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Background: Enterocytozoon bieneusi is a zoonotic pathogen widely distributed in animals and humans. It can cause diarrhea and even death in immunocompromised hosts. Approximately 800 internal transcribed spacer (ITS) genotypes have been identified in E. bieneusi. Farmed foxes and raccoon dogs are closely associated to humans and might be the reservoir of E. bieneusi which is known to have zoonotic potential. However, there are only a few studies about E. bieneusi genotype identification and epidemiological survey in foxes and raccoon dogs in Henan and Hebei province. Thus, the present study investigated the infection rates and genotypes of E. bieneusi in farmed foxes and raccoon dogs in the Henan and Hebei provinces. Result: A total of 704 and 884 fecal specimens were collected from foxes and raccoon dogs, respectively. Nested PCR was conducted based on ITS of ribosomal RNA (rRNA), and then multilocus sequence typing (MLST) was conducted to analyze the genotypes. The result showed that infection rates of E. bieneusi in foxes and raccoon dogs were 18.32% and 5.54%, respectively. Ten E. bieneusi genotypes with zoonotic potential (NCF2, NCF3, D, EbpC, CHN-DC1, SCF2, CHN-F1, Type IV, BEB4, and BEB6) were identified in foxes and raccoon dogs. Totally 178 ITS-positive DNA specimens were identified from foxes and raccoon dogs and these specimens were then subjected to MLST analysis. In the MLST analysis, 12, 2, 7 and 8 genotypes were identified in at the mini-/ micro-satellite loci MS1, MS3, MS4 and MS7, respectively. A total of 14 multilocus genotypes were generated using ClustalX 2.1 software. Overall, the present study evaluated the infection of E. bieneusi in foxes and raccoon dogs in the Henan and Hebei province, and investigated the zoonotic potential of the E. bieneusi in foxes and raccoon dogs. Conclusions: These findings expand the geographic distribution information of E. bieneusi' host in China and was helpful in preventing against the infection of E. bieneusi with zoonotic potential in foxes and raccoon dogs. [ABSTRACT FROM AUTHOR]

Toxoplasma gondii infects almost all warm‐blooded animals, including humans and non‐human primates. Many cases of T. gondii infection in non‐human primates have been reported worldwide. In this study, 15 monkeys were collected from zoos in Henan Province between 2016 and 2019. A modified agglutination test (MAT) (cut‐off: 1:8) showed that 46.7% (7/15) of the heart juices had T. gondii IgG antibody transformation. One viable T. gondii strain was successfully isolated from the myocardium of a rhesus monkey by bioassay in mice. This strain was designated as TgMonkeyCHn1. The DNA of T. gondii tachyzoites was obtained using cell cultures, and the genotype of this strain was determined by PCR‐RFLP with 10 markers and the virulence genes ROP5 and ROP18. The genotype and ROP18/ROP5 (3/6) of TgMonkeyCHn1 did not match any known genotypes. In addition, the TgMonkeyCHn1 formed low number of tissue cysts and was non‐lethal to mice. To our knowledge, this is the first T. gondii strain isolated from Old World monkeys. Rhesus monkey is a new host record for T. gondii. [ABSTRACT FROM AUTHOR]

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Background: Captive wild animals in zoos infected with Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi, and Blastocystis sp. can be sources of zoonotic infections and diseases. Therefore, to investigate the distribution of these pathogens in captive wild animals of zoos in Henan, China, a total of 429 fresh fecal samples were collected from six zoos in Henan, China. The infection rates of Cryptosporidium spp., G. duodenalis, E. bieneusi, and Blastocystis sp. were determined by PCR analysis of corresponding loci. Positive results for Cryptosporidium (C. parvum and C. hominis) were subtyped based on the (gp60) gene. Results: The overall prevalence was 43.1% (185/429), and the prevalence of Cryptosporidium, Giardia duodenalis, Enterocytozoon bieneusi, and Blastocystis sp. were 2.8% (12/429), 0.5% (2/429), 20.8% (89/429), and 19.1% (82/429), respectively. Five Cryptosporidium species, namely, C. hominis, C. parvum, C. muris, C. andersoni, and C. macropodum, were identified in this study. Cryptosporidium parvum was further subtyped as IIdA19G1. Two Giardia duodenalis assemblages (A and E) were also identified. A total of 20 Enterocytozoon bieneusi genotypes were detected, including 18 known (BEB6, D, HND-1, CD7, SDD1, Henan-IV, KIN-1, CHK1, Peru8, Henan-V, CHG11, CHG-1, CHS9, CHG21, Type-IV, CHC9, CM5, and CHB1) and 2 novel genotypes (CHWD1 and CHPM1). A total of nine subtypes of Blastocystis sp. (ST1, ST2, ST3, ST5, ST6, ST7, ST10, ST13, and ST14) were identified in captive wild animals in zoos in the present study. Cryptosporidium andersoni, nine Enterocytozoon bieneusi genotypes, and five Blastocystis subtypes were here first identified in new hosts. Conclusions: Our study has expanded the host ranges of these four pathogens. The data indicate that animals in zoos can commonly be infected with these four zoonotic pathogens, and animals in zoos are potential sources of zoonotic infections in humans. [ABSTRACT FROM AUTHOR]

