Clonal spread of Escherichia coli O101: H9-ST10 and O101: H9-ST167 strains carrying fosA3 and bla CTX-M-14 among diarrheal calves in a Chinese farm, with Australian Chroicocephalus as the possible origin of E. coli O101: H9-ST10.

During a 2018 antimicrobial resistance surveillance of Escherichia coli isolates from diarrheal calves in Xinjiang Province, China, an unexpectedly high prevalence (48.5%) of fosfomycin resistance was observed. This study aimed to reveal the determinants of fosfomycin resistance and the underlying transmission mechanism. Polymerase chain reaction (PCR) screening showed that all fosfomycin-resistant E. coli carried the fosA3 gene. Pulsed-field gel electrophoresis (PFGE) and southern blot hybridization revealed that the 16 fosA3-positive isolates belonged to four different PFGE patterns (i.e., A, B, C, D). The fosA3 genes of 11 clonally related strains (pattern D) were located on the chromosome, while others were carried by plasmids. Whole-genome and long-read sequencing indicated that the pattern D strains were E. coli O101: H9-ST10, and the pattern C, B, and A strains were O101: H9-ST167, O8: H30-ST1431, and O101: H9 with unknown ST, respectively. Among the pattern C strains, the bla CTX-M-14 gene was co-localized with the fosA3 gene on the F18: A-: B1 plasmids. Interestingly, phylogenetic analysis based on core genome single nucleotide polymorphisms (cgSNPs) showed that the O101: H9-ST10 strains were closely related to a Australian-isolated Chroicocephalus-origin E. coli O101: H9-ST10 strain producing CTX-M-14 and FosA3, with a difference of only 11 SNPs. These results indicate possible international dissemination of the high-risk E. coli clone O101: H9-ST10 by migratory birds.

[The preparation of oligonucleotide chips for detecting BMPR-IB gene polymorphism in sheep].

BMPR-IB gene increases ovulation rate and litter size as a major gene in Chinese Merino, because of 746 A to G mutation. The aim of this study was to make oligonucleotide chips to determine single nucleotide polymorphism (SNP) in BMPR-IB gene. The oligonucleotide chips were manufactured by using six sequence-specific oligonucleotide probes derived from polymorphic regions in the mutation and spotting the probes by microarrayer onto the aldehyde modified glass slides. The 746 A to G mutation was detected with ovine blood in reaction cells of chips. The results were identified by designed the Arraydoctor 2.0 software, in accordance with those of restriction fragment length polymorphisms (RFLP). The results showed the oligonucleotide chips are a parallel, accurate and effective way for screening prolificacy in molecular-assisted selection(MAS)of sheep.

[Detection of major gene on litter size in sheep].

The current study was designed to detect SNPs within BMP15 and BMPR-IB gene and investigate the effect of the genes on sheep litter size. Four sheep lines, HU-Yang, Chinese Merino monotocous, Chinese Merino multiparous for wool production and Chinese Merino multiparous for mutton production, were used in this study. Litter sizes were recorded for each ewe in the four lines. Primers for BMP15 and BMPR-IB gene were designed from database sheep sequence and polymorphisms were detected by PCR-RFLP method. The results showed that there was no polymorphism with BMP15 gene among the four lines, and there was an A / G SNP with BMPR-IB gene at base 746 among the four lines. Three types of genotype (BB, B+ and ++), based on A / G locus, were found within each line. The frequencies of genotypes were significantly different among the lines (P<0.001), with BB genotype primarily existing in HU-Yang, ++ genotype in Chinese Merino monotocous line, and B+ genotype in Chinnese Merino multiparous lines. The A / G mutation influence significantly the sheep litter sizes, and the BB and B+ ewes had significant higher litter sizes than ++ ewes. The results of present study showed simultaneously that the genotype of BMPR-IB was a perfect predictor of the sheep litter sizes. These results intensively indicated that BMPR-IB is a major gene to affect litter size in sheep, and could be used as the molecular genetic marker to select litter size in sheep.