First genetic characterization of Toxoplasma gondii infection in Arctic foxes (Vulpes lagopus) in China.

Toxoplasma gondii can infect virtually all warm-blooded animals including foxes. However, little is known of the molecular epidemiology and genotypes of T. gondii infecting foxes in China. Therefore, the present study characterized T. gondii genotypes in foxes in China for the first time. During November 2014 to October 2015, brain tissue samples collected from 264 Arctic foxes (Vulpes lagopus) in Jilin, Heilongjiang and Shandong provinces were used to detect the T. gondii B1 gene by a semi-nested PCR, and the positive samples were genotyped at 10 nuclear loci (i.e., SAG1, alternative SAG2, 5'-and 3'-SAG2, SAG3, L358, BTUB, c22-8, GRA6, c29-2, PK1) and an apicoplast locus (Apico) by multi-locus PCR-RFLP technology. Twenty-one (7.96%) samples from 264 foxes were positive for T. gondii B1 gene. T. gondii infection in male and female foxes was 7.14% and 8.70%, respectively. The highest infection rate (11.86%) was detected in foxes from Shandong, followed by foxes from Jilin (6.49%) and Heilongjiang (2.90%). Two genotypes (ToxoDB#9 and ToxoDB#10) were identified. This is the first genetic characterization of T. gondii from foxes in China, which provides basic data for the surveillance and control of T. gondii infection in foxes, other animals and humans.

First report of Cryptosporidium canis in farmed Arctic foxes (Vulpes lagopus) in China.

BACKGROUND: Cryptosporidium is an important genus of enteric zoonotic parasites, which can infect a wide range of animals including foxes. Little information is available concerning the prevalence and molecular characterisation of Cryptosporidium spp. in farmed Arctic foxes (Vulpes lagopus) in China. Thus, the objective of the present study was to investigate the prevalence of Cryptosporidium spp. in Arctic foxes in China using nested PCR amplification of the small subunit ribosomal RNA (SSU rRNA) gene. FINDINGS: The overall prevalence of Cryptosporidium spp. in Arctic foxes was 15.9 % (48/302), with 12.9 % in male (18/139) and 18.4 % in female (30/163) foxes, respectively. The prevalence in different farms varied from 0 to 31.43 %. The prevalence of infection in different age groups varied from 14.1 % to 19.0 %. Foxes from Hebei Province (7.8 %, 11/141) had a significantly lower Cryptosporidium spp. prevalence than those from Heilongjiang Province (22.9 %, 16/70) and Jilin Province (23.1 %, 21/91) (P= 0.0015). Sequence analysis of the SSU rRNA gene indicated that all the 48 isolates represented C. canis. CONCLUSIONS: This is the first report of C. canis infection in farmed Arctic foxes in China, which also provides foundation data for preventing and controlling Cryptosporidium infection in foxes, other animals and humans.

Prevalence, risk factors and multilocus genotyping of Enterocytozoon bieneusi in farmed foxes (Vulpes lagopus), Northern China.

BACKGROUND: Microsporidiosis is a common disease in animals and humans around the world. Enterocytozoon bieneusi is the most common microsporidian species in humans. Many animal species may be a potential source of human microsporidiosis. However, information concerning prevalence and genotypes of E. bieneusi infection in farmed foxes (Vulpes lagopus) is scarce. Therefore, the present study examined prevalence, risk factors and genotypes of E. bieneusi in farmed foxes in northern China using a genetic approach. RESULTS: Of 302 fecal samples from farmed foxes, 37 (12.25%, 95% CI 8.55-15.95) were PCR-positive for E. bieneusi, and the prevalence was highly associated with the farming mode in that foxes raised outdoors (26.03% positive, 95% CI 18.91-33.15) had a significantly higher E. bieneusi prevalence than those raised indoors. Eleven internal transcribed spacer (ITS) genotypes were identified among the positive samples: four known E. bieneusi genotypes (Peru 8, Types IV, CHN-DC1 and D) and seven novel genotypes (NCF1-NCF7). Genotype NCF2 was the commonest (n = 13) and was found in five farms across three provinces (Jilin, Heilongjiang and Hebei). All genotypes belonged to phylogenetic group 1. Multilocus sequence typing (MLST) analyses revealed additional diversity. CONCLUSIONS: These findings indicate the presence of zoonotic E. bieneusi infection in farmed foxes in northern China. This is also the first report of genotypes Peru8, CHN-DC1 and Type IV, and seven novel genotypes (NCF1-NCF7) in farmed foxes by ITS combining with microsatellite and minisatellite markers for the first time. The results will provide baseline data for preventing and controlling E. bieneusi infection in farmed foxes, other animals and humans.

