| Cryptosporidium and Enterocytozoon bieneusi is typical zoonotic intestinal pathogens.In this study,molecular biology techniques were used to detect the prevalence of two intestinal pathogens of broilers in some areas of Zhejiang Province,and their species and subtypes/genotypes were identified by sequence comparative analysis,and their genetic and evolutionary characteristics were analyzed.(1)This study used polymerase chain reaction to detect and sequence 812 chicken feces DNA samples based on SSU rRNA gene loci,812 chicken fecal DNA samples from 11 sampling sites in Zhejiang Province were detected by polymerase chain reaction method and sequence analysis was performed.The results showed that the total infection rate of Cryptosporidium in this study was 6.3%(51/812),5 of the 11 sampling sites were positive,and the infection rate was between 2.3% and 33.0%.In this study,the infection rate of two sampling sites in Hangzhou was high(33.0% and 16.1%),and no Cryptosporidium infection was found in the two sampling sites in Ningbo.The difference in Cryptosporidium infection rate among sampling points was significant(P≤0.01).The Cryptosporidium infection rates of broilers of different days of age were: The age of 30 days,0%(0/20),30 ～ 60 days of age 10.6%(38/358),60 ～ 90 days of age 1.1%(4/378);At 90 days of age,the infection rate was 16.1%(9/56).No significant correlation was found between the infection rate and the age of chickens,and the difference in the Cryptosporidium infection rate was very significant among the groups of each age(P≤0.01).Cryptosporidium baileyi(n=49)and Cryptosporidium meleagridis(n=2)were detected at the SSU rRNA gene locus.The positive samples of Cryptosporidium meleagridis were subtyped based on the gp60 gene locus,and the two sequences were completely consistent,both of which were subtype IIIb A24G1R1.The results indicate that Cryptosporidium infection exists in Zhejiang broilers,and it is necessary to strengthen feeding management and pay more attention to the prevention and control of parasites and intestinal pathogens.(2)This study used polymerase chain reaction to detect and sequence 812 chicken feces DNA samples based on ITS sequences.The results showed that in this study,the total infection rate of Enterocytozoon bieneusi was 1.7%(14/812),8 of 11 sampling sites were positive,and the infection rate was between 1.0%and 5.6%,among which the infection rate was the highest at one sampling site in Taizhou,and no Enterocytozoon bieneusi infection was found at two sampling sites in Ningbo.There was no significant difference in the infection rate of Enterocytozoon bieneusi among all sampling sites(P=0.11).The infection rates of Enterocytozoon bieneusi in broilers of different days of age were: The age of 30,0%(0/20),30 ～ 60 days of age 0.6%(2/358),60 ～ 90 days of age and 2.6%(10/378);The infection rate of broilers at 90 days of age was 3.6%(2/56).The infection rate was positively correlated with the increase in age of broilers,and there was no significant difference in the infection rate of Enterocytozoon bieneusi among all groups of age(P=0.10).Four genotypes of Enterocytozoon bieneusi were identified by sequence analysis,belonging to three different groups,group 1: D(n=2);group 2: BEB6(n=4),J(n=2);group 11: Pt Eb IX(n=5).The results showed that the infection rate of Enterocytozoon bieneusi in broilers in Zhejiang was low,but there was a public health risk.In conclusion,the infection rates of Cryptosporidium and Enterocytozoon bieneusi of broilers in some areas of Zhejiang are low.Cryptosporidium baileyi was the dominant species,Pt Eb IX was the dominant genotype in Enterocytozoon bieneusi.Results to study the endemic situation of poultry Cryptosporidium and Enterocytozoon bieneusi in China,the distribution characteristics of subtypes/genotypes and the relationship between genetic evolution provided relevant data support. |
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