Dicrocoelium dendriticum. This result was confirmed with PCR and sequencing, and in one case, the DNA of Fasciola hepatica was also demonstrated. In addition, oocysts of Cystoisospora canis, a Cystoisospora ohioensis-like sp. and Eimeria stiedai, as well as sporocysts of a Sarcocystis sp. were also detected. All samples were negative for Neospora caninum and Toxoplasma gondii. In conclusion, no evidence was found for the infection of BARF-fed dogs and cats with parasites that are usually associated with this diet and considered as clinico-pathological risk factors for these pets themselves (e.g., N. caninum, T. gondii). However, fluke eggs (probably originating from ruminants) and oocysts of E. stiedai (from rabbit liver in the food) were demonstrated as pseudoparasites. These species are usually not considered among parasite-associated risks of BARF-feeding, implying that other animals living near BARF-fed pets are neglected in this context. However, where intermediate hosts of D. dendriticum occur in urban areas, BARF-feeding may indirectly affect later other dogs and cats. It was also shown here that BARF-feeding may contribute to the contamination of the environment with E. stiedai oocysts, thus increasing the risks of biliary coccidiosis in nearby living pet rabbits that would otherwise not have access to oocysts of E. stiedai. [ABSTRACT FROM AUTHOR]

Background Human microsporidiosis presents as an important and rapidly emerging opportunistic infection. However, the exact burden of this infection especially in the pediatric population of Northern India remains unknown. In this study, we investigated the prevalence of microsporidia among human immunodeficiency virus (HIV)-positive and HIV-negative pediatric patients who presented with diarrhea.

Introduction: Cryptosporidium, Cystoisospora, and Giardia duodenalis are gastrointestinal protozoa parasites that cause diarrhea in various animals. However, information regarding the detection and phylogenetic characterization of gastrointestinal protozoa parasites in cats is limited throughout South Korea. Therefore, this study aimed to determine the detection and identify subspecies of gastrointestinal protozoa parasites in cats from South Korea. Methods: A total of 290 fecal samples were collected from stray, companion, and shelter cats in six provinces. Cryptosporidium, Cystoisospora, and G. duodenalis were identified by PCR. All positive samples were subtyped by PCR and sequencing of gp60, ITS-1, tpi, bg, and gdh. Results: The overall detection of gastrointestinal protozoan parasitic infection was 17.93%. G. duodenalis was the most prevalent, with 7.93%, followed by Cystoisospora spp. (7.24%) and Cryptosporidium spp. (4.48%). In addition, C. felis (n=10), C. parvum (n=2), C. ryanae (n=1), Cystoisospora felis (n=14), Cystoisospora suis (n=5), Cystoisospora ohioensis (n=1), Cystoisospora spp. were identified in subspecies analysis of positive samples. C. felis showed a significant association with diarrhea (7.81%) and living condition (6.04%), and Cystoisospora felis in diarreha (9.38%) according to detection. Through phylogenetic analysis of the tpi, bg, and gdh genes from 23 G. duodenalispositive samples, it was confirmed that the samples of present study belonged to assemblage A, B, C, and D. Discussion: South Korean cats have a high rate of gastrointestinal protozoan parasites infection with cat-specific Cryptosporidium and Cystoisospora, which are associated with living conditions and diarrhea symptoms. Moreover, zoonotic and other animal-specific subtype of protozoan parasites have been detected in cat feces. [ABSTRACT FROM AUTHOR]

Background Human microsporidiosis presents as an important and rapidly emerging opportunistic infection. However, the exact burden of this infection especially in the pediatric population of Northern India remains unknown. In this study, we investigated the prevalence of microsporidia among human immunodeficiency virus (HIV)-positive and HIV-negative pediatric patients who presented with diarrhea. Methods A total of 263 children were recruited consisting of 98 HIV seropositive with diarrhea and 165 HIV seronegative but with diarrhea. Morning stool samples were collected and both direct and formol ether concentrated samples were examined for the presence of intestinal parasites. The modified acid-fast staining was done for coccidian parasites and trichrome stain for microsporidia detection. Further, the species were detected using a real-time polymerase chain reaction (PCR) targeting a conserved region of the small ribosomal subunit rRNA gene of Enterocytozoon bieneusi , Encephalitozoon hellem , Encephalitozoon intestinalis , and Encephalitozoon cuniculi. Results Overall, one or more parasites were detected in 52.04% (51/98) of HIV seropositive and 53.33% (88/165) of seronegative children (p = 0.8391). However, coccidian parasites were detected in a significantly huge number of HIV seropositive children (21.43% [21/98]) as compared with HIV seronegative children (4.24% [7/165]). Microsporidial DNA could be detected in HIV seropositive with diarrhea children (17.35% [17/98]) by PCR. A significant correlation between low CD4 count (≤ 200/μL) and intestinal parasite positivity could be established. Conclusion Microsporidia is a significant cause of diarrhea in HIV seropositive pediatric patients and should be kept in mind as one of the differential diagnoses in such patients. [ABSTRACT FROM AUTHOR]

