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Background: Cryptosporidium and Giardia are well-known important intestinal zoonotic pathogens that can infect various hosts and cause diarrhoeal diseases. We aimed to determine the epidemiological prevalence and molecular characterization of Cryptosporidium and Giardia species in Himalayan marmot (Marmota himalayana, class Marmota) in the Qinghai Tibetan Plateau Area of Qinghai Province, Northwest China. Methods: Overall, 243 Himalayan marmot fecal samples were collected in 2017 and in 2019 and a two-step nested PCR technique was performed to amplify the fragments of the SSU rRNA gene of Cryptosporidium and 18S ribosomal RNA gene of Giardia. Molecular characterization of Cryptosporidium was performed with the primary primers NDIAGF2 and N-DIAGR2, the secondary primers CPB-DIAGF and CPB-DIAGR. Similarly, molecular characterization of Giardia was used the first primers Gia2029 and Gia2150c, the secondary primers RH11 and RH4. The positive PCR products were sequenced and the sequences were processed by Clustal Omega and BLAST. Phylogenetic analysis was achieved by NJ method in MEGA. Results: The infection rate of Cryptosporidium spp. and G. duodenalis was 4.9% (12/243) and 0.8% (2/243) in M. himalayana, respectively. Cryptosporidium spp. are characterized as novel genotypes Cryptosporidium marmot genotype I (n=3) and Cryptosporidium marmot genotype II (n=9); G. duodenalis assemblage A (n=2) was found in M. himalayana. Conclusion: This is the first report of Cryptosporidium spp. and G. duodenalis infections in M. himalayana in Qinghai of Northwest China. The results indicate the existence of Cryptosporidium species and G. duodenalis infections that may have a potential public health significance. [ABSTRACT FROM AUTHOR]

Background: Cryptosporidiosis and giardiasis are significant parasitic diseases shared between humans and domestic animals. Due to the close contact between humans and domestic animals in rural areas, it is important to consider the potential transmission of zoonotic parasites from infected domestic animals to humans. This investigation aimed to determine the prevalence and molecular characteristics of Cryptosporidium spp. and Giardia duodenalis in domestic animals and villagers. Methods: A total of 116 fecal samples from villagers and 686 fecal samples from domestic animals in Heilongjiang Province, China, were analyzed for two parasites using nested polymerase chain reaction (PCR) targeting various genetic loci and DNA sequence analysis of the PCR products. Results: By sequence analysis of the SSU rRNA gene, the prevalence of Cryptosporidium in humans was 0.9% (1/116), with one species of C. parvum (n = 1) detected; among domestic animals, the prevalence was 2.6% (18/686), with five species identified: C. suis (n = 7) and C. scrofarum (n = 7) in pigs, C. meleagridis (n = 1) in chickens, C. andersoni (n = 1) in cattle, and C. canis (n = 2) in foxes. C. parvum and C. canis were further subtyped as IIdA19G1 and XXa4 on the basis of gp60 gene. Regarding G. duodenalis, based on the SSU rRNA, bg, gdh, and tpi genes, the prevalence in domestic animals was 5.1% (31/608), with three assemblages identified: A (n = 1) in pigs, D (n = 1) in foxes, and E (n = 27) in geese, cattle, pigs, ducks, and sheep, along with mixed infection of A + E (n = 1) in one pig and B + E (n = 1) in one sheep. No G. duodenalis was detected in humans (0/116). Conclusions: The present results show that no overlap of subtypes between animals and villagers was found in Cryptosporidium spp. and G. duodenalis, indicating a minor role of domestic animals in infecting humans in this population. However, the presence of zoonotic protozoa in domestic animals highlights the need for special attention to high-risk individuals during close contact with domestic animals. [ABSTRACT FROM AUTHOR]

Simple Summary: Giardia duodenalis is a common zoonotic intestinal protozoan and causes significant economic losses and potential risks to public health. China is among the several major producers of donkeys worldwide, with approximately 2.6 million donkeys being reared in 2019, but studies on G. duodenalis infection in donkeys were limited. Thus, in order to better understand the molecular epidemiology of G. duodenalis in donkeys in China, 815 fecal samples were collected from donkeys in three representative geographical locations in Shanxi Province, North China, to determine the presence and genotypes of G. duodenalis based on three established loci (tpi, bg, and gdh). A total of 137 out of 815 fecal samples were detected as G. duodenalis-positive with an overall prevalence of 16.81%. Among the detected assemblages A, B, and E, the most predominant assemblage was B in the three study areas. Three MLGs (MLG-novel-1 to 3) were identified with multilocus genotypes (MLGs) analysis and validated by a phylogenetic tree. This is the first report of G. duodenalis infection in donkeys in Shanxi Province, which not only enhances our understanding of the genetic diversity of G. duodenalis in donkeys in China but also provides essential baseline data for the prevention and control of G. duodenalis infection donkeys in the study areas. Giardia duodenalis is a ubiquitous flagellated protozoan, causing significant economic losses to animal husbandry and posing threats to public health. China ranks the world's sixth largest major producer of donkeys, rearing approximately 2.6 million donkeys in 2019, but limited investigation of G. duodenalis prevalence has been conducted in the past, and it is yet to be known whether donkeys in Shanxi Province are infected with G. duodenalis. In the present study, a total of 815 fecal samples collected from donkeys in representative regions of Shanxi Province, North China, were examined for G. duodenalis using nested PCR. Then, the assemblages and multilocus genotypes (MLGs) were examined based on three established loci: namely, β-giardin (bg), triosephosphate isomerase (tpi), and glutamate dehydrogenase (gdh). The overall prevalence of G. duodenalis in donkeys in Shanxi Province was 16.81% (137/815). The region was identified as the main risk factor for the observed difference in G. duodenalis prevalence in donkeys among the three study areas (χ2 = 21.611, p < 0.001). Assemblages A, E, and B were identified, with the latter as the predominant assemblage. Three MLGs (MLG-novel-1 to 3) were formed based on sequence variation among the three loci. The present study reveals the presence of G. duodenalis in donkeys in Shanxi Province, North China, for the first time, which not only enriches the data on the distribution of G. duodenalis in donkeys in China but also provides useful baseline data for planning control strategies against G. duodenalis infection in the sampled areas. [ABSTRACT FROM AUTHOR]

Giardia duodenalis is a gastrointestinal protozoan ubiquitous in nature. It is a confirmed zoonotic pathogen, and cattle are considered a source of giardiasis outbreaks in humans. This study aimed to evaluate the prevalence and multilocus genotype (MLG) of G. duodenalis in dairy cattle in Central Inner Mongolia. This study was based on the small subunit ribosomal RNA (SSU rRNA), glutamate dehydrogenase (gdh), triosephosphate isomerase (tpi), and beta-giardin (bg) genes of G. duodenalis. DNA extraction, polymerase chain reaction (PCR), and sequence analysis were performed on 505 dairy cattle fecal samples collected in 2021 from six sampling sites and four age groups in Central Inner Mongolia to determine the prevalence and MLG distribution of G. duodenalis. The PCR results of SSU rRNA revealed that the overall prevalence of G. duodenalis was 29.5% (149/505) and that the overall prevalence of the diarrhea and nondiarrhea samples was 31.5% (46/146) and 28.5% (103/359), respectively; the difference was not significant (p > 0.05). SSU rRNA sequence analysis revealed that G. duodenalis assemblage E (91.1%, 133/146) was primarily detected and that assemblage A (8.9%, 13/146) was detected in 13 samples. The G. duodenalis—positive samples were PCR amplified and sequenced for gdh, tpi, and bg, from which 38, 47, and 70 amplified sequences were obtained, respectively. A combination of G. duodenalis assemblages A and E were detected in seven samples. Multilocus genotyping yielded 25 different assemblage E MLGs, which formed six subgroups. To the best of our knowledge, this is the first report regarding G. duodenalis infection in dairy cattle in Inner Mongolia, China. This study revealed that Inner Mongolian cattle pose a risk of giardiasis transmission to humans and that the distribution of local cattle G. duodenalis assemblage E MLGs is diverse. The findings of this study can bridge the knowledge gap in the molecular epidemiological investigation of giardiasis in Central Inner Mongolia. [ABSTRACT FROM AUTHOR]

