Cryptosporidium Spp Research Articles

Assessment of a Multiplex PCR for the Simultaneous Diagnosis of Intestinal Cryptosporidiosis and Microsporidiosis: Epidemiologic Report from a French Prospective Study

Microsporidiosis and cryptosporidiosis are associated with chronic diarrhea in immunocompromised patients. The objectives of this study were to: i) assess a multiplex quantitative PCR assay targeting Cryptosporidium spp and the microsporidian Enterocytozoon bieneusi and Encephalitozoon spp, and ii) provide an update on the epidemiology of these pathogens. A prospective study was conducted from January 2017 to January 2019. Performance of the assay was assessed, and all cryptosporidia and microsporidia isolates were genotyped. The sensitivity of the multiplex PCR method reached 1 copy/μL for each targeted pathogen. The sensitivity of co-proantigen testing in the diagnosis of cryptosporidiosis was 73%. The sensitivity of microscopy in the diagnosis of cryptosporidiosis was 64%, and microsporidiosis, 50%. Among the 456 patients included, 14 were positive for Cryptosporidium spp (4 different species); 5, for E. bieneusi; and 2, for Encephalitozoon intestinalis. The overall prevalence of cryptosporidia was 3.1%, and of microsporidia, 1.5%; in kidney transplant recipients (n= 82), corresponding values were 7.3% and 2.4% (6 and 2 patients), respectively. Two cases of E. intestinalis infection were diagnosed in children who had traveled to the tropics. This study is the first to assess a multiplex quantitative PCR method for the simultaneous diagnosis of intestinal microsporidiosis and cryptosporidiosis. The highest prevalences of both pathogens were observed in kidney transplant recipients.

Experimental infection of cats with Cystoisospora felis

Background Cystoisospora felis is a common parasite of cats and is diagnosed by fecal flotation, but false‐negative results can be common.Hypothesis/ObjectivesTo experimentally inoculate cats with C. felis oocysts, to compare fecal flotation and polymerase chain reaction (PCR) results, and to describe any clinical signs consistent with infection.AnimalsSix cats.Methods Cystoisospora felis oocysts were identified morphologically from feces of a naturally infected kitten with diarrhea, sporulated oocysts (5000) were inoculated to 6 cats that were negative for fecal parasites by fecal flotation and by a fluorescent antibody assay (FA) for Giardia spp. and Cryptosporidium spp. Cats were observed daily for the presence of clinical signs consistent with infection. Fecal samples were evaluated by fecal flotation and FA up to 3 times per week post inoculation (PI) to Day 27. Thirty‐six samples collected before inoculation and from Days 8, 10, 13, 15, and 20 PI were assayed using an internal transcribed spacer 1 (ITS1) PCR that amplifies DNA of C. felis.ResultsAll cats were negative for C. felis by both assays before inoculation. All cats shed C. felis oocysts by Day 10 PI, oocysts were not detected by fecal flotation after Day 15 PI. Cystoisospora felis DNA was amplified from 24/36 (66.6%) fecal samples from 6/6 (100%) of the cats. Oocysts were not detected by fecal flotation in 4 of the samples that were positive for C. felis DNA by PCR. Clinical signs were not recognized in any of the study cats.Conclusions and Clinical ImportanceFecal flotation is a convenient assay for detection of C. felis but could occasionally give false‐negative results when compared to this ITS1 PCR.

Human Astrovirus and Cryptosporidium Co-infection among Children with Gastroenteritis in Diyala Governorate

Background: Gastroenteritis is the second leading cause of death in children under five years old. The highest mortality from diarrheal diseases in children lived crowded areas were poor sanitary and hygienic conditions found. Objective: To determine the co-infections between human astrovirus and Cryptosporidium spp. among children with gastroenteritis below 5 years of age in the Diyala governorate. Patients and Methods: A cross-sectional study was carried out for 100 patients with acute gastroenteritis who attended the Emergency Department of Pediatrics in Al-Batool Teaching Hospital for Maternity and Pediatric in Baqubah city, during the period from July 2019 to February 2020. Real time-PCR was used to detect human astrovirus while enzyme- linked immunosorbent assay was used to detect Cryptosporidium spp. A specific formula sheet was used to collect demographic data such as age, gender, mother education, and type of milk feeding. Results: Out of 100 samples, 14 samples were positive for human astrovirus and 38 samples were positive for Cryptosporidium. The infection rate is more common in males (9 cases) (25cases) rather than females, and in age group less than years 11(78.58%) and 34(89.47%) respectively. Artificial feeding showed higher frequencies of infection than others in the study population. Co-infection astrovirus and Cryptosporidium found in four patients, all of whom were males aged under one-year and the difference was statistically significant with all parameters. Conclusion: A high proportion of positive cases for human astrovirus and Cryptosporidium spp. were notes in males and the age group less than one year’s co-infection showed high frequency in the study population. Keywords: Gastroenteritis, human astrovirus, Cryptosporidium, co-infection

Frequency of Cryptosporidiosis in Children having Persistent Diarrhea.