Immunocompromised patients with human immunodeficiency virus (HIV) infection are prone to multiple infections, of which parasitic infections are an important cause. Parasitic protozoal infections - both by common and rare protozoa are documented in such patients. Here, we report a rare and interesting case of five protozoal infections affecting a single HIV-infected person at the same time of initial presentation. A 64-years-male came to us with complaints of chronic diarrhea for 6 months. He was investigated and found to be positive for HIV I. His stool examination revealed cysts of Entameba histolytica and Giardia lamblia and oocysts of Cryptosporidium species and Cystoisospora species. His toxoplasma IgG was also positive in high titer. The patient was medically diagnosed and was treated with medications as clinically prescribed - antiretroviral therapy was initiated and he was discharged in due course. A total of five protozoal infections were documented affecting a single person - newly diagnosed immunocompromised male, which by sheer qualitative count of patient case histories, indeed is a rare case reported in the medical literature. [ABSTRACT FROM AUTHOR]

Abstract Introduction Cats (Felis catus) are the only felines that live in close contact with humans. Since cats can act as vectors, carriers, reservoirs and definitive hosts of many gastrointestinal (GI) parasites, parasitic assessment could contribute to their survival and well‐being. Aims The current study aimed to assess the diversity and prevalence of GI parasites in domestic and feral cats from Ratnanagar in Chitwan in Central Nepal. Methods A total of 107 fresh faecal samples of cats (90 household cats and 17 feral cats) of varied ages and sex were collected and transported to the laboratory. The copromicroscopic examination was carried out following direct wet mount, formalin‐ethyl acetate sedimentation, saturated salt flotation, acid‐fast staining and sporulation techniques. Furthermore, associated risk factors were evaluated to ascertain the predictor of risks for parasitic acquisition. Results The current study revealed an overall 95.3% prevalence rate with a 100% rate in feral cats and 94.4% in household cats. Altogether, 18 (17 known and one unknown) different species of GI parasites were reported with the helminths (95.3%; 11 species) and the protozoa (55.1%; seven species). Besides age and sex, outdoor lifestyle, absence or unknown history of medication and hunting behaviour of the felines are the predictors of risk. Furthermore, mixed infection was comparatively higher than single infection in the faecal samples. Conclusions Cats harbour a higher prevalence and greater diversity of GI parasites, and parasitism varies with age and sex. This finding can be essential for veterinarians and public health authorities for strategic treatment and for assessing the zoonotic transmission of the parasites from these felines. Importantly, an effective medication strategy for cats and owners is recommended.

Introduction: Cats (Felis catus) are the only felines that live in close contact with humans. Since cats can act as vectors, carriers, reservoirs and definitive hosts of many gastrointestinal (GI) parasites, parasitic assessment could contribute to their survival and well‐being. Aims: The current study aimed to assess the diversity and prevalence of GI parasites in domestic and feral cats from Ratnanagar in Chitwan in Central Nepal. Methods: A total of 107 fresh faecal samples of cats (90 household cats and 17 feral cats) of varied ages and sex were collected and transported to the laboratory. The copromicroscopic examination was carried out following direct wet mount, formalin‐ethyl acetate sedimentation, saturated salt flotation, acid‐fast staining and sporulation techniques. Furthermore, associated risk factors were evaluated to ascertain the predictor of risks for parasitic acquisition. Results: The current study revealed an overall 95.3% prevalence rate with a 100% rate in feral cats and 94.4% in household cats. Altogether, 18 (17 known and one unknown) different species of GI parasites were reported with the helminths (95.3%; 11 species) and the protozoa (55.1%; seven species). Besides age and sex, outdoor lifestyle, absence or unknown history of medication and hunting behaviour of the felines are the predictors of risk. Furthermore, mixed infection was comparatively higher than single infection in the faecal samples. Conclusions: Cats harbour a higher prevalence and greater diversity of GI parasites, and parasitism varies with age and sex. This finding can be essential for veterinarians and public health authorities for strategic treatment and for assessing the zoonotic transmission of the parasites from these felines. Importantly, an effective medication strategy for cats and owners is recommended. [ABSTRACT FROM AUTHOR]