Simple Summary: Giardia duodenalis is a world-widely distribution intestinal protozoan parasite that can infect human and a broad range of mammals. It can cause a number of clinical symptoms including diarrhoea, abdominal pain, vomiting, severe dehydration and anemia etc. in animals and humans. Eight genetic assemblages (A to H) have been identified in G. duodenalis based on molecular analysis. In this study, the infection rates and genotypes of G. duodenalis in swine are investigated in Fujian province, southeastern China. G. duodenalis assemblages E was found in pigs in the present study. These results provide epidemiological data for giardiasis prevention, control and public health. Giardia duodenalis, an intestinal parasite, is widely distributed in humans and various animals, such as pigs, cattle and cats. The clinical symptoms of giardiasis are characterized as including abdominal pain, acute or chronic diarrhea, and bloating and weight loss in humans and animals, leading to public and veterinary health problems worldwide. However, the prevalence and genotypes of G. duodenalis in pigs in Fujian Province, southeastern China, have not been reported. In the present study, 725 fecal samples were collected from six cities (Fuqing, Putian, Nanping, Longyan, Sanming, Zhangzhou) in Fujian Province and analyzed for G. duodenalis prevalence and genotypes using nested PCR targeting the beta-giardin (bg), glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi) genes. The results shown that total occurrence rate of G. duodenalis was 26.9% (195/725) in pigs, with significant differences in the prevalence among different regions (χ2 = 86.508, p < 0.05) and groups (χ2 = 12.748, p < 0.05). 195, 11 and 6 samples were detected at the bg, tpi and gdh loci, respectively. Each one belonged to a subtype of assemblage E and was analyzed using sequences obtained in this study. Based on phylogenetic analyses of sequences from the three genetic loci, only one MLG E1 was found. The results indicated that pigs may present a potential zoonotic risk of spreading G. duodenalis infection from animals to humans in this area. The findings of the present study also provide basic data for the prevention and control of G. duodenalis infection in pigs and humans in China. [ABSTRACT FROM AUTHOR]

Background: Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi can cause important intestinal diseases in ruminants. However, data on the distribution of these three protozoan pathogens in Tibetan sheep are limited. Methods: We collected 761 fecal samples from Tibetan sheep across four seasons in Qinghai Province, China, and screened the samples for Cryptosporidium spp., G. duodenalis and E. bieneusi using PCR-based sequence analysis of the genes encoding 18S ribosomal RNA, triosephosphate isomerase and the internal transcribed spacer, respectively. Results: The positivity rates of Cryptosporidium spp., G. duodenalis and E. bieneusi in Tibetan sheep were 3.68% (28/761 samples), 1.58% (12/761) and 6.44% (49/761), respectively. Four species of Cryptosporidium were identified: C. xiaoi (n = 13 samples), C. ubiquitum (n = 8), C. bovis (n = 6) and C. ryanae (n = 1). Two G. duodenalis assemblages, namely the A (n = 2 samples) and E (n = 10) assemblages, were detected. Five zoonotic E. bieneusi genotypes were found: BEB6 (n = 21 samples), COS-I (n = 14), CHS3 (n = 11) and CGS1 (n = 2) from group 2, and PIGEBITS5 (n = 1) from group 1. Geographic differences in the distribution of E. bieneusi, and seasonal differences for all the three protozoan pathogens were noted. Conclusions: Our results elucidate the prevalence and genetic diversity of these three pathogens in Tibetan sheep across different regions and seasons, including zoonotic pathogens such as C. ubiquitum, C. ryanae, G. duodenalis assemblage A and five genotypes of E. bieneusi. [ABSTRACT FROM AUTHOR]

Background: Few studies have molecularly characterized the potential zoonotic protozoa, Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in sheep and goats in China, therefore total 472 fecal samples were collected from eight provinces and infection rates of three protozoa were determined by PCR analysis of corresponding loci. All PCR positive samples were sequenced to identify the genotype. Results: The overall infection rates for Cryptosporidium, G. duodenalis, and E. bieneusi were 1.9% (9/472), 20.6% (97/472), and 44.5% (210/472), respectively. C. xiaoi (n = 5), C. ubiquitum (n = 3), and C. anderson (n = 1) were identified in goats. 97 G. duodenalis strains were successfully detected, and assembly E (n = 96) and assembly A (n = 1) were identified. Two novel G. duodenalis multilocus genotype (MLGs) were identified, with one belonging to subgroup AI and the other to subgroup E5. Nine known genotype (BEB6, CD6, CHC8, CHG3, CHG5, Peru6, CHG1, CHG2, and COS-I) and four new genotype (CHG26, CHG27, CHG28, and CHS18) were identified in E. bieneusi, with CHG3 dominant in this group. Conclusions: The present results highlight the role of sheep and goats as reservoir hosts for this three gastrointestinal pathogens. In summary, we provided a platform for more detailed research on genotyping or subtyping intestinal pathogens to better understand their risks and modes of transmission. [ABSTRACT FROM AUTHOR]

Giardia duodenalis is a flagellated parasitic microorganism that parasitizes in the intestines of humans and animals. Although asymptomatic infections commonly exist in both humans and animals, some enteric symptoms have been reported in immunocompromised individuals, posing a threat to public health. Children could be infected with G. duodenalis through an environment contaminated by infective animals. Thus, the investigation of the prevalence and genotypes of G. duodenalis in zoo animals is important. In this study, 672 fecal samples of 113 species of animals, including non-human primates, artiodactyla, perissodactyla, proboscidian, marsupial, birds, carnivora, and rodents, were collected from three zoos in Hangzhou city, Dalian city, and Suzhou city in China. The samples were screened for the positivity of G. duodenalis by nested PCR based on the β-giardin (bg), triosephosphate isomerase (tpi), and glutamate dehydrogenase (gdh) gene loci. The overall G. duodenalis prevalence was 10.6% (71/672). The prevalence in non-human primates, artiodactyla, perissodactyla, proboscidian, marsupial, birds, carnivora, and rodent was 6.9% (10/144), 9.0% (12/133), 17.1% (6/35), 0% (0/6), 8.7% (2/23), 13.3% (28/211), 6.7% (7/105), and 40.0% (6/15), respectively. The region and category were considered risk factors for G. duodenalis infection in zoo animals (p < 0.001). Additionally, four genotypes of G. duodenalis were identified in zoo animals, including assemblage E (n = 46), assemblage A (n = 18), assemblage B (n = 6), and assemblage D (n = 1). The assemblages A, B, D, and E are also genotypes observed in humans and other animals. Among the sequences obtained in our study, one multilocus genotype (MLG) of the sub-assemblage AI was observed within assemblage A. Furthermore, three MLGs were detected within assemblage B. These findings reveal G. duodenalis genetic variability in zoo animals in three cities in China and suggest that zoo animals could be a potential source of human infection with G. duodenalis. [ABSTRACT FROM AUTHOR]

Cryptosporidium spp. and Giardia duodenalis are commonly detected intestinal protozoa species in humans and animals, contributing to global gastroenteritis spread. The present study examined the prevalence and zoonotic potential of Cryptosporidium spp. and G. duodenalis in Himalayan marmots and Alashan ground squirrels in China's Qinghai-Tibetan Plateau area (QTPA) for the first time. Four hundred ninety-eight intestinal content samples were collected from five counties of QTPA of Gansu province, China. All samples were examined for Cryptosporidium spp. and G. duodenalis by PCR amplification. The resultant data were statistically analyzed by chi-square, Fisher's test and Bonferroni correction using SPSS software 25. 0. Cryptosporidium positive samples were further subtyped through analysis of the 60-kDa glycoprotein (gp60) gene sequence. A total of 11 and 8 samples were positive for Cryptosporidium spp. and G. duodenalis, respectively. Prevalence of Cryptosporidium spp. and G. duodenalis were 2.5% (10/399) and 1.5% (6/399) in Himalayan marmots, 1.0% (1/99) and 2.0% (2/99) in Alashan ground squirrels, respectively. Sequence analysis confirmed the presence of C. rubeyi (n = 2), ground squirrel genotype II (n = 7), chipmunk genotype V (n = 1) and horse genotype (n = 1). The horse genotype was further subtyped as novel subtype VIbA10. G. duodenalis zoonotic assemblages A (n = 1), B (n = 6), E (n = 1) were identified in the present study. This is the first study to identify Cryptosporidium spp. and G. duodenalis in Himalayan marmots and Alashan ground squirrels, suggesting the potential zoonotic transmission of the two pathogens in QTPA. [ABSTRACT FROM AUTHOR]

The zoonotic protozoa parasites Cryptosporidium spp. and Giardia duodenalis infect a wide range of hosts, including humans. Pigs are reservoir hosts for Cryptosporidium spp. and G. duodenalis, which can transmit cryptosporidiosis and giardiasis to humans and other animals. The current study sought to investigate the infection rates and species/genotypes of Cryptosporidium spp. and G. duodenalis in pigs in Hubei of China. The nested PCR and sequence analyses of the small ribosomal subunit RNA (SSU rRNA) gene for Cryptosporidium spp. and the b-giardin (bg) gene for G. duodenalis was utilized to screen for the infection of those parasites in a total of 826 fresh fecal samples. Both Cryptosporidium spp. and G. duodenalis infection rates were 0.97% (8/826). Of the genotyped positive isolates, 6/8 (75%) were C. scrofarum and 2/8 (25%) were C. suis, while two zoonotic species G. duodenalis assemblage E and assemblage A were also detected in 7/8 (87.5%) isolates and 1/8 (12.5%) isolates, respectively. The findings suggest that both of those parasites in pig in intensive farms of Hubei province, China, pose a potential public health risk. [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]