Cryptosporidium spp., Enterocytozoonbieneusi, and Giardiaduodenalis are common enteric pathogens that are capable of infecting humans and animals. Total of 1,005 fecal samples from captive pet birds were collected from seven locations in Henan Province, China. The results demonstrated that 9.9% (99/1,005) of the captive birds were infected with one of these three pathogens. Enterocytozoonbieneusi was the most prevalent species among the birds (45/1,005, 4.5%) followed by G. duodenalis (33/1,005, 3.3%) and Cryptosporidium spp. (21/1,005, 2.1%). Five Cryptosporidium species were identified, namely, C. baileyi (10), C. galli (5), C. meleagridis (4), C. andersoni (1), and C. parvum (1). Two known E. bieneusi genotypes were identified: Peru 6 (44) was identified in pigeons (34) and European turtle doves (10); whereas, the genotype PtEb I (1) was only identified in a pigeon. Only G. duodenalis assemblage E (33) was identified in some pet birds. To the best of our knowledge, this study is the first to undertake the molecular identification of G. duodenalis in birds in China. The identification of potentially zoonotic species/genotypes of the pathogens suggests that exposure to the excreta of these birds, either directly or via food and water, may pose a threat to human health. [ABSTRACT FROM AUTHOR]

Background: Enterocytozoon bieneusi is a parasite that infects humans and a wide range of other animals. The large migratory waterfowl, the whooper swan (Cygnus cygnus), travels through many cities during its migration and can spread parasites. Despite receiving increasing attention worldwide, there have been no reports of E. bieneusi infection occurring in C. cygnus. Therefore, this study aims to assess the prevalence and genetic characteristics of E. bieneusi in C. cygnus in Sanmenxia, China. Methods: Altogether, 467 fresh fecal samples were collected in the Swan Wetland Park in Sanmenxia, China. Genomic DNA was extracted from fresh fecal samples (n = 467) and E. bieneusi was identified by nested PCR amplification of the internal transcribed spacer (ITS) region. ITS-positive sequences were aligned and phylogenetically analyzed to determine the genotypes of E. bieneusi. Results: The overall prevalence of E. bieneusi in C. cygnus was 7.49% (35/467). Sequencing of the 35 positive samples revealed eight known genotypes (EbpA, EbpC, Henan-III, Henan-IV, BEB6, CD9, Peru6 and PtEb IX) and three novel genotypes (CSW1, CSW2 and CSW3). The phylogenetic tree constructed from the ITS sequences showed that seven genotypes (Peru6, EbpA, EbpC, Henan-III, CSW3, Henan-IV and CSW1) clustered within the zoonotic Group 1 while the remaining novel genotype CSW2 clustered within Group 5. Conclusions: To our knowledge, this is the first report of E. bieneusi in C. cygnus. Of public health significance, our results suggest that migratory C. cygnus might play an important role in the water-borne transmission of E. bieneusi. Effective strategies will be necessary to control E. bieneusi infection in C. cygnus, other animals and humans. [ABSTRACT FROM AUTHOR]