Objective:Globally childhood diarrheal diseases continue to be the second leading cause of death. Cryptosporidium spp are important intestinal parasites that cause diarrhea in humans and animals particularly in developing countries. This investigation was carried out to find out the frequency of cryptosporidiosis in children presenting with persistent diarrhea.Methods:Two hundred stool samples were collected in this descriptive cross-sectional study conducted at Microbiology Department, Combined Military Hospital, Lahore Pakistan between the months of July to Dec 2014. Children aged five years to 12 years who presented with persistent diarrhea were included in the study. Stool specimens were processed using the modified acid-fast staining method, and microscopically examined for Cryptosporidium infection.Results:The average age of study participants was 7.95 with a standard deviation of 2.21 years. Among the participants 66% were males whereas 34% were females. Twenty eight percent had presence of oocysts in stool samples.Conclusions:The frequency of Cryptosporidiosis among children with persistent diarrhea was 28%. This high frequency indicates that this population is uniquely susceptible to infection. It also highlights the need for education about hygiene, accurate diagnosis, and treatment of Cryptosporidiosis. There is also a need for additional studies regarding the occurrence of this pathogen.

Performance and operational feasibility of two diagnostic tests for cryptosporidiosis in children (CRYPTO-POC): a clinical, prospective, diagnostic accuracy study

SummaryBackgroundCryptosporidiosis is a common cause of diarrhoea in young children (aged younger than 24 months) in low-resource settings but is currently challenging to diagnose. Light-emitting diode fluorescence microscopy with auramine-phenol staining (LED-AP), recommended for tuberculosis testing, can also detect Cryptosporidium species. A lateral-flow test not requiring refrigerator storage (by contrast with most immunochromatographic lateral-flow assays) has also recently been developed for Cryptosporidium spp detection. We aimed to evaluate the diagnostic accuracy and operational feasibility of LED-AP and the lateral-flow test strip for cryptosporidiosis in children.MethodsWe did a prospective diagnostic accuracy study in two health-care facilities in Ethiopia, in a consecutive series of children younger than 5 years of age with diarrhoea (three or more loose stools within the previous 24 h) or dysentery (at least one loose stool with stains of blood within the previous 24 h). Stool samples were tested for Cryptosporidium spp by LED-AP and the lateral-flow test strip; accuracy of each test was estimated by independent and blind comparison with a composite reference standard comprising quantitative immunofluorescent antibody test (qIFAT), ELISA, and quantitative PCR (qPCR). Quantitative cutoff values for diarrhoea-associated infection were established in an embedded case-control substudy, with cases of cryptosporidiosis coming from the 15 districts in and around Jimma and the eight districts surrounding Serbo, and community controls without diarrhoea in the previous 48 h recruited by weekly frequency matching by geographical district of the household, age group, and enrolment week.FindingsStool samples from 912 children with diarrhoea or dysentery and 706 controls from the case-control substudy were tested between Dec 22, 2016, and July 6, 2018. Estimated reference-standard cutoff values for cryptosporidiosis positivity were 2·3 × 105 DNA copies per g of wet stool for qPCR, and 725 oocysts per g for qIFAT. LED-AP had a sensitivity for cryptosporidiosis of 88% (95% CI 79–94; 66 of 75 samples) and a specificity of 99% (98–99; 717 of 726 samples); the lateral-flow test strip had a sensitivity of 89% (79–94; 63 of 71 samples) and a specificity of 99% (97–99; 626 of 635 samples).InterpretationLED-AP has high sensitivity and specificity for cryptosporidiosis and should be considered as a dual-use technology that can be easily integrated with existing laboratory infrastructures in low-resource settings. The lateral-flow test strip has similar sensitivity and specificity and provides an alternative that does not require microscopy, although purchase cost of the test strip is unknown as it is not yet available on the market.FundingNorwegian Research Council GLOBVAC fund, The Bill & Melinda Gates Foundation, Norwegian Society for Medical Microbiology, University of Bergen, and Vestfold Hospital Trust.