Background: Giardia duodenalis is a common parasitic diarrheal agent in humans, especially in developing countries. The aim of this study was to investigate the prevalence and multilocus genetic characterization of G. duodenalis in patients with diarrhea and animals in northeastern China, and to assess the epidemiological role of animals in the transmission of human giardiasis. Methods: A total of 1739 fecal specimens from 413 diarrheal patients and 1326 animals comprising 16 mammal species were collected in Heilongjiang Province of China and screened for G. duodenalis by PCR and sequencing of the SSU rRNA gene. All G. duodenalis-positive specimens were subtyped by PCR and sequencing of the bg, tpi, and gdh genes. To detect additional mixed infections of different assemblages, assemblage A/B/E-specific PCRs were performed to amplify the tpi gene. Results: Sequence analysis of the SSU rRNA gene determined the prevalence of G. duodenalis (5.81%, 24/413) in diarrheal patients, with a peak in minors aged 5–17 years, and identified assemblages A and B. MLG-AII and MLG-B1 were obtained based on concatenated nucleotide sequences of the bg, tpi, and gdh genes, with MLG-AII being identical to a cat-derived isolate reported previously. By sequence analysis of the SSU rRNA gene, G. duodenalis was detected in 214 (16.14%) animals belonging to 11 mammal species, with the prevalence ranging from 1.69 to 53.85%, and assemblages A to G were identified. Sequence analysis of the bg, tpi, and gdh genes from 46 specimens produced 31 MLGs, including MLG-AI (n = 1), MLG-B2–B8 (n = 18), and MLG-E1–E23 (n = 27). Conclusions: The finding of G. duodenalis in diarrheal patients enhances consciousness of detecting G. duodenalis in clinical practice and emphasizes the importance of health education in local inhabitants, especially in the age group of 5–17 years. The identification of seven assemblages (A to G) and 33 MLGs reveals genetic heterogeneity of G. duodenalis in the investigated areas. Due to insufficient homology data on the zoonotic transmission of G. duodenalis, the precise epidemiological role that animals play in the transmission of human giardiasis needs to be assessed by more large-scale molecular epidemiological investigations of local humans and animals. [ABSTRACT FROM AUTHOR]

Giardia duodenalis is the underlying cause of a significant number of outbreaks of gastrointestinal illness in humans and animals worldwide. The purpose of this study was to elucidate the prevalence and genetic diversity of G. duodenalis in captive alpine musk deer (Moschus chrysogaster) in China. A total of 202 fecal samples were collected from three farms in Gansu Province, China. Identification of G. duodenalis was conducted by nested PCR targeting the genes coding for SSU rRNA, β-giardin (bg), glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi). The overall prevalence of G. duodenalis in captive alpine musk deer in surveyed area was 19.3% (39/202). Two G. duodenalis genetic assemblages were identified, namely assemblage A and E. Mixed genotype infections (A+E) were found in 15.4% (6/39) of positive samples. Multilocus genotyping (MLG) analysis of G. duodenalis isolates revealed six novel assemblage A MLGs formed by two newly-described MLG-subtypes which belonged to sub-assemblage AI. To the best of our knowledge, this is the first report on MLG of G. duodenalis isolates in captive alpine musk deer in China. The presence of zoonotic assemblages and sub-assemblages of G. duodenalis in deer species suggests that these animals may potentially act as a reservoir of this protozoan for humans. [ABSTRACT FROM AUTHOR]

Background: Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi are important zoonotic protists in humans and animals around the world, including nonhuman primates (NHPs). However, the prevalence, genetic identity and zoonotic potential of these pathogens in wild NHPs remain largely unclear. Methods: A total of 348 fecal samples were collected from wild NHPs at four locations in Yunnan, southwestern China, and analyzed for these pathogens using nested PCR targeting various genetic loci and DNA sequence analysis of the PCR products. The zoonotic potential of the pathogens was assessed by comparing the genetic identity of the pathogens in these animals with that previously reported in humans. Results: Altogether, two (0.6%), 25 (7.2%) and 30 (8.6%) samples were positive for Cryptosporidium sp., G. duodenalis and E. bieneusi, respectively. The Cryptosporidium sp. identified belonged to C. parvum subtype IIdA20G1. Both assemblages A (n = 3) and B (n = 22) were identified among G. duodenalis-positive animals. Five genotypes in zoonotic Group 1 were identified within E. bieneusi, including Type IV (n = 13), D (n = 7), Peru8 (n = 6), MMR86 (n = 2) and HNFS01 (n = 2). All genotypes and subtypes identified are known human pathogens or phylogenetically related to them. Conclusions: Data from this study suggest a common occurrence of zoonotic genotypes of G. duodenalis and E. bieneusi in wild NHPs in southwestern China. [ABSTRACT FROM AUTHOR]

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Copyright of Parasite (1252607X) is the property of EDP Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Knowledge gaps in the exposure parameters for recreational water activities make quantitative risk assessment related to water recreation difficult. Therefore, the annual exposure frequency and single exposure duration for the recreational water activities of residents from ten cities in the North and South of China were investigated. Questionnaire interviews were carried on recreational water activities comprising swimming (SW), boating (BA), playing in interactive fountains (PF), and watching fountains (WF). Quantitative microbial risk assessment for the exposure of urban residents to Cryptosporidium and Giardia was also performed. For the four recreational water activities, the participation rates of urban residents in SW and WF were higher than the others. For SW and BA, the mean annual exposure frequency and single exposure duration for males were significantly higher than those for females. PF and WF showed the opposite. The annual exposure frequency for above 35-year-old residents was higher than that for young residents (18–35 years). However, the single exposure duration for young residents was highest in SW, BA, and PF. The mean annual exposure frequency and single exposure duration for North China residents were higher than those for South China residents in all recreational water activities, except for SW. Overall, the annual exposure frequency and single exposure duration in recreational water activities for all urban residents followed a lognormal distribution. In the four recreational water activities, the total annual infection risk of male exposure to Cryptosporidium was 1.0 × 10-2, with the confidence intervals between 95 and 5% of [4.3 × 10-4, 3.7 × 10-2], whereas that for females was 6.8 × 10-3 and [4.2 × 10-4, 2.4 × 10-2]. Also, the annual infection risk of males to Giardia was 8.8 × 10-3 and [5.1×10-4, 3.2×10-2], and that of females was 5.3 × 10-3 and [4.0 × 10-4, 1.8 × 10-2]. These results demonstrated that SW and PF made the highest contribution to the total annual infection risk. Sensitivity analysis highlighted that the characterization of exposure parameters plays a critical role in health risk assessment, which may provide a scientific basis for recreational water quality standards formulation. [ABSTRACT FROM AUTHOR]

Background: The waterborne pathogens Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi and Cyclospora cayetanensis can cause intestinal diseases in humans. An understanding of their occurrence and transport in the environment is essential for accurate quantitative microbial risk assessment. Methods: A total of 238 influent samples were collected from four wastewater treatment plants (WWTPs) and 88 samples from eight sewer locations in Guangzhou, China. PCR-based tools were used to detect and genetically characterize Cryptosporidium spp., G. duodenalis and E. bieneusi. Eimeria spp. and Cyclospora spp. were also analyzed to assess the sources of Cryptosporidium spp., G. duodenalis and E. bieneusi in wastewater. Results: The overall occurrence rates in the WWTP and sewer samples were 14.3% (34/238) and 13.6% (12/88) for Cryptosporidium spp., 55.5% (132/238) and 33.0% (29/88) for G. duodenalis, 56.3% (134/238) and 26.1% (23/88) for E. bieneusi and 45.4% (108/238) and 47.7% (42/88) for Eimeria spp., respectively. Altogether, 11 Cryptosporidium species and genotypes, six G. duodenalis genotypes, 11 E. bieneusi genotypes and four C. cayetanensis were found, together with the presence of nine Eimeria species. The common occurrence of Cryptosporidium rat genotype IV, C. muris and Eimeria papillata and E. nieschulzi suggested that rodents were significant sources of the enteric pathogens detected in the wastewater samples. Conclusions: While the dominant Cryptosporidium spp. detected in the raw wastewater sampled in this study are not pathogenic to humans, the widely detected G. duodenalis assemblage A and E. bieneusi genotypes D and Type IV are well-known zoonotic pathogens. Further studies are needed to monitor the occurrence of these waterborne pathogens in WWTPs to better understand their transmission and environmental transport in China. [ABSTRACT FROM AUTHOR]