Enterocytozoon bieneusi is one of the most frequently diagnosed Microsporidia of humans and most animals. However, there is no information on E. bieneusi infection of pigs in Tibet and Henan, China. In this study, 1,190 fecal samples were collected from pigs in Tibet and Henan and screened for the presence of E. bieneusi. The overall prevalence of E. bieneusi infection was 54.2% (645/1,190), with differences in prevalence observed among geographical areas, ages, and pig breeds. Moreover, 10 E. bieneusi genotypes were identified based on internal transcribed spacer region genotyping, including eight known genotypes (EbpC, EbpA, CHG19, CHC5, Henan‐III, I, D, and H) and two novel genotypes (XZP‐I and XZP‐II). Multilocus sequence typing revealed 18, 7, 17, and 13 genotypes at minisatellite/microsatellite loci MS1, MS3, MS4, and MS7, respectively. Strong linkage disequilibrium (LD) and few numbers of recombination events, suggest a clonal structure of the E. bieneusi population examined in this study. The low pairwise genetic distance (FST) and gene flow (Nm) values indicated limited gene flow in the E. bieneusi population from different hosts, with phylogenetic, structure, and median‐joining network analyses all indicating the existence of host and geographical isolation. The identification of isolates belonging to nine human‐pathogenic genotypes indicates that pigs play an important role in the dissemination of E. bieneusi, improving our present understanding of E. bieneusi epidemiology in the studied region. [ABSTRACT FROM AUTHOR]

Giardia duodenalis is a common and widespread intestinal protozoan parasite of both humans and animals. Previous epidemiological and molecular studies have identified Giardia infections in different animals and humans, but only limited information is available about the occurrence and genotypes of Giardia in cattle in China. In this study, we determined the occurrence of giardiasis and genetically characterized G. duodenalis in dairy cattle in Henan Province, central China. The overall prevalence of G. duodenalis was 7.2% (128/1777) on microscopic analysis, with the highest infection rate (22.7%) in calves aged less than 1 month. G. duodenalis assemblages and subtypes were identified with multilocus genotyping based on the SSU rRNA, β-giardin (bg), glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi) genes. Two assemblages were detected in the successfully sequenced samples: assemblage A (n = 58), assemblage E (n = 21), with a mixed E and A assemblage (n = 2). Four novel subtypes of the gdh gene and seven of the bg gene were found among the G. duodenalis assemblage E isolates. Using the nomenclature for the multilocus genotype (MLG) model, nine novel multilocus genotypes E (MLGs E1–E9) and three MLGs A (a novel subtype AI, previously detected subtype AII-1, and a combination of both) were identified. MLG AII-1 identified in this study may be an important zoonotic subtype. The dairy cattle in Henan are a potential public health concern. [ABSTRACT FROM AUTHOR]

Few data are available on the molecular characterization of Cryptosporidium spp. in chickens and ducks in China. In this study, 2579 faecal samples from 46 chicken farms and eight Pekin duck farms in 21 prefectures in Henan Province were examined. The overall infection rate of Cryptosporidium was 10.6% (163/1542) in layer chickens (10 out of 17 farms), 3.4% (16/473) in broilers (five out of 29 farms), and 16.3% (92/564) in Pekin ducks (four out of eight farms), respectively. The highest infection rates were observed in 31-day-old to 60-day-old layer chickens (24.6%) and 11-day-old to 30-day-old Pekin ducks (40.3%). The season of highest prevalence in chickens was spring (15.6%) and the lowest was winter (P<0.01). One hundred and eighty-seven Cryptosporidium-positive samples were analysed by polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis of the small subunit rRNA gene, and 55 were further analysed by DNA sequencing of the PCR products. Two Cryptosporidium species were identified: Cryptosporidium baileyi (184/187) on 15 chicken farms and four duck farms, and Cryptosporidium meleagridis (3/187) on three layer chicken farms. C. baileyi was the predominant Cryptosporidium species, found in all age groups of chickens and all Cryptosporidium-positive ducks examined, whereas C. meleagridis was only identified in 31-day-old to 120-day-old layer chickens. Considering the large size of the chicken industry and the close contact between chickens and humans, and that C. meleagridis is the third most common Cryptosporidium parasite in humans, then C. meleagridis could potentially become an emerging zoonosis in some areas in China. [ABSTRACT FROM AUTHOR]

The distribution and public health significance of Cryptosporidium species/genotypes in pigs differ among geographic areas and studies. To characterize the prevalence of cryptosporidiosis in pigs in Henan, China, a total of 1,350 fecal samples from 14 farms in ten prefectures in Henan Province were examined. The overall prevalence of Cryptosporidium was 8.2% (111/1,350), with the highest infection rate (79/383 or 20.6%) in 1-2-month-old piglets and the lowest infection rates in 3-6-month-old pigs. Cryptosporidium-positive samples from 108 animals were analyzed by polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis of the small subunit rRNA gene, and 35 were further analyzed by DNA sequencing of the PCR products. Two Cryptosporidium species/genotype were identified, including Cryptosporidium suis (94/108) and the Cryptosporidium pig genotype II (14/108). C. suis infection was more common in younger piglets whereas the pig genotype II was relatively common in older pigs. These findings suggest that pigs are not a major source of zoonotic Cryptosporidium in the study area. [ABSTRACT FROM AUTHOR]