Background: Cystoisospora spp. is considered a potential pathogen to cause diarrhoea in cats, and the disease occurs clinically, especially in young, weak and immunocompromised cats. The present study aimed to evaluate the clinical utility and diagnostic roles of neutrophil-to-lymphocyte ratio (NLR) and monocyte-to-lymphocyte (MLR) ratios in diarrheal cats infected with Cystoisospora spp. Materials, Methods & Results: Twenty-five cats with diarrhea due to infection caused by Cystoisospora spp. and 15 healthy cats were included in this study. The anamnesis, clinical and laboratory findings of all cats were recorded. A clinical severity score was generated for diarrheal cats, which included activity, appetite, presence of vomit, and faecal consistency. Also, cats infected with Cystoisospora spp. were classified as systemic inflammatory response syndrome (SIRS) positive (+) and SIRS (-) group according to the presence of at least 3 of the SIRS criteria. The Cystoisospora spp. oocysts were diagnosed by direct smear and simple faecal flotation. Cats found positive for other parasitic and protozoal agents by faeces examination were excluded from the study. Complete blood counts (CBC) were performed with an automated blood cell counter for all cats. Blood smears were prepared (with May-Grunwald Giemsa) from samples, and the CBC results were confirmed microscopically (obj.100x). White blood cell (WBC), neutrophil, lymphocyte and monocyte counts were recorded directly from the CBC. The neutrophil-to-lymphocyte ratio was calculated as the ratio of the absolute neutrophil count to the absolute lymphocyte count. The monocyte-to-lymphocyte was calculated as the absolute monocyte count ratio to the absolute lymphocyte count. White blood cell (P = 0.015), neutrophil (P = 0.001), monocyte (P = 0.033), NLR (P = 0.000) and MLR (P = 0.042) of diarrheal cats infected with Cystoisospora spp. were significantly higher than in the healthy group. There was no significant difference between groups in the lymphocyte counts (P > 0.05). SIRS (+) group had statistically significantly higher WBC (P = 0.014), neutrophil counts (P = 0.000), NLR (P = 0.000) and MLR (P = 0.037) than the healthy group. Also, neutrophil counts and NLR of the SIRS (+) group were statistically significantly higher than the SIRS (-) group (P = 0.032; P = 0.001, respectively). However, there was no significant difference SIRS (+), SIRS (-), and healthy groups regarding lymphocyte and monocyte counts (P > 0.05). The clinical severity of the disease positively correlated with NLR and MLR. Also, the best cut-off value of NLR to predict SIRS was > 1.67, with 92.86% sensitivity and 84.62% specificity. MLR was not a useful predictor for SIRS. Discussion: NLR and MLR are parameters derived from complete blood count. In recent years, they have been used in human and veterinary medicine as a biomarker in the diagnosis, prognosis and treatment follow-up of various diseases and provide valuable information about inflammation. In cats, there are limited studies on NLR, and to our knowledge, there are no studies on MLR. The current study suggests that NLR and MLR may be used to determine the severity of the inflammatory condition in diarrheal cats infected with Cystoisospora spp., and NLR is a useful marker for predicting SIRS status in these cats. In addition, this study also will contribute to larger-scale studies in the future. [ABSTRACT FROM AUTHOR]

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Introduction: Intestinal coccidian parasites known to cause opportunistic infections have increased since the past years and are often indistinguishable from other forms of communityacquired diarrhoea, which suggests a need for proper and rapid diagnostic techniques to recover and identify the organism. Aim: To investigate if Auramine Rhodamine (AR) staining is a sensitive and time conserving staining technique in contrast to modified acid fast stain. Materials and Methods: A descriptive observational study was conducted at Nizam’s Institute of Medical Sciences, Hyderabad, India, over a period of three months (i.e., from July to September 2019). A total of 100 stool samples received by the Department of Microbiology were subjected to macroscopic and microscopic examination by saline mount, iodine mount, kinyoun acid fast stain, modified AR staining. Results: In the present study the prevalence of coccidian infections were observed to be 6%. Coccidian parasites reported were Cryptosporidium species and Cystoisospora species immunocompromised patients were found to be more prone to infections with coccidian parasites. The AR stain showed 100% agreement with modified acid fast stain. Conclusion: This study concluded that the AR stain is a rapid and better stain than kinyoun acid-fast stain for the detection of coccidian parasites.

Background: Several gastrointestinal parasites that infect cats pose potential health threats for humans and animals. The present study is the first to report gastrointestinal (GIT) parasites in feces of stray cats from Gharbia governorate, Egypt. Findings were combined with those published in the earlier surveys from various Egyptian governorates, and various meta-analyses were conducted to underline the parasitic zoonoses from cats in Egypt. Results: Out of 143 samples tested in Gharbia, 75 (52.4%) were found infected with 13 different parasites. Co-infections were observed in 49.3% of positives. Several parasites were detected, e.g., Toxocara cati (30.0%), Toxascaris leonina (22.4%), hookworms (8.4%), taeniids (4.2%), Strongyloides spp. (2.1%), Physaloptera spp. (2.1%), Alaria spp. (1.4%) and Dipylidium caninum (0.7%). Opisthorchis-like eggs were found in a single sample being the first report from cats in Africa. Oocysts of 4 coccidian parasites were identified, and a few Toxoplasma gondii-like oocysts were detected in 2 samples (1.4%). Results of the meta-analysis illustrated that occurrence of T. gondii oocysts in feces of cats from Egypt may have been overestimated in earlier studies; 1432 cats have been tested and displayed a 5 times higher pooled prevalence (11.9%) than the published global pooled prevalence for T. gondii oocysts in cats. This overestimation might have occurred because some small-sized oocysts that belong to other coccidian parasites were mis-identified as T. gondii. Toxocara cati had a high pooled prevalence (22.5%) in cats from Egypt, which is even greater than the published pooled prevalence in cats globally; however, several reports from Egypt have neglected the role of T. cati in human toxocarosis. Dipylidium caninum displayed also a high prevalence (26.7%). Conclusion: Several zoonotic parasite species have been found in stray cats from Egypt, raising concerns about the risks to the Egyptian human population as well as environmental contamination. Prompt surveillance supervised by the government and accompanied by data dissemination will be helpful for developing effective control strategies. [ABSTRACT FROM AUTHOR]

