| Toxoplasma gondii is an opportunistic pathogenic parasite common in warm-blooded animals and humans,it can infect not only terminal host cats and felines,but also intermediate hosts such as dogs,horses and birds.It caused serious harm,mainly resulting in infected animal abortion,stillbirth,birth defects and other reproductive hazards.T.gondii can also infect humans,causing severe illness in people with weakened immune systems.Neospora caninum is an intracellular protozoa,can infect many domestic and wild animals,people with low immunity may become opportunistic hosts.At present,only dogs and canids are the final hosts of N.caninum.Animals infected with N.caninum often cause fetal abortion and inflammation of the heart,brain and liver.It does the most harm to cows,at least 40% of all cow miscarriage cases worldwide are caused by N.caninum.As intermediate hosts for T.gondii and N.caninum in wild birds,information about the infection of both parasites is limited.This study randomly collected 239 wild birds from yiyang city,hunan province,China,including 38 portions of chestnut bunting,44 portions of tree pipit,26 portions of yellow-breasted bunting and 131 portions of tree sparrow.T.gondii and N.caninum were detected from DNA of brain tissue of wild birds:(1)Detection of T.gondii infection: In this study,genomic DNA of brain tissue was extracted and B1 gene was detected by Mn-PCR-RFLP method,the results showed that 13 of 239 wild birds were T.gondii positive(5.51%).Our results suggest that wild birds can carry and possibly transmit T.gondii.(2)Detection of N.caninum infection: In this study the NC-5 gene and the internal transcriptional spacer 1(ITS-1)region of the ribosomal region of N.caninum were targeted.DNA of wild bird brain was detected by specific PCR.The results showed that among 239 wild birds,the infection rate of N.caninum was 15.5%(37/239);Among them,the pipit is 20.5%(9/44),the tree sparrow is 18.3%(24/131),the chestnut bunting is 7.9%(3/38),and the yellow-breasted bunting is 3.8%(1/26);Phylogenetic relationship analysis showed that isolates N.caninum from different hosts and geographical sources had genetic diversity,it can be further divided into two distinct groups.This is the first time that N.caninum has been found in wild birds in China,providing important data for the future research and control of N.caninum.(3)Genotyping of T.gondii: In this experiment,genetic markers were used to genotype 13 positive samples obtained from B1 gene detection,genetic markers include SAG1,SAG2(5’+3’ SAG2,alter.SAG2),SAG3,BTUB,GRA6,c22-8,c29-2,L358,PK1,and Apico.Result display,in 13 samples was positive,all sites have 3 samples have been finished genotyping and identified as ToxoDB#10.The molecular prevalence and genetic characteristics of T.gondii in wild birds in hunan province have been reported for the first time.The results of this study suggest that wild birds are an important potential source of T.gondii and N.caninum infection in other animals,there is also a risk of mixed infection with T.gondii and N.caninum,it fills in the blank of related research on T.gondii genetic information and enriches experimental data for the prevention and treatment of two parasitic diseases and further research. |
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