First report of Toxoplasma gondii Seroprevalence in Farmed Arctic Foxes (Vulpes lagopus) in Eastern and Northeastern China.

Until now, no information on Toxoplasma gondii seroprevalence in Arctic foxes (Vulpes lagopus) was available in China. A serological survey was undertaken to assess T. gondii seroprevalence in farmed Arctic foxes in eastern and northeastern China. Antibodies to T. gondii were examined in 1346 farmed Arctic foxes using the modified agglutination test (MAT). A total of 113 (8.39%, 95% confidence interval [CI] 6.91-9.87) serum samples were positive to T. gondii at a 1:25 cutoff. Seroprevalence of T. gondii infection in male Arctic foxes was 8.68% (95% CI 6.75-10.6), which was higher than that in the female Arctic foxes (7.95%, 95% CI 5.65-10.26). The prevalence in polar foxes was 7.07% (95% CI 5.14-8.99), which was lower than that in the blue foxes (9.75%, 95% CI 7.49-11.99). T. gondii seroprevalence in Arctic foxes in Heilongjiang, Jilin, Liaoning, and Shandong Provinces was 9.85% (95% CI 5.75-13.95), 9.21% (95% CI 5.54-12.87), 7.37% (95% CI 5.22-9.51), and 8.68% (95% CI 5.66-11.70), respectively. There were no statistically significant differences between T. gondii seroprevalence and morphs, sex, or regions of Arctic foxes in logistic regression analysis (p < 0.05). The results of the present survey indicated that T. gondii infection in farmed Arctic foxes is prevalent in China. To our knowledge, this is the first report of T. gondii seroprevalence in Arctic foxes in China.

Molecular detection and genotypic characterization of Toxoplasma gondii in wild waterfowls in Jilin Province, Northeastern China.

Toxoplasma gondii can infect almost warm-blooded animals, including humans. Limited information about T. gondii infection in wild waterfowls is available in China. The present study was conducted to determine prevalence and genotype T. gondii infection in 11 wild waterfowl species in Jilin Province, northeastern China. A total of 249 wild waterfowls were sampled between April and July 2013 from Jilin Province, and the tissue samples were collected for the detection of T. gondii by a semi-nested PCR targeting the B1 gene. The positive samples were genotyped at 11 genetic markers (SAG1, 5'- and 3'-SAG2, alternative SAG2, SAG3, BTUB, GRA6, L358, PK1, c22-8, c29-2, and Apico) using multilocus polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technology. The overall prevalence of T. gondii in the wild waterfowls was 7.2% (18/249, 95% confidence interval [CI] 4.0-10.4), with the highest prevalence (22.0%, 95% CI 10.5-33.5) in Anas formosa, followed by Anas platyrhynchos (20.0%, 95% CI 6.0-44.0), Falcated teal (12.5%, 95% CI 0.0-35.4), and Fulica atra (4.0%, 95% CI 0.0-11.7). Of 18 positive samples, only 2 samples (TgWfjl1 and TgWfjl2) were genotyped completely, and one genotype, namely ToxoDB #9, was revealed. The result of this survey has implications for better understanding of the genetic diversity of T. gondii in China. This is the first report of prevalence and genotypic characterization of T. gondii in wild waterfowls in northeastern China.

Prevalence and genotyping of Toxoplasma gondii in naturally-infected synanthropic rats (Rattus norvegicus) and mice (Mus musculus) in eastern China.

BACKGROUND: Synanthropic rats and mice share the same environment with humans and play an important role in epidemiology of toxoplasmosis; however, there is limited information about prevalence and genetic characterization of Toxoplasma gondii in synanthropic rats and mice in China. FINDINGS: In the present study, the prevalence and genetic characterization of T. gondii naturally infected synanthropic rodents (Rattus norvegicus and Mus musculus) were investigated in the urban area of Xuzhou city, Eastern China between June 2013 and August 2014. DNA from the brain of each animal was prepared and screened by specific PCR assay targeting 35-fold repeated B1 gene (B1-PCR). PCR positive DNA samples were further genotyped by multi-locus PCR-RFLP. Overall, out of 123 synanthropic rodents, 29 samples were positive by B1 gene-targeted PCR (23.6%). Of these, 7 out of 31 (22.3%) M. musculus were positive, whereas the positive rate of R. norvegicus was 23.9% (22/92). Multi-locus PCR-RFLP analysis reveals that seven PCR-positive samples were completely genotyped and they were identified as type China 1 (ToxoDB# 9). CONCLUSION: To our knowledge, this is the first report of molecular detection and genetic characterization of T. gondii infection in synanthropic rodents in Eastern China. The results of the present study showed a high infection pressure of T. gondii exists in the environment and synanthropic rodents infected by T. gondii may be an important source of infection for cats and other animals.