Cyclosporiasis has historically been underdiagnosed due to a lack of clinical recognition and challenges in laboratory detection. Microscopic detection of Cyclospora requires either the modified Kinyoun's acid-fast or safranin stains, which are not part of the standard ova-and-parasite examination. Alternatively, the use of fluorescence microscopy for the detection of Cyclospora due to the organism's autofluorescent properties has been an available tool in the field of clinical parasitology. However, it is unknown how ultraviolet (UV) fluorescence performs compared to permanent stains in routine diagnostic laboratory settings. In this study, we present a direct head-to-head comparison of UV fluorescence against modified acid-fast (MAF) stains using 50 (35 Cyclospora positive, 5 Cystoisospora positive, and 10 Cystoisospora negative) blinded, concentrated stool specimens. Five technically competent medical laboratory technologists of varying years of experience in clinical parasitology at a large reference laboratory independently read each of the MAF slides and UV wet mounts. Overall, the performance of UV fluorescence was similar to MAF slides with a higher agreement of 88.8% (95% CI 86.3%-91.3%) compared to 85.2% (95% CI 84.2%-86.2%), respectively. A higher false-negative rate was associated with MAF slides (MAF: 11.6%, UV: 6.8%). However, UV fluorescence was found to have a higher proportion of false positives (UF: 3.2%, MAF: 2%) and misidentified coccidia. Discrepant results regardless of methods were more frequently observed with technologists who have less experience in parasite morphology. This study is the first to document evidence to support the increased sensitivity and utility of UV fluorescence to improve laboratory diagnosis of cyclosporiasis., Importance: This study is important as there is a dearth of studies in our field of clinical parasitology that investigate and establish performance characteristics of classic and newer, non-molecular methods. While these studies may not seem as heavy hitting as some new technologies described in our related disciplines, for our field, such studies are long overdue and critically lacking.

Abstract Background Fecal examinations in pet cats and dogs are key components of routine veterinary practice; however, their accuracy is influenced by diagnostic methodologies and the experience level of personnel performing the tests. The VETSCAN IMAGYST system was developed to provide simpler and easier fecal examinations which are less influenced by examiners’ skills. This system consists of three components: a sample preparation device, an automated microscope scanner, and analysis software. The objectives of this study were to qualitatively evaluate the performance of the VETSCAN IMAGYST system on feline parasites (Ancylostoma and Toxocara cati) and protozoan parasites (Cystoisospora and Giardia) and to assess and compare the performance of the VETSCAN IMAGYST centrifugal flotation method to reference centrifugal and passive flotation methods. Methods To evaluate the diagnostic performance of the scanning and algorithmic components of the VETSCAN IMAGYST system, fecal slides were prepared by the VETSCAN IMAGYST centrifugal flotation technique with pre-screened fecal samples collected from dogs and cats and examined by both an algorithm and parasitologists. To assess the performance of the VETSCAN IMAGYST centrifugal flotation technique, diagnostic sensitivity and specificity were calculated and compared to those of conventional flotation techniques. Results The performance of the VETSCAN IMAGYST algorithm closely correlated with evaluations by parasitologists, with sensitivity of 75.8–100% and specificity of 93.1-100% across the targeted parasites. For samples with 50 eggs or less per slide, Lin’s concordance correlation coefficients ranged from 0.70 to 0.95 across the targeted parasites. The results of the VETSCAN IMAGYST centrifugal flotation method correlated well with those of the conventional centrifugal flotation method across the targeted parasites: sensitivity of 65.7–100% and specificity of 97.6–100%. Similar results were observed for the conventional passive flotation method compared to the conventional centrifugal flotation method: sensitivity of 56.4–91.7% and specificity of 99.4–100%. Conclusions The VETSCAN IMAGYST scanning and algorithmic systems with the VETSCAN IMAGYST fecal preparation technique demonstrated a similar qualitative performance to the parasitologists’ examinations with conventional fecal flotation techniques. Given the deep learning nature of the VETSCAN IMAGYST system, its performance is expected to improve over time, enabling it to be utilized in veterinary clinics to perform fecal examinations accurately and efficiently.

Microtubules and specialized microtubule-containing structures are assembled from tubulins, an ancient superfamily of essential eukaryotic proteins. Here, we use bioinformatic approaches to analyze features of tubulins in organisms from the phylum Apicomplexa. Apicomplexans are protozoan parasites that cause a variety of human and animal infectious diseases. Individual species harbor one to four genes each for α- and β-tubulin isotypes. These may specify highly similar proteins, suggesting functional redundancy, or exhibit key differences, consistent with specialized roles. Some, but not all apicomplexans harbor genes for δ- and ε-tubulins, which are found in organisms that construct appendage-containing basal bodies. Critical roles for apicomplexan δ- and ε-tubulin are likely to be limited to microgametes, consistent with a restricted requirement for flagella in a single developmental stage. Sequence divergence or the loss of δ- and ε-tubulin genes in other apicomplexans appears to be associated with diminished requirements for centrioles, basal bodies, and axonemes. Finally, because spindle microtubules and flagellar structures have been proposed as targets for anti-parasitic therapies and transmission-blocking strategies, we discuss these ideas in the context of tubulin-based structures and tubulin superfamily properties.