• Pet cats are host Cryptosporidium spp. and Giardia duodenalis to humans. • The prevalence of Cryptosporidium spp. and Giardia duodenalis was 0.8 % and 2.0 %. • Cryptosporidium spp. IIdA19G1 subtype was first detected in pet cats in Henan. • Identification of four Giardia duodenalis assemblages in pet cats: A1, C, D, and F. • The infected pet cats were more likely to experience emaciation symptoms. Cryptosporidium spp. and G. duodenalis often infect humans, cats, and other mammals, causing diarrhea and being responsible for numerous outbreaks of waterborne and foodborne infections worldwide. The rapid increase in the number of pet cats poses a substantial public health risk. However, there were few reports about the infection of Cryptosporidium spp. and G. duodenalis infections in pet cats in Henan Province, central China. Thus, to understand the prevalence and genetic distribution of Cryptosporidium spp. and G. duodenalis in pet cats, and to evaluate the zoonotic potential, possible transmission routes and public health implications of isolates, fecal samples (n = 898) were randomly collected from pet cats in 11 cities in Henan Province, central China. Nested PCR based on the SSU rRNA gene and bg gene was used to the prevalence of Cryptosporidium spp. and G. duodenalis , respectively. The prevalence was 0.8 % (7/898) and 2.0 % (18/898) for Cryptosporidium spp. and G. duodenalis respectively. Additionally, the Cryptosporidium spp. positive isolates were identified as C. parvum subtype IIdA19G1 by gp60 gene. In the present study, the IIdA19G1 subtype was discovered in pet cats for the first time in China, enriching the information on the host type and geographical distribution of Cryptosporidium spp. in China. For G. duodenalis, a total of 18 G. duodenalis positive samples were identified, belonging to four assemblages: a zoonotic assemblage A1 (4/898), three host-specific assemblages C (8/898), D (5/898), and F (1/898). Interestingly, we found that pet cats infected with Cryptosporidium spp. and G. duodenalis are more likely to experience emaciation symptoms compared to the negative group. More importantly, the prevalence of Cryptosporidium spp. and G. duodenalis detected in the present study were low, but the subtype IIdA19G1 of Cryptosporidium spp. and the assemblages A1, C, D, and F of G. duodenalis have the potential for zoonotic transmission. Thus, we should focus on preventing and controlling the risk of cross-species transmission that may occur in pet cats in Henan Province. [Display omitted] [ABSTRACT FROM AUTHOR]

Tick-borne diseases (TBDs) impose a significant constraint to livestock production world widely. In this paper, we presented a case of TBD in a cattle farm in Henan, China. 35 blood samples (7 samples sent by veterinarian, 28 samples gathered by our colleagues) were collected from ill, surviving and asymptomatic cattle and microscopic observation and PCR assays were conducted to characterize the pathogens. Genus Ixodes feeding on these cattle were collected and identified. Theileria annulata -like and Anaplasma marginale -like pathogens were observed in the blood smears stained with Giemsa staining under microscope. Furthermore, 5 out of 7 cattle blood samples were found to be positive for T. annulata by PCR. In the 28 blood specimens, three were positive for T. annulata , while A. marginale DNA was detected in nine blood DNA samples. Besides, 56 ticks feeding on cattle were collected from this farm and were all identified as Rhipisephalus microplus , meanwhile, 10 of them were found to be positive for A. marginale . In addition, phylogenetic analysis of the msp4 gene sequences of A. marginale obtained in this study showed that the isolate from cattle (KX840009) fell in the same clade with that of R . microplus (KX904527), sharing 100% similarity. To the best of our knowledge, this is the first confirmed report of outbreak of theileriosis/anaplasmosis in cattle farms in Henan, China. [ABSTRACT FROM AUTHOR]

Highlights: [•] The overall prevalence of Cryptosporidium in goats was 3.48% (44/1256). [•] C. ubiquitum, C. andersoni and C. xiaoi were identified. [•] The zoonotic XIIa subtype 2 of C. ubiquitum was found. [•] C. ubiquitum and C. andersoni were the first identified species in goats. [•] Age-associated distribution of Cryptosporidium was noticed in goats. [ABSTRACT FROM AUTHOR]