Isolation and Genetic Characterization of Cryptosporidium from Captive Wildlife of India

Background: Cryptosporidiosis is an emerging zoonotic protozoan disease caused by Cryptosporidium spp. The infection was reported worldwide from domestic animals and humans, including wild animals. From India, no such reports were published on Cryptosporidium infection in captive wildlife. Hence, a pilot study was conducted to report the occurrence of Cryptosporidium infection in captive wildlife of India. Methods: Faecal samples (n=788) were collected from 127 captive wildlife species of three zoological parks viz., Sri Venkateswara Zoological Park (SVZP), Tirupati (n=242); Indira Gandhi Zoological Park (IGZP), Visakhapatnam (n=218); Nehru Zoological Park (NZP), Hyderabad (n=328) and screened for Cryptosporidium infection. Preliminary screening of faecal samples was done for the detection of Cryptosporidium oocysts by modified Ziehl-Neelsen (mZN) staining method and the test positives were confirmed by nested PCR targeting 18S rRNA gene. Nested PCR amplicons were sequenced for determining the Cryptosporidium species. The resultant data were statistically analyzed by Fisher/Chi square, Fisher Exact test using SPSS software v 17.0.Result: In mZN staining method, 7.23 percent of isolates were found to be positive for Cryptosporidium and the highest rate of infection was detected in wildlife at NZP, Hyderabad (8.23%), followed by SVZP, Tirupati (7.44%) and IGZP, Visakhapatnam (5.50%). Cryptosporidium positive faecal samples by mZN staining were further confirmed by nested PCR and positive amplicons were sequenced for determination of Cryptosporidium species. Genetic characterization revealed five species viz., Cryptosporidium parvum; C. ryanae, C. suis, C. muris and Cryptosporidium avian genotype III. The study conclude that, Cryptosporidium infection was prevalent in the captive wildlife from the zoological parks of India and species variation was marked among the wildlife. Based on the available literature, the current study is the first of its kind on the prevalence of Cryptosporidium in captive wildlife from India.

Detection of Intestinal Parasites and Genetic Characterization of Enterocytozoon bieneusi in Hedgehogs from China.

Microsporidia are a group of unicellular and opportunistic intestinal parasites in which Enterocytozoon bieneusi is a frequent species causing microsporidial infections in humans. Many domesticated and wild animals have been shown to be hosts of E. bieneusi and other microsporidia. The role of hedgehogs in the ecology of microsporidia is unclear; therefore, we investigated the prevalence and genetic diversity of E. bieneusi, Cryptosporidium, and Blastocystis spp. in hedgehogs (Erinaceus amurensis) collected from Hubei Province in Central China. PCR amplification of the internal transcribed spacer region of the ribosomal DNA indicated that 9.8% (4/41) hedgehogs were positive to E. bieneusi, but none (0/41) was positive to Cryptosporidium and Blastocystis spp. Phylogenetic analysis showed the strains detected from the hedgehogs belong to four novel genotypes (EA1-EA4), which were most closely related to type IV of group 1c. This study demonstrated that hedgehogs are hosts of E. bieneusi and may play a role in the transmission of E. bieneusi to humans in the process of being caught and slaughtered.

Asymptomatic Intestinal Parasitic Infestations among Children Under Five Years in Selected Communities in the Ho Municipality, Ghana.

The study investigated intestinal parasitic infestations (IPIs) and possible risk factors associated with asymptomatic children under five (5) years in five (5) selected communities in the Ho Municipality. The study design was cross- sectional, with a simple random sampling technique involving 150 asymptomatic children under 5 years from 5 selected communities (Klave, Hoe, Freetown, Dave and Godokpe) in the Ho Municipality. A questionnaire was used to obtain socio-demographics and other relevant parameters. Direct wet preparation, formol-ether concentration and Modified ZN staining techniques were used for the identification of intestinal parasites from participants' stool samples. The Fisher's exact test and binary logistic regression analysis were used to determine the difference in IPIs proportions and assess the risk factors associated with IPIs respectively. The overall IPIs cases was 14% (21/150). Cryptosporidium spp was most predominant [5.3% (8/150)], followed by Entamoeba spp [3.3% (5/150)], Cyclospora cayetenensis [2.7% (4/150)], Ascaris lumbricoides [1.3% (2/150)], Giardia lamblia [0.7% (1/150)] and Strongyloides stercoralis [0.7% (1/150)]. Children in rural communities (23.4%) recorded significantly higher case rate compared to those in urban communities (9.8%0), (p=0.04). Lower educational attainment of mother [OR=0.55, 95% CI (0.37 - 0.83), p-value = 0.015] and residence in rural communities [OR = 0.53, 95% CI (0.33-0.88)], p-value = 0.025] were significantly associated with IPIs. Asymptomatic IPIs are quite prevalent among children under 5 years in the Ho Municipality. The study thus recommends active sensitization programs for parents/guardians on preventive measures and school health programs should be instituted in rural communities.