Background: Fecal examinations in pet cats and dogs are key components of routine veterinary practice; however, their accuracy is influenced by diagnostic methodologies and the experience level of personnel performing the tests. The VETSCAN IMAGYST system was developed to provide simpler and easier fecal examinations which are less influenced by examiners' skills. This system consists of three components: a sample preparation device, an automated microscope scanner, and analysis software. The objectives of this study were to qualitatively evaluate the performance of the VETSCAN IMAGYST system on feline parasites (Ancylostoma and Toxocara cati) and protozoan parasites (Cystoisospora and Giardia) and to assess and compare the performance of the VETSCAN IMAGYST centrifugal flotation method to reference centrifugal and passive flotation methods. Methods: To evaluate the diagnostic performance of the scanning and algorithmic components of the VETSCAN IMAGYST system, fecal slides were prepared by the VETSCAN IMAGYST centrifugal flotation technique with pre-screened fecal samples collected from dogs and cats and examined by both an algorithm and parasitologists. To assess the performance of the VETSCAN IMAGYST centrifugal flotation technique, diagnostic sensitivity and specificity were calculated and compared to those of conventional flotation techniques. Results: The performance of the VETSCAN IMAGYST algorithm closely correlated with evaluations by parasitologists, with sensitivity of 75.8–100% and specificity of 93.1-100% across the targeted parasites. For samples with 50 eggs or less per slide, Lin's concordance correlation coefficients ranged from 0.70 to 0.95 across the targeted parasites. The results of the VETSCAN IMAGYST centrifugal flotation method correlated well with those of the conventional centrifugal flotation method across the targeted parasites: sensitivity of 65.7–100% and specificity of 97.6–100%. Similar results were observed for the conventional passive flotation method compared to the conventional centrifugal flotation method: sensitivity of 56.4–91.7% and specificity of 99.4–100%. Conclusions: The VETSCAN IMAGYST scanning and algorithmic systems with the VETSCAN IMAGYST fecal preparation technique demonstrated a similar qualitative performance to the parasitologists' examinations with conventional fecal flotation techniques. Given the deep learning nature of the VETSCAN IMAGYST system, its performance is expected to improve over time, enabling it to be utilized in veterinary clinics to perform fecal examinations accurately and efficiently. [ABSTRACT FROM AUTHOR]

Introduction: Despite antiretroviral therapy, it is still common to find the presence of opportunistic infections such as enteroparasites, which cause a big impact on the health and economy of patients.Objetive: To determine the frequency, etiology and factors associated with enteroparasitosis in patients with HIV at a reference hospital in Lambayeque, Peru.Material and Methods: A prospective observational study was conducted in diarrheic patients with HIV infection in a hospital in Lambayeque from January to June, 2016. The study was a census. Both the collection and transport as well as the analysis of the samples followed the recommendations of the Manual of procedures for the diagnosis of human parasitosis of the National Institute of Health.Results: The meadian age was 28 years, the male sex predominated, and the percentage of parasitism was 73.08%. Patients with viral load >1000 were 69.2% and patients with HAART were 55.8%. Blastocystis hominis was the most frequent parasite (30.77%), followed by Cryptosporidium spp. and Cystoisospora belli with 21.2% and 11.5%, respectively. The bivariate analysis between viral load and HAART with parasitism, demonstrated an association with parasitism.Conclusions: Enteroparasitosis is frequent in patients with HIV and diarrhea, predominating opportunistic infections over non opportunistic ones. Keywords: Parasitosis; Cystoisospora; HIV; HAART; CD4; Blastocystis hominis

Multiple groups have recently reported involvement of the gallbladder mucosa of immunocompetent patients by cystoisospora organisms. However, this has recently been disproved with the support of molecular and ultrastructural studies. Here we present a summary of these events, recounting how this pseudo-Cystoisospora epidemic began and ended. This review also highlights the important role played by ancillary techniques in supplementing the morphologic diagnosis of pathogens. [ABSTRACT FROM AUTHOR]

Twenty-four fecal samples were collected from captive amur hedgehogs (Erinaceus amurensis) in Zhengzhou, China. Based on morphological and molecular analysis, the overall prevalence of Cystoisospora was 62.5% (15/24). These samples contained two types of coccidian oocysts, including C. rastegaievae (50.0%, 12/24) and a new species named C. yuensis n. sp. (12.5%, 3/24). Sporulated oocysts (n = 30) of C. yuensis n. sp. are ovoid, (20.6 ± 1.4) μm × (20.9 ± 0.9) μm, with a shape index (length/width) of 1.0 and a smooth and bi-layered oocyst wall, 1.3 μm thick (outer layer 0.8 μm, inner 0.5 μm). A polar granule is present, but micropyle cap, micropyle, and oocyst residuum are absent. The sporocysts are ovoid-shaped, (9.3 ± 0.6) μm × (8.5 ± 1.1) μm, with a shape index (length/width) of 1.1. Stieda, substieda bodies, and refractile bodies are absent. Residuum is scattered and distributed around the entire sporocysts. At the 18S rRNA locus, C. yuensis n. sp. exhibited the highest identity to C. timoni (99.3%) from a slender-tailed meerkat. It has 98.0% identity at the 28S rRNA locus and 99.3% at the ITS locus. Based on morphological and molecular data, this isolate is a new species of Cystoisospora. Additionally, we have provided data on the prevalence of C. rastegaievae in China and sequences of the 18S rRNS, 28S rRNA, and ITS loci. [ABSTRACT FROM AUTHOR]

