Identification Of Cryptosporidium Research Articles

Frequency and Molecular Identification of Cryptosporidium in Adult Prim'Holstein Dairy Cattle Farms in the North of France.

Cryptosporidium apicomplexan protozoa are ubiquitous intracellular agents affecting humans and animals. In particular, bovine cryptosporidiosis is recognized as endemic worldwide. However, epidemiological investigations remain limited in France regarding the burden of these parasites in cattle. To improve our understanding of the epidemiology of cryptosporidiosis, the main aim of this study was to determine the frequency and the genetic diversity of Cryptosporidium in adult Prim'Holstein dairy cattle farms in the north of France. Fecal specimens were collected from 1454 non-diarrheic and non-pregnant animals (nulli-, primi-, or multiparous) throughout 20 farms in an area of 110 km around Lille. For Cryptosporidium species identification, nested PCR followed by sequence and phylogenetic analyses were used. The overall frequency of Cryptosporidium spp. in-fection was 30.00% (C.I. 95%: 12.83-54.33) in farms and 0.89% (C.I. 95%: 0.498-1.57) at the individual level. In primi- or multiparous cows, only C. andersoni was found. C. ryanae, C. bovis/xiaoi and C. andersoni were detected in heifers. The phylogenetic tree confirmed that analyzed sequences were grouped with known reference sequences reported in dairy cattle. Further studies on the cumulative prevalence, risks factors and pathogenicity are needed to give a more accurate assessment of the impact of Cryptosporidium infection in dairy cattle in France.

Detection and Identification of Cryptosporidium in Raw Urban Wastewaters Entering the Bu-shehr Coastal Area by Nested-PCR

Detection and Identification of Cryptosporidium in Raw Urban Wastewaters Entering the Bu-shehr Coastal Area by Nested-PCR

Molecular characteristics and zoonotic potential of enteric protists in domestic dogs and cats in Egypt.

Domestic dogs and cats can be a source of human infection by a wide diversity of zoonotic pathogens including parasites. Genotyping and subtyping tools are useful in assessing the true public health relevance of canine and feline infections by these pathogens. This study investigated the occurrence, genetic diversity, and zoonotic potential of common diarrhea-causing enteric protist parasites in household dogs and cats in Egypt, a country where this information is particularly scarce. In this prospective, cross-sectional study a total of 352 individual fecal samples were collected from dogs (n = 218) and cats (n = 134) in three Egyptian governorates (Dakahlia, Gharbeya, and Giza) during July-December 2021. Detection and identification of Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi, and Blastocystis sp. were carried out by PCR and Sanger sequencing. Basic epidemiological variables (geographical origin, sex, age, and breed) were examined for association with occurrence of infection by enteric protists. The overall prevalence rates of Cryptosporidium spp. and G. duodenalis were 1.8% (95% CI: 0.5-4.6) and 38.5% (95% CI: 32.0-45.3), respectively, in dogs, and 6.0% (95% CI: 2.6-11.4) and 32.1% (95% CI: 24.3-40.7), respectively, in cats. All canine and feline fecal samples analyzed tested negative for E. bieneusi and Blastocystis sp. Dogs from Giza governorate and cats from Dakahlia governorate were at higher risk of infection by Cryptosporidium spp. (p = 0.0006) and G. duodenalis (p = 0.00001), respectively. Sequence analyses identified host-adapted Cryptosporidium canis (n = 4, one of them belonging to novel subtype XXe2) and G. duodenalis assemblages C (n = 1) and D (n = 3) in dogs. In cats the zoonotic C. parvum (n = 5) was more prevalent than host-adapted C. felis (n = 1). Household dogs had a limited (but not negligible) role as source of human giardiasis and cryptosporidiosis, but the unexpected high frequency of zoonotic C. parvum in domestic cats might be a public health concern. This is the first molecular-based description of Cryptosporidium spp. infections in cats in the African continent to date. Molecular epidemiological data provided here can assist health authorities and policy makers in designing and implementing effective campaigns to minimize the transmission of enteric protists in Egypt.

