Adaptive Evolution of Escherichia coli to an α-Peptide/β-Peptoid Peptidomimetic Induces Stable Resistance.

Antimicrobial peptides (AMPs) and synthetic analogues thereof target conserved structures of bacterial cell envelopes and hence, development of resistance has been considered an unlikely event. However, recently bacterial resistance to AMPs has been observed, and the aim of the present study was to determine whether bacterial resistance may also evolve against synthetic AMP analogues, e.g. α-peptide/β-peptoid peptidomimetics. E. coli ATCC 25922 was exposed to increasing concentrations of a peptidomimetic (10 lineages), polymyxin B (10 lineages), or MilliQ water (4 lineages) in a re-inoculation culturing setup covering approx. 500 generations. All 10 lineages exposed to the peptidomimetic adapted to 32 × MIC while this occurred for 8 out of 10 of the polymyxin B-exposed lineages. All lineages exposed to 32 × MIC of either the peptidomimetic or polymyxin B had a significantly increased MIC (16-32 ×) to the selection agent. Five transfers (≈ 35 generations) in unsupplemented media did not abolish resistance indicating that resistance was heritable. Single isolates from peptidomimetic-exposed lineage populations displayed MICs against the peptidomimetic from wild-type MIC to 32 × MIC revealing heterogeneous populations. Resistant isolates showed no cross-resistance against a panel of membrane-active AMPs. These isolates were highly susceptible to blood plasma antibacterial activity and were killed when plasma concentrations exceeded ≈ 30%. Notably, MIC of the peptidomimetic against resistant isolates returned to wild-type level upon addition of 25% plasma. Whole-genome sequencing of twenty isolates from four resistant lineages revealed mutations, in murein transglycosylase D (mltD) and outer-membrane proteins, which were conserved within and between lineages. However, no common resistance-conferring mutation was identified. We hypothesise that alterations in cell envelope structure result in peptidomimetic resistance, and that this may occur via several distinct mechanisms. Interestingly, this type of resistance result in a concomitant high susceptibility towards plasma, and therefore the present study does not infer additional concern for peptidomimetics as future therapeutics

Introgression of Chromosome 3Ns from Psathyrostachys huashanica into Wheat Specifying Resistance to Stripe Rust

Wheat stripe rust is a destructive disease in the cool and humid wheat-growing areas of the world. Finding diverse sources of stripe rust resistance is critical for increasing genetic diversity of resistance for wheat breeding programs. Stripe rust resistance was identified in the alien species Psathyrostachys huashanica, and a wheat- P. huashanica amphiploid line (PHW-SA) with stripe rust resistance was reported previously. In this study, a P. huashanica 3Ns monosomic addition line (PW11) with superior resistance to stripe rust was developed, which was derived from the cross between PHW-SA and wheat J-11. We evaluated the alien introgressions PW11-2, PW11-5 and PW11-8 which were derived from line PW11 for reaction to new Pst race CYR32, and used molecular and cytogenetic tools to characterize these lines. The introgressions were remarkably resistant to CYR32, suggesting that the resistance to stripe rust of the introgressions thus was controlled by gene(s) located on P. huashanica chromosome 3Ns. All derived lines were cytologically stable in term of meiotic chromosome behavior. Two 3Ns chromosomes of P. huashanica were detected in the disomic addition line PW11-2. Chromosomes 1B of substitution line PW11-5 had been replaced by a pair of P. huashanica 3Ns chromosomes. In PW11-8, a small terminal segment from P. huashanica chromosome arm 3NsS was translocated to the terminal region of wheat chromosomes 3BL. Thus, this translocated chromosome is designated T3BL-3NsS. These conclusions were further confirmed by SSR analyses. Two 3Ns-specific markers Xgwm181 and Xgwm161 will be useful to rapidly identify and trace the translocated fragments. These introgressions, which had significant characteristics of resistance to stripe rust, could be utilized as novel germplasms for wheat breeding

Optimal in‐feed amino acid ratio for laying hens based on deletion method

A total of 56 Hy-line W-36 hens from 28 to 30 weeks were used on nitrogen balance (NB) trial to estimate daily N maintenance requirements (NMR) and the genetic potential for daily N retention (NRmaxT). The treatments consisted of six graded levels of nitrogen in the diets (N1 = 8; N2 = 16; N3 = 24; N4 = 32; N5 = 40; and N6 = 48 g N/kg of feed), formulated using the dilution technique. The regression analyses between nitrogen intake and excretion were performed to fit the exponential function and to determine the NMR = 292 mg/BWkg0.67, which was applied for further calculation of NRmaxT = 1,883 mg/BWkg0.67. A second NB trial was conducted, and a total of 96 Hy-line W-36 hens were used in the same period to estimate the ideal amino acid ratio (IAAR). Twelve treatments with eight replicates and one bird per cage were used. A balanced diet (BD) was formulated to meet the IAAR and the requirement of other nutrients for pullets. The limiting diets were formulated diluting BD with cornstarch and refilled with synthetic AAs and other feed ingredients, except for the AA under study. In each trial, the data of nitrogen intake, excretion, deposition and retention were obtained in a NB trial. The IAAR determined by Goettingen approach was Lys 100, Met+Cys 88, Trp 21, Thr 69, Arg 109, Val 90, Ile 75, Leu 127, Phe+Tir 110, Gly+Ser 73 and His 29%. The IAAR determined by Louvain approach was Lys 100, Met+Cys 88, Trp 21, Thr 69, Arg 104, Val 91, Ile 78, Leu 121, Phe+Tir 119, Gly+Ser 77 and His 29%.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Univ Estadual Paulista, Fac Ciencias Agr & Vet, Dept Anim Sci, Sao Paulo, BrazilEvonik Ind, Dept Anim Nutr Res, Hanau, GermanyUniv Gottingen, Anim Nutr, Gottingen, GermanyUniv Estadual Paulista, Fac Ciencias Agr & Vet, Dept Anim Sci, Sao Paulo, BrazilFAPESP: 2013/25761-4FAPESP: 2016/00536-