Background: Giardia duodenalis is an important opportunistic zoonotic intestinal protozoon, which could parasitize yaks. However, a few studies have been conducted on the seasonal infection of G. duodenalis in yaks in China. Methods: Overall, 1,027 fecal samples were collected from yaks of two age groups in seven cities of Qinghai Province, China at four seasons between May 2016 and Sep 2017. The prevalence and assemblages were analyzed by nested PCR and multilocus sequence typing (MLST). Results: The overall prevalence of G. duodenalis was 2.04% (21/1027) based on triose phosphate isomease (tpi) locus. No significant differences in prevalence of the organism in yaks were found among different sampling areas. Additionally, same result was also presented in different seasons. However, there was statistically significant difference between young yaks within 6 months (8.33%, 4/48) and adult yaks over 6 months (1.73%, 17/979). The assemblage A recognized as a zoonotic assemblage (n=3) was found in yaks (>6 months) from Xining, while assemblage E (n=18) was detected from yaks in six cities. There were 5, 2 and 3 G. duodenalis subtypes detected positive at the tpi, the β-giardin (bg), and the glutamate dehydrogenase (gdh) loci, with 2, 2 and 3 novel subtypes, respectively. Three samples were successfully sequenced at all three loci, forming 1 assemblages A multilocus genotype (MLG) and 2 assemblages E MLGs, not reported. Conclusion: This study indicated a zoonotic potential of G. duodenalis in yaks from Qinghai Province and provides basic information about the epidemiology of G. duodenalis. [ABSTRACT FROM AUTHOR]

Giardia lamblia (syn Giardia duodenalis) is an important protozoan parasite that can cause enterocyte damage and loss of brush border of the epithelial cells in the intestine, resulting in shortening of microvilli and altered epithelial barrier function. Many animals have been detected as the hosts of the G. lamblia. However, the information on the epidemiology and molecular detection of G. lamblia in dairy calves and sika deer in northeastern China is limited. To investigate the prevalence and genotypes of dairy calves and sika deer in northeastern China, a total of 321 fecal samples from dairy calves in Heilongjiang Province and 818 fecal samples from sika deer in four provinces (Jilin Province, Heilongjiang Province, Liaoning Province, and Inner Mongolia Autonomous Region) in China were conducted by PCR methods, between September 2017 and April 2018. The overall prevalence of G. lamblia in dairy calves in Heilongjiang Province and sika deer in the four provinces was 4.98% (16/321) and 0.61% (5/818), respectively. In this study, the point prevalence of Giardia spp. in different factor groups was dissimilar. A total of 16 Giardia spp. positive samples in dairy calves were identified as assemblage E based on the triosephosphate isomerase (tpi), β-giardine (bg), and glutamate dehydrogenase (gdh) genes. Furthermore, two positive samples of assemblage A and three positive samples of assemblage E were identified with gdh and bg genes in the sika deer. Assemblage A was zoonotic genotype of G. lamblia, and assemblage E was identified as the predominant assemblage in dairy calves and sika deer. This study reported the prevalence and genotypes of G. lamblia in dairy calves in Heilongjiang Province and sika deer in four provinces in China. These results provided basic information to understand the epidemiology of G. lamblia in dairy calves and sika deer in China. [ABSTRACT FROM AUTHOR]

Cryptosporidium spp., Enterocytozoon bieneusi, and Giardia duodenalis are zoonotic pathogens commonly found in the intestinal tract of mammalian hosts including livestock and humans. The prevalence of these eukaryote microorganisms in domestic animals and their interaction with intestinal microbiota are not yet fully recognized. We analyzed the intestinal microbiota composition with metagenomics and functional characterization with Cluster of Orthologous (COG) in Bactrian camels, which were raised on Qinghai-Tibet Plateau, Northwest China. Thus, fecal samples were collected from the animals to determine the parasite infection and the profile of microbiota. Analysis of intestinal microbiota at genus level revealed important features of interaction between parasites infection and bacterial community. Coprococcus and Prevotella were more abundant while Akkermansia had lower relative abundance with E. bieneusi infection. Bacteria of Akkermansia, Lactococcus, Oxalobacter, Sphaerochaeta, Paludibacter, Fibrobacter, Anaerovibrio, Pseudomonas, Mogibacterium, Pseudoramibacter_Eubacterium, YRC22, Flexispira, SMB53, AF12, and Roseburia genera were found under-presented and Oscillospira genus over-presented when G. duodenalis infection was present. Meanwhile, Cryptosporidium spp. and E. bieneusi co-infected animals showed lower relative abundance of Allobaculum, Rikenella, Shuttleworthia, Epulopiscium, Bilophila, Dorea, Fibrobacter, and TG5. Results demonstrate important interaction between the intestinal parasites and microbiota, and provide informative link for understanding the co-evolution of zoonotic pathogens and bacteria in domestic animals. [ABSTRACT FROM AUTHOR]

Cryptosporidium and Giardia are important intestinal zoonotic pathogens that can infect various hosts and cause diarrhoeal diseases. There are few reports of the epidemiological prevalence and molecular characterization of Cryptosporidium and Giardia in wild birds around Qinghai Lake and in the surrounding areas on the Qinghai-Tibetan Plateau, Northwest China. Therefore, the aim of this study was to determine the Cryptosporidium spp. and Giardia duodenalis genotypes and their epidemiological prevalence in wild birds by PCR amplification. To our knowledge, this is the first report of a variety of Cryptosporidium spp. and G. duodenalis infections in wild birds from that area, with overall prevalence rates of 8.98% (61/679) and 3.39% (23/679), respectively. Furthermore, PCR sequencing confirmed the presence of Cryptosporidium baileyi (n = 3), Cryptosporidium parvum (n = 58), and G. duodenalis assemblage B (n = 19) and E (n = 4) in wild birds from the areas around Qinghai Lake. The results of the present study demonstrated the wide distribution of Cryptosporidium and Giardia among wild birds, which has potential public health significance. Moreover, the study findings also provided useful molecular epidemiological data for monitoring and investigating the two parasitic protozoa in wild animals and surrounding environments. [ABSTRACT FROM AUTHOR]

The protozoan parasite Giardia duodenalis is known to infect humans and a wide range of animals globally. However, no studies on G. duodenalis infection in Bactrian camels have been reported. In the present study, in order to examine the prevalence and genetic diversity of G. duodenalis in Bactrian camels, 852 fecal samples were collected from 24 sampling sites in three geographical areas (Gansu province, Inner Mongolia, and Xinjiang Uygur autonomous regions) of northwestern China, and subjected to multilocus sequence typing (MLST) analysis targeting the 18S rRNA, β-giardin (bg), glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi) genes. About 84 fecal samples tested positive for Giardia infection, with an overall prevalence of 9.8%, including three samples from camel calves with diarrhea. Significant differences (χ2 = 80.7, df = 2, P < 0.01) in the prevalence were found in Bactrian camels belonging to three geographical areas, with the highest (33.3%) in Gansu province and the lowest (4.2%) in Xinjiang Uygur autonomous region. Furthermore, significantly different prevalences (χ2 = 34.2, df = 2, P < 0.01) were revealed among age groups, with the highest (35.7%) in camels aged 3 to 6 years old, and the lowest (7.5%) in camels aged > 6 years old. Sequence analysis identified two assemblages, including zoonotic assemblage A and ungulate-adapted assemblage E, with the latter as the dominant G. duodenalis assemblage in each age group and at all sampling sites having positive samples except Hotan. Genetic variations were detected among G. duodenalis isolates in these camels, and eight, three, and seven haplotypes were identified at loci bg, gdh, and tpi, respectively, forming two multilocus genotypes (MLGs) of zoonotic assemblage A and one MLG of assemblage E. To the best of our knowledge, this is the first report on G. duodenalis infection in Bactrian camels, and the data indicate that G. duodenalis have a broad host range. [ABSTRACT FROM AUTHOR]

Cryptosporidium and Giardia are well-known parasitic protozoans responsible for waterborne and foodborne diarrhoeal diseases. However, data are not available on market vegetables contaminated with Cryptosporidium and Giardia in China. In the present study, 642 different vegetable samples were collected from Xining City street vendors in the Qinghai Province to study the Cryptosporidium and Giardia contamination rates via PCR and sequence analyses. Cryptosporidium spp. and Giardia duodenalis were detected in 16 (2.5%) and 73 (11.4%) samples, respectively. Two species of Cryptosporidium, C. parvum (n = 11) and C. andersoni (n = 5), were identified. G. duodenalis assemblage B was identified in almost all positive samples (n = 72), except one sample that contained G. duodenalis assemblage E. We report on the rate of Cryptosporidium and Giardia contamination in vegetables for the first time from the Qinghai Tibetan Plateau Area (QTPA) in China. [ABSTRACT FROM AUTHOR]

