Past, current, and potential treatments for cryptosporidiosis in humans and farm animals: A comprehensive review.

The intracellular protozoan parasite of the genus Cryptosporidium is among the leading causes of waterborne diarrheal disease outbreaks throughout the world. The parasite is transmitted by ingestion of infective oocysts that are highly stable in the environment and resistant to almost all conventional disinfection methods and water treatments. Control of the parasite infection is exceedingly difficult due to the excretion of large numbers of oocysts in the feces of infected individuals that contaminate the environment and serve as a source of infection for susceptible hosts including humans and animals. Drug development against the parasite is challenging owing to its limited genetic tractability, absence of conventional drug targets, unique intracellular location within the host, and the paucity of robust cell culture platforms for continuous parasite propagation. Despite the high prevalence of the parasite, the only US Food and Drug Administration (FDA)-approved treatment of Cryptosporidium infections is nitazoxanide, which has shown moderate efficacy in immunocompetent patients. More importantly, no effective therapeutic drugs are available for treating severe, potentially life-threatening cryptosporidiosis in immunodeficient patients, young children, and neonatal livestock. Thus, safe, inexpensive, and efficacious drugs are urgently required to reduce the ever-increasing global cryptosporidiosis burden especially in low-resource countries. Several compounds have been tested for both in vitro and in vivo efficacy against the disease. However, to date, only a few experimental compounds have been subjected to clinical trials in natural hosts, and among those none have proven efficacious. This review provides an overview of the past and present anti-Cryptosporidium pharmacotherapy in humans and agricultural animals. Herein, we also highlight the progress made in the field over the last few years and discuss the different strategies employed for discovery and development of effective prospective treatments for cryptosporidiosis.

Molecular identification and genetic characteristics of Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi in human immunodeficiency virus/acquired immunodeficiency syndrome patients in Shanghai, China.

BACKGROUND: Opportunistic infections are a ubiquitous complication in human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) patients. Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi are common opportunistic intestinal pathogens in humans. In China, despite the number of HIV/AIDS patients being extremely large, only a few studies have investigated opportunistic infections caused by intestinal pathogens in this patient population. The aims of this study were to elucidate the occurrence and genetic characteristics of Cryptosporidium spp., G. duodenalis, and E. bieneusi in HIV/AIDS patients. METHODS: We collected fecal specimens from 155 HIV/AIDS patients (one from each patient). All of the specimens were examined for the presence of the pathogens by genotyping using polymerase chain reaction and sequencing of the small subunit ribosomal RNA gene for Cryptosporidium spp.; the triosephosphate isomerase, ß-giardin and glutamate dehydrogenase genes for G. duodenalis; and the internal transcribed spacer region of the rRNA gene for E. bieneusi. The Cryptosporidium-positive specimens were further subtyped by polymerase chain reacion and sequencing of the 60-kDa glycoprotein gene. RESULTS: Six (3.9%), three (1.9%), and eight (5.2%) HIV/AIDS patients were positive for Cryptosporidium spp., G. duodenalis, and E. bieneusi, respectively. No statistical differences were observed in occurrence rate between the groups by gender, clinical symptom (diarrhea), and CD4+ cell count. Four Cryptosporidium species were identified: Cryptosporidium hominis (n = 2), Cryptosporidium parvum (n = 1), Cryptosporidium meleagridis (n = 1), and Cryptosporidium andersoni (n = 2). Furthermore, two C. hominis subtypes (IeA12G3T3 and IaA28R4) were detected. Three G. duodenalis-positive specimens were successfully amplified and sequenced at the triosephosphate isomerase and ß-giardin loci, which led to the identification of assemblages C and B, respectively. Seven genotypes (D, Type IV, EbpC, Peru11, EbpD, A, and I) were identified in E. bieneusi-positive specimens. CONCLUSIONS: Our findings should increase awareness of AIDS-related opportunistic intestinal pathogens, and indicate the need for routine examination in clinical practice for the detection of Cryptosporidium spp., G. duodenalis, and E. bieneusi. Homology analyses of the three intestinal pathogens at the nucleotide and/or amino acid levels indicated their zoonotic potential.

Cryptosporidium parvum and gp60 genotype prevalence in dairy calves worldwide: a systematic review and meta-analysis.