Abstract There is great diversity in swine coccidia, which are responsible for causing intestinal disorders ranging from sporadic diarrhea to severe cases of hemorrhagic enteritis. Thus, determining the species of coccidia that affect the animals of a region and associating them with the characteristics of the farms become extremely important. The objective of this study was to determine the prevalence of coccidia parasites in pigs reared in a family farming production system in the Semiarid Region of the State of Paraíba, Northeast Brazil. Fecal samples for analysis were collected from 187 pigs on 51 farms. For morphological analysis, 1,590 sporulated oocysts were used. The prevalence of oocysts in fecal samples was 56.6% (106/187). The most prevalent species were Eimeria suis (21.9%), followed by Eimeria neodebliecki (16.6%), Eimeria perminuta (14.9%), Eimeria polita (12.8%), Eimeria debliecki (10.6%), Eimeria porci (10.1%), Cystoisospora suis (3.7%), Eimeria scabra (1.6%) and Eimeria cerdonis (0.5%). It can be concluded that pigs from the Semiarid Region of the State of Paraíba were parasitized by a diversity of coccidia species, mainly of the genus Eimeria, and predominantly presented with mixed infections occurring in the subclinical form.

Wild ungulates have expanded their geographical range across Europe and Portugal is no exception. Despite the known benefits associated with the increase of these populations (e.g. increased prey for wild carnivores), the negative impacts also need to be taken into account (e.g. damages in agriculture and forestry, ungulate–vehicle collisions). Additionally, their role as reservoirs of zoonotic agents has gained scientific relevance due to the potential human health risks, impact on livestock and food safety. In northeast Portugal, Montesinho Natural Park, three species of ungulates occur in sympatry, the wild boar Sus scrofa, the red deer Cervus elaphus and the roe deer Capreolus capreolus. Considering their close association with humans and livestock, it is essential to understand their role as reservoirs of infectious diseases, namely as vectors for parasitic infections. In order to achieve this, 112 fresh faecal samples were collected to assess, by means of coprological analyses, their parasite diversity, prevalence and mean intensity. In total, 88 (78.60%, ±69.81–85.76) samples were infected with at least one parasite species. Parasite prevalence was different among the three species, with the red deer showing higher prevalence values (83.6%), then the wild boar (80.2%) and the roe deer (46.7%). The results have revealed that these species carry parasites that not only represent a health problem for domestic ruminants and domestic pigs (e.g. Muellerius sp., Trichostrongylidae, Strongylidae, Metastrongylus sp., Moniezia benedeni, Eimeria spp. and Cystoisospora sp.) but they can also pose a potential public health risk (e.g. Balantidium coli). The implementation of surveillance programs must include regular monitoring protocols of wild ungulates. [ABSTRACT FROM AUTHOR]

Objectives: There is recently reported increased prevalence of Isospora organisms in cholecystectomy specimens from immunocompetent patients, especially in acalculous cholecystectomies. We performed an ultrastructural and molecular evaluation of these specimens.Methods: From 28 gallbladders with intraepithelial inclusions, two specimens with diffuse involvement of the gallbladder epithelium were analyzed by electron microscopy. Polymerase chain reaction was performed on five samples for the ITS2 region of C belli and eukaryotic 18S region. The 18S products were sequenced by next-generation sequencing.Results: Electron microscopic analysis showed cytoplasmic condensations leading to vacuole formation. In contrast with true C belli, there were no identifiable organelles or organization. None of these cases showed amplified products other than human on molecular analysis.Conclusions: Electron microscopic analysis demonstrates that the inclusions are condensed cytoplasmic material and not true organisms. [ABSTRACT FROM AUTHOR]

These updated guidelines from the Infectious Diseases Community of Practice of the American Society of Transplantation review the diagnosis, prevention, and management of intestinal parasites in the pre‐ and post‐transplant period. Intestinal parasites are prevalent in the developing regions of the world. With increasing travel to and from endemic regions, changing immigration patterns, and the expansion of transplant medicine in developing countries, they are increasingly recognized as a source of morbidity and mortality in solid‐organ transplant recipients. Parasitic infections may be acquired from the donor allograft, from reactivation, or from de novo acquisition post‐transplantation. Gastrointestinal multiplex assays have been developed; some of the panels include testing for Cryptosporidium, Cyclospora, Entamoeba histolytica, and Giardia, and the performance is comparable to conventional methods. A polymerase chain reaction test, not yet widely available, has also been developed to detect Strongyloides in stool samples. New recommendations have been developed to minimize the risk of Strongyloides donor‐derived events. Deceased donors with epidemiological risk factors should be screened for Strongyloides and recipients treated if positive as soon as the results are available. New therapeutic agents and studies addressing the optimal treatment regimen for solid‐organ transplant recipients are unmet needs. [ABSTRACT FROM AUTHOR]