Limited data are available on infection rates and genetic identity of Enterocytozoon bieneusi and Giardia duodenalis in horses and donkeys. In this study, 865 fecal specimens were collected from donkeys (n = 540) and horses (n = 325) in three provinces and autonomous regions in northern China during 2015–2019. Enterocytozoon bieneusi was detected and genotyped by PCR and sequence analyses of the ribosomal internal transcribed spacer (ITS) and G. duodenalis was detected and genotyped by PCR and sequence analyses of the β-giardin, glutamate dehydrogenase, and triosephosphate isomerase genes. The overall infection rates of E. bieneusi and G. duodenalis were 21.9% (118/540) and 11.5% (62/540) in donkeys, and 7.4% (24/325) and 2.8% (9/325) in horses, respectively. These differences in infection rates of E. bieneusi and G. duodenalis between donkeys and horses were significant (χ2 = 30.9, df = 1, P < 0.0001; χ2 = 20.4, df = 1, P < 0.0001, respectively). By age, the 28.9% infection rate of E. bieneusi in donkeys under 6 months was significantly higher than that in animals over 6 months (6.0%; χ2 = 35.2, df = 1, P < 0.0001). In contrast, donkeys of 6–12 months had higher infection rate (35.9%) of G. duodenalis than donkeys under 6 months (9.9%; χ2 = 22.1, df = 1, P < 0.0001) and over 12 months (8.7%; χ2 = 17.3, df = 1, P < 0.0001). In horses, animals of > 12 months had significantly higher infection rate (31.1%) of E. bieneusi than horses under 6 months (3.4%; χ2 = 29.4, df = 1, P < 0.0001) and 6–12 months (3.8%; χ2 = 26.1, df = 1, P < 0.0001). Twenty genotypes of E. bieneusi were detected, including six known ones and 14 new genotypes. Among them, nine genotypes in 45% E. bieneusi–positive specimens belonged to the zoonotic group 1. Similarly, three G. duodenalis assemblages were detected, including A (in 2 horses and 30 donkeys), B (in 6 horses and 29 donkeys), and E (in 1 horse); three donkeys had coinfections of assemblages A and B. The assemblage A isolates identified all belong to the sub-assemblage AI. These results indicate that unlike in other farm animals, there is a common occurrence of zoonotic E. bieneusi and G. duodenalis genotypes in horses and donkeys. [ABSTRACT FROM AUTHOR]

Giardia duodenalis is a common enteric protozoan that infects a range of hosts including humans and other mammals. Multilocus genotyping of G. duodenalis in captive non-human primates (NHPs) from zoos in China is limited. In this study, we evaluated 302 NHP fecal samples collected from 32 different NHP species. The primates were from 12 zoos distributed across eight provinces and two municipalities (Chongqing and Beijing) of China. The overall infection rate was 8.3% (25/302). The six G. duodenalis-positive zoos and their infection rates were: Suzhou Zoo (40.0%, 4/10), Yangzhou Zoo (22.2%, 2/9), Dalian Zoo (16.7%, 4/24), Chengdu Zoo (12.8%, 6/47), Guiyang Forest Wildlife Zoo (12.1%, 7/58), and Changsha Zoo (4.7%, 2/43). Molecular analysis of three loci, beta-giardin (bg), triose phosphate isomerase (tpi), and glutamate dehydrogenase (gdh), showed high genetic heterogeneity, and seven novel subtypes (BIII-1, MB10-1, WB8-1, B14-1, MB9-1, DN7-1, and BIV-1) were detected within assemblage B. Additional analysis revealed 12 different assemblage B multilocus genotypes (MLGs), one known MLG and 11 novel MLGs. Based on phylogenetic analysis, 12 assemblage B MLGs formed two main clades, MLG-SW (10–12, 18) and MLG-SW (13, 14, 16, 17), the other four MLG-SW (15, 19, 20, 21) were scattered throughout the phylogenetic tree in this study. Using multilocus genotyping, this study expands our understanding of the occurrence of Giardia infection and genetic variation in Giardia in captive non-human primates from zoos in China. [ABSTRACT FROM AUTHOR]

Giardia duodenalis is a zoonotic intestinal parasite infecting humans and mammals worldwide. In this study, we evaluated the prevalence of G. duodenalis in racehorses in China and genetically characterized it. In total, 621 fecal samples were collected from racehorses at 17 equestrian clubs in 15 cities in China. Forty-eight (7.7%) animals from 11 equestrian clubs were positive for G. duodenalis of assemblages A (n = 10), B (n = 36), and E (n = 2), based on the small subunit ribosomal RNA (SSU rRNA) gene. Statistically significant differences in the prevalence of this parasite were detected among the different equestrian clubs (χ2 = 49.55, df = 16, p < 0.01), whereas no significant differences were detected according to age (χ2 = 0.64, df = 1, p > 0.05) or sex (χ2 = 1.41, df = 2, p > 0.05). The G. duodenalis-positive samples were further subtyped based on three other genes, which identified 5, 4, and 4 genotypes at the triose phosphate isomerase (tpi), glutamate dehydrogenase (gdh), and β-giardin (bg) loci, respectively. Subassemblage BIV was the predominant genotype. A phylogenetic analysis of the concatenated sequences of subassemblage BIV showed that the multilocus genotypes from the horses were genetically different from those of humans and nonhuman primates, indicating the evolution of host separation in G. duodenalis subassemblage BIV. Our study extends our understanding of the transmission of G. duodenalis between animals and humans. [ABSTRACT FROM AUTHOR]

Giardia duodenalis is a zoonotic parasitic protist and poses a threat to human and animal health. This study investigated the occurrence of G. duodenalis infection in post-weaned calves from Sichuan province, China. Faecal samples were collected from a total of 306 post-weaned calves (3–12 months old) from 10 farms, including 4 intensive feeding farms and 6 free-ranging farms. The overall infection rate of G. duodenalis was 41.2% (126/306) based on the PCR results at any of the three genetic loci: beta-giardin (bg), triose-phosphate isomerase (tpi) and glutamate dehydrogenase (gdh) genes. Giardia duodenalis assemblages E (n = 115, 91.3%), A (n = 3, 2.4%), and A mixed with E (n = 8, 6.3%) were identified among the 126 positive specimens. Multilocus sequence typing of G. duodenalis revealed 34 assemblage E multilocus genotypes (MLGs), 1 assemblage A MLG and 7 mixed assemblage (A and E) MLGs. The eBURST data showed a high degree of genetic diversity within assemblage E MLGs. The phylogenetic tree revealed that MLG E3 was the primary MLG subtype in Sichuan province and also the most widely distributed in China. [ABSTRACT FROM AUTHOR]

To assess the prevalence and molecular characteristics of Cryptosporidium spp., Giardia duodenalis , and Enterocytozoon bieneusi in grazing adult sheep from Xinjiang Uygur Autonomous Region, China, 318 fecal samples were collected and screened for the presence of these parasites by polymerase chain reaction. The overall infection rate for the three pathogens was 13.5% (43/318), with observed individual infection rates of 0.9% (3/318), 7.5% (24/318), and 6.3% (20/318) for Cryptosporidium spp., G. duodenalis , and E. bieneusi , respectively. Three Cryptosporidium species were identified amongst the samples, including C. xiaoi (n = 1), C. ubiquitum (n = 1), and C. parvum (n = 1), with gp60 -based subtyping analysis identifying C. parvum as subtype IIdA15G1 and C. ubiquitum as subtype XIIa. Eight E. bieneusi genotypes were identified based on internal transcribed spacer region sequencing, including six known (BEB6, CHG1, CHG3, CHS3, CHS8, and COS-I) and two novel (designated XJS1 and XJS2) genotypes. All G. duodenalis -positive samples were identified as assemblage E based on small subunit rRNA (n = 24) and gdh (n = 10) gene sequence analysis. These data support the occurrence of host adaptation by Cryptosporidium spp., G. duodenalis , and E. bieneusi in sheep, and the zoonotic risk may posed by these parasites in Xinjiang, China. Unlabelled Image • Sheep have played a role in the spread of C. parvum and C. ubiquitum in Xinjiang, China • Identified Eight E. bieneusi genotypes that belonged to cattle host-specific Group 2. • All the G. duodenalis -positive specimens belonged to assemblage E. • Two novel E. bieneusi genotypes were named as XJS1 and XJS2. [ABSTRACT FROM AUTHOR]

Giardia duodenalis is an important zoonotic intestinal protozoan of animals and humans. We collected 450 faecal specimens from four age groups (pre-weaned piglets, weaned piglets, juveniles, adults) of Zangxiang pigs from Shaanxi and Qinghai provinces, to investigate the prevalence and genetic diversity of G. duodenalis at the β-giardin (bg), triosephosphate isomerase (tpi) and glutamate dehydrogenase (gdh) loci using nested PCRs in the present study. A total of 28 faecal samples were positive for presence of G. duodenalis, with an overall prevalence of 6.2%. Giardia duodenalis was detected in pigs from all age groups and in both investigated provinces. Significant differences (P < 0.0001) in prevalence were observed among the four age groups with prevalence decreasing with age. Sequence analysis indicated existence of genetic diversity of G. duodenalis isolates from Zangxiang pigs, with 4, 2 and 4 haplotypes at the bg, tpi and gdh loci, respectively. Two assemblages were identified, including the zoonotic assemblage B and assemblage E, with the latter as the predominant assemblage found in both locations and all age groups except adults. The present study expanded the host range of G. duodenalis and provided fundamental data for controlling G. duodenalis infection in Zangxiang pigs. [ABSTRACT FROM AUTHOR]