Cryptosporidium is a significant zoonotic pathogen that often occurs in dairy cattle. We conducted a systematic review and meta-analysis of the prevalence of Cryptosporidium parvum infection in dairy calves worldwide to help improve global animal husbandry and public policy implementation. Published articles were obtained from PubMed and Web of Science from January 1, 2000 to December 31, 2021. The prevalence of C. parvum infection in dairy calves was estimated using a random effects model, and the sources of heterogeneity were explored using meta-regression. In total, 118 datasets were included in the final quantitative analysis. The results showed that the global prevalence of C. parvum in dairy calves was 21.9% (7755/42,890; 95% confidence interval: 19.9-23.9%). C. parvum infection was high in pre-weaned dairy calves (24.9%, 6706/29,753) and diarrhea dairy calves (33.6%, 1637/6077). In countries with low dairy stocking density (<10 cows/farm), the prevalence of C. parvum in dairy calves was also relatively low (15.2%, 1960/16,584). Three subtype families [IIa (72.2%, 2293/3177), IId (27.4%, 872/3177), and IIl (0.4%, 12/3177)] were detected in dairy calves globally from selected studies. C. parvum IIa was the dominant zoonotic subtype. In the IIa subtype family of C. parvum, the proportions of subtypes from high to low (top nine) were IIaA15G2R1 (32.4%, 742/2293), IIaA18G3R1 (11.8%, 271/2293), IIaA13G2R1 (8.2%, 187/2293), IIaA16G1R1 (6.4%, 147/2293), IIaA20G1R1 (3.5%, 81/2293), IIaA16G3R1 (3.4%, 78/2293), IIaA17G2R1 (2.7%, 62/2293), IIaA18G1R1 (2.5%, 58/2293), and IIaA15G1R1 (2.4%, 56/2293). In the IId subtype family of C. parvum, the proportions of subtypes (top four) were IIdA19G1 (36.0%, 314/872), IIdA15G1 (27.3%, 238/872), IIdA20G1 (16.2%, 141/872), and IIdA14G1 (13.0%, 113/872). Furthermore, IId is commonly found in China (771/872). The study results indicated that the IIa subtype family is globally prevalent, while IId is found in Asia, Europe, and Africa and IIl is only found in Europe. Diarrhea in dairy calves is associated with C. parvum infection and a significantly higher prevalence is observed in diarrheic calves. Age and stock density are two significant risk factors in the prevalence of C. parvum in dairy calves. The prevention and control of this zoonosis in dairy calves should receive greater attention, especially in regions with a high degree of intensive dairy farming.

Role of Cryptosporidium spp in Development of Colorectal Cancer.

BACKGROUND: Colorectal cancer is one of the most common malignancies in humans. About 20% of the cancer incidence was attributed to infectious agents highlighting the association between infectious agents and the development of cancers. It has been suspected that Cryptosporidium spp. infection may be correlated with colon adenocarcinoma.  Aim: investigate the percentage of cryptosporidiosis among colorectal cancer patients. SUBJECTS: 100 patients were recruited from Medical Research Institute, Alexandria University. METHODS: Fresh stool specimens were collected, homogenized and examined using direct wet mount and by permanent staining of faecal smears using Modified ZN staining. Molecular detection by PCR amplification of Cryptosporidium COWP gene. RESULTS: Significantly higher proportion of colorectal cancer patients (32.5%, 42.5%) tested positive by MZN and ELISA respectively compared to only 3.3% and 5% of positive MZN and ELISA among control group. Also, positive PCR was detected among higher proportion of colorectal cancer patients (47.5%) and only 5% of control group. Odds of colorectal cancer is 19 times among positive cases of Cryptosporidium by PCR than those without proven infection by PCR (OR 19.12; 95% CI 4.82-75.99). Comparison of the assessment of Cryptosporidium infection made by two techniques produces a kappa value of 0.770, and .759 respectively between NZN, ELISA and PCR as a gold standard, suggesting a good agreement between the two techniques and PCR. This value of kappa is significantly different from zero, K.770, p<0.001 for MZN and K.759, p<.001 for ELISA. Specificity of MZN (100%) is higher than that of ELISA (96.2%) and both reported higher specificity than sensitivity denoting that both tests are good positive to rule in the presence of infection at 40% prevalence. CONCLUSION: Cryptosporidium infection is significantly higher among cancer colon patients reinforcing that it might be considered as a likely risk factor for the development cancer colon.

Occurrence and Molecular Characterization of Cryptosporidium Infection in HIV/Aids Patients in Algeria.

The estimated prevalence rate of adults living with HIV infection in MENA is one of the lowest in the world. To date, no data on the genetic characteristics of Cryptosporidium isolates from HIV/AIDS patients in Algeria were available. This study aimed to identify Cryptosporidium species and subtype families prevalent in Algerian HIV-infected patients and contribute to the molecular epidemiology mapping of Cryptosporidium in the MENA region. A total of 350 faecal specimens from HIV/AIDS patients were analysed using microscopy, and a Cryptosporidium infection was identified from 33 samples, with 22 isolates successfully sequencing and confirming species and subtypes. Based on sequence analysis, 15 isolates were identified as C. parvum with family subtypes IIa (n = 7) and IId (n = 8), while five were identified as C. hominis (family subtypes Ia (n = 2) and Ib (n = 3)) and two as C. felis. The C. parvum subtype families IIa and IId predominated, suggesting potential zoonotic transmission. More extensive sampling of both humans and farm animals, especially sheep, goats and calves, as well as a collection of epidemiological data are needed for a better understanding of the sources of human C. parvum infections in Algeria.