Molecular identification of Cryptosporidium, Giardia, and Blastocystis from stray and household cats and cat owners in Tehran, Iran

Cryptosporidiosis, giardiasis, and blastocystosis are among the most important parasitic diseases common between humans and cats. In addition, there are concerns about the possible transmission of zoonotic parasites from infected cats to humans. Hence, we investigated the molecular epidemiology of Cryptosporidium spp., Giardia duodenalis, and Blastocystis sp. in stray and household cats and cat owners. Our study was performed on 132, 33, and 33 fecal samples of stray and household cats, as well as cat owners in Tehran, Iran. Cryptosporidium spp. was identified using a nested PCR targeting the small subunit ribosomal RNA gene (SSU rRNA) and sequencing the internal amplified fragments. Furthermore, to perform multilocus genotyping of G. duodenalis, the ß-giardin (bg), glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi) genes were amplified to assess the DNA of G. duodenalis in the fecal samples of cats and cat owners. In addition, Blastocystis was detected by targeting the SSU rRNA gene, and the subtypes of Blastocystis were determined via the sequencing of amplicons. Cryptosporidium felis and Cryptosporidium canis were detected in seven stray cats (5.3%) and one household cat (3%). The bg gene of G. duodenalis was amplified and successfully sequenced in two (1.5%) stray cats and revealed assemblages F and B of G. duodenalis. Sequencing and phylogenic analysis of SSU rRNA gene nucleotide sequences of Blastocystis detected ST5 and ST10 in stray cats (1.5%), ST1 in household cats (9.1%), and ST1, ST2, ST3, and ST7 in owners (30.3%). The low prevalence of Cryptosporidium, Giardia and Blastocystis in cats and the presence of species/assemblages/subtypes with limited zoonotic potential indicate that cats had a minor role in their owners' infection in the investigated population. However, the presence of zoonotic protozoa in cats suggests the necessity of special attention to high-risk individuals during close contact with cats. Therefore, it is recommended that veterinarians, physicians, and urban managers plan to prevent, control, or treat these parasites to help the urban community live healthily alongside cats.

Molecular identification of Cryptosporidium baileyi in Muscovy ducks (Cairina moschata domesticus) in free-range production systems

ABSTRACT: Cryptosporidiosis is considered an infection with impact on animal health. It has been associated with high morbidity and mortality rates, leading to significant economic losses to the poultry industry. This study investigated the presence of Cryptosporidium spp. in domestic ducks of family Anatidae (Cairina moschata) from two rustic commercial aviaries located in the city of Rio de Janeiro, Brazil. A total of 315 fecal samples were collected from domestic ducks in two different areas (N=186 in area A and N=129 in area B). The microscopic analysis was conducted using a sugar centrifugal flotation technique for the identification of Cryptosporidium spp. oocysts, followed by PCR/sequencing analyses of the partial sequence of the 18S rDNA gene to determine the Cryptosporidium species. Of the 315 samples collected, only 10 (186/5.38%) from area A were positive for Cryptosporidium. The nucleotide sequence and phylogenetic analyses identified that all samples were identical (100%) and belonged to Cryptosporidium baileyi species, which is closely related to gastric species and of importance in animal health.

CRYPTOSPORIDIUM SPP. IN PET DWARF WINTER WHITE RUSSIAN HAMSTERS (PHODOPUS SUNGORIS SUNGORIS) IN CHINA.

Cryptosporidium spp. have been identified in a wide range of hosts, such as humans and domestic and wild animals, while less information about the prevalence of Cryptosporidium spp. in pet hamsters is documented. A total of 351 dwarf winter white Russian hamsters' fecal specimens were collected from 6 pet markets from the cities of Luzhou and Ziyang in Sichuan province in the southwestern part of China. The prevalence of Cryptosporidium spp. determined with nested-PCR amplification of the partial small-subunit (SSU) rRNA gene was 39.32% (138/351). The highest prevalence of Cryptosporidium spp. was in pet market 5 (79.49%, 62/78), followed by pet market 6 (38.64%, 17/44). The lowest prevalence of Cryptosporidium spp. was observed in pet market 3 (14.89%, 7/47). Statistically significant differences in the prevalence of Cryptosporidium spp. were observed among different pet markets (χ2 = 76.386, df = 5, P < 0.05), and a further post hoc test revealed that only pet market 5 was significantly different from other pet markets. Molecular analysis showed that 4 different Cryptosporidium species or genotypes were identified: Cryptosporidium parvum (n = 127), Cryptosporidium chipmunk genotype III (n = 6), Cryptosporidium andersoni (n = 4), and Cryptosporidium wrairi (n = 1). The identification of Cryptosporidium spp. was further tested with the 60-kDa glycoprotein (GP60) gene, and the positive rate was 29.7% (41/138). This is the first molecular report on Cryptosporidium spp. infection in dwarf winter white Russian hamsters in China. With C. parvum and C. andersoni being identified in both humans and pet hamsters, these findings suggest that pet hamsters may be potential reservoirs of zoonotic Cryptosporidium species and subtypes.