The occurrence of Cryptosporidium and Giardia species in slaughter, sewage and river waters of the Qinghai Tibetan Plateau Area (QTPA), China, was investigated. A total of 456 samples were collected from different locations in the QTPA to study the contamination rates of Cryptosporidium spp. and Giardia via PCR and subsequent sequence analysis. Ten samples were Cryptosporidium positive, and 97 were Giardia positive, as confirmed by PCR amplification of the SSU rRNA gene. The percentages of positive Cryptosporidium and Giardia detection were 2.2% (10/456) and 21.3% (97/456), respectively. Cryptosporidium was detected in only sewage and river waters. Six species of Cryptosporidium were identified: Cryptosporidium hominis (n = 5), C. andersoni (n = 1), C. environmental (n = 1), C. struthionis (n = 1), C. canis (n = 1), and C. parvum (n = 1). G. duodenalis assemblage A was identified in almost all positive samples (n = 96), and one sample harboured G. duodenalis assemblage E. The results suggest that Cryptosporidium and Giardia species circulate through the aqueous environment and different hosts. Therefore, we strongly recommend that the local government and health authorities in China undertake control measures to reduce the contamination of water sources by these protozoa to protect the health of humans and animals. [ABSTRACT FROM AUTHOR]

The cat is a reported reservoir for several zoonotic pathogens, including Cryptosporidium spp., Giardia duodenalis, and Blastocystis sp. These parasites represent a significant, but often neglected, threat to humans and animals. Furthermore, Tritrichomonas foetus has been described inhabiting the digestive tract of cats, and may be causative agents of gastrointestinal symptoms. However, scant data are available concerning the molecular epidemiology of these parasites in domestic cats in China. This study examined fecal samples from domestic cats in Eastern China to unravel the molecular epidemiology of four protozoans. Of the 346 samples examined, 47 (13.6%) were positive for the detected pathogens, including 8 (2.3%), 5 (1.4%), 2 (0.6%), and 35 (10.1%) samples positive for Cryptosporidium spp., G. duodenalis, Blastocystis sp., and T. foetus, respectively. Co-infection with Cryptosporidium spp. and T. foetus was detected in three cats, no other mixed infections were observed. No age, sex or fecal condition predisposition was observed with any of the four pathogens. The species/assemblages/subtypes/genotypes were C. felis, Assemblage A and F, ST1, and cat genotype for Cryptosporidium spp., G. duodenalis, Blastocystis sp., and T. foetus detected in this study, respectively. The presence of zoonotic species/assemblages/subtypes/genotypes poses a threat to public health. These findings provide useful information for the design of prevention and control strategies to reduce the burden of protozoal infections in cats. [ABSTRACT FROM AUTHOR]

Sewage treatment plants (STPs) are one of the sources of pathogens discharged into surface water. An investigation was carried out over the duration of 12 months in Henan Province, China, to evaluate the health influence of municipal wastewater effluent discharge on water quality of the receiving water. A discharge-based quantitative microbial risk assessment (QMRA) was employed, taking into account the vegetables consumption habits of the Chinese, population subgroups with different immune statuses and ages, to evaluate the incremental disease burden from agricultural irrigation and swimming exposure scenarios associated with increased concentration of the protozoan Cryptosporidium and/or Giardia in the receiving river. The results shown that all the STP influent samples contained Cryptosporidium and Giardia with average density of 142.31 oocysts/L and 1187.06 cysts/L, respectively. The QMRA results demonstrated that the estimated additional health burdens due to discharged effluent for both parasites were slightly violated the threshold of 10 −6 DALYs per person per year set by WHO. Mitigation measures should be planned and executed by season since more disease burdens were borne during hot season than other seasons. The sensitivity analysis highlighted the great importance of stability of STP treatment process. This study provides useful information to improve the safety of surface water and deduce the disease burden of the protozoa in Henan Province and other region inside and outside China. [ABSTRACT FROM AUTHOR]

Giardia duodenum (G. duodenalis) can cause giardiasis and infect a variety of hosts. So far, there have been no detailed data regarding the positive rate of G. duodenalis in sheep and goats in China. Here, a systematic literature review was carried out to investigate the epidemiology of G. duodenalis in sheep and goats in China. To perform the meta-analysis, the databases CNKI, VIP, WanFang, PubMed, Web of science and ScienceDirect were employed for screening studies related to the prevalence of G. duodenalis in sheep and goats in China. The total prevalence of G. duodenalis in sheep and goats was estimated to be 7.00% (95% CI: 4.00–10.00). In the age subgroup, the prevalence of G. duodenalis in sheep and goats of >12 months (11.29%; 95% CI: 8.08–14.97) was higher than that in sheep and goats of ≤12 months (7.57%; 95% CI: 3.95–12.24). An analysis based on seasons showed that the prevalence of G. duodenalis in sheep and goats was higher in summer (11.90%; 95% CI: 0.50–35.05) than that in other seasons. The prevalence of G. duodenalis in sheep and goats after 2016 was 8.57% (95% CI: 5.34–11.79), which was higher than others. The highest prevalence of G. duodenalis in sheep and goats was 13.06% (95% CI: 6.26–19.86) recorded in Southwestern China. The prevalence of Giardia infection in sheep (7.28%; 95% CI: 2.30–14.73) was higher than that in goats (5.43%; 95% CI: 2.73–8.98). The NOAA's National Center for Environmental Information (https://gis.ncdc.noaa.gov/maps/ncei/cdo/monthly) was used to extract relevant geoclimatic data (latitude, longitude, elevation, temperature, precipitation, humidity, and climate). By analyzing the data of each subgroup, it was shown that region, genetype, and climate were potential risk factors for giardiasis prevalence in sheep and goats. Based on the analysis of common factors and geographical factors, it is recommended to strengthen effective management measures (e.g. ventilation and disinfection in warm and humid areas) and formulate relevant policies according to local conditions. Breeders should strengthen the detection of G. duodenalis in sheep and goats, customize corresponding control measures according to the diet and living habits of sheep and goats, and strengthen the protection of sheep and lamb calves, so as to reduce the incidence rate and reduce the economic loss of China's animal husbandry. • Giardia duodenalis is common prevalent among sheep and goats in China. • The pooled Giardia duodenalis prevalence in sheep and goats was estimated as 7.00% (1132/16,347). • The prevalence of Giardia duodenalis in sheep and goats was unevenly in different regions. • This is the meta-analysis of the Giardia duodenalis prevalence in sheep and goats in China for the first time. [ABSTRACT FROM AUTHOR]

Objective: To investigate the prevalence and genotypes of Cryptosporidium spp. and Giardia lamblia in dogs and cats from a pet hospital in Shanghai Municipality., Methods: A total of 145 fresh fecal samples were collected from pet dogs and cats in a pet hospital in Shanghai Municipality during the period from November 2021 to June 2022, including 99 dog fecal samples and 46 cat fecal samples. The small subunit ribosomal ribonucleic acid ( SSU rRNA ) gene of Cryptosporidium and the triose phosphate isomerase ( TPI ) gene of G. lamblia were amplified using nested PCR assay, and the positive amplification products were sequenced from both directions. The sequence assembly was performed using the software Clustal X 2.1, and sequence alignment was conducted using BLAST. A phylogenetic tree was created with the Neighbor-Joining method using MEGA 11.0 to identify parasite species or genotype., Results: The overall prevalence of Cryptosporidium and G. lamblia was 20.00% (29/145) in 145 pet dog and cat fecal samples, with the prevalence of 0.69% (1/145) and 19.31% (28/145) in Cryptosporidium and G. lamblia , respectively. G. lamblia was only detected in dog fecal samples, with prevalence of 18.18% (18/99), while the detection rates of Cryptosporidium and G. lamblia were 2.17% (1/46) and 21.74% (10/46) in cat fecal samples. Nucleotide sequence analysis showed that one Cryptosporidium positive sample was characterized as C. felis , and 28 G. lamblia positive samples were all characterized as Giardia assemblage A, which showed 100% sequence homology with human isolates of Giardia . Phylogenetic analysis revealed that the sequences obtained in this study belonged to the same branch with the reported Giardia assemblage A., Conclusions: Cryptosporidium and G. lamblia infection was prevalent in pet dogs and cats from the study pet hospital in Shanghai Municipality, and there is a zoonotic risk for the species and genotype. Intensified surveillance of Cryptosporidium and Giardia infection is recommended in pets and their owners, and improved management of pet keeping is required.