Molecular identification and antiprotozoal activity of silver nanoparticles on viability of Cryptosporidium parvum isolated from pigeons, pigeon fanciers and water.

Cryptosporidium is a protozoan that causes acute gastroenteritis, abdominal pain, and diarrhea in many vertebrate species, including humans, animals and birds. A number of studies have reported the occurrence of Cryptosporidium in domestic pigeons. Thus, this study aimed to identify Cryptosporidium spp. in samples collected from domestic pigeons, pigeon fanciers, and drinking water, as well as to investigate the antiprotozoal activity of biosynthesized silver nanoparticles (AgNPs) on the viability of isolated Cryptosporidium parvum (C. parvum). Samples were collected from domestic pigeons (n = 150), pigeon fanciers (n = 50), and drinking water (n = 50) and examined for the presence of Cryptosporidium spp. using microscopic and molecular techniques. The antiprotozoal activity of AgNPs was then assessed both in vitro and in vivo. Cryptosporidium spp. was identified in 16.4% of all examined samples, with C. parvum identified in 5.6%. The highest frequency of isolation was from domestic pigeon, rather than from pigeon fanciers or drinking water. In domestic pigeons, there was a significant association between Cryptosporidium spp. positivity and pigeon's age, droppings consistency, housing, hygienic and heath conditions. However, Cryptosporidium spp. positivity was only significantly associated with pigeon fanciers' gender and heath condition. The viability of C. parvum oocysts was reduced using AgNPs at various concentrations and storage times in a descending manner. In an in vitro study, the highest reduction in C. parvum count was observed at the AgNPs concentration of 1000 µg/mL after a 24 h contact time, followed by the AgNPs concentration of 500 µg/mL after a 24 h contact time. However, after a 48 h contact time, a complete reduction was observed at both 1000 and 500 µg/mL concentrations. Overall, the count and viability of C. parvum decreased with increasing the AgNPs concentration and contact times in both the in vitro and in vivo studies. Furthermore, the C. parvum oocyst destruction was time-dependent and increased with increasing the contact time at various AgNPs concentrations.

Molecular epidemiology of Cryptosporidium species in Kpong and its environs, Ghana.

BACKGROUND: Cryptosporidium is a ubiquitous enteric protozoan pathogen infecting humans, domestic animals, and wildlife worldwide. It is a waterborne pathogen with recognized zoonotic potential and a definite cause of diarrhea and nutritional disorders in institutional and community settings. One challenge facing the world's supply of clean drinking water is contamination from feces and soil. It has been established that small quantities of oocysts, the infective stage, can cause human disease. Also, their resistance to chlorination and other water treatment procedures has been demonstrated. Kpong, a community in the Lower Manya Krobo Municipality of the Eastern Region of Ghana, is one of the primary sources of water supply to Accra, the capital city of Ghana. Being able to determine the effectiveness of water treatment processes and identifying sources of contamination of this pathogen in our water bodies is thus of public health importance. The study aimed to conduct molecular epidemiology of Cryptosporidium spp. in the Lower Manya Krobo Municipality. METHODOLOGY/PRINCIPAL FINDINGS: A total of 230 samples, 180 fecal samples from cattle and 50 water samples (tap water and well water) were collected from the following communities: Kpong, Akwernor, Ablotsi, Nuaso, and Atua, all in the Lower Manya Krobo Municipality. Samples were screened for Cryptosporidium by microscopy and PCR. The 18S rRNA gene was amplified by nested polymerase chain reaction (PCR), and the final product was sequenced. The prevalence of Cryptosporidium from the fecal samples was estimated as 10% (18/180) by microscopy, while all 50 water samples were negative. However, PCR gave the prevalence of Cryptosporidium as 47.8% (86/180) for fecal samples and 20% (10/50) for water samples. Based on the 18S rRNA gene, three sequenced samples showed high homology to C. parvum species. The phylogenetic analysis confirmed this as these sequences clustered with C. parvum sequences from other countries. CONCLUSION/SIGNIFICANCE: Cryptosporidium parvum was identified as the persistent species in the study communities. This outcome supports the evidence that domesticated animals serve as potential reservoirs of zoonotic transmission of cryptosporidiosis. The persistence of cryptosporidiosis in cattle indicates its presence in the human population. In addition, the presence of Cryptosporidium parvum in the wells makes it alarming and necessary to consider a holistic approach such as One Health Strategies to identify and control cases in humans.

