Genetic characteristics of Giardia duodenalis from sheep in Inner Mongolia, China.
ABSTRACT: Giardia duodenalis is an important zoonotic pathogen for both human and animal health. Although there have been reports on G. duodenalis infections in animals all over the world, information regarding the prevalence and genetic characteristics of G. duodenalis in sheep in Inner Mongolia, China, is limited. In this study, 209 sheep fecal specimens were collected in this autonomous region. We established that the prevalence of G. duodenalis was 64.11% (134/209), as determined using nested PCR detection and sequences analysis of the small subunit ribosomal RNA (SSU rRNA) gene. Based on the beta-giardin (bg) locus, the glutamate dehydrogenase (gdh) locus, and the triose phosphate isomerase (tpi) locus to study genetic characteristics, both assemblages A (2.99%, 4/134) and E (97.01%, 130/134) were found. Five novel nucleotide sequence of assemblage E were detected, two at the bg locus, two at the gdh locus, and one at the tpi locus. Multilocus genotyping yielded four assemblage E and two assemblage A multilocus genotypes (MLGs), including four novel assemblage E MLGs and one novel assemblage A MLG. Results of this study indicated that G. duodenalis was highly prevalent in sheep in Inner Mongolia. This study is the first to use the multilocus genotyping approach to identify G. duodenalis in sheep from this region.
Project description:BACKGROUND:Giardia duodenalis is an important intestinal protozoan infecting both humans and animals, causing significant public health concern and immeasurable economic losses to animal husbandry. Sheep and goats have been reported as common reservoirs of G. duodenalis, but only a limited amount of information is available for native breeds of these small ruminants in China. The present study investigated the prevalence and multilocus genotypes of G. duodenalis in black-boned sheep and goats, two important native breeds in Yunnan Province, southwestern China. METHODS:Fecal samples were collected from 336 black-boned goats and 325 black-boned sheep from five counties (Meishui, Shanshu, Shilin, Yongsheng and Nanping) of Yunnan Province and the genomic DNA was extracted from these feces. The prevalence of G. duodenalis was determined by the nested PCR targeting the ?-giardin (bg) gene. The assemblages and multilocus genotypes (MLGs) were investigated based on analyses of three genetic loci, i.e. bg, glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi). RESULTS:Giardia duodenalis infection was detected in both black-boned sheep and goats, and the prevalence of G. duodenalis in black-boned sheep (21.8%, 71/325) was significantly higher (?2?=?36.63, df?=?1, P?<?0.001) than that in black-boned goats (4.8%, 16/336). Significant differences in prevalence were also observed in goats and sheep from different counties (?2?=?39.83, df?=?4, P?<?0.001) and age groups (?2?=?97.33, df?=?3, P?<?0.001). Zoonotic assemblage A and animal-specific assemblage E were identified in both black-boned sheep and goats with the latter as the predominant assemblage. Based on sequences obtained from the three genetic loci (bg, gdh and tpi), 16 MLGs were obtained in black-boned sheep and goats, including 15 MLGs in assemblage E and one MLG in assemblage A. CONCLUSIONS:Our results not only extended the host range of G. duodenalis, but also revealed high genetic variations in G. duodenalis assemblages. The findings of the present study also provide baseline data for preventing and controlling G. duodenalis infection in black-boned sheep and goats in Yunnan Province.
Project description:Giardia duodenalis and Cryptosporidium spp. are common gastrointestinal protozoa in mammals. Many studies have been conducted on the distribution of G. duodenalis and Cryptosporidium spp. genotypes in sheep and cattle. However, in China, information about molecular characterization and genetic analysis of G. duodenalis and Cryptosporidium spp. in goats is limited. In this study, 342 fecal samples from adult goats were collected from 12 farms in Sichuan Province, China. The occurrence of G. duodenalis and Cryptosporidium spp. in adult goats was 14.9% (51/342) and 4.7% (16/342), respectively. All G. duodenalis were identified as assemblage E, with two novel genotypes (assemblages E17 and E18) being detected at the beta-giardin (bg) locus. Based on three loci-beta-giardin (bg), triose phosphate isomerase (tpi), and glutamate dehydrogenase (gdh)-multilocus sequence typing revealed three novel multilocus genotypes (MLGs) of assemblage E (MLG-E1, E2, E3 (sc)). Small Subunit (SSU) rRNA-based PCR identified two Cryptosporidium species, namely C. xiaoi (11/16) and C. suis (5/16). This study is not only the first to report C. suis infection in adult goats in China but is also the first to use the MLG approach to identify G. duodenalis in adult goats.
Project description:BACKGROUND:Giardia duodenalis is a flagellated parasite that causes diarrhea in humans and other animals. Although G. duodenalis is found in companion animals worldwide, information regarding the prevalence and genetic characteristics of G. duodenalis in pet chipmunks in China is limited. The present study therefore aimed to investigate the prevalence and genotypes of G. duodenalis in pet chipmunks in Sichuan province, southwestern China, as well as to assess zoonotic potential of revealed assemblages. RESULTS:A total of 279 fecal samples were collected from pet chipmunks in seven pet shops and one breeding facility in Sichuan province, southwestern China. The prevalence of G. duodenalis was 8.6% (24/279), as determined by nested PCR detection of the beta giardin (bg) gene. Giardia duodenalis assemblages and subtypes were determined using multilocus genotyping of the bg, triosephosphate isomerase (tpi), and glutamate dehydrogenase (gdh) loci. Two assemblages were identified: potentially zoonotic assemblage A (54.2%, 13/24) and rodent-specific assemblage G (45.8%, 11/24). A total of 24, 17 and 17 sequences of the bg, gdh and tpi loci, respectively, were successfully obtained, which formed four, four and three subtypes, respectively. Moreover, four assemblage A (MLGs A1-A4) and three assemblage G (MLGs G1-G3) multilocus genotypes were identified. CONCLUSIONS:To our knowledge, this is the first study that investigated G. duodenalis in pet chipmunks in China. Detection of assemblage A in pet chipmunks and in previous studies in humans suggests a possible role of chipmunks as a reservoir for human giardiasis in Sichuan Province, China.
Project description: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.
Project description: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, ?-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.
Project description:Giardia duodenalis is an important protozoan parasite. It is an established zoonotic pathogen and dairy calves have been implicated as one of the most important sources of human infection. This study was conducted to assess the prevalence and multilocus genotyping of G. duodenalis in dairy calves in the Xinjiang Uyghur Autonomous Region, northwestern China.A total of 514 fresh fecal samples were randomly collected from dairy calves in 15 farms in Xinjiang, 13.4 % (69/514) tested positive for G. duodenalis by polymerase chain reaction (PCR) detection of the small subunit ribosomal RNA (SSU rRNA) gene, with the prevalence being 9.7 % (23/237) and 16.6 % (46/277) in pre- and post-weaned calves, respectively. Sequence analysis of the SSU rRNA gene predominantly detected G. duodenalis assemblage E (92.8 %, 64/69), whereas assemblage A was identified in five samples (7.2 %, 5/69). All G. duodenalis-positive samples were assayed with PCR followed by sequencing at ?-giardin (bg), glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi) genes, and 29, 37 and 33 sequences were obtained, respectively. The presence of mixed G. duodenalis assemblage A and E was detected in only one sample. Multilocus genotyping yielded 15 multilocus genotypes (MLGs), one new assemblage A MLG, and 14 assemblage E MLGs. All assemblage E MLGs identified here differed genetically from those of cattle from Henan Province, Central China.Our data indicate that G. duodenalis is a common parasite in dairy calves in Xinjiang, China, and calves appear to be a reservoir of G. duodenalis that is infectious to humans. The differences in the distribution of G. duodenalis assemblage E MLGs from cattle were likely to be because of geographical segregation.
Project description:Giardia duodenalis is a common human and animal pathogen. It has been increasingly reported in wild and captive non-human primates (NHPs) in recent years. However, multilocus genotyping information for G. duodenalis infecting NHPs in southwestern China is limited. In the present study, the prevalence and multilocus genotypes (MLGs) of G. duodenalis in captive NHPs in southwestern China were determined. We examined 207 fecal samples from NHPs in Sichuan and Guizhou provinces, and 16 specimens were positive for G. duodenalis. The overall infection rate was 7.7%, and only assemblage B was identified. G. duodenalis was detect positive in northern white-cheeked gibbon (14/36, 38.9%), crab-eating macaque (1/60, 1.7%) and rhesus macaques (1/101, 0.9%). Multilocus sequence typing based on beta-giardin (bg), triose phosphate isomerase (tpi) and glutamate dehydrogenase (gdh) revealed nine different assemblage B MLGs (five known genotypes and four novel genotypes). Based on a phylogenetic analysis, one potentially zoonotic genotype of MLG SW7 was identified in a northern white-cheeked gibbon. A high degree of genetic diversity within assemblage B was observed in captive northern white-cheeked gibbons in Southwestern China, including a potentially zoonotic genotype, MLG SW7. To the best of our knowledge, this is the first report using a MLGs approach to identify G. duodenalis in captive NHPs in Southwestern China.
Project description:<h4>Background</h4><i>Giardia duodenalis</i> is a common human intestinal parasite worldwide, and the causative agent of diarrhea, with the severity of disease ranging from asymptomatic to intense and debilitating infection. <i>G. duodenalis</i> is known to consist of eight genetically distinct assemblages, named from A to H. No data available on the genotypes and genetic diversity of <i>G. duodenalis</i> circulating in Qatar.<h4>Methods</h4>We genotyped 54 human <i>Giardia</i> isolates, collected from asymptomatic immigrants in Qatar, using a multilocus genotyping (MLGs) tool. We also investigated relationships between the subjects' genotypes and their demographic data.<h4>Results</h4>Genomic DNA from 54 isolates were tested by PCR and sequence analysis at three loci: <i>glutamate dehydrogenase</i> (<i>gdh</i>), <i>β-giardin</i> (<i>bg</i>) and <i>triose phosphate</i> (<i>tpi</i>)). Assemblage A was identified in nine (16.67%), assemblage B in thirty (55.55%), and a mixture of assemblages A+B in fifteen (27.78%) isolates. All assemblage A isolates, genotyped in different loci, were assigned to sub-assemblage AII, and six of them had MLGs AII-1 while one new MLG was identified in two isolates. Sequences of assemblage B isolates have high level of genetic diversity and high presence of heterogeneous peaks, especially within the <i>gdh</i> gene. No significant associations between genotypes and the immigrants' demographic data were found due to the extensive number of new variants.<h4>Conclusions</h4>MLGs was used herein to genotype 54 immigrant <i>Giardia</i> isolates. The high level of genetic variability found in our isolates hampered MLGs determination, more investigations are now required to consolidate our findings, and to enable a comprehensive understanding of the diversity within <i>G. duodenalis</i> assemblage B isolates.
Project description:<h4>Background</h4><i>Giardia duodenalis</i> is an important opportunistic zoonotic intestinal protozoon, which could parasitize yaks. However, a few studies have been conducted on the seasonal infection of <i>G. duodenalis</i> in yaks in China.<h4>Methods</h4>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).<h4>Results</h4>The overall prevalence of <i>G. duodenalis</i> 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 <i>G. duodenalis</i> 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.<h4>Conclusion</h4>This study indicated a zoonotic potential of <i>G. duodenalis</i> in yaks from Qinghai Province and provides basic information about the epidemiology of <i>G. duodenalis</i>.
Project description:Giardia duodenalis is an important zoonotic parasite which can parasitize in the intestines of humans and various animals. However, the information about the prevalence and genetic diversity of G. duodenalis in goats in China is limited. It is yet to be known whether Yunnan black goats, a unique goat breed in subtropical Yunnan province, southwestern China, are infected with G. duodenalis. Thus, a total of 907 fecal samples were collected from Yunnan black goats in five regions in Yunnan province, to estimate the prevalence and genotypes of G. duodenalis using a PCR-based approach. The G. duodenalis prevalence is 4.2% (38/907) in Yunnan black goats by nested amplification of the ?-giardin (bg) gene, and the genotypes are identified as assemblage E, with 5 novel subtypes (E11-E15). Multilocus sequence typing revealed that 11, 18, and 38 samples were amplifiable on tpi (triose phosphate isomerase), gdh (glutamate dehydrogenase), and bg locus, respectively, and identified three novel multilocus genotypes (MLGs): MLGE9-MLGE11. To our knowledge, this is the first report of G. duodenalis prevalence and genotypes in Yunnan black goats in China, which extended the host range of G. duodenalis and provided basic data for controlling G. duodenalis infection in Yunnan black goats.