Cryptosporidium and Giardia (major causes of diarrhea) are widely distributed in Chinese source waters and threaten human health. A new spatially explicit GloWPa-TGR-Crypt-Giar C1 model is presented to simultaneously estimate mean monthly (oo)cyst concentrations in surface and ground waters in the Three Gorges Reservoir (TGR) watershed. A quantitative risk assessment of protozoal infections considered different source waters, transmission pathways, regions, susceptible subpopulations, and drinking water treatments. Monthly mean Cryptosporidium oocyst and Giardia cyst concentrations ranged between 0.5-19.3 oocysts/10 L and 0.2-5.0 cysts/10 L in surface water, respectively, and 0.007-0.3 oocysts/10 L and 0.002-0. 2 cysts/10 L in groundwater. The cumulative disease burdens attributable to cryptosporidiosis and giardiasis were, respectively, 5.77×10 -5 DALYs (disability-adjusted life years/person/year) and 4.63×10 -6 DALYs in urban areas, and 6.35×10 -4 DALYs and 8.84×10 -5 DALYs in rural areas, which were much higher than the reference risk level recommended by the World Health Organization ([Formula: see text] DALYs). The annual burden associated with consuming surface water was calculated to be 3.84×10 -4 DALYs for Cryptosporidium and [Formula: see text] DALYs for Giardia, whereas consuming groundwater entailed the lower burdens (1.26×10 -5 and 3.50×10 -6 DALYs, respectively). Most DALYs were a consequence of consumption of directly supplied surface water. Fifty percent of the health burden was carried by immunodeficiency with HIV. Children (0-4 years) were more likely to have an individual disease burden than adults (15-64 years). Males were more susceptible than females. Improving sanitation through adequate ozone and microfiltration treatment should be considered when attempting to reduce disease burden. Sensitivity analysis highlighted the importance of reducing (oo)cyst loads to protect the watershed. The methodology and results described will help in evaluating and reducing the burden of protozoal infection associated with surface and ground waters in the TGR and similar watersheds., (© 2022. The Author(s).)

Cryptosporidiosis, microsporidiosis, and giardiasis contribute significantly to the high burden of zoonotic diarrhea worldwide. Goats constitute an important species in animal agriculture by providing cashmere wool, meat, and dairy products for human consumption. However, zoonotic pathogens with the potential to cause morbidity and to degrade production have been reported frequently in goats recently. The present study examined 629 fecal specimens from goats, including 315 cashmere goats, 170 dairy goats and 144 meat goats, in multiple cities of Shaanxi and Henan provinces, northwestern and central China, to investigate the infection rate and species/assemblages/genotypes of Giardia duodenalis , Cryptosporidium spp. and Enterocytozoon bieneusi . Of these samples, 274 (43.6%) were positive for three zoonotic pathogens, including 80 (12.7%), 104 (16.5%) and 179 (28.5%) for G. duodenalis , Cryptosporidium spp. and E. bieneusi , respectively. Infections with G. duodenalis , Cryptosporidium spp. and E. bieneusi existed in meat, dairy and cashmere goats, with the highest infection rate of each pathogen being observed in meat goats. DNA sequencing of the SSU rRNA gene from 104 Cryptosporidium -positive specimens revealed existence of Cryptosporidium xiaoi , and the zoonotic parasites Cryptosporidium parvum and Cryptosporidium ubiquitum . Genotyping of G. duodenalis based on the triosephosphate isomerase (TPI) gene identified parasites from zoonotic assemblage A in four cashmere goats and the animal-adapted assemblage E in a group of 76 goats that included cashmere, dairy and meat animals. Polymorphisms in the ribosomal internal transcribed spacer characterized E. bieneusi genotype CHG1 and a novel genotype named as SX1 in both dairy and cashmere goats, genotypes CHS7 and COSI in meat goats, the genotype CHG2 in dairy goats, and the human-pathogenic genotype BEB6 in dairy and meat goats. This is the first detailed study to compare infection rate of the zoonotic protozoan pathogens in cashmere, dairy and meat goats in China. Our research discovered Cryptosporidium spp. and E. bieneusi infections, each with zoonotic potential in meat goats, and G. duodenalis and Cryptosporidium spp. in cashmere goats raising a significant public health concern. [ABSTRACT FROM AUTHOR]

Giardia duodenalis is an important protozoan parasite that is known to be zoonotic. To assess the potential zoonotic transmission of giardiasis from dogs and to identify genetic diversity of G. duodenalis in dog populations, we examined the infection rate and genotypes of G. duodenalis in both pet dogs (from pet dog farms, pet shops, pet hospitals, pet markets) and stray dogs of different ages in Henan Province, China. A total of 940 fresh fecal specimens were collected from 2007 to 2013 in Henan Province. The overall infection rate of G. duodenalis was 14.3% (134/940) as determined by microscopy, with the highest infection rate (17.3%) observed in dogs from shelters. Young dogs were more likely to be infected with G. duodenalis than adult dogs, and G. duodenali s cysts were found more frequently in diarrheic dogs. All G. duodenalis -positive isolates were characterized at the triose phosphate isomerase ( tpi ), glutamate dehydrogenase ( gdh ), and β-giardin ( bg ) loci, and 37, 51, and 48 sequences were obtained, respectively. The dog-specific assemblages C and D were identified using multi-locus sequence analysis. Six novel sequences of the tpi locus, one novel sequence of the gdh locus and two novel sequences of the bg locus were detected among the G. duodenalis assemblage C isolates, while two novel sequences of the gdh locus were found among the G. duodenalis assemblage D isolates. Our data indicate that G. duodenalis is a common parasite and cause of diarrheal disease in dogs in Henan Province. However, there was no evidence for zoonotic G. duodenalis assemblages in the study population. [ABSTRACT FROM AUTHOR]

The aim of this study was to carry out a survey for the presence ofGiardia duodenalisinfection in canine using ELISA and PCR and to identify risk factors for infection. Samples from 318 dogs’ feces living in nine cities in China were used in the present study. Each sample was tested for the presence ofG. duodenalis-specific antigens using ELISA and 197 out of 318 samples were further examined for the presence ofG. duodenalisusing PCR. The overall rate of canines infected with giardiasis in the present study was 16.04 % and 15.22 % using ELISA and PCR, respectively. No significant difference was found between sex andGiardiapositivity. Young dogs (up to one year) and living in communities were identified as risk factors for infection by multivariate logistic regression analysis. In conclusion, giardiasis in dogs was present in nine cities in China; as risk factors, young dogs (up to one year) and living in communities were of great significance.Giardia-infected canine should be treated for hygienic management to prevent transmission of the infection from dog to human. [ABSTRACT FROM PUBLISHER]

Giardia duodenalis is a common protozoan that colonizes and reproduces in the small intestine, causing giardiasis. This parasite infects a wide range of vertebrate hosts, including humans, domestic animals and wildlife. It has been suggested that chinchillas ( Chinchilla lanigera ) kept as domestic pets are potential reservoirs for the zoonotic transmission of G. duodenalis . In this study, 140 chinchilla samples from four cities in China were examined to determine the prevalence of G. duodenalis . Thirty-eight (27.1%) chinchillas were found to be positive for G. duodenalis . The prevalence of infection was analyzed in relation to collection site, age and sex. Molecular characterization was also carried out on the 38 chinchilla samples to determine common genotypes. G. duodenalis assemblages A and B were identified in the chinchilla samples by analysis of the small subunit ribosomal RNA ( ssur RNA ) gene. Genotyping at the subtype level using multiple genes (glutamate dehydrogenase ( gdh ), triose phosphate isomerase ( tpi ) and β-giardin ( bg ) genes) determined that the majority of assemblage A isolate sequences were identical to subtype AI. Assemblage B isolates showed variability among the nucleotide sequences belonging to subtype BIV. This is the first report of G. duodenalis in chinchillas from China. As subtype AI and BIV are associated with human infection, G. duodenalis in chinchillas should be regarded as zoonotic. [ABSTRACT FROM AUTHOR]

Non-human primates (NHPs) are commonly infected with Cryptosporidium spp. and Giardia duodenalis . However, molecular characterisation of these pathogens from NHPs remains scarce. In this study, 2,660 specimens from 26 NHP species in China were examined and characterised by PCR amplification of 18S rRNA, 70 kDa heat shock protein ( hsp70 ) and 60 kDa glycoprotein ( gp60 ) gene loci for Cryptosporidium ; and 1,386 of the specimens by ssrRNA, triosephosphate isomerase ( tpi ) and glutamate dehydrogenase ( gdh ) gene loci for Giardia . Cryptosporidium was detected in 0.7% (19/2660) specimens of four NHP species including rhesus macaques (0.7%), cynomolgus monkeys (1.0%), slow lorises (10.0%) and Francois’ leaf monkeys (6.7%), belonging to Cryptosporidium hominis (14/19) and Cryptosporidium muris (5/19). Two C. hominis gp60 subtypes, IbA12G3 and IiA17 were observed. Based on the tpi locus, G. duodenalis was identified in 2.2% (30/1,386) of specimens including 2.1% in rhesus macaques, 33.3% in Japanese macaques, 16.7% in Assam macaques, 0.7% in white-headed langurs, 1.6% in cynomolgus monkeys and 16.7% in olive baboons. Sequence analysis of the three targets indicated that all of the Giardia -positive specimens belonged to the zoonotic assemblage B. Highest sequence polymorphism was observed at the tpi locus, including 11 subtypes: three known and eight new ones. Phylogenetic analysis of the subtypes showed that most of them were close to the so-called subtype BIV. Intragenotypic variations at the gdh locus revealed six types of sequences (three known and three new), all of which belonged to so-called subtype BIV. Three specimens had co-infection with C. hominis (IbA12G3) and G. duodenalis (BIV). The presence of zoonotic genotypes and subtypes of Cryptosporidium spp. and G. duodenalis in NHPs suggests that these animals can potentially contribute to the transmission of human cryptosporidiosis and giardiasis. [ABSTRACT FROM AUTHOR]