Detection and Molecular Identification of Cryptosporidium Species Among Children with Malignancies

Cryptosporidiosis represents a major health problem worldwide particularly among children. Its diagnosis is still difficult and demands sensitive methods. In Egypt, there is little documentation of infection among children with malignancies. This work was designed to study the infection rate of Cryptosporidium among children with malignancies, compare the performance of modified Ziehl-Neelsen (MZN) stain with nested polymerase chain reaction (PCR) and identify the species subtypes of positive cases. The study was conducted on 100 children with malignancies (leukemia, lymphoma and solid tumors), below 10 years of age, from El-Shatby hospital, Alexandria University. After obtaining the informed consent, their stool samples were collected and examined microscopically following MZN stain for the diagnosis of Cryptosporidium spp. All samples were then subjected to nested PCR. Restriction fragment length polymorphism (RFLP) targeting the Cryptosporidium oocyst wall protein (COWP) gene was applied to positive cases, using restriction enzyme RsaI for digestion of nested PCR products. Out of the 100 examined children, MZN detected higher positive cases compared to nested PCR. Six cases (6%) were diagnosed positive by MZN stain, three of which (3%) were concordantly positive by nested PCR. All positives were among children with acute lymphoblastic leukemia (ALL). Fair agreement was found between the two tests (K = 0.36). Genotyping results revealed that positive samples were of Cryptosporidium parvum (C. parvum) type II. Low Cryptosporidium infection rate was detected among children with malignancies. MZN diagnosed more positive cases compared to nested PCR. C. parvum type II was the identified species among the examined children. Further optimization of PCR steps is needed.

Longitudinal detection of Cryptosporidium spp. in 1-10-week-old dairy calves on a farm in Xinjiang, China.

Cryptosporidiosis is an important cause of morbidity and mortality in the cattle industry and leads to severe economic losses. Fecal samples were collected from 25 dairy calves once a week for 10 weeks for continuous longitudinal detection of Cryptosporidium spp. Cryptosporidium spp. were detected via nested PCR amplification of the ribosomal small subunit RNA gene, followed by restriction fragment length polymorphism analysis with enzymes SspI and MboII to identify the species. PCR results indicated that all calves were infected with Cryptosporidium spp. at least once, with an average overall prevalence rate of 52.0% (130/250). One-week-old calves had the highest occurrences of Cryptosporidium infection (96.0%), 2-week-old calves (80.0%) had the second highest, and calves with watery diarrhea also had a higher occurrence of infection (92.3%). Four Cryptosporidium species, C. parvum, C. bovis, C. ryanae, and C. andersoni, were identified, with C. parvum being the most common. Forty-eight C. parvum isolates were further subtyped via nested PCR amplification of the 60-kDa glycoprotein gene, and all were identified as subtype IIdA15G1. The results demonstrated that C. parvum mainly infects dairy calves which are younger than 3 weeks old.

Assessing risk factors associated with prevalence of canine gastrointestinal parasitic zoonoses in Andhra Pradesh, India

Prevalence and risk factors associated with gastrointestinal parasites in dogs from central coastal districts of Andhra Pradesh, India was estimated by coprological examination. On examination of faecal samples of 1,907 dogs by saturated zinc sulphate floatation, sedimentation technique and modified Ziehl-Nielsen staining, an overall prevalence of 52.2% was observed. Single species infections were more common (49.7%) than mixed infections (2.5%). A total of 11 different species were detected. The most prevalent species was Ancylostoma canis followed by Toxocara canis, Cystoisospora ohioensis, Dipylidium caninum, Toxascaris leonina, Capillaria aerophila, Diphyllobothrium latum, Cystoisospora canis, Entamoeba spp., Trichuris vulpis and Cryptosporidium spp. The overall prevalence was high in stray dogs (65.9%). The prevalence of A. caninum (31.6%) was significant in pet dogs. Significant relationship between the season and prevalence of parasites was observed, the infection being more prevalent during rainy season. Binary logistic regression analysis identified age, season and the living condition of dogs and lack of zoonotic awareness of owners as risk factors for different parasitic infection.

