The diagnosis and molecular epidemiology investigation of avian hepatitis E in Shandong province, China.

BACKGROUND: Avian hepatitis E virus (HEV) is the pathogenic agent of big liver and spleen disease (BLS) and of hepatitis-splenomegaly syndrome (HSS) in chickens, which have caused economic losses to the poultry industry in China. In this study, 18 samples of BLS chickens were collected to reveal the molecular epidemiological characteristics of avian HEV in the province of Shandong, China. RESULTS: Gross and microscopic lesions of clinical samples were observed; then, virology detection and genetic analysis of avian HEV were performed. The results showed that there was significant swelling and rupture in the liver and that the spleen was enlarged. Microscopic lesions demonstrated obvious hemorrhage in the liver, with infiltration of heterophilic granulocytes, lymphocytes, and macrophages, as well as the reduction of lymphocytes in the spleen. Eleven of the 18 samples were positive for avian HEV, with a positive rate of 61.11%. More importantly, all avian HEV-positive samples were mixed infections: among these, the mixed infections of avian HEV and chicken infectious anemia virus (CIAV) and avian HEV and fowl adenovirus (FAdV) were the most common. Furthermore, the genetic evolution analysis showed that all avian HEV strains obtained here did not belong to the reported 4 genotypes, thus constituting a potential novel genotype. CONCLUSIONS: These results of this study further enrich the epidemiological data on avian HEV in Shandong, prove the genetic diversity of avian HEV in China, and uncover the complex mixed infections of avian HEV clinical samples.

Prevalence and multilocus sequence typing of Clostridium perfringens isolated from different stages of a duck production chain.

Clostridium perfringens (C. perfringens) is a zoonotic microorganism and rarely reported in duck production chain. This study aimed to investigate prevalence, serotype distribution, antibiotic resistance and genetic diversity of C. perfringens at different stages of a duck production chain. In total, 319 samples were collected from a large-scale rearing and slaughter one-stop enterprise in Weifang, China, of which 42.95% of samples were positive for C. perfringens. All isolates were genotype A. Cpe and cpb2 genes were found in 2.54% and 24.87% of the isolates, respectively. Antimicrobial susceptibility testing revealed that 55.47% of the isolates resistant to at least 5 classes of commonly used antibiotics. Multilocus sequence typing (MLST) results showed that 65 representative isolates were divided into 47 sequences types (STs), 33.85% of them were included into four clonal complexes (CC). Some of isolates from breeding and slaughtering stages were distributed in the same CC or ST, indicating duck products may be contaminated by C. perfringens originated from the breeding stage. Part of duck isolates were distributed in the same CC as human isolates and systemically close with human isolates. The high contamination rates of duck products, the isolates with multi-drug antibiotic resistance or the cpe gene, and the close relationship between strains from human and ducks, indicated potential public health risks, not only control measures at slaughtering stage but also at rearing stage should be considered to reduce this risks.

Prevalence and characterization of Clostridium perfringens isolated from different chicken farms in China.

Clostridium perfringens (C. perfringens) is a common pathogenic microorganism present in nature, which can cause animal and human diseases, such as necrotizing enteritis (NE) in poultry. Little is known about the current prevalence status of C. perfringens from poultry farms of different types and regions in China. From December 2018 to August 2019, we investigated the prevalence, genotype distribution and drug resistance of C. perfringens from Guangdong, Pingyin, Tai'an and Weifang. A total of 622 samples were collected and processed for C. perfringens isolation, among which 239 (38.42%) samples were determined to be positive for C. perfringens. A total of 312 isolates of C. perfringens were recovered (1-5 strains were isolated for each positive sample), and 98.72% of the isolates were identified as type A, while the others were type F. Antimicrobial susceptibility testing revealed that 47.71% of the isolates were resistant to at least five classes of commonly used antibiotics. Multilocus sequence typing (MLST) showed that 74 representative isolates were divided into 63 sequence types (STs), and the Simpson's diversity index (Ds) of the STs for the five farms was 0.9799. 37.84% of the isolates were classified into seven clonal complexes (CC1-CC7), and the isolates from the same farm were more concentrated in the minimum spanning tree. In addition, some cloaca isolates and feed isolates were distributed in the same ST or CC; this result indicates that the C. perfringens in chicken can come from the environment (feed etc.).