American variants of Cryptosporidium hominis: Over-sexed and over here?

A new study in this issue of Cell Host & Microbe from Huang et al. provides important insights into the global epidemiology of human-infectious Cryptosporidium and mechanisms leading to the rapid emergence and rise to dominance of new, possibly more virulent, parasite strains.

Health risks of Cryptosporidium and Giardia in the application of surface water and septic tank effluent in Chinese agriculture: Impact on cancer patients identified by quantitative microbial risk assessment.

The protozoa Cryptosporidium and Giardia are major causes of diarrhea and are commonly found on vegetables in China. They pose a health risk, particularly to immunocompromised individuals, including cancer patients. A quantitative microbial risk assessment of Chinese data evaluated the risks of Cryptosporidium and Giardia exposure arising from the application of surface water and septic tank effluent to agricultural land. Exposure via agricultural produce consumption (consumers) and agricultural practices (farmers) was considered for subpopulations of cancer patients and immunocompetent people in urban and rural areas, and risk mitigation scenarios were modelled. The cumulative disease burdens attributable to cryptosporidiosis and giardiasis were, respectively, 9.68×10-6 and 5.57×10-5 disability-adjusted life years per person per year (DALYs pppy) for immunocompetent people, and 3.14×10-5 and 1.51×10-4 DALYs pppy for cancer patients. Cancer patients were approximately three times more likely to have an individual disease burden than immunocompetent people. The disease burden was higher for consumers than farmers, and higher in rural areas than urban areas (all exceeding the maximum recommended by the World Health Organization). The highest burdens were in provinces of high population, such as Henan, Guangdong, and Sichuan, while the burden associated with human and livestock fecal effluent application was higher than with surface water irrigation. Of the three vegetables studied, lettuce posed the greatest risk, followed by bok choy, while cucumber posed the least risk. Risk mitigation scenario analysis showed that pre-treatment of surface water and feces, and appropriate post-harvest handling of vegetables, including disinfection, cooking, and adequate surface heat treatment (75 °C for 60 s), should be considered when attempting to reduce disease burdens. The methodology and findings of this study are useful for evaluating and reducing the burden of Cryptosporidium and Giardia infections associated with agricultural irrigation and fertilization practices, particularly on cancer patients.

Identification and Evaluation of Cryptosporidium Species from New York City Cases of Cryptosporidiosis (2015 to 2018): a Watershed Perspective.

Watersheds that supply residents with drinking water have the potential for contamination with Cryptosporidium oocysts. To evaluate any potential similarities between Cryptosporidium species previously found in the New York City (NYC) watershed and those causing disease in NYC, the species were identified in stool specimens from residents with cryptosporidiosis. Genetic analysis was performed on 628 positive stool samples collected from NYC residents between 2015 and 2018 to determine the species present. A total of 547 samples yielded positive results by real-time PCR. Of these samples, 512 (93.6%) were identified to the species level, with 94.7% positive for either Cryptosporidium hominis or Cryptosporidium parvum (56.4% and 38.5%, respectively), including one coinfection. Less common Cryptosporidium species identified included C. felis, C. canis, C. ubiquitum, C. meleagridis, and a Cryptosporidium sp. chipmunk genotype. Results were evaluated and compared to species and genotypes of Cryptosporidium previously identified from stormwater collected within the NYC watershed. While there was overlap with some of the rare species found in case specimens, the prevalence and distribution of species did not suggest a connection between sources previously identified in the watershed and the species causing human cases of cryptosporidiosis in NYC residents. IMPORTANCE It is important to identify the species causing human cryptosporidiosis in a population in order to investigate possible sources or routes of contamination. Many species of Cryptosporidium are host-adapted and therefore have the potential to be tracked back to specific sources that can subsequently be managed. There has been no evidence to suggest that the water supply has ever been a source of cryptosporidiosis cases in NYC, and since 2013, the New York City Department of Environmental Protection has further reduced the risk of disease through the use of ultraviolet treatment to inactivate any Cryptosporidium present in the source water. However, as one of the largest unfiltered water supplies in the country, it is important to evaluate watershed sources for potential impacts to public health. In this unique study, species of Cryptosporidium causing disease in NYC residents were identified and compared with previously identified species from the watershed.

Molecular characterization and prevalence of Cryptosporidium spp. in sheep and goats in western Inner Mongolia, China.