Background Cryptosporidium spp., Enterocytozoon spp., Giardia spp. and Cyclospora spp. are important intestinal protozoan parasites causing diarrhea in humans, livestocks and wildlife and have a significant impact on public health. No reports exist about simultaneous prevalence rates or genotyping data of these four parasites in outpatients from China. Methods Fecal specimens from 252 diarrhea patients in a pediatric clinic (n = 169) and an intestinal clinic (n = 83) of a hospital in Shanghai, China, were collected between October 2012 and March 2013. All samples were examined for the presence of the four parasites by using molecular methods. Results In total, 76/252(30.16%) patients were positive for at least one intestinal parasite, of which Cryptosporidium spp., Enterocytozoon bieneusi and Giardia intestinalis were detected by nested PCR in 34 (13.49%), 34 (13.49%) and 17 (6.75%) of the fecal specimens, respectively. Sequence analysis showed that all Cryptosporidium-positive specimens were C. andersoni and that most G. intestinalis- positive patients were infected by assemblage C, which is usually found in canids, while only one sample was from assemblage B. Eight patients were co-infected with Cryptosporidium spp. and Enterocytozoon, while one was coinfected with Cryptosporidium and Giardia. Conclusions The patients infected with Cryptosporidium and Enterocytozoon bieneusi had higher infection rates in winter than in spring in this area. Data indicated that C. andersoni is the fourth major Cryptosporidium species infecting humans in addition to C. hominis, C. parvum and C. meleagridis. Our study also revealed a short-term outbreak of cryptosporidiosis and microsporidiosis and sporadic cases of giardiasis that occurred among humans in Shanghai, China. [ABSTRACT FROM AUTHOR]

Micromammals have historically been recognized as highly contentious species in terms of the maintenance and transmission of zoonotic pathogens to humans. Limited information is currently available on the epidemiology and potential public health significance of intestinal eukaryotes in wild micromammals. We examined 490 faecal samples, grouped into 155 pools, obtained from 11 micromammal species captured in 11 Spanish provinces for the presence of DNA from Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi and Blastocystis sp. The presence of Leishmania spp. was investigated in individual spleen samples. All micromammal species investigated harboured infections by at least one eukaryotic parasite, except Apodemus flavicollis, Myodes glareolus, Sorex coronatus and Sciurus vulgaris, but the sample size for these host species was very low. Cryptosporidium spp. was the most prevalent species found (3.7%, 95% confidence interval [CI]: 2.2-5.7), followed by G. duodenalis (2.8%, 95% CI: 1.6-4.6) and E. bieneusi (2.6%, 95% CI: 1.4-4.3). All pooled faecal samples tested negative for Blastocystis sp. Leishmania infantum was identified in 0.41% (95% CI: 0.05-1.46) of the 490 individual spleen samples analysed. Sequence analyses allowed the identification of Cryptosporidium andersoni (5.9%), C. ditrichi (11.7%), C. muris (5.9%), C. parvum (5.9%), C. tyzzeri (5.9%), rat genotypes CR97 (5.9%) and W19 (5.9%), vole genotypes V (11.7%) and VII (5.9%) and Cryptosproridium spp. (35.3%) within Cryptosporidium (n = 17). Known genotypes C (66.7%) and Peru11 (25.0%) and a novel genotype (named MouseSpEb1, 8.3%) were detected within E. bieneusi (n = 12). None of the G. duodenalis-positive samples could be genotyped at the assemblage level. Molecular data indicate that wild micromammals were primarily infected by rodent-adapted species/genotypes of eukaryotic pathogens and thereby have a limited role as a source of human infections. The presence of ruminant-adapted species C. andersoni along with finding C. parvum is indicative of an overlap between domestic/peri-domestic and sylvatic transmission cycles of these agents., (© 2022 Wiley-VCH GmbH.)

Highlights: [•] The police and farm dogs have been infected with Giardia duodenalis in Shenyang. [•] Significantly higher infection rates of farm dogs than police dogs were seen. [•] The high occurrence of potentially zoonotic subtype AI-1 in dogs is of public health concern. [Copyright &y& Elsevier]

Background: Over 200 cryptosporidiosis outbreaks have been reported, but little is known if other enteric pathogens were also involved in some of these outbreaks. Recently, an outbreak of cryptosporidiosis linked to poor hygiene by two Cryptosporidium hominis subtypes occurred in a pediatric hospital ward (Ward A) in China, lasting for more than 14 months. In this study, the concurrence during the outbreak of three other enteric pathogens with a similar transmission route, Giardia duodenalis, Enterocytozoon bieneusi, and Clostridium difficile, was assessed. Methods/Principal Findings: The occurrence of G. duodenalis, E. bieneusi, and C. difficile in 78 inpatients from Ward A and 283 and 216 inpatients from two control wards (Wards C and D) in the same hospital was examined using molecular diagnostic tools. Significantly higher infection rates were found in children in Ward A for all study pathogens than in Wards C and D (P<0.01): 9.5% versus 1.4% and 0% for G. duodenalis, 10.8% versus 2.8% and 3.7% for E. bieneusi, and 60.8% versus 37.8% and 27.8% for C. difficile, respectively. These differences were mostly seen in children ≤12 months. Enteric pathogen-positive children in Ward A (31/58 or 53.4%) were more likely to have mixed infections than those in Ward C (4/119 or 3.4%) or D (5/68, 7.4%; P<0.01). Having cryptosporidiosis was a risk factor for G. duodenalis (OR = 4.3; P = 0.08), E. bieneusi (OR = 3.1; P = 0.04), and C. difficile (OR = 4.7; P<0.01) infection. In addition, a lower diversity of G. duodenalis, E. bieneusi, and C. difficile genotypes/subtypes was observed in Ward A. Conclusions/Significance: Data from this study suggest that multiple pathogens were concurrently present during the previous cryptosporidiosis outbreak. Examination of multiple enteric pathogens should be conducted when poor hygiene is the likely cause of outbreaks of diarrhea. Author Summary: The transmission of Giardia duodenalis, Enterocytozoon bieneusi, and Clostridium difficile is poorly understood in developing countries despite their wide occurrence. Because they are transmitted by the same fecal-oral route as Cryptosporidium, in this study, we have examined the occurrence of these enteric pathogens in children during a cryptosporidiosis outbreak in a pediatric hospital in China. Using molecular diagnostic tools, we have detected significantly higher infection rates of these enteric pathogens in the outbreak ward than in two control wards in the same hospital. We have also shown a much higher occurrence of these pathogens in children having cryptosporidiosis than those having no cryptosporidiosis. We have demonstrated that the genetic diversity of enteric pathogens is much lower in the outbreak ward than in control wards. Therefore, other enteric pathogens are concurrently present during the cryptosporidiosis outbreak, and examinations for multiple enteric pathogens should be conducted when poor hygiene is considered the likely cause of outbreaks of diarrhea. [ABSTRACT FROM AUTHOR]

Abstract: The Three Gorges Reservoir (TGR) is the biggest lake in the world and a major water source in China. There is no information about occurrence and impact of Cryptosporidium and Giardia on the aquatic ecosystem. 61 surface water samples from 23 monitoring sites and 5 treated effluent samples were collected and analyzed. Cryptosporidium oocysts and Giardia cysts were found, respectively, in 86.4% and 65.2% of a total of 66 water samples, with high concentrations in treated effluent. The mean percent recovery was 29.14% for oocysts and 34.86% for cysts. A seasonal pattern was observed, with positive samples for Cryptosporidium more frequent in flood period and positive samples for Giardia more frequent in impounding period. Counts of enterococci, fecal coliforms and total coliforms, and turbidity were significantly associated with Cryptosporidium concentration in backwater (water in a main river which is backed up by the Three Gorges Dam) areas of tributaries but not Giardia. High associations were also found between oocyst and cyst in backwater areas of tributaries and cities. The risks of infection and illness due to water consumption in four different exposure routes were estimated. The results showed that swimming in the TGR has the highest infection risk with 1.39 × 10−3 per time (95% confidence interval (CI): 0.05–600.3 × 10−5) for Cryptosporidium and 2.08 × 10−4 per time (95% CI: 0.05–878.87 × 10−6) for Giardia, while directly drinking unboiled tap water treated with the conventional process has the highest morbidity with 524.98 per 100,000 population per year (95% CI: 10.35–2040.26) for Cryptosporidium and 5.89 per 100,000 population per year (95% CI: 0.08–22.67) for Giardia. This study provides new useful information for drinking water plants, health care workers and managers to improve the safety of tap water and deduce the risk of surface water contamination in China. [Copyright &y& Elsevier]