Prevalence and multilocus sequence typing of Clostridium perfringens isolated from 4 duck farms in Shandong province, China.

Clostridium perfringens is an important zoonotic microorganism. The present study was undertaken to investigate prevalence, serotype distribution, antibiotic resistance, and genetic diversity of C. perfringens isolates from 4 duck farms in Shandong, China. In total, 424 samples of cloacal swabs and environment were collected from 3 commercial meat-type duck farms in Tai'an, Liaocheng, and Weifang and one breeder duck farm in Liaocheng between December 2018 and June 2019, of which, 207 (48.82%) samples were determined to be positive for C. perfringens; a total of 402 isolates of C. perfringens were recovered, all of which were identified as type A; 30.85% of the isolates were positive for cpb2 gene; and cpe gene was found in 0.5% of the isolates. Antimicrobial susceptibility testing revealed that some of the isolates exhibited high antibiotic resistance, and 39.14% of the isolates were resistant to at least 5 classes of commonly used antibiotics. Multilocus sequence typing analysis showed that 85 representative isolates encompassed 54 different sequences types (STs), clustered in 5 clonal complexes (CCs) and 40 singletons. ST3, the most common ST in 54 STs, constituting 15.29% of all isolates, was also the most prevalent ST of isolates from the Liaocheng breeder duck farm (farm 3). CC1, the most prolific CC, containing 15.29% of the analyzed isolates, was the popular subtype of isolates from Liaocheng meat duck farm (farm 2). Although all the isolates belong to type A, the genetic diversity varied greatly in different regions; the Simpson's Diversity Index of STs for Liaocheng, Tai'an, and Weifang were 0.5941, 0.9198, and 0.9627, respectively. Some of cloacal isolates and environmental isolates were distributed in the same ST or CC, indicating close genetic relationship between cloacal isolates and environmental isolates. A portion of the strains from humans and ducks was found to be phylogenetically close. The close relationship between strains from humans and ducks, the high antibiotic resistance of C. perfringens, and the cpe-positive isolates indicated potential public health risks.

Transmission identification of Escherichia coli aerosol in chicken houses to their environments using ERIC-PCR.

In order to study E. coli aerosol spreading from chicken houses to their surrounding air, air samples, including indoor and outdoor air (upwind 10 and 50 m as well as downwind 10, 50, 100, 200 and 400 m away) of 5 chicken houses were collected using six-stage Andersen microbial samplers and Reuter-Centrifugal samplers (RCS). E. coli concentrations (CFU/m(3) air) collected from different sampling sites were calculated. E. coli strains from chicken feces samples were also isolated. Furthermore, the enterobacterial repetitive intergenic consensus (ERIC)-PCR method was applied to amplify the isolated E. coli strain DNA samples. Through the genetic similarity analyses of the E. coli obtained from different sampling sites, the spreading of bioaerosol from animal houses to the ambient air was characterized. The results showed that the isolated E. coli concentrations in indoor air (9-63 CFU/m(3)) in 5 chicken houses were higher than those in upwind and downwind air, but there were no significant differences between the indoor and downwind sites 10 m away from all the 5 houses (P>0.05). The phylogenetic tree indicated that a part of the E. coli (34.1%) isolated from indoor air had 100% similarity with those isolated from feces, and that most of E. coli isolated (54.5%) from downwind at 10, 50, 100 or even 200 m had 100% similarity with those isolated from indoor air or feces too. But those isolated from upwind air had a lower similarity (73%-92%) with corresponding strains isolated from indoor air or feces. Our results suggested that some strains isolated from downwind air and indoor air originated in the chicken feces, but most of isolates obtained from upwind air samples did not come from the chicken feces or indoor air. Effective hygienic measures should be taken in animal farms to prevent or minimize downwind spreading of microorganism aerosol.