Cryptosporidium spp. are zoonotic intestinal parasites that infect fish, birds, reptiles and mammals. Cryptosporidium spp. are common cause of diarrhea. In this study, a total of 1032 fecal samples were collected from the rectums of sheep and goats. The samples were analyzed using nested polymerase chain reaction (nested PCR) based on the small subunit ribosomal RNA (SSU rRNA) gene of Cryptosporidium spp. The average infection rate of Cryptosporidium spp. was 2.23% (n = 23), and three Cryptosporidium species were identified, namely Cryptosporidium ubiquitum (8/23), Cryptosporidium andersoni (5/23) and Cryptosporidium xiaoi (10/23). Subtyping of C. ubiquitum and C. xiaoi was carried out by DNA sequence analysis of the 60-kDa glycoprotein (gp60) gene. Eight C. ubiquitum isolates were identified as zoonotic subtype XIIa. Nine C. xiaoi isolates were identified as subtypes XXIIIc (n = 1), XXIIIf (n = 3) and XXIIIg (n = 5). Subtype XXIIIg was first found in Chinese sheep. C. ubiquitum subtype XIIa was found in both sheep and goats, suggesting that sheep and goats are important sources of C. ubiquitum infections.

First report on occurrence and genotypes of Enterocytozoon bieneusi, Cryptosporidium spp. and Cyclospora cayetanensis from diarrheal outpatients in Ningbo, Southeast China.

Enterocytozoon bieneusi, Cryptosporidium spp. and Cyclospora cayetanensis are three important zoonotic pathogens which were a major cause of foodborne or waterborne intestinal diseases in humans and animals. However, very little data about occurrence and genotypes of the three parasites in Ningbo in the south wing of the Yangtze River Delta, China, which is important for a tourist city. In the present study, molecular characterization of E. bieneusi, C. cayetanensis and Cryptosporidium spp. in fecal samples from 489 diarrheal outpatients were carried out. As a result, a total of 35 (7.16%, 35/489) and three (0.61%, 3/489) samples were positive for E. bieneusi and C. cayetanensis respectively. No Cryptosporidium-positive sample or mixed-infections were detected. Four known E. bieneusi genotypes (Type IV, D, I and CHN4) and 8 novel genotypes (NBH1-NBH8) were identified with type IV was the dominant genotype (n = 14), followed by genotypes D (n = 5), NBH8 (n = 5) and NBH7 (n = 3). The remaining genotypes were found in one sample each, and these genotypes were belonged to the previously described high-potential zoonotic group 1. One novel sequence named NBC315, and the other two sequences (NBC30 and NBC370) identical with the reported sequence were detected. Therefore, the existence and importance of zoonotic potential of E. bieneusi and C. cayetanensis in diarrheal outpatients in Ningbo indicates the public health threats, and more investigations should be carried out in human populations, animals and other environmental sources from the One Health perspective.

Detection of unusual Cryptosporidium parvum subtype in patients with gastrointestinal cancer in Egypt.

While the importance of cryptosporidiosis in immunocompromised persons is well known, the prevalence of Cryptosporidium spp. in cancer patients is not clear. The current study was designed to assess the occurrence and genetic characteristics of Cryptosporidium spp. in patients with gastrointestinal (GI) cancer in Egypt. Stool samples were collected from 100 patients with GI malignancies and 20 healthy individuals without any GI manifestations (control group). They were screened by microscopy and the immunochromatographic RIDA®QUICK Cryptosporidium kit. Subtyping of Cryptosporidium spp. was conducted by sequence analysis of the glycoprotein 60 (gp60) locus. Sociodemographic, environmental data and information on GI symptoms, cancer types, and clinical treatment were obtained via a questionnaire. By microscopy and RIDA®QUICK, only 7% (7/100) of GI cancer patients were positive for Cryptosporidium, compared with 40% (40/100) by gp60 nPCR. No positives were obtained from the control group. Male sex (P = 0.02) and younger age (P = 0.004) were major Cryptosporidium risk factors for infection. The occurrence of Cryptosporidium was also significantly more frequent (P = 0.003) in watery stool samples. Sequence analysis of the gp60 amplicons (~ 400 bp) identified a novel C. parvum subtype with nine TCA repeats and eleven ACATCA repeats. A formal subtype designation could not be made due to the short sequence length. More studies should be conducted to verify the common occurrence of this unusual C. parvum subtype and establish its genetic identity.

Zoonotic potential of guinea pigs: Outbreak of cryptosporidiosis combined with chlamydiosis in a breeding guinea pig herd.