Coccidia of the genus Isospora , their origin, taxonomy, and host specificity have been discussed for many years. The crucial point in question being the division of the genus, based on distinct evolutionary history and the presence/absence of the Stieda body, into the genera Isospora (Eimeriidae) parasitizing mainly birds and reptiles, and Cystoisospora (Sarcocystidae) parasitizing mammals. The description of the majority of Isospora species from rodents is based solely on the oocysts found in their faeces. Some of them have been described with the presence of the Stieda body, some without it, and, simultaneously, for all the described species the molecular data are entirely lacking. This study reveals the origin of isosporan oocysts found in faeces of bank voles based on morphological analyses, phylogenetic analyses, and experimental infections. Morphological analyses showed the presence of the Stieda body complex on sporocysts. Phylogenetic analyses demonstrated close phylogenetic relationships between Isospora from bank voles and avian isosporans. Experimental inoculations of bank voles with sporulated oocysts of Isospora did not result in the production of unsporulated oocysts. Hence, these organisms should be considered pseudoparasites of the bank voles/rodents (probably originating from avian Isospora species). [ABSTRACT FROM AUTHOR]

Introduction: Cryptosporidium , Cystoisospora , and Giardia duodenalis are gastrointestinal protozoa parasites that cause diarrhea in various animals. However, information regarding the detection and phylogenetic characterization of gastrointestinal protozoa parasites in cats is limited throughout South Korea. Therefore, this study aimed to determine the detection and identify subspecies of gastrointestinal protozoa parasites in cats from South Korea., Methods: A total of 290 fecal samples were collected from stray, companion, and shelter cats in six provinces. Cryptosporidium , Cystoisospora , and G. duodenalis were identified by PCR. All positive samples were subtyped by PCR and sequencing of gp60 , ITS-1 , tpi , bg , and gdh ., Results: The overall detection of gastrointestinal protozoan parasitic infection was 17.93%. G. duodenalis was the most prevalent, with 7.93%, followed by Cystoisospora spp. (7.24%) and Cryptosporidium spp. (4.48%). In addition, C. felis (n=10), C. parvum (n=2), C. ryanae (n=1), Cystoisospora felis (n=14), Cystoisospora suis (n=5), Cystoisospora ohioensis (n=1), Cystoisospora spp. were identified in subspecies analysis of positive samples. C. felis showed a significant association with diarrhea (7.81%) and living condition (6.04%), and Cystoisospora felis in diarreha (9.38%) according to detection. Through phylogenetic analysis of the tpi, bg, and gdh genes from 23 G. duodenalispositive samples, it was confirmed that the samples of present study belonged to assemblage A, B, C, and D., Discussion: South Korean cats have a high rate of gastrointestinal protozoan parasites infection with cat-specific Cryptosporidium and Cystoisospora, which are associated with living conditions and diarrhea symptoms. Moreover, zoonotic and other animal-specific subtype of protozoan parasites have been detected in cat feces., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Yun, Moon, Lee, Kang, Ku and Hwang.)

Abstract In this study, we provide the first description of Cystoisospora infection in Asian small-clawed otters (Aonyx cinereus). In July 2017, three juvenile otters recently imported from the Republic of Indonesia showed severe diarrhea and were diagnosed with coccidial infection; two of them eventually died. Fecal examination revealed the presence of numerous oocysts. Sporulated oocysts showed typical Cystoisospora features, measuring 24.6 ± 1.6 (22.0–27.0) × 21.8 ± 1.4 (19.0–25.0) μm, with an oocyst length/width ratio of 1.1 ± 0.1 (1.0–1.3). Each sporocyst contained four sporozoites in a head-to-tail arrangement. The Stieda body was absent, and the sporocyst residuum was present. These morphological characteristics differentiated this species from the other valid Cystoisospora species described from mustelids. Molecular analysis was conducted at two loci: the nuclear 18S ribosomal RNA and mitochondrial cytochrome c oxidase subunit I genes. The 18S sequence showed high similarity with canine Cystoisispora ohioensis (1-bp difference, 1422/1423 [99.9%]). At the cytochrome c oxidase subunit I gene locus, the sequence from otters was identical to that of feline Cystoisospora rivolta (847/847 [100%]). Phylogenetic analyses using concatenated data demonstrated that Cystoisospora sp. from otters and C. rivolta grouped together in the same Cystoisospora clade. Based on these data, we concluded that Cystoisospora sp. detected from otters appeared to be highly similar to C. rivolta. Graphical abstract Image 1 Highlights • We characterized Cystoisospora infection in three Asian small-clawed otters. • Two otters showed anorexia, green diarrhea, and vomiting. • Fecal examination revealed numerous oocysts with typical Cystoisospora features. • Sporulated oocysts were morphologically differentiated from those of other Cystoisospora spp. from mustelids. • Molecular and morphological features were highly similar to feline C. rivolta. [ABSTRACT FROM AUTHOR]