CRISPR/Cas12a-powered immunosensor suitable for ultra-sensitive whole Cryptosporidium oocyst detection from water samples using a plate reader

Waterborne pathogens, such as Cryptosporidium parvum, pose a major threat to public health globally, and this requires screening of drinking and environmental water for low number of contaminating microbes. However, current detection approaches generally require trained experts with sophisticated instruments, and are not suitable for large-scale screening and rapid outbreak response. Recent advances in ultrasensitive CRISPR/Cas-based biosensing continue to expand the range of detectable molecular targets, however single microbes could not be directly detected so far, especially in environmental samples. Here, we report an ultrasensitive CRISPR/Cas12a-powered immunosensing method suitable for microbial detection which links antibody-based recognition with CRISPR/Cas12a-based fluorescent signal amplification through an antibody-DNA conjugate. This approach is shown here to detect whole 4 µm size Cryptosporidium parvum oocysts with a linear range from 6.25 – 1600 oocysts/mL, at a maximum sensitivity of single oocyst per sample. Its potential to apply to various complex sample matrices has also been demonstrated. After sample dilution by factor of 10, we were able to detect 10 oocysts from a back-wash mud samples from water treatment plate. This method uses the same experimental setup (plate reader) as a conventional ELISA assay thus reducing the need for microscopy-based identification of Cryptosporidium, which represents the gold-standard but requires high level expertise and time-consuming manual counting. This work highlights the potential of CRISPR/Cas-based biosensing for water quality assessment and ultrasensitive whole pathogen detection.

Molecular Identification of Cryptosporidium spp. and Giardia spp. in Wild Birds in Southeastern Brazil

Introduction: The identification of Cryptosporidium spp., Giardia spp. and Enterocytozoon bieneusi in birds is relevant since these animals can act as disseminators of these parasites to humans through environmental contamination. The aim of this study was to determine the molecular occurrence of Cryptosporidium spp. and Giardia spp. in wild birds in southeastern Brazil and genetically characterize the isolates obtained. Methods: A total of 256 fecal samples were collected from 172 captive and 84 free-living wild birds. The DNA extracted was subjected to nested-PCR and semi-nested PCR analysis for amplification of fragments of the 18S rDNA and gdh genes of Cryptosporidium spp. and Giardia spp., respectively. Results: With respect to Cryptosporidium spp., the overall occurrence was 3.91%. Of samples from captive wild birds, six (3.49%) were positive: two waterfowl and four non-aquatic birds. Among the samples from free-living wild birds, four (4.76%) were positive, all non-aquatic birds. Regarding Giardia spp., the overall occurrence was 3.1%. Of samples from captive wild birds, four (2.32%) were positive, all waterfowl; of the samples from free-living wild birds, four (4.76%) were positive for the parasite, all non-aquatic birds. Conclusions: The presence of C. meleagridis and G. duodenalis assemblage B suggests that epidemiological studies involving wild birds and humans are needed to better understand the impact of avian cryptosporidiosis and giardiasis on avian health and their possible implications for public health.