INTRODUCTION: In a guinea pig herd with 26 breeding animals, several individuals of all age categories died (16/26) after three animals had been newly introduced from another herd. Furthermore, the population suffered of apathy, anorexia, severe weight loss and conjunctivitis, as well as abortions and stillbirths. At the same time, the owner experienced a SARS-CoV-2 infection with pneumonia, which was confirmed by taking a PCR test. Chlamydia caviae was detected from the conjunctiva and vagina/uterus in one juvenile animal together with an intestinal Cryptosporidium wrairi infection. Oocysts were found histologically in the small intestine, which was confirmed by PCR. C. wairi is a parasite adapted to guinea pigs with zoonotic potential, which causes diarrhoea with frequent deaths in larger guinea pig herds. C. caviae is also a zoonotic pathogen and often the cause of conjunctivitis, pneumonia and abortions in guinea pigs and can lead to upper respiratory tract disease, conjunctivitis but also severe pneumonia in humans. The increased death cases and the clinical signs could be traced back to an infection with Cryptosporidium wrairi, complicated by a co-infection of C. caviae. We suspect that the abortions were caused by C. caviae, but since the population was treated with various antibiotics effective against chlamydial infections, it was no longer possible to verify this by PCR testing. Unfortunately, more animals succumbed and finally only two animals of the originally 26 were left. With this case report, we would like to point out to veterinarians that guinea pigs can be an important source of zoonotic infections for various pathogens, especially since they are popular pets and often come into close contact with children where hygiene might not always be strictly followed.

INTRODUCTION: Dans un groupe de cobayes de 26 animaux reproducteurs, plusieurs individus de toutes les catégories d'âge sont morts (16/26) après l'introduction de trois animaux provenant d'un autre groupe. En outre, la population a souffert d'apathie, d'anorexie, de perte de poids sévère et de conjonctivite ainsi que d'avortements et de mortinatalité. La présence de Chlamydia caviae a pu être détectée dans la conjonctive et le vagin/utérus d'un animal juvénile, ainsi qu'une infection intestinale à Cryptosporidium wrairi. Des oocystes ont été trouvés histologiquement dans l'intestin grêle, ce qui a été confirmé par PCR. C. wairi est un parasite adapté aux cobayes avec un potentiel zoonotique, qui provoque des diarrhées avec des morts fréquentes dans les grands groupes de cobayes. C. caviae est également un agent pathogène zoonotique et est souvent à l'origine de conjonctivites, de pneumonies et d'avortements chez les cobayes ; il peut entraîner des maladies des voies respiratoires supérieures, des conjonctivites mais aussi des pneumonies graves chez l'homme. L'augmentation des cas de décès et les signes cliniques pourraient être attribués à une infection par Cryptosporidium wrairi, compliquée par une co-infection par C. caviae. Nous soupçonnons que les avortements ont été causés par C. caviae, mais comme la population a été traitée avec divers antibiotiques efficaces contre les infections à chlamydia, il n'était plus possible de le vérifier par des tests PCR. Malheureusement, d'autres animaux ont succombé et il ne restait finalement que deux animaux sur les 26 d'origine. Avec ce rapport de cas, nous aimerions attirer l'attention des vétérinaires sur le fait que les cochons d'Inde peuvent être une source importante d'infections zoonotiques pour divers pathogènes, d'autant plus qu'il s'agit d'animaux de compagnie populaires qui sont souvent en contact étroit avec des enfants avec lesquels l'hygiène n'est pas toujours strictement respectée.

Molecular evaluation of Cryptosporidium spp. among breeding calves of Lorestan province Western Iran.

BACKGROUND: Cryptosporidium spp. are opportunistic intestinal protozoans with global distribution and are of great importance as zoonotic protozoans are common to humans and domestic animals, including cattle and calves. Identification and detection of parasite species using precise methods including molecular methods can be an effective step in treating and controlling parasites. OBJECTIVES: This study aimed to investigate the prevalence of Cryptosporidium among breeding calves of Khorramabad city, Lorestan province, Western Iran, using PCR. METHODS: The faecal samples were taken from 181 healthy and diarrhoeal calves and after the Ziehl Neelsen Acid-fast staining and microscopic evaluation, the genomic DNA was extracted for molecular evaluations. To detect Cryptosporidium species, specific primers targeting the SAM-1 gene of Cryptosporidium and a commercial master mix were used for PCR. RESULTS: Out of 181 faecal samples of breeding calves in Khorramabad city, 9 samples (5%) were positive for Cryptosporidium spp. using the PCR method. Statistical analysis of the data showed that there was no significant statistical relationship between Cryptosporidium infection of the calves and variables of age, breed, type of water consumption, clinical signs of diarrhoea, and sampling location, while parasite infection had a significant relationship with calf gender so that all Cryptosporidium positive samples were from male calves (p ≤ 0.05). CONCLUSIONS: Considering the presence of Cryptosporidium infection, the region's traditional grazing system, and the close relationship between livestock and humans, there is a possibility of human infection in the region. So preventive measures such as periodic animal testing with sensitive and accurate diagnostic techniques including PCR, pharmacological treatment of livestock, water hygiene and the use of industrial grazing instead of traditional grazing to improve the hygiene of food consumed by livestock are recommended.

