Prevalence and genetic diversity of bovine viral diarrhea virus in dairy herds of China.

To determine the nationwide prevalence and genetic diversity of bovine viral diarrhea virus (BVDV) in China, 92 dairy farms with more than 500 animals in 19 provinces of China were surveyed in 2017. At each farm, ear notch samples from calves less than six months old and bulk tank milk (BTM) samples were collected. A total of 901 ear notch samples and 329 BTM samples from 183 tanks were sampled. A total of 20 (20/901, 2.22 %) ear notch samples from 10 (10/92, 10.86 %) farms tested positive for BVDV by IDEXX Antigen Point-of-Care (POC) Test kit and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). In addition, 80 of 183 (80/183, 43.7 %) BTM samples from 43 (43/92, 46.7 %) farms were identified as positive by qRT-PCR. The RNA of positive and suspect samples identified by qRT-PCR was subjected to 5'- untranslated region (UTR) amplification by nested RT-PCR and then sequenced. A total of 119 sequences were obtained and phylogenetic analysis of these 5'-UTR sequences revealed the presence of eight different subgenotypes of BVDV-1 including 1a (n = 37, 31.09 %), 1b (n = 5, 4.20 %), 1c (n = 34, 28.57 %), 1d (n = 2, 1.68 %), 1m (n = 25, 21.01 %), 1q (n = 6, 5.04 %), and two unknown subgenotypes which were tentatively typed as "BVDV-1v" (n = 8, 6.72 %) and "BVDV-1w" (n = 2, 1.68 %), respectively. BVDV-1a, 1c, and 1m were the dominant strains, collectively accounting for 80.67 % (96/119) of all sequences. Phylogenetic analysis based on selected N-terminal autoprotease (Npro) sequences confirmed the classification of the 5'-UTR sequences. In conclusion, the prevalence of BVDV persistent infection in dairy cattle was high and genetic diversity was high and increasing, revealing a serious threat to the health of cattle in China and highlighting the need for BVDV control.

Phenotypic and genotypic characterization of antimicrobial resistance profiles in Streptococcus dysgalactiae isolated from bovine clinical mastitis in 5 provinces of China.

Bovine mastitis is among the most prevalent and costly diseases of dairy animals and is caused by a variety of bacterial pathogens including Streptococcus dysgalactiae. However, comprehensive studies reporting the prevalence and antimicrobial resistance profiles of S. dysgalactiae isolated from bovine mastitis are scarce. Therefore, this study was to investigate the occurrence of S. dysgalactiae associated with bovine clinical mastitis, to assess their antimicrobial resistance profiles, and to analyze the phenotypic and genotypic profiling of resistant isolates. In total, 1,180 milk samples were collected from dairy cows with clinical mastitis belonging to 74 commercial dairy herds located in 14 provinces of China from January 2014 to May 2016. Overall S. dysgalactiae isolates were recovered from 88 (7.5%) of the mastitic milk samples. The antimicrobial susceptibility of these isolates was tested against 8 antimicrobial agents by using minimum inhibitory concentrations. Results showed that 82 (93.2%) isolates expressed resistance to more than one antimicrobial agent. Antimicrobial resistance was highest against kanamycin (89.8%), sulfonamide (83.0%), and streptomycin (58.0%), which can be attributed to the intrinsic resistance for most of Streptococcus spp. against those antimicrobial substances. Strikingly, 30 (34.1%) and 12 (13.6%) isolates were found resistant to cephalexin and ceftriaxone, respectively. BlaTEM, ermB, and tetM were the most prevalent resistance genes. All isolates carried at least one of all tested resistance genes. Also, 1.1, 12.5, 18.2, 36.4, and 31.8% of isolates were positive for at least one tested resistance gene in 1, 2, 3, 4, or 5 classes of antimicrobials. Survival analysis showed a significant association between ermB and survival of the S. dysgalactiae isolates at increasing erythromycin concentrations. No other statistically significant associations were observed between the phenotypic and genotypic resistance profiles. This study concludes a considerable prevalence of S. dysgalactiae associated with bovine mastitis in dairy herds of China and these isolates exhibited high resistance rates to tested antimicrobials, coupled with high occurrence of resistance genes. Both the prevalence of S. dysgalactiae and their antimicrobial resistance profiles strongly varied among dairy herds, demonstrating the need for antimicrobial susceptibility surveillance at the herd level to ensure optimal therapeutic results.

Incidence of clinical mastitis and distribution of pathogens on large Chinese dairy farms.

Knowledge of the incidence of clinical mastitis (CM) and the distribution of pathogens involved is essential for development of prevention and control programs as well as treatment protocols. No country-wide study on the incidence of CM and the distribution of pathogens involved has been conducted in China. Core objectives of this study were, therefore, to determine the cumulative incidence of CM and the distribution of pathogens causing CM on large Chinese (>500 cows) dairy farms. In addition, associations between the distribution of CM pathogens and bedding materials and seasonal factors were also investigated. Bacterial culture was done on a total of 3,288 CM quarter milk samples from 161 dairy herds (located in 21 provinces) between March 2014 and September 2016. Additional data, including geographical region of herds, herd size, bedding types, and number of CM cases during the last month, were also recorded. Mean cumulative incidence of CM was 3.3 cases per 100 cows per month (range = 1.7 to 8.1). The most frequently isolated pathogens were Escherichia coli (14.4%), Klebsiella spp. (13.0%), coagulase-negative staphylococci (11.3%), Streptococcus dysgalactiae (10.5%), and Staphylococcus aureus (10.2%). Streptococcus agalactiae was isolated from 2.8% of CM samples, whereas Streptococcus uberis were isolated from 2.1% of samples, and 15.8% of 3,288 samples were culture-negative. Coagulase-negative staphylococci, E. coli, and other Enterobacter spp. were more frequently isolated in the northwest than the northeast or south of China. Streptococcus dysgalactiae, other streptococci, and Strep. agalactiae were more frequently isolated in winter (October-March), whereas E. coli and Klebsiella spp. were mostly isolated in summer (April-September). Streptococcus dysgalactiae was more often isolated from CM cases of herds using sand bedding, whereas Klebsiella spp. and other streptococci were more common in herds using organic bedding. The incidence of CM and distribution of pathogens differed among herds and better mastitis management is needed. Furthermore, geography, bedding materials, and season should be included when designing mastitis control and prevention schemes for Chinese dairies.