Transfer of Cryptosporidium oocysts from feeder rodents to neonate snakes and its impact on parasite identification methods used by zoological institutions

Parasites are found in a variety of species and can cause issues in animal health and welfare. One parasite in particular, Cryptosporidium, is a protozoan that impacts the gastrointestinal and respiratory tract of its hosts, which includes humans, rodents, cattle, birds, fish, and reptiles. In squamates, specifically snakes, Cryptosporidium serpentis has been reported to cause lethargy, weight loss, and mortality. Another species, Cryptosporidium parvum, causes infections in mice, humans, and cattle but is nonpathogenic in snakes. Common identification methods, such as ELISA antigen testing, Acid-fast (AF) staining, and Immunofluorescent antibody (IFA) testing, are frequently used because of their low cost and simplicity; however, these can appear positive for both C. parvum and C. serpentis. Polymerase chain reaction (PCR) is also used for Cryptosporidium identification and is one of the only methods that easily differentiates between Cryptosporidium species. Differentiating between these species is necessary as zoological protocols recommend euthanizing or permanently isolating squamates infected with C. serpentis. Rodents are a staple of snake diets in human care, so it is crucial to understand how their diet could impact infection identification. Additionally, immunocompetent mice infected by C. parvum are known to show no clinical signs of infection, so feeder rodent companies may be unaware that their rodents are infected. The purpose of this study is to show how consuming an infected prey item could cause a false positive using common Cryptosporidium identification methods, specifically IFA, and the importance of using species-specific identification methods such as PCR.

First Molecular Characterization of Cryptosporidium spp. in Patients Living with HIV in Honduras.

Cryptosporidiosis is one of the most important causes of gastroenteritis in the world, especially in low- and middle-income countries. It is caused by the Apicomplexan parasite Cryptosporidium spp., and mainly affects children and immunocompromised people, in whom it can pose a serious threat to their health, or even be life threatening. In Honduras, there are no data on parasite species or on molecular diversity or Cryptosporidium subtypes. Therefore, a cross-sectional study was conducted between September 2019 and March 2020 for the molecular identification of Cryptosporidium spp. in 102 patients living with HIV who attended a national hospital in Tegucigalpa. Stool samples were analyzed by direct microscopy, acid-fast stained smears, and a rapid lateral flow immunochromatographic test. All samples that tested positive were molecularly analyzed to identify the species and subtype of the parasite using three different markers: gp60, cowp, and 18Sr. PCR products were also sequenced. Four out of 102 samples (3.92%) were positive for Cryptosporidium parvum, and all were assigned to subtype IIa. These findings suggest a possible zoonotic transmission in this population.

Multi-spatial contamination of environmental aquatic matrices with Cryptosporidium: a climate, health, and regulatory framework for the Philippines

BackgroundCryptosporidium is a waterborne global pathogen causing diarrhea primarily in infants and immunocompromised individuals. The Philippines is a tropical country susceptible to the influences of climate change and water crises. To date, the country has no existing epidemiologic data, regulation, or strategy for monitoring Cryptosporidium in freshwater systems. We, therefore, endeavored to provide evidence on the multi-spatial contamination of Cryptosporidium in environmental aquatic matrices using low-cost, user-friendly, and sustainable strategies and submit implications on the presence of Cryptosporidium in freshwater systems in a climate, health, and regulatory framework.ResultsHere, we present the microscopic detection of Cryptosporidium oocysts in low-volume (50 mL) environmental samples of surface water (SW), sediments (BW), and substrate-associated biofilm (SAB) and in 1 L bulk SW investigated by PCR. The multi-spatial distribution of Cryptosporidium oocysts in the low-volume (50 ml) aquatic matrices based on microscopy was highest at 69% (20/29) in SW and lowest at 50% (13/26) in BW. Immunofluorescence technique provided the highest microscopic positivity rate with 59% (17/29), 38% (10/26), and 50% (10/20) detection in SW, BW, and SAB, respectively. The detection and identification of Cryptosporidium in 1 L bulk SW by PCR and sequence analysis was recorded in total at 21% (6/29) in sampling sites where the differential identification of C. parvum, C. hominis, and Cryptosporidium spp. was 7% (2/29), 10% (3/29), and 3% (1/29), respectively.ConclusionsWe report the microscopical and first molecular epidemiologic data of Cryptosporidium from the most significant environmental freshwater systems in the Philippines. The presence of the two main human and animal pathogenic species C. parvum and C. hominis from the largest lakes and major water reservoirs in the country calls for sustainable solutions in safeguarding the quality of freshwater resources in a climate, health, and regulatory approach.