Gastrointestinal parasites in young dogs and risk factors associated with infection.

Young dogs are particularly susceptible to infections with endoparasites. The occurrence of endoparasites was investigated in young dogs from Central Germany between July 2020 and July 2022. In total, 386 fecal samples originating from 171 dogs were examined for the prevalence of endoparasites using a combined flotation- and sedimentation technique and conventional PCR. Overall, in 41.2% (159/386) of the examined samples, endoparasites were detected. The most frequently occurring endoparasites were Giardia duodenalis (29%), Cryptosporidium spp. (9.1%), Cystoisospora spp. (7.3%), and Toxocara canis (6%). Sequencing of G. duodenalis positive samples showed that most infections belonged to the host-specific assemblages C (38.4% (43/112)) and D (35.7% (40/112)). The zoonotic assemblage A was identified in 8% (9/112) of the samples. Moreover, mixed infections were observed as follows: C/D in 5 (4.5%), D/A in 4 (3.6%), and C/A in 3 (2.7%) samples. All assemblage A infections were assigned to the potentially zoonotic subassemblage AI. Co-infections of G. duodenalis and Cryptosporidium spp. were observed in 3.1% (12/386) of the samples. Analyzing several host factors for their potential association with endoparasitic infection, the origin of dogs, as well as the living environment were identified as the main risk factors for infection with endoparasites. Overall, this study shows a high infection rate with endoparasites, especially G. duodenalis, in young dogs from Germany. The results of this study contribute to further insight into the distribution and potential risk factors associated with endoparasitic infections, as well as the zoonotic potential these parasites may present.

Cross-genera amplification and identification of Colpodella sp. with Cryptosporidium primers in fecal samples of zoo felids from northeast China / Amplificação cruzada de gêneros e identificação de Colpodella sp. com iniciadores de Cryptosporidium em amostras fecais de zoofilia do nordeste da China

Abstract The protozoans include many intracellular human pathogens. Accurate detection of these pathogens is necessary to treat the diseases. In clinical epidemiology, molecular identification of protozoan is considered a more reliable and rapid method for identification than microscopy. Among these protozoans, Cryptosporidium considered being one of the important water-borne zoonotic pathogens and a major cause of a diarrheal disease named cryptosporidiosis in humans, domestic animals, and wild animals. This study was aimed to identify Cryptosporidium in zoo felids (N= 56) belonging to different zoo of China, but accidentlly Colpodella was encountered in the zoo felids sample and phylogenetic data confirmed this unexpected amplification from fecal samples using two-step nested-PCR. Phylogenetic analysis revealed the fact about the specific primers used previously by many researchers and cross-genera amplification. We came to know that genetically sequenced amplicon gives more accurate identification of species. This study suggests more investigation on Colpodella which has been neglected previously but gains the attention of researchers after identified from humans and animals and has been known to correlate with neurological symptoms in patients.

Resumo Os protozoários incluem muitos patógenos humanos intracelulares. A detecção acurada desses patógenos é necessária para tratar as doenças. Na epidemiologia clínica, a identificação molecular de protozoários é considerada o método de identificação mais confiável e rápido do que a microscopia. Entre esses protozoários, o Cryptosporidium é considerado um dos importantes patógenos zoonóticos transmitidos pela água e uma das principais causas de uma doença diarreica denominada criptosporidiose em humanos, animais domésticos e selvagens. Este estudo teve como objetivo identificar Cryptosporidium em zoofelídeos (N = 56) pertencentes a diferentes zoológicos da China, mas acidentalmente Colpodella foi encontrada na amostra de zoofelídeos e os dados filogenéticos confirmaram essa amplificação inesperada de amostras fecais usando nested-PCR em duas etapas. A análise filogenética revelou o fato sobre os primers específicos usados ​​anteriormente por muitos pesquisadores e a amplificação entre gêneros. Ficamos sabendo que o amplicon sequenciado geneticamente fornece uma identificação mais acurada das espécies. Este estudo sugere mais investigação sobre Colpodella, que foi negligenciada anteriormente, mas ganha a atenção dos pesquisadores depois de identificada em humanos e animais e é conhecida por se correlacionar com sintomas neurológicos em pacientes.

Global prevalence and risk factors of Cryptosporidium infection in Equus: A systematic review and meta-analysis.

