Virulence properties of Campylobacter jejuni are enhanced by displaying a mycobacterial TlyA methylation pattern in its rRNA

Campylobacter jejuni is a bacterial pathogen that is generally acquired as a zoonotic infection from poultry and animals. Adhesion of C. jejuni to human colorectal epithelial cells is weakened after loss of its cj0588 gene. The Cj0588 protein belongs to the type I group of TlyA (TlyA^I) enzymes, which 2′‐O‐methylate nucleotide C1920 in 23S rRNA. Slightly longer TlyA^II versions of the methyltransferase are found in actinobacterial species including Mycobacterium tuberculosis, and methylate not only C1920 but also nucleotide C1409 in 16S rRNA. Loss of TlyA function attenuates virulence of both M. tuberculosis and C. jejuni. We show here that the traits impaired in C. jejuni null strains can be rescued by complementation not only with the original cj0588 (tlyA ^I) but also with a mycobacterial tlyA ^II gene. There are, however, significant differences in the recombinant phenotypes. While cj0588 restores motility, biofilm formation, adhesion to and invasion of human epithelial cells and stimulation of IL‐8 production in a C. jejuni null strain, several of these properties are further enhanced by the mycobacterial tlyA ^II gene, in some cases to twice the original wild‐type level. These findings strongly suggest that subtle changes in rRNA modification patterns can affect protein synthesis in a manner that has serious consequences for bacterial pathogenicity.     

Causalities of war: The connection between type VI secretion system and microbiota

Microbiota niches have space and/or nutrient restrictions, which has led to the coevolution of cooperation, specialisation, and competition within the population. Different animal and environmental niches contain defined resident microbiota that tend to be stable over time and offer protection against undesired intruders. Yet fluxes can occur, which alter the composition of a bacterial population. In humans, the microbiota are now considered a key contributor to maintenance of health and homeostasis, and its alteration leads to dysbiosis. The bacterial type VI secretion system (T6SS) transports proteins into the environment, directly into host cells or can function as an antibacterial weapon by killing surrounding competitors. Upon contact with neighbouring cells, the T6SS fires, delivering a payload of effector proteins. In the absence of an immunity protein, this results in growth inhibition or death of prey leading to a competitive advantage for the attacker. It is becoming apparent that the T6SS has a role in modulating and shaping the microbiota at multiple levels, which is the focus of this review. Discussed here is the T6SS, its role in competition, key examples of its effect upon the microbiota, and future avenues of research.     

Dark microbial CO2 fixation in temperate forest soils increases with CO2 concentration

Dark, that is, nonphototrophic, microbial CO₂ fixation occurs in a large range of soils. However, it is still not known whether dark microbial CO₂ fixation substantially contributes to the C balance of soils and what factors control this process. Therefore, the objective of this study was to quantitate dark microbial CO₂ fixation in temperate forest soils, to determine the relationship between the soil CO₂ concentration and dark microbial CO₂ fixation, and to estimate the relative contribution of different microbial groups to dark CO₂ fixation. For this purpose, we conducted a ¹³C‐CO₂ labeling experiment. We found that the rates of dark microbial CO₂ fixation were positively correlated with the CO₂ concentration in all soils. Dark microbial CO₂ fixation amounted to up to 320 µg C kg^?1 soil day^?1 in the Ah horizon. The fixation rates were 2.8–8.9 times higher in the Ah horizon than in the Bw1 horizon. Although the rates of dark microbial fixation were small compared to the respiration rate (1.2%–3.9% of the respiration rate), our findings suggest that organic matter formed by microorganisms from CO₂ contributes to the soil organic matter pool, especially given that microbial detritus is more stable in soil than plant detritus. Phospholipid fatty acid analyses indicated that CO₂ was mostly fixed by gram‐positive bacteria, and not by fungi. In conclusion, our study shows that the dark microbial CO₂ fixation rate in temperate forest soils increases in periods of high CO₂ concentrations, that dark microbial CO₂ fixation is mostly accomplished by gram‐positive bacteria, and that dark microbial CO₂ fixation contributes to the formation of soil organic matter.     

Brenneria alni,causal agent bark canker of Alnus subcordata

In 2014, bark cankers were observed on Caucasian alder (Alnus subcordata) trees in Iran. The disease was characterized by a dark watery liquid often exuding from longitudinal cankers in the bark of the tree trunks which stained the surface. Symptomatic tissue from A. subcordata was sampled from a number of sites in the Mazandaran province. Isolations were performed on nutrient agar supplemented with sucrose (SNA) and yielded bacterial colonies that were uniform, round and whitish. The bacterial strains isolated from alder trees in Iran were similar to Brenneria alni based on phenotypic and genotypic (nucleotide sequences of the 16S rRNA, and housekeeping genes gyrB, and infB) characteristics. The pathogenicity of the representative strains was determined by inoculating stem pieces of A. subcordata. All tested strains caused longitudinal necrotic lesions 30 days after inoculation and were re-isolated from this tissue. To our knowledge, this is the first report of the occurrence of B. alni in Iran, and on A. subcordata globally.     