Cryptosporidium bollandi n. sp. (Apicomplexa: Cryptosporidiiae) from angelfish (Pterophyllum scalare) and Oscar fish (Astronotus ocellatus)

The species name Cryptosporidium bollandi n. sp. is proposed for Cryptosporidium piscine genotype 2 based on morphological, biological and molecular characterisation. Phylogenetic analyses of 18S rRNA (18S) sequences revealed that C. bollandi n. sp. was most closely related to piscine genotype 4 (5.1% genetic distance) and exhibited genetic distances of 10.0%, 12.2% and 25.2% from Cryptosporidium molnari, Cryptosporidium huwi and Cryptosporidium scophthtalmi, respectively. At the actin locus, C. bollandi n. sp. was again most closely related to piscine genotype 4 (6.8% genetic distance) and exhibited 15.5% (C. molnari), 18.4% (C. huwi), 22.9% (C. scophthalmi) and up to 27.5% genetic distance from other Cryptosporidium spp. (Cryptosporidium felis). Phylogenetic analysis of concatenated 18S and actin sequences showed that C. bollandi n. sp. exhibited 12.9% (C. molnari) to 21.1% (C. canis) genetic distance from all other Cryptosporidium spp. Genetic data as well as previous histological analysis clearly supports the validity of C. bollandi n. sp. as a separate species.

Water security and detection of Cryptosporidium spp. in samples from the Guarapiranga Reservoir, São Paulo

Water security depends, among other factors, on the hydrological cycle, quantity and quality of water. Several microorganisms that pose risks to human and animal health are waterborne, such as the protozoan Cryptosporidium spp. The parasitological diagnosis of Cryptosporidium spp. in biological material and environmental samples can be accomplished through several techniques that include optical microscopy with analysis of fresh material or through the use of concentration techniques, indirect immunofluorescence and molecular biology that allows differentiation between species. Oocysts of Cryptosporidium spp. are eliminated through feces from parasitized hosts and released into drinking water reservoirs through untreated sewage. Therefore, its proper identification is necessary when causing human gastrointestinal changes. The objective of this work is to analyze the presence of oocysts of Cryptosporidium spp. in recreational areas of the Guarapiranga Resevoir located in the state of São Paulo, Brazil. Water samples were collected and concentrated between July and November and analyzed through the technique of Ziehl Neelsen. Samples showed positive results for Cryptosporidium spp. and cyanobacteria, which are commonly found in the aquatic biota. The latter can be found in large quantities in eutrophic environments due to high amount of organic matter released in water bodies. Oocysts were observed in 60% of the analyzed samples and the distribution between positive and negative samples was not dependent on the month of sampling, but showed statistically significant differences according to the sampling site.

Prevalence of Cryptosporidiosis in Ostriches from Central and South Parts of Iraq

The prevalence of ostrich’s cryptosporidiosis was determined for the first time in central and south parts of Iraq to study the effects of age, sex, and months on the infection rate and to record the morphological characterization of Cryptosporidium spp in ostriches. A total of 200 ostriches fecal samples were examined by traditional methods for the detection of the parasite. The total infection rate was 11% (22/200), and the highest infection rate was 12.26% (13/106) in chicks ( 9 months), while the lowest infection rate was 9.57% (9/94) recorded in the adult (> 9 months). The study included seven provinces in the central and south parts of Iraq, Wasit, Baghdad, Babylon, Diyala, Karbala, Al-Najaf, and Al-Qadisiyah. The highest infection rate was reported in Al-Najaf (23.53%) compared with the lowest rate (0%) in Wasit province. March reported an infection rate of 50%, while the lowest rate (0%) was registered during summer months (June, July, and August) with a significant difference (P≤0.05) among months of the study. Morphologically, the study indicated the widespread of Cryptosporidium parasites in ostriches in both central and south areas of Iraq.

