Phage-encoded sigma factors alter bacterial dormancy

By entering a reversible state of reduced metabolic activity, dormant microorganisms are able to tolerate suboptimal conditions that would otherwise reduce their fitness. Dormancy may also benefit bacteria by serving as a refuge from parasitic infections. Here we focus on dormancy in the Firmicutes, where endospore development is transcriptionally regulated by the expression of sigma factors. A disruption of this process could influence the survivorship and reproduction of phages that infect spore-forming hosts with implications for coevolutionary dynamics. Here, we characterized the distribution and diversity of sigma factors in nearly 3,500 phage genomes. Homologs of sporulation-specific sigma factors were identified in phages that infect spore-forming hosts. Unlike sigma factors required for phage reproduction, the sporulation-like sigma factors were non-essential for lytic infection. However, when expressed in the spore-forming Bacillus subtilis, sigma factors from phages activated the bacterial sporulation gene network and reduced spore yield. Our findings suggest that the acquisition of host-like transcriptional regulators may allow phages to manipulate a complex and ancient trait in one of the most abundant cell types on Earth.

Optimization of an Economical Medium Composition For The Coculture of Clostridium Butyricum And Bacillus Coagulans

Clostridium butyricum is a spore-forming probiotic existing in the intestines of humans and animals which can promote the enhancement of beneficial bacteria and maintain intestinal microecological balance. However, it is difficult to improve the production level of C. butyricum by conventional fermentation process. In this study, a co-fermentation process of C. butyricum DL-1 and Bacillus coagulans ZC2-1 was established to improve the viable counts and spore yield of C. butyricum, and the formula of coculture medium was optimized by flask fermentation. The results showed that the optimum medium composition is bran 10 g/L, corn steep powder 15 g/L, peptone 15 g/L, K2HPO4 1 g/L and MnSO4 0.5 g/L. Cultured stationarily in the optimal medium for 36 h, the number of viable bacteria of C. butyricum DL-1 reached 1.5×108 CFU/mL and the spore forming rate was 92.6%. The results revealed an economical and effective medium composition for the coculture of C. butyricum and B. coagulans. The co-fermentation process established in this study provides a new fermentation mode for the industrial production of other absolute anerobic bacteria.

Effects of Temperature and Culture Media Composition on Sporulation, Mycelial Growth, and Antifungal Activity of Isaria javanica pf185

The fungal isolate Isaria javanica pf185 has potential as a mycopesticide because it demonstrates insecticidal activity against the green peach aphid and antifungal activity against Colletotrichum gloeosporioides. For commercialization of this isolate, determination of the optimal and least expensive culture conditions is required; however, these data are not currently available. This study describes the conditions for optimal development of conidia and production of metabolites for the biocontrol of the fungal pathogen. The optimal culture conditions were examined using cultures on solid agar and liquid media. High growth temperature enhanced spore formation but reduced antifungal activity in both solid and liquid media. The highest spore yield was obtained in a medium containing glucose as a carbon source and yeast extract as a nitrogen source. Soybean powder and wheat bran were effective nitrogen sources that promoted spore production and antifungal activity of the isolate. These results revealed the basic, cost-effective growth media for commercial production of a biopesticide with insecticidal and antifungal properties for use in integrated pest management.

Fructose and Trehalose Selectively Enhance In Vitro Sporulation of Paenibacillus larvae ERIC I and ERIC II Strains

Paenibacillus larvae is a Gram-positive bacterium, the spores of which are the causative agent of the most destructive brood disease of honeybees, American foulbrood (AFB). Obtaining viable spores of pathogen strains is requisite for different studies concerning AFB. The aim of this work was to investigate the effects of five saccharides that may naturally occur in higher amounts in bee larvae on in vitro sporulation of P. larvae. The effect of individual saccharides at different concentrations on spore yields of P. larvae strains of epidemiologically important ERIC genotypes was examined in Columbia sheep blood agar (CSA) and MYPGP agar media. It was found that fructose in ERIC I and trehalose in ERIC II strains at concentrations in the range of 0.5–2% represent new sporulation factors that significantly enhanced the yields of viable spores in both media, mostly in a concentration-dependent manner. The enhancements in spore yield were mainly caused by improvements of the germination ability of the spores produced. Glucose, maltose and sucrose at 1% or 0.5% concentrations also supported sporulation but to a lower extent and not in all strains and media. Based on the knowledge gained, a novel procedure was proposed for the preparation of viable P. larvae spores with supposed improved quality for AFB research.

A spore quality–quantity tradeoff favors diverse sporulation strategies in Bacillus subtilis

Quality–quantity tradeoffs govern the production of propagules across taxa and can explain variability in life-history traits in higher organisms. A quality–quantity tradeoff was recently discovered in spore forming bacteria, but whether it impacts fitness is unclear. Here we show both theoretically and experimentally that the nutrient supply during spore revival determines the fitness advantage associated with different sporulation behaviors in Bacillus subtilis. By tuning sporulation rates we generate spore-yield and spore-quality strategists that compete with each other in a microscopic life-cycle assay. The quality (yield) strategist is favored when spore revival is triggered by poor (rich) nutrients. We also show that natural isolates from the gut and soil employ different life-cycle strategies that result from genomic variations in the number of rap-phr signaling systems. Taken together, our results suggest that a spore quality–quantity tradeoff contributes to the evolutionary adaptation of sporulating bacteria.

Artificial Sporulation Induction (ASI) by kinA Overexpression Affects the Proteomes and Properties of Bacillus subtilis Spores

To facilitate more accurate spore proteomic analysis, the current study focuses on inducing homogeneous sporulation by overexpressing kinA and assesses the effect of synchronized sporulation initiation on spore resistance, structures, the germination behavior at single-spore level and the proteome. The results indicate that, in our set up, the sporulation by overexpressing kinA can generate a spore yield of 70% within 8 h. The procedure increases spore wet heat resistance and thickness of the spore coat and cortex layers, whilst delaying the time to spore phase-darkening and burst after addition of germinant. The proteome analysis reveals that the upregulated proteins in the kinA induced spores, compared to spores without kinA induction, as well as the ‘wildtype’ spores, are mostly involved in spore formation. The downregulated proteins mostly belong to the categories of coping with stress, carbon and nitrogen metabolism, as well as the regulation of sporulation. Thus, while kinA overexpression enhances synchronicity in sporulation initiation, it also has profound effects on the central equilibrium of spore formation and spore germination, through modulation of the spore molecular composition and stress resistance physiology.

Productivity, pathogenicity, host range, and spore mass-propagation of local strain of Mattesia sp. isolated from insect cadavers of certain stored grain pests in Egypt

Average yields of Mattesia spores (spore productivity) had varied from a minimum yield (0.17 × 107 spores) for Laemophloeus turcicus adult to a maximum yield (7.46 × 107 spores) for Plodia interpunctella larva. Comparatively, the highest increase in Mattesia spore yield, recorded from P. interpunctella larva (7.46 × 107 spores) over the lowest one, estimated for L. turcicus adult (0.17 × 107 spores), was nearly 44-fold. The increase in Mattesia spore yields that calculated from the other hosts (P. interpunctella pupa or moth; Galleria mellonella larva; Rhyzopertha dominica adult; Sitophilus zeamais), over that estimated for L. turcicus adult, was less than 10-