Genotyping and identification of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi from free–range Tibetan yellow cattle and cattle–yak in Tibet, China

To assess the prevalence and molecular characteristics of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in natural grazing local breed cattle, 513 fecal samples from Tibetan yellow cattle and cattle–yaks were tested for these pathogens. Cryptosporidium, G. duodenalis and E. bieneusi in Tibetan yellow cattle prevalence were 0.7% (3/442), 3.8% (17/442) and 2.5% (11/442), respectively. Cryptosporidium bovis (n = 3), G. duodenalis assemblages A (n = 2) and E (n = 15) were identified based on sequence analysis. Among three E. bieneusi genotypes, I (n = 7), EbpC (n = 2) and CHC8 (n = 2) detected, EbpC was classified into Group 1, which has a significant zoonotic importance, whereas genotypes I and CHC8 belonged to Group 2. None of these pathogens was identified in cattle–yaks. The presence of zoonotic C. bovis, assemblage A and EbpC indicates Tibetan yellow cattle may be a potential spread source of intestinal pathogens with a zoonotic transmission risk. The relationships between natural free–range mode and the prevalence or genetic diversity of these pathogens need be confirmed in further studies.

Molecular identification of Cryptosporidium spp. in pet snakes in Beijing, China.

Few reports of Cryptosporidium spp. in snakes in China have been published. To determine the infection rate and document the presence of Cryptosporidium in pet snakes using molecular methods, 273 fecal samples were collected from eight species of pet snakes from 13 pet households in Beijing, China, and were examined by PCR amplification of the small subunit ribosomal RNA gene. Cryptosporidium was detected from 17 of 273 (6.2%) samples, and nine out of 13 households tested positive for Cryptosporidium with a range of 3.3 to 33.3% among households showing significant difference (p < 0.01). The infection rate of Cryptosporidium for females and males was 6.5% (13/201) and 5.6% (4/72), respectively, showing no significant difference (p > 0.05). Six out of eight pet snake species tested positive for Cryptosporidium with a range of 4.2 to 9.1% among species, showing no significant difference (p > 0.05). Two Cryptosporidium species were identified: Cryptosporidium serpentis in 10 samples and Cryptosporidium varanii in seven samples. No zoonotic Cryptosporidium species occur in our study populations.

Prevalence and Molecular Genotyping of Cryptosporidium Spp. in Diarrheic Patients from Bandar Abbas City, Southern Iran

Background: Cryptosporidium species are recognized as one of the most important gastrointestinal pathogens of humans and livestock. Objectives: This study aimed to determine the prevalence and sub-genotypes of Cryptosporidium spp. among diarrheic patients in Bandar Abbas City, Iran. Methods: Diarrheic fecal samples were collected from 170 patients in three hospitals of Bandar Abbas, Iran, from October 2018 to May 2019. Initial parasitological identification of Cryptosporidium spp. was performed by modified Ziehl-Neelsen (ZN) staining. For molecular analysis, the positive specimens and the suspected ones of Cryptosporidium spp. were evaluated by sequence analysis of the 60-kDa glycoprotein gene (gp60). The collected data were analyzed using SPSS software and the relationship between the variables and the presence of Cryptosporidium spp. assessed by the chi-square test. To assess the degree of agreement between PCR and ZN staining, Cohen’s kappa-index was applied. Results: Of the 170 diarrheic patients, 98 (57.6%) were male, and 72 (42.4%) were female. Prevalence of Cryptosporidium spp. by parasitological examination was 1.8% (3/170). However, using PCR, Cryptosporidium spp. was detected in 12% (6/50) of the positive microscopically samples (3 samples) and 47 suspected specimens. Sequence analysis of the gp60 gene showed that all of the positive isolates were Cryptosporidium parvum in which all subtypes belonged to allele family IId. Two distinct nucleotide sequences obtained from this study were deposited in GenBank under the accession numbers MN820453 and MN820454. Conclusions: The predominance of C. parvum (subtype family IId) in this study emphasizes the importance of zoonotic Cryptosporidium transmission in Bandar Abbas, Southern Iran.