Evaluation of Different Oocyst DNA Extraction Methods for Cryptosporidium spp. Research in Environmental Samples.

The major problem of the PCR method for the search of protozoan cysts/oocysts in environmental samples is the presence of inhibitors. DNA extraction methods capable of removing inhibitory substances of environmental origin and recovering the DNA are decisive for the efficiency of PCR. This study aimed to compare the efficiency of different DNA extraction methods for the search by Cryptosporidium oocysts in water samples by molecular methods. DNA extraction from water samples was performed using four different methods. Two methods use a chaotropic buffer to extract DNA and promote the selective binding of DNA to a silica membrane (GuSCN-silica and GFX Kit). The other method is based on the lysis and digestion of the samples in buffer and proteinase K, adsorption of impurities by an "InhibitEX" insertion matrix and purification of the DNA by a silica column (QIAamp Kit). The fourth method uses ionic and non-ionic detergents and proteinase K, to solubilize and separate the DNA from proteins, and a paramagnetic resin for DNA purification in the presence of high concentrations of guanidine ions (MAGNEX DNA Kit). Nested-PCR was performed, and the Cryptosporidium SSU rDNA gene amplified. The results demonstrated that MAGNEX and GFX commercial kits showed higher sensitivity, with detection of up to 100 oocysts/mL and 104 oocysts/mL respectively. In conclusion, this study confirmed that for low-DNA environmental samples, extraction methods should include an efficient oocyst wall breaking step, and showed that the best Cryptosporidium DNA extraction methods are those that use paramagnetic resins.

Prevalence and molecular characterization of Cryptosporidium spp. in pigs in Xinjiang, China

Cryptosporidium spp. are distributed ubiquitously worldwide, and pigs are considered as one of the main reservoir hosts. Eight hundred one porcine fecal specimens were collected from seven intensive pig farms in Xinjiang Uygur Autonomous Region, China. Cryptosporidium spp. were screened via PCR amplification of the small ribosomal subunit RNA gene, and 143 specimens (17.9%, 143/801) from all seven farms tested positive for Cryptosporidium spp. Cryptosporidium prevalence in the pigs differed significantly among farms (p < 0.01). The highest Cryptosporidium spp. prevalence in post-weaned pigs was 39.5% (111/281), followed by fattening pigs (23.2%, 30/129), pre-weaned pigs (1.2%, 2/169), and sows (0/222). Significant differences were observed between age groups (p < 0.01). C. suis was the predominantly identified species (62.9%, 90/143), followed by C. scrofarum (35.7%, 51/143), and C. parvum (1.4%, 2/143). Two C. parvum specimens were subtyped by analyzing the 60-kDa glycoprotein (gp60) gene sequences and were identified as IIdA14G1 and IIdA15G1. To our knowledge, this is the first report of C. parvum infection in pigs in China. The identification of three Cryptosporidium species, including zoonotic C. parvum in pigs in Xinjiang raises concern for the health of both swine animals and personnel in the pig industry.

Quantifying public health risks from exposure to waterborne pathogens during river bathing as a basis for reduction of disease burden.

A Quantitative Microbial Risk Assessment (QMRA) technique was applied to assess the public health risk from exposure to infectious microorganisms at bathing areas of three rivers in Bangladesh. The QMRA assessed the probability of illness due to the accidental ingestion of river water impacted by untreated sewage. The simplified QMRA was based on average concentrations of four reference pathogens Escherichia coli (E. coli) O157:H7, Cryptosporidium spp, norovirus and rotavirus relative to indicator bacterium E. coli. Public health risk was estimated as the probability of infection and illness from a single exposure of bathers. The risks of illness were ranged from 7 to 10% for E. coli O157:H7, 13 to 19% for Cryptosporidium, 7 to 10% for norovirus and 12 to 17% for rotavirus. The overall risk of illness at the rivers was slightly higher in children (9-19%) compared to adults (7-16%). The risks of illness in individuals exposed to the river bathing were unacceptably high, exceeding the USEPA acceptable risk of 3-6 illnesses per hundred bathing events. This study gives a basis for reducing the burden of disease in the population by applying appropriate risk management. Findings and methods of this study will be helpful for other countries with similar socio-economic and geographic settings.

