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Enterocytozoon bieneusi, an obligate intracellular microsporidian parasite, can infect humans and a wide variety of animals worldwide. However, information on the prevalence and molecular characterization of E. bieneusi in pet rats and guinea pigs is lacking. In this study, 325 fecal samples were collected from 152 pet fancy rats and 173 pet guinea pigs purchased from pet shops in Henan and Shandong provinces. The prevalence of E. bieneusi was 11.2% (17/152) in pet fancy rats and 20.2% (35/173) in pet guinea pigs. Genotypes D (n = 12), Peru11 (n = 3), S7 (n = 1) and SCC-2 (n = 1) were identified in pet fancy rats, and genotype S7 (n = 30) and a novel genotype PGP (n = 5) were identified in pet guinea pigs. The ITS sequence and its phylogenetic analysis showed that the novel genotype PGP was distinctly different; it exhibited less than 50% similarity to the reference sequences, and did not cluster with any of the known E. bieneusi genotype groups, forming a unique branch between groups 6 and 7. These data suggest that this is a new E. bieneusi genotype group. This is the first report of E. bieneusi infection in pet fancy rats and pet guinea pigs worldwide. The identification of zoonotic genotypes D, Peru11, and S7 suggests that pet fancy rats and guinea pigs can be potential sources of human microsporidiosis.

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Neospora caninum infection is a significant cause of abortion in cattle. We investigated the tissue distribution of N. caninum in aborted bovine fetuses and dam blood samples by a nested PCR assay, and compared the nested PCR with ELISA in the diagnosis of N. caninum infection. In total, 26 aborted fetuses and 813 blood samples were collected from 8 dairy herds in Beijing (n=212) and Tianjin (n=601), China. Fifteen fetuses (57.7%) were tested N. caninum-positive by the nested PCR. N. caninum DNA was detected from the brain of 52%, kidneys of 22%, skeletal muscle of 18%, and heart of 4% of the aborted fetuses. The PCR-positive cases (55%, 11/20) were higher than seropositive cows (40%, 8/20) in a subset of 20 fetuses, but the PCR results of blood samples of the 20 cows were all negative. The seroprevalence of the 813 samples was 15.5% (43.4% of samples from Beijing, 5.7% of samples from Tianjin), compared to the PCR-positive blood samples of 0.9%. Our study showed that the nested PCR is a valuable diagnostic tool for the primary diagnosis of N. caninum in aborted fetuses, while ELISA is the preferred assay for testing blood samples collected from cows. The two assays are complementary in determining whether abortions are associated with N. caninum infection in cattle.

Abortion in dairy cattle causes considerable economic losses to the dairy industry. Aborted fetuses and samples from the corresponding aborting dams from 12 dairy herds in Beijing were tested for 9 abortifacient infectious pathogens by PCR between 2008 and 2010. From a total of 80 abortion cases collected during this period, infectious agents were detected in 45 (56.3%) cases, 22 (48.9%) of which represented co-infections with two or three infectious agents. The detected pathogens included infectious bovine rhinotracheitis virus (36.3%) andNeospora caninum(31.3%), followed by bovine viral diarrhoea virus (7.5%),Brucella abortus(6.3%),Tritrichomonas foetus(5%) andToxoplasma gondii(1.3%).Campylobacter fetus, Coxiella burnetiiandChlamydophila psittaciwere not detected in any abortion case. Findings from this study indicated that infectious bovine rhinotracheitis virus andNeospora caninumwere the main potential causes of abortions in Beijing dairy herds, whereas the bacterial pathogens were not, in contrast to reports from other countries. This is the first study to test nine abortifacient infectious agents by PCR at the same time, and it is also the first time to report the involvement of a variety of infectious agents in bovine abortion cases in China.

Immunity to Eimeria is species-specific, and chickens with immunity to one species of Eimeria remain susceptible to other Eimeria species. This presents a major challenge in the development of effective vaccines against multiple Eimeria species. In this study, we cloned the antigenic epitope of a tachyzoite surface protein gene of Eimeria tenella, a tachyzoite surface protein gene of Eimeria acervulina and the gametocyte protein gene of Eimeria maxima, and constructed prokaryotic and eukaryotic plasmids carrying the multi-epitope antigenic gene. Immunization of chickens with the multivalent DNA and protein conferred partial protection against infection by the three Eimeria species, as shown by increased CD4+ T lymphocytes in the intestinal mucosa, decreased oocyst excretion and intestinal lesions, and increased body weight gain compared with non-immunized controls. The DNA prime-protein boost immunization schedule induced greater cellular immunity and protection from Eimeria infection than immunization with DNA or protein alone. Our findings demonstrated that DNA prime-protein boost immunization with a multivalent vaccine could stimulate protective immunity against challenge infection of multiple Eimeria species. This work provides a promising step towards DNA-protein vaccination against multiple species of pathogens.

The prevalence and genotype of Toxoplasma gondii infection in dogs in Henan Province, Central China was investigated. A total of 125 blood samples were collected from pet dogs during April to June 2013, and all samples were examined by indirect hemagglutination antibody test (IHA) and nested PCR. The overall T. gondii prevalence in pet dogs was 24.0% (30/125), with 20.8% (26/125) in IHA and 10.4% (13/125) in PCR, respectively. No statistical associations were found between animal gender and age and the prevalence of T. gondii infection. Thirteen positive DNA samples were genotyped using 11 PCR-RFLP markers, including SAG1, (3’+5’) SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico. Of these, only 2 samples were genotyped with complete data for all loci, and a novel genotype (type III at SAG3 and GRA6 loci, and type I at other loci) was identified. This is the first report of genetic characterization of T. gondii infection in dogs in China.