Abstract: The prevalence of Cryptosporidium spp. was investigated in scale quail (Coturnix coturnix japonica) farms in Henan Province, China between September 2006 and August 2007. One thousand eight hundred and eighteen fecal samples from 47 quail farms in five areas were collected for the examination of Cryptosporidium oocysts. The overall prevalence of Cryptosporidium was 13.1% (95% CI 13.1±1.6%) (29 of 47 farms), with 72–100-day-old quails having the highest prevalence (23.6%, 95% CI 23.6±2.6%) (χ 2 =64.91; ρ <0.01). The highest prevalence was observed in autumn (21.8%, 95% CI 21.8±3.1%) and the lowest in winter (χ 2 =74.83; ρ <0.01). Two hundred and thirty-nine Cryptosporidium-positive samples were analyzed by PCR-restriction fragment length polymorphism (RFLP) analysis of the small subunit (SSU) rRNA gene, and 42 were further analyzed by DNA sequencing of the PCR products. Two Cryptosporidium species were identified, Cryptosporidium baileyi in 237 birds on 29 farms, and potentially zoonotic Cryptosporidium meleagridis in only two birds on two farms. These findings may suggest that quails are not a major source of zoonotic Cryptosporidium in the study area. [Copyright &y& Elsevier]

Abstract: Cryptosporidium and Giardia infections are common causes of diarrhea worldwide. To better understand the transmission of human cryptosporidiosis and giardiasis in Henan, China, 10 Cryptosporidium-positive specimens and 18 Giardia-positive specimens were characterized at the species/genotype and subtype levels. Cryptosporidium specimens were analyzed by DNA sequencing of the small subunit rRNA and 60kDa glycoprotein genes. Among those genotyped, nine belonged to C. hominis and one C. felis, with the former belonging to three subtype families: Ia, Ib, and Id. The three Ib subtypes identified, IbA16G2, IbA19G2, and IbA20G2, were very different from the two common Ib subtypes (IbA9G3 and IbA10G2) found in other areas of the world. The distribution of Giardia duodenalis genotypes and subtypes was assessed by sequence analysis of the triosephosphate isomerase (tpi) gene. The assemblages A (eight belonging to A-I and four A-II) and B (belonging to six new subtypes) were found in 12 and six specimens, respectively. More systematic studies are needed to understand the transmission of Cryptosporidium and G. duodenalis in humans in China. [ABSTRACT FROM AUTHOR]

Enterocytozoon bieneusi is a zoonotic pathogen of the phylum Microspora that infects humans as well as a variety of animal species worldwide. While molecular epidemiologic studies have characterized this parasite in various hosts, isolates from many susceptible hosts have not yet been examined. In this study, E. bieneusi was isolated from 47 Pere David's deer ( Elaphurus davidianus ) in Henan, China and characterized via PCR analysis of the internal transcribed spacer (ITS) gene. E. bieneusi was detected in 16 out of 47 (34.0%) fecal specimens examined. Sequence analysis of the ITS revealed six known genotypes: type IV (4), EbpC (4), EbpA (4), BEB6 (2), COS-I (1), and COS-II (1). Of these, type IV, EbpC, and EbpA are known to cause human microsporidiosis worldwide, whereas the remaining genotypes are generally specific to ruminants. The present study indicated that Pere David's deer are naturally infected with E. bieneusi , predominantly with zoonotic genotypes, and may pose a risk for human transmission. [ABSTRACT FROM AUTHOR]