Bovine Cryptosporidiosis and the Egyptian Diarrheic Buffalo Calves

The negative impact of diarrhea on livestock health is well known, Cryptosporidium, is one of the protozoan that causes diarrhea in calves especially buffalo calves. Some species of Cryptosporidium represent a zoonotic hazard. This study aimed to distinguish the potential species of Cryptosporidium in affected buffalo calves and evaluate a modified technique to improve the molecular detection and identification of Cryptosporidium. Twenty buffalo calves suffered from diarrhea were enrolled in the study. The enrollment criteria depended on the results of Ziehl–Neelsen stain. Sugar floatation technique was performed followed by oocyst concentration to form a pellet for DNA extraction. Multiplex PCR was performed for identification and differentiation of Cryptosporidium Spp. The results showed a mixed infection in 4 samples; the most common type of Cryptosporidium affecting the examined buffalo calves was C.parvum (10), followed by C.bovis (7 samples) and C.andersoni (6 samples) while no C.reyne was detected in the examined samples. The zoonotic type, C.parvum was found in 50% of the total affected animals. The current study detects three Cryptosporidium spp. namely C.parvum, C. andersoni, and C.bovis linked to diarrhea in the studied buffalo calves. Mixed infection with more than one species of Cryptosporidium was present. Standard detection of oocyte in fecal samples using modified Ziehl-Neelsen stain is a simple way for diagnosis of Cryptosporidium. However, a more sophisticated method is recommended to detect and differentiate the zoonotic species in calves, as they symbolize a crucial source of human infection.

Risk factors for sporadic cryptosporidiosis: A systematic review and meta-analysis

Cryptosporidium spp. is an important cause of gastrointestinal disease worldwide, responsible for 69 million cases of illness in 2016. Information on the sources and transmission pathways of human cryptosporidiosis results mainly from outbreak investigations.A systematic review and a meta-analysis of case-control and cohort studies were performed to determine the main risk factors associated with sporadic cryptosporidiosis. Suitable scientific articles were identified through a systematic literature search and subjected to a methodological quality assessment. From each study, odds ratio (OR) measures were extracted or calculated, as well as study characteristics such as population type, design, type of model and risk factor hierarchy. Mixed-effects meta-analysis models were adjusted by population type to appropriate data partitions.From 1985 identified references, the quality assessment stage was passed by 57 cohort and case-control studies focusing on sporadic cryptosporidiosis. The eligible studies were conducted between 1983 and 2016 and provided 568 OR categorized for meta-analysis.This meta-analysis identified travel, immunocompromising conditions, contact with infected humans, waterborne transmission (contact with recreational waters, wastewater, and consumption of untreated drinking water), contact with animals and food consumption as the relevant risk factors for sporadic cryptosporidiosis. With regards to food exposures, consumption of meat, dairy products (raw milk) and dishes consumed outside home were found significantly associated with cryptosporidiosis. The consumption of poorly washed fruits and vegetables significantly increases ORs. This meta-analysis reveals that some potential sources of Cryptosporidium such as shellfish or vegetables are under-investigated.Future case-control studies for sporadic cryptosporidiosis should include population at risk, and investigate other potential sources in relation to the genotype and the subtype of Cryptosporidium spp.

Cryptosporidiosis in HIV-Positive Patients, Bandar Abbas, Iran: A Cross-Sectional Study

Background: Cryptosporidiosis is a parasitic disease caused by a small protozoan of the genus coccidian. It is important in the medical and veterinary area, since it is a zoonotic parasite. Transmission of fecal-oral infections occurs through direct and indirect contact with food and contaminated drinks. Many HIV-positive individuals live in the areas where intestinal parasites are hyperendemic. Objectives: Given the significance of the study associated with opportunistic infectious diseases in this group of patients, we decided to study the prevalence of the parasite in those individuals. Methods: In this descriptive cross-sectional study, we collected a single fecal specimen of 133 HIV-positive patients seeking treatment from the center for Behavioral Disease Counseling of Bandar Abbas, Iran. Stool samples examined by direct, formalin-ether concentration method and modified Ziehl-Neelsen stain for Cryptosporidium spp. Results: The mean age of the patients was 42.15 (ranged 12 - 82 years). No oocyst of cryptosporidium spp. or other intestinal coccidian was found in any of the samples. Only four species including Giardia lamblia, Hymenolepis nana, and both Entamoeba coli and Blastocystis hominis were found in three patients, which indicated less diversity and low frequency of parasitic infection in this study. Conclusions: Monitoring and evaluating the therapeutic process of the patients lead to less infectious diseases, especially those of opportunistic parasites.