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.

Prevalence and genotypic identification of Cryptosporidium spp. and Enterocytozoon bieneusi in captive Asiatic black bears (Ursus thibetanus) in Heilongjiang and Fujian provinces of China

BackgroundCryptosporidium and Enterocytozoon bieneusi are two important pathogens with zoonotic potential that cause enteric infections in a wide range of hosts, including humans. Both are transmitted from animals to humans by direct contact or through contaminated equipment. Bears are frequently found in Chinese zoos as ornamental animals as well as farmed as commercial animals, and are therefore in close contact with zoo- or farm-keepers, but the prevalence and zoonotic potential of Cryptosporidium and E. bieneusi in bears is poorly understood. In this study, we aimed to provide data on the occurrence and genetic diversity of Cryptosporidium and E. bieneusi in Asiatic black bears from Heilongjiang and Fujian, China. From May 2015 to December 2017, 218 fresh fecal specimens were collected from captive Asiatic black bears in Heilongjiang (n = 36) and Fujian (n = 182), China. Cryptosporidium and E. bieneusi were examined by PCR amplification of the partial small subunit of ribosomal DNA (SSU rDNA) and the internal transcribed spacer (ITS) region of rDNA, respectively. C. andersoni-positive isolates were subtyped through PCR analysis of the four minisatellite/microsatellite (MS1, MS2, MS3 and MS16) loci.ResultsThe overall prevalence of Cryptosporidium and E. bieneusi were 2.4% (4/218) and 6.4% (14/218), respectively, with 2.8% (1/36) and 22.2% (8/36) in the Heilongjiang Province, and 1.6% (3/182) and 3.3% (6/182) in the Fujian Province. Sequence analysis confirmed the presence of Cryptosporidium species: C. andersoni (n = 3) and a genotype termed Cryptosporidium rat genotype IV (n = 1). All three identified C. andersoni belonged to the MLST subtype A4, A4, A4, A1. Two known E. bieneusi genotypes D (n = 4) and SC02 (n = 10) were identified, both of which belong to zoonotic Group 1.ConclusionsThis is the first report of C. andersoni and Cryptosporidium rat genotype IV in bears. The discovery of the zoonotic potential of E. bieneusi genotype D in bears highlights its significant zoonotic potential and potential threat to human health.

Prevalence and genotypic identification of Cryptosporidium in free-ranging and farm-raised donkeys (Equus asinus asinus) in Xinjiang, China.

The prevalence and zoonotic potential of Cryptosporidium in donkeys is poorly understood. Here, 680 fecal specimens were collected from 178 free-ranging and 502 farmed donkeys in Xinjiang, China. Cryptosporidium was identified using PCR amplification of the small subunit of ribosomal DNA. Cryptosporidium-positive isolates were subtyped using PCR analysis of the 60 kDa glycoprotein gene (gp60). The overall prevalence of Cryptosporidium was 2.4% (16/680), with 3.2% (16/502) in farmed donkeys and 0% (0/178) in free-ranging donkeys. Cryptosporidium hominis (n = 13), C. parvum (n = 1) and Cryptosporidium horse genotype (n = 2) were identified. The C. hominis isolates belonged to two subtypes, IkA16 (n = 9) and IkA16G1 (n = 4). The subtype of C. parvum was IIdA15G1, whereas the two Cryptosporidium horse genotype isolates were of subtype VIaA15G4. The predominance of C. hominis in donkeys suggests that these animals are infected through human contact.

Prevalence and genotyping identification of Cryptosporidium in adult ruminants in central Iran