Intoduction: Cryptosporidiosis is a zoonotic disease caused by Cryptosporidium infection with the main symptom of diarrhea. The present study performed a metaanalysis to determine the global prevalence of Cryptosporidium in Equus animals. Methods: Data collection was carried out using Chinese National Knowledge Infrastructure (CNKI), VIP Chinese journal database (VIP), WanFang Data, PubMed, and ScienceDirect databases, with 35 articles published before 2021 being included in this systematic analysis. This study analyzed the research data through subgroup analysis and univariate regression analysis to reveal the factors leading to high prevalence. We applied a random effects model (REM) to the metadata. Results: The total prevalence rate of Cryptosporidium in Equus was estimated to be 7.59% from the selected articles. The prevalence of Cryptosporidium in female Equus was 2.60%. The prevalence of Cryptosporidium in Equus under 1-year-old was 11.06%, which was higher than that of Equus over 1-year-old (2.52%). In the experimental method groups, the positive rate detected by microscopy was the highest (10.52%). The highest Cryptosporidium prevalence was found in scale breeding Equus (7.86%). The horses had the lowest Cryptosporidium prevalence (7.32%) among host groups. C. muris was the most frequently detected genotype in the samples (53.55%). In the groups of geographical factors, the prevalence rate of Cryptosporidium in Equus was higher in regions with low altitude (6.88%), rainy (15.63%), humid (22.69%), and tropical climates (16.46%). Discussion: The search strategy use of five databases might have caused the omission of some researches. This metaanalysis systematically presented the global prevalence and potential risk factors of Cryptosporidium infection in Equus. The farmers should strengthen the management of young and female Equus animals, improve water filtration systems, reduce stocking densities, and harmless treatment of livestock manure.

Cryptosporidium spp. and Blastocystis spp. coinfection as a reason of an acute diarrhea in a young healthy veterinary Polish student - Case report.

OBJECTIVES: Cryptosporidium belongs to enteric parasites responsible for prolonged symptoms in the gastrointestinal tract, both in immunocompetent and immunocompromised individuals. One of the risk factors of infection is contact with an infected person or animals (cattle). The case is described of a young man admitted to the Department of Tropical and Parasitic Diseases of the Medical University in Poznan, Poland, because of watery diarrhea with high fever and in whom symptomatic treatment did not produce any improvement. MATERIAL AND METHODS: A 21-year -old male was examined and his epidemiologic history obtained. Primary blood test, anti- Toxocara IgG (ELISA) and anti-Toxoplasma gondii IgG and IgM (ELISA) were performed. PCR detected 16 enteropathogens in a stool sample. Microscopic parasitic stool examination was also performed based on Ziehl-Neelsen method, which allowed the assessment of the presence of cryptosporidium life stages. RESULTS: Epidemiology data provided information that the patient was a veterinary student who therefore had many contacts with domestic animals. Multiplex PCR detected a genetic material of Cryptosporidium. The result was confirmed with repeated positive direct stool examinations which gave the evidence of Cryptosporidium spp. oocysts and vacuolar forms of Blastocystis spp.. CONCLUSIONS: 1) Cryptosporidium is responsible for watery diarrhea in healthy individuals. 2) Contact with animals (cattle) is a potential risk factor for infection. 3) Protozoan co-infection should be treated to shorten the symptomatic period and to avoid post-infection complications. 4) Different diagnostic methods increase the possibility to establish appropriate diagnosis.

Distribution of Cryptosporidium parvum subgenotypes in pre-weaned calves in Germany.

Cryptosporidium parvum is a main cause of severe diarrhea in young calves leading to economic loss and animal suffering. Little is known about the epidemiology of the genetic subtypes that may differ in their infectiousness and zoonotic potential. The present study analyzes the distribution of Glycoprotein 60 (GP60) variants in C. parvum positive samples obtained from young calves in Germany where individual fecal samples of 441 pre-weaned calves collected from 98 German dairy herds were evaluated for cryptosporidiosis (Göhring et al., 2014). DNA was extracted for Restriction Fragment Length Polymorphism (RFLP) analysis from 268 samples that were positive for Cryptosporidium by coproantigen EIA originating from 91 farms confirming C. parvum as the only species occurring. GP60 subtype analysis revealed 12C. parvum subtypes, all belonging to the zoonotic allele family IIa. The most widespread subtype was IIaA15G2R1 (71.4%). Other subtypes occurred sporadically: IIaA16G2R1, IIaA16G3R1 and IIaA17G2R1 (6.2%), IIaA13G1R2 (3.5%), IIaA14G1R1 (3.1%), IIaA14G2R1 (1.3%), IIaA17G1R1 (0.9%) and IIaA13G2R1, IIaA16G1R2, IIaA17G1R2 and IIaA24G1R1 (0.4%). Altogether a high diversity of subgenotypes was found with IIaA15G2R1 clearly dominating irrespective of geographic region. All subgenotypes are considered zoonotic underscoring that pre-weaned calves are likely to be a reservoir for zoonotic C. parvum and thus may be a risk for animal-to-human transmission.