Highlights • Coccidiosis is important as cause of mortality in camels (Camelus dromedarius and C. bactrianus). • Eimeria cameli develops in the lamina propria of the small intestine; only sexual stages were identified. • Cystisospora orlovi causes severe coccidiosis of the large intestines of nursing camels; only sexual stages are known. Abstract Domesticated Old World camels (Camelus dromedarius and C. bactrianus) are important for the economy of several countries in Asia, Africa, and the Arabian Peninsula, and coccidiosis is important as a cause of mortality in juvenile camels. There is confusion concerning the species of coccidian parasites in camels and their life cycles. The objective of the present paper is to review biology of the Eimeria and Cystoisospora species in camels. The following conclusions were drawn. Although five species of Eimeria; E. cameli, E. rajasthani, E. dromedarii, E. bactriani , and E. pellerdyi were named from camels, only E. cameli, E. rajasthani, E. dromedarii have been consistently found in numerous surveys and they are morphologically distinct. We consider E. pellerdyi and E. bacterini as species enquirende/ not valid. E. cameli oocysts are distinctive, dark brown and up to 108 μm long. Its gametogonic stages and oocysts are present in the lamina propria of small intestines; only sexual stages have been confirmed. The remaining species of Eimeria (E. rajasthani and E. dromedarii) in camels are <40 μm long and their endogenous stages are unknown. There is one valid species of Cystoisospora , C. orlovi in camels and is associated with severe disease in young camels, both pastoral and stall fed camels. Camels as young as nine days old can develop severe diarrhea and can die before oocysts are detected in feces. Lesions and endogenous stages are confined to the large intestine. The main lesion is hemorrhagic, diphtheroid to hemorrhagic colitis-associated with sexual stages; asexual stages are unknown. Oocysts are rarely excreted by adult camels, and in low numbers. Therefore, infection in very young camels remains unexplained. [ABSTRACT FROM AUTHOR]

This guideline is intended as an aid in the design, implementation and interpretation of studies for the assessment of drug efficacy against Eimeria in cattle and sheep, Cystoisospora in pigs and dogs, and Cryptosporidium in cattle. It deals with the most important aspects of how to conduct both experimental and field studies for dose determination, dose confirmation and assessment of field effectiveness. Also, guidance on the selection of animals, diagnostic techniques, statistical evaluation and methods for the preparation, maintenance and use of parasites is provided. The specific management conditions that may influence the course of natural infections and consequently determine treatment schemes are mentioned and suggestions for best practice in sampling and evaluation of data prior to conducting of efficacy studies are given. The guideline is also intended to assist investigators in carrying out specific studies, provide relevant information for registration authorities involved in the decision-making process, assist in the approval of anticoccidial drugs in the target species, and facilitate the world-wide adoption of standard procedures. Although currently not implemented, issues of drug resistance testing and alternative methods for drug testing are also discussed as future issues in drug testing against mammalian coccidia. [ABSTRACT FROM AUTHOR]

Relatively little is known about protozoan parasites in African animals. Here we investigated the occurrence of protozoan parasites in mammals from South Africa. Oocysts of protozoan parasites were detected in 13 of 56 (23%) fecal samples using conventional microscopic examination methods. Cryptosporidium spp. and Cystoisospora spp. were detected in eight (14%) and five (9%) samples, respectively. Mixed parasitic infection of Cryptosporidium spp. and Cystoisospora spp. was recorded in banded mongoose ( Mungos mungo). Cryptosporidium spp. was detected for the first time in cheetah ( Acinonyx jubatus), spotted hyena ( Crocuta crocuta), and African polecat ( Ictonyx striatus). Antibodies to Toxoplasma gondii and Neospora caninum were not detected by enzyme-linked immunosorbent assay in any of 32 sera tested. We detected T. gondii by PCR in tissues of five of 243 (2%) animals: domestic dog ( Canis lupus familiaris), gerbil ( Gerbilliscus spp.), greater kudu ( Tragelaphus strepsiceros), honey badger ( Mellivora capensis), and white-tailed mongoose ( Ichneumia albicauda). Our isolation of T. gondii from white-tailed mongoose and honey badger was a unique finding. All tissue samples were negative for N. caninum. The study increases our knowledge on the occurrence of protozoan parasites in populations of wild and domestic animals in South Africa. [ABSTRACT FROM AUTHOR]

Cystoisospora is responsible for morbidity in immunocompromised patients. PCR is sensitive for diagnosing Cystoisospora; however, it needs reevaluation for differential molecular diagnosis of cystoisosporiasis. We aimed at evaluating melting curve analysis (MCA) after real-time PCR (qPCR) in diagnosis and genotyping of Cystoisospora as an alternative to conventional PCR. We included 293 diarrheic stool samples of patients attending the Department of Clinical Oncology and Nuclear Medicine of Cairo University Hospitals, Egypt. Samples were subjected to microscopy, nested PCR (nPCR) and qPCR targeting the internal transcribed spacer 2 region (ITS2) of the ribosomal RNA (r RNA) gene followed by melting temperatures (Tms) analysis and comparing the results to PCR-RFLP banding patterns. Using microscopy and ITS2-nPCR, 3.1% and 5.8% of cases were Cystoisospora positive, respectively, while 10.9% were positive using qPCR. Genotyping of Cystoisospora by qPCR-MCA revealed 2 genotypes. These genotypes matched with 2 distinct melting peaks with specified Tms at 85.8°C and 88.6°C, which indicated genetic variation among Cystoisospora isolates in Egypt. Genotype II proved to be more prevalent (65.6%). HIV-related Kaposi sarcoma and leukemic patients harbored both genotypes with a tendency to genotype II. Genotype I was more prevalent in lymphomas and mammary gland tumors while colorectal and hepatocellular tumors harbored genotype II suggesting that this genotype might be responsible for the development of cystoisosporiasis in immunocompromised patients. Direct reliable identification and differentiation of Cystoisospora species could be established using qPCR-Tms analysis which is useful for rapid detection and screening of Cystoisospora genotypes principally in high risk groups. [ABSTRACT FROM AUTHOR]