Toxoplasmosis is one of the most common foodborne diseases in the world. The objective of this study was to determine Toxoplasma gondii infection in lambs from Henan province, China. A total of 166 lamb hearts were collected from 2017 to 2019. T. gondii infection was determined by the Modified Agglutination Test (MAT) using heart juice of lambs. 11 isolates (TgSheepCHn3 - TgSheepCHn13) were obtained from samples with MAT titers ≥1:100. The rate of T. gondii isolation increased with antibody titer against T. gondii (P < 0.05). No isolate was obtained from samples with titer 1:25 and 1:50, suggesting the cut-off titer for MAT is better set at 1:100. With cut-off value of 1:100, IgG antibodies to T. gondii were found in 25.3% (42/166) of the lambs by MAT. T. gondii parasite was not found in IHC and HE-stained tissue sections of lamb hearts (0/166). Sixty-seven heart tissues with ≥1:25 MAT titers were subjected to acid pepsin digestion and detected T. gondii by PCR. Only 7.5% (5/67) of DNA amplified products were found in heart tissues by the primer TOX5/TOX8. Brain tissue cysts were observed in all mice infected with the 11 isolates at day 60 post infection, suggesting these isolates are non-lethal to mice. PCR-RFLP analysis revealed that 7 isolates belonged to ToxoDB#2, 4 isolates belonged to ToxoDB#4. This is the first isolation of ToxoDB#2 and ToxoDB#4 from lambs in China. Interestingly, none of these isolates belongs to the ToxoDB#9 that is common in China. Our results suggest that the genetic diversity and population structure of T. gondii from China maybe more abundant and magical than previous speculation. • A total of 25.3% (42/166) lambs showed sera conversion of T. gondii antibodies (cut-off titer for MAT: 1:100). • 11 viable T. gondii isolates were obtained from lambs, ToxoDB genotype #2 and #4 were identified. • The rate of isolation T. gondii increased with antibody titer against T. gondii in lamb (P < 0.05). [ABSTRACT FROM AUTHOR]

Toxoplasma gondii is widely distributed and can infect many species of warm-blooded animals, including swine. This study aimed to evaluate the prevalence of T. gondii in swines from the central of China. A total of 2798 samples, including 305 hearts, 2086 diaphragms, and 407 sera were collected from different swine in Henan Province, China. The modified agglutination test was used to detect antibodies against T. gondii in sera from jugular vein blood and heart blood (cut-off: 1:25), diaphragm juice (cut-off: 1:10). T. gondii DNA was screened from the digestive fluids of all diaphragm tissue samples and seropositive hearts, and attempt to isolate viable T. gondii strain by bioassay in mice. A total of 9.94% (278/2798) swine tested positive for T. gondii antibodies. Region, but not gender, was associated with T. gondi seropositivity in swine. T. gondii nucleic acid was not found in the tissue digestive fluids (2090 swines). Three groups of mice showed T. gondii antibodies after having been bioassayed with diaphragm samples (n = 81, which came from 2090 swine). No viable T. gondii strain was isolated from muscle of swine. This is the first large-scale survey T. gondi infection in swine from the central of China. Overall, the prevalence of viable T. gondii in swine was low. Nevertheless, T. gondii infection is present in swine from the central of China. Consumers may acquire T. gondii infection from ingestion of raw or undercooked pork. [ABSTRACT FROM AUTHOR]

Cryptosporidium spp., Giardia duodenalis , Cyclospora cayetanensis , and Enterocytozoon bieneusi are known etiological agents of self-limiting diarrhea, chronic disorders, and severe debilitating illnesses in humans, particularly children and patients with immunodeficiency diseases. To assess the pathogen carriage status of raw vegetables and fruits and the potential transmission routes of the aforementioned parasites in Henan province, China, a total of 1099 vegetables and fruits samples (21 items) were purchased and collected from agricultural farms or open markets. Cryptosporidium spp., E. bieneusi , C. cayetanensis and G. duodenalis were screened by employing polymerase chain reaction (PCR) amplification of species-specific genes. Three kinds of human pathogenic agent (E. bieneusi , C. cayetanensis and C. parvum) were identified on the surfaces of the vegetables and fruits (3.7%, 41/1099). E. bieneusi was found in 3.5% (38/1099) of the samples, whereas C. cayetanensis and C. parvum were only identified in two (0.2%) and one (0.1%) of the vegetable and fruit samples, respectively. No G. duodenalis contamination was detected in the present study. In total, 12 different E. bieneusi ITS genotypes (eight known and four novel) were detected, of which the ten (EbpA, CM8, CHG19, EbpC, CTS3, Henan-IV, and CHV1 to CHV4) that occurred in 20 samples (20/38, 52.6%) clustered into the previously described high potential zoonotic group 1 in the phylogenetic analysis. The remaining two known genotypes (BEB8 and CD6) detected in 18 samples (18/38, 47.4%) belonged to group 2. That C. cayetanensis , C. parvum and some E. bieneusi genotypes have been reported in humans, highlights the possible risk of foodborne related disease outbreaks. • Three types of protozoan were identified on surface of vegetables and fruits. • These pathogens had previously been reported in human and animals stool. • A risk of parasitic illnesses associated with fresh fruits and vegetables consumption. [ABSTRACT FROM AUTHOR]