BackgroundApicomplexan parasites of the genus Cryptosporidium infect a wide range of animal species as well as humans. Cryptosporidium spp. can cause life threatening diarrhea especially in young animals, children, immunocompromised patients and malnourished individuals. Asymptomatic cryptosporidial infections in animals can also occur, making these animals potential reservoirs of infection.MethodsIn the present study, a molecular survey of Cryptosporidium spp. in ruminants that were slaughtered for human consumption in Yazd Province, located in central Iran was conducted. Faeces were collected per-rectum from 484 animals including 192 cattle, 192 sheep and 100 goats. DNA was extracted from all samples and screened for Cryptosporidium by PCR amplification of the 18S rRNA gene. Positives were Sanger sequenced and further subtyped by sequence analysis of the 60 kDa glycoprotein (gp60) locus.ResultsIn total, Cryptosporidium spp. were detected in 22 animals: C. andersoni and C. bovis in seven and two cattle faecal samples, respectively, C. ubiquitum in five sheep, and C. xiaoi in six sheep and two goat samples, respectively. To our knowledge, this study provides for the first time, molecular information concerning Cryptosporidium species infecting goats in Iran, and is also the first report of C. ubiquitum and C. xiaoi from ruminants in Iran.ConclusionThe presence of potentially zoonotic species of Cryptosporidium in ruminants in this region may suggest that livestock could potentially contribute to human cryptosporidiosis, in particular among farmers and slaughterhouse workers, in the area. Further molecular studies on local human populations are required to more accurately understand the epidemiology and transmission dynamics of Cryptosporidium spp. in this region.

Prevalence and species identification of Cryptosporidium spp. in the newborn dairy calves from Muang District, Khon Kaen Province, Thailand.

Aim:This study aims to determine the prevalence of Cryptosporidium spp. infection and to identify the species of Cryptosporidium spp. in newborn dairy calves between December 2016 and March 2017 in Muang District, Khon Kaen Province, Thailand.Materials and Methods:A total of 200 fecal samples from newborn dairy calves of the ages 1 day up to 28 days were collected and the presence of Cryptosporidium oocysts was examined microscopically using the modified Kinyoun’s acid-fast staining technique. Then, Cryptosporidium species were identified using nested polymerase chain reaction amplification of 18S rRNA gene and sequencing.Results:The modified Kinyoun’s acid-fast staining revealed the presence of Cryptosporidium oocysts in 51% (102/200). Sequence analysis of the 18S rRNA gene identified two species, namely, Cryptosporidium bovis (n=11) and Cryptosporidium ryanae (n=11) and one isolated strain could not be identified.Conclusion:This study indicated that newborn dairy calves aging up to 4 weeks were highly infected with Cryptosporidium spp., and the infection mostly occurred in diarrheic dairy calves. This is the first report of Cryptosporidium in dairy calves in Khon Kaen Province and the results provide baseline information for further studies and control of Cryptosporidium infection in dairy calves in the study area.

The Prevalence of Cryptosporidium among Children Hospitalized because of Gastrointestinal Symptoms and the Efficiency of Diagnostic Methods for Cryptosporidium.

Cryptosporidium is a parasite that causes watery diarrhea among both children and adults. However, because many physicians do not routinely ask for Cryptosporidium diagnostic test, cryptosporidiosis prevalence is likely underestimated. The current study investigated the prevalence of cryptosporidiosis among children admitted to the hospital with gastrointestinal symptoms. Stool sample was collected from each child and subjected to routine microbiological culture. Cryptosporidium presence was tested by three different methods: real-time PCR (RT-PCR), quick antigen, and microscopic examination with acid-fast staining. Each positive specimen was further tested with BioFire FilmArray Multiplex PCR (bioMérieux SA, Marcy-l'Etoile, France) to determine that Cryptosporidium is the only pathogen in the sample. Demographic and epidemiological data were collected from the patients' medical records. Of 291 patient stool samples, nine were positive for Cryptosporidium hominis or Cryptosporidium parvum. The average age of the nine Cryptosporidium-positive cases was 2.3 years, lower than the average age of the study population (4.2 years). Of the positive cases, 66.7% were Arabs and 33.3% were Jews. The main complaint of children with Cryptosporidium was diarrhea. Regarding the laboratory methods for Cryptosporidium identification, of the positive samples, 100% (9/9) were identified by RT-PCR, 88% (8/9) were positive by antigen test, and only 67% (6/9) were positive by microscopic examination with acid-fast staining. Because of the low incidence of Cryptosporidium among patients in our facility, the prevalence of cryptosporidiosis still cannot be established. Nevertheless, among the other pathogens found in stool samples, Cryptosporidium is the second leading cause of hospitalization due to gastrointestinal disease in children in our area.