Characterization of NDM-5-producing Enterobacteriaceae isolates from retail grass carp ( Ctenopharyngodon idella) and evidence of bla NDM-5-bearing IncHI2 plasmid transfer between ducks and fish.

We aimed to characterize NDM-5-producing Enterobacteriaceae from aquatic products in Guangzhou, China. A total of 196 intestinal samples of grass carp collected in 2019 were screened for carbapenemase genes. Characterization of bla NDM-5 positive isolates and plasmids was determined by antimicrobial susceptibility testing, conjugation experiments, Illumina HiSeq, and Nanopore sequencing. One Citrobacter freundii and six Escherichia coli strains recovered from seven intestinal samples were verified as bla NDM-5 carriers (3.57%, 7/196). The bla NDM-5 genes were located on the IncX3 ( n=5), IncHI2 ( n=1), or IncHI2-IncF ( n=1) plasmids. All bla NDM-5-bearing plasmids were transferred by conjugation at frequencies of ~10 -4-10 -6. Based on sequence analysis, the IncHI2 plasmid pHNBYF33-1 was similar to other bla NDM-5-carrying IncHI2 plasmids deposited in GenBank from Guangdong ducks. In all IncHI2 plasmids, bla NDM-5 was embedded in a novel transposon, Tn 7051 (IS 3000-ΔIS Aba125-IS 5-ΔIS Aba125- bla NDM-5- ble MBL- trpF- tat-∆ dct-IS 26-∆ umuD-∆IS Kox3-IS 3000), which was identical to the genetic structure surrounding bla NDM-5 found in some IncX3 plasmids. The IncHI2-IncF hybrid plasmid pHNTH9F11-1 was formed by homologous recombination of the bla NDM-5-carrying IncHI2 plasmid and a heavy-metal-resistant IncF plasmid through ∆Tn 1721. To the best of our knowledge, this is the first report on the characterization of bla NDM-5-bearing plasmids in fish in China. The IncHI2 plasmid pHNBYF33-1 may be transmitted from ducks, considering the common duck-fish freshwater aquaculture system in Guangdong. Tn 7051 is likely responsible for the transfer of bla NDM-5 from IncX3 to IncHI2 plasmids in Enterobacteriaceae, resulting in the expansion of transmission vectors of bla NDM-5.

Zinc Supplementation Forms Influenced Zinc Absorption and Accumulation in Piglets.

The study aimed at determining the effect of different zinc (Zn) supplementation forms on Zn accumulation, activities of Zn-containing enzymes, gene expression of metallothionein (MT), and Zn transporters in piglets. Eighteen piglets were randomly divided into three groups: (a) a basal diet supplemented with 150 mg/kg Zn from Zn methionine (Zn-Met) in the feed (Zn-Met group), (b) a basal diet supplemented with 150 mg/kg Zn from Zn sulfate (ZnSO4) in the feed (ZnSO4, feed group), and (c) a basal diet supplemented with the same dose of Zn as in ZnSO4,feed group but in water (ZnSO4, water group). The results showed that Zn-Met added in feed and ZnSO4 dissolved in drinking water significantly improved (p < 0.05) the Zn concentration in liver and jejunum and the apparent digestibility of Zn in comparison with the ZnSO4 added in feed. In addition, dietary Zn supplementation as Zn-Met significantly increased (p < 0.05) the activity of alkaline phosphatase (AKP) in the jejunum of piglets in comparison with the ZnSO4, feed group. Furthermore, the Zn-Met and ZnSO4, water groups showed an improved total superoxide dismutase activity (T-SOD) in the ileum as compared to the ZnSO4, feed group. Meanwhile, the qPCR and western blot results showed that Zn-Met and ZnSO4 dissolved in drinking water increased the expression of MT in the jejunum in comparison with the ZnSO4 added in the piglets' feed. However, different Zn supplementation forms had no effect on the mRNA expressions of Zip4 and ZnT1 transporters. In conclusion, Zn-Met added in feed and ZnSO4 dissolved in drinking water had higher bioavailability in piglets.