Coordinated regulation of anthranilate metabolism and bacterial virulence by the GntR family regulator MpaR in Pseudomonas aeruginosa

The GntR family regulators are widely distributed in bacteria and play critical roles in metabolic processes and bacterial pathogenicity. In this study, we describe a GntR family protein encoded by PA4132 that we named MpaR (M vfR-mediated P QS and a nthranilate r egulator) for its regulation of Pseudomonas quinolone signal (PQS) production and anthranilate metabolism in Pseudomonas aeruginosa. The deletion of mpaR increased biofilm formation and reduced pyocyanin production. RNA sequencing analysis revealed that the mRNA levels of antABC encoding enzymes for the synthesis of catechol from anthranilate, a precursor of the PQS, were most affected by mpaR deletion. Data showed that MpaR directly activates the expression of mvfR, a master regulator of pqs system, and subsequently promotes PQS production. Accordingly, deletion of mpaR activates the expression of antABC genes, and thus, increases catechol production. We also demonstrated that MpaR represses the rhl quorum-sensing (QS) system, which has been shown to control antABC activity. These results suggested that MpaR function is integrated into the QS regulatory network. Moreover, mutation of mpaR promotes bacterial survival in a mouse model of acute pneumonia infection. Collectively, this study identified a novel regulator of pqs system, which coordinately controls anthranilate metabolism and bacterial virulence in P. aeruginosa.     

Misannotations of the genes encoding sugar N‐formyltransferases

Tens of thousands of bacterial genome sequences are now known due to the development of rapid and inexpensive sequencing technologies. An important key in utilizing these vast amounts of data in a biologically meaningful way is to infer the function of the proteins encoded in the genomes via bioinformatics techniques. Whereas these approaches are absolutely critical to the annotation of gene function, there are still issues of misidentifications, which must be experimentally corrected. For example, many of the bacterial DNA sequences encoding sugar N‐formyltransferases have been annotated as l ‐methionyl‐tRNA transferases in the databases. These mistakes may be due in part to the fact that until recently the structures and functions of these enzymes were not well known. Herein we describe the misannotation of two genes, WP_088211966.1 and WP_096244125.1, from Shewanella spp. and Pseudomonas congelans, respectively. Although the proteins encoded by these genes were originally suggested to function as l ‐methionyl‐tRNA transferases, we demonstrate that they actually catalyze the conversion of dTDP‐4‐amino‐4,6‐dideoxy‐d ‐glucose to dTDP‐4‐formamido‐4,6‐dideoxy‐d ‐glucose utilizing N¹⁰‐formyltetrahydrofolate as the carbon source. For this analysis, the genes encoding these enzymes were cloned and the corresponding proteins purified. X‐ray structures of the two proteins were determined to high resolution and kinetic analyses were conducted. Both enzymes display classical Michaelis–Menten kinetics and adopt the characteristic three‐dimensional structural fold previously observed for other sugar N‐formyltransferases. The results presented herein will aid in the future annotation of these fascinating enzymes.     

鸡肠道微生物演替与早期定植的研究进展

鸡肠道中寄生着数量庞大且复杂多样的微生物,对宿主的生长发育和健康十分重要,既影响着饲料消化、营养物质吸收,又参与了宿主肠道形态和免疫系统的调控。深入了解鸡肠道微生物区系的时空变化及早期定植特点,将有助于提出新的肠道微生态干预策略,应用于生产。就鸡肠道微生物组成和演替、早期微生物区系建立及调控等方面进行综述,并总结了一些最新研究进展。     

乙醇对瘤胃液接种稻秸的体外发酵产物及细菌群落结构的影响

旨在研究乙醇对山羊瘤胃液与水稻秸秆厌氧共培养的影响。利用频繁传代的体外发酵技术和高通量测序方法,分析了短链脂肪酸(SCFA)产量和细菌群落的变化。结果表明,经体外培养传代8次的稻秸发酵液的总短链脂肪酸产量显著高于瘤胃液(P<0.01);与未添加乙醇的稻秸发酵液相比,添加乙醇显著提高了乙酸、戊酸和己酸的比例,降低了丙酸和丁酸的比例(P<0.01),总SCFA产量及异丁酸和异戊酸比例无显著差异。与瘤胃液相比,稻秸发酵液的拟杆菌门(Bacteroidetes)相对丰度下降,厚壁菌门(Firmicutes)相对丰度升高(P<0.05),且添加乙醇显著提高了厚壁菌门和放线菌门(Actinobacteria)的相对丰度(P<0.05);添加乙醇使双歧杆菌属(Bifidobacterium)、未定性的毛螺菌属(unidentified Lachnospiraceae)、产琥珀酸菌属(Succiniclasticum)、脱硫弧菌属(Desulfovibrio)和未定性的梭菌属(unidentified Clostridiales)的相对丰度显著升高(P<0.05)。乙醇使稻秸发酵液的显著性差异物种(Biomarker)增加;稻秸发酵液与瘤胃液亲缘关系较近,而添加乙醇显著改变了细菌区系;短链脂肪酸比例在稻秸发酵液细菌群落多样性中具有重要作用。研究表明,体外频繁传代和添加乙醇可以提高稻秸发酵液的乙酸、戊酸和己酸产量,乙醇改变了稻秸发酵液的细菌群落结构。