The role of food chain in antimicrobial resistance spread and One Health approach to reduce risks.

The incidence of antimicrobial resistance (AMR) is rapidly spreading worldwide. It is depleting the repertoire of antibiotics in use but the pace of development of new antibiotics is stagnant for decades. Annually, millions of people are killed by AMR. This alarming situation urged both scientific and civil bodies to take steps to curb AMR as a top priority. Here we review the various sources of AMR in the environment, especially focusing on the food chain. Food chain inculcates pathogens with AMR genes and serves as a conduit for its transmission. In certain countries, the antibiotics are more used in livestock than in humans. It is also used in agriculture crops of high value products. The indiscriminate use of antibiotics in livestock and agriculture increased rapid emergence of AMR pathogens. In addition, in many countries nosocomial settings are spewing AMR pathogens, which is a serious health hazard. Both the developed and low and middle income countries (LMIC) face the phenomenon of AMR. Therefore, a comprehensive approach for monitoring all sectors of life is required to identify the emerging trend of AMR in environment. AMR genes' mode of action must be understood to develop strategies to reduce risk. The new generation sequencing technologies, metagenomics and bioinformatics capabilities can be resorted to quickly identify and characterize AMR genes. The sampling for AMR monitoring can be done from multiples nodes of the food chain as envisioned and promoted by the WHO, FAO, OIE and UNEP under the One Health approach to overcome threat of AMR pathogens.

Fusion of the N-terminal 119 amino acids of RelA with the CTD domain render growth inhibitory effects of the latter, (p)ppGpp-dependent.

The guanosine nucleotide derivatives ppGpp and pppGpp are central to the remarkable capacity of bacteria to adapt to fluctuating environments and metabolic perturbations. They are synthesized by two proteins, RelA and SpoT in E. coli and the activities of each of the two enzymes are highly regulated for homeostatic control of intracellular (p)ppGpp levels. Characterization of the mutant studied here indicates that moderate level expression of RelA appreciably reduces growth of cells wherein the basal levels of (p)ppGpp are higher than in the wild type without elevating the levels further. Consistent with this result, a large part of the growth inhibition effect is reproduced by overexpression of RelA NTD-CTD fusion lacking the (p)ppGpp synthesis function. A null mutation in relA abolishes this growth inhibitory effect suggesting its requirement for basal level synthesis of (p)ppGpp. Accordingly, increase in the (p)ppGpp levels in the relA1 mutant by spoT202 mutation largely restored the growth inhibitory effects of overexpression of RelA NTD-CTD fusion. Expression of this construct consisting of 119 amino acids of the N-terminal hydrolytic domain (HD) fused in-frame with the CTD domain (±TGS domain) renders the growth inhibitory effects (p)ppGpp-responsive-inhibited growth only of spoT1 and spoT202 relA1 mutants. This finding uncovered an hitherto unrealized (p)ppGpp-dependent regulation of RelA-CTD function, unraveling the importance of RelA NTD-HD domain for its regulatory role. An incremental rise in the (p)ppGpp levels is proposed to progressively modulate the interaction of RelA-CTD with the ribosomes with possible implications in the feedback regulation of the (p)ppGpp synthesis function, a proposal that accounts for the nonlinear kinetics of (p)ppGpp synthesis and increased ratio of RelA:ribosomes, both in vitro as well as in vivo.