Molecular detection of Blastocystis in black bears and sika deer in northern China.

Protists of the Blastocystis genus are distributed worldwide and can infect a range of hosts. However, data concerning Blastocystis infection are limited for sika deer and are not available for black bears. Therefore, in the present study, a total of 312 black bears (Ursus thibetanus) from Heilongjiang Province and 760 sika deer (Cervus nippon) from four different northern Chinese provinces were investigated. Blastocystis infection in these animals was detected via PCR amplification of the small subunit rRNA gene in fecal samples. The prevalence of Blastocystis infection in black bears and sika deer was 14.4% (45/312 positive samples) and 0.8% (6/760 positive samples), respectively. Young black bears (18.3%) had a significantly higher Blastocystis prevalence than adult bears (9.1%). The prevalence of Blastocystis was significantly higher in black bears raised outdoors (24.6%) than in bears raised indoors (12.2%). Blastocystis-positive sika deer were only found in Jilin Province (1.3%, 6/480). Female sika deer (0%, 0/61) had a significantly lower Blastocystis prevalence than males (0.9%, 6/699). Sanger sequencing was used to determine the small subunit rRNA gene sequences of the Blastocystis-positive PCR products. A neighbor-joining phylogenetic tree based on the small subunit rRNA gene sequences showed that only Blastocystis subtype (ST)1 was identified in black bears, whereas ST10 and ST14 were found in sika deer. This is the first report of Blastocystis ST1 infection in black bears. These findings also extend the distribution information of Blastocystis subtypes, which will provide a foundation for further study of Blastocystis in different hosts in China.

Molecular detection of Cryptosporidium spp. and Enterocytozoon bieneusi in Longjiang Wagyu cattle in Northeastern China.

Cryptosporidium spp. and Enterocytozoon bieneusi are two important zoonotic pathogens that can cause diarrhea and other gastrointestinal illnesses in humans and animals. However, the prevalence and genotype of the parasites in Longjiang Wagyu cattle in Heilongjiang Province, Northeast China have not been reported. In the present study, a total of 423 fecal samples of Longjiang Wagyu cattle collected from different farms in Heilongjiang Province, Northeast China, were examined for Cryptosporidium spp. and E. bieneusi using nested PCR. The overall infection rates for Cryptosporidium spp. and E. bieneusi were 6.38% (n = 27) and 7.09% (n = 30), respectively. The prevalence in different age groups ranged from 3.80% (95% confidence interval (CI) 1.01-6.59) to 8.36% (95% CI 4.83-11.90) for Cryptosporidium spp. and 5.97% (95% CI 2.52-9.43) to 7.94% (95% CI 4.49-11.40) for E. bieneusi. By analyzing the DNA sequences of the small subunit (SSU) rRNA gene, two Cryptosporidium species were detected in this study, namely C. parvum (n = 25) and C. ryanae (n = 2). The IIdA20G1 subtype was further identified by using the 60-kDa glycoprotein (gp60) gene of C. parvum. E. bieneusi was identified using three known sequences through the analysis of internal transcribed spacer (ITS) sequences: J (n = 23), I (n = 5), and BEB4 (n = 2), and all belonged to group 2. The results indicated that some of the Cryptosporidium species and E. bieneusi genotypes identified in Longjiang Wagyu cattle in the study areas might have zoonotic potential.

Molecular detection of Cryptosporidium and Enterocytozoon bieneusi in dairy calves and sika deer in four provinces in Northern China.

The protistan pathogens Cryptosporidium and Enterocytozoon bieneusi can cause significant intestinal diseases in animals and humans. However, limited information is available regarding prevalence and molecular characterization of Cryptosporidium and E. bieneusi in ruminants in Northern China. In this study, the overall prevalence of Cryptosporidium and E. bieneusi was 19.3% (62/321) and 28.97% (93/321) in dairy calves and 1.10% (9/818) and 13.57% (111/818) in sika deer (Cervus nippon) in four provinces in Northern China, respectively. The prevalence of Cryptosporidium and E. bieneusi in different factor groups was various. Five Cryptosporidium species/genotypes were identified, of which C. parvum, C. ryanae, C. bovis, and C. andersoni were only found in dairy calves, and only Cryptosporidium deer genotype was found in sika deer. Moreover, J, I, and BEB4 ITS genotypes of E. bieneusi were found in dairy calves, and six known genotypes (JLD-III, JLD-IX, JLD-VII, EbpC, BEB6, and I) and ten novel genotypes (namely LND-I and JLD-XV to JLD-XXIII) were found in sika deer in this study. Cryptosporidium parvum and E. bieneusi genotype J were identified as the predominant species/genotypes in dairy calves, whereas the predominance of Cryptosporidium spp. and E. bieneusi in sika deer was Cryptosporidium deer genotype and BEB6, respectively. The present study reported the prevalence and genotypes of Cryptosporidium and E. bieneusi in dairy calves and sika deer in four provinces in northern China. The present findings also suggest that investigated dairy calves and sika deer may play an important role in the transmission of E. bieneusi and Cryptosporidium to humans and other animals, and also in an effort to better understand the epidemiology of these enteric pathogens in China.

Detection of Point Prevalence and Assemblages of Giardia spp. in Dairy Calves and Sika Deer, Northeast China.

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), ß-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.

Molecular detection and genotyping of Toxoplasma gondii in free-ranging pigs in Northeastern China.

Pig is the well-known intermediate host of T. gondii, a ubiquitous and obligate intracellular zoonotic pathogen. However, information about prevalence and genotypes of T. gondii infection in free-ranging pigs is scarce. Therefore a total of 186 hilar lymph nodes specimens were collected from free-ranging pigs from rural regions from Jilin (n = 119) and Liaoning (n = 67) provinces, northeastern China, and were investigated from T. gondii infection by semi-nested PCR of the B1 gene. Positive testing samples were genotyped by using polymerase chain reaction followed by restriction-fragment length polymorphism technology (PCR-RFLP) applied to 11 loci. The overall prevalence of T. gondii in investigated free-ranging pigs was 18.3% (34/186), with 16.4% (11/67) in Liaoning Province and 19.3% (23/119) in Jilin Province. Moreover, two genotypes, namely ToxoDB #9 and ToxoDB #10, were detected in investigated pigs. The present study showed a high T. gondii prevalence in free-ranging compared to farmed pigs as reported in previous studies by others, raising a major public health concern. It is essential to establish efficient strategies to prevent and control T. gondii infection in free-ranging pigs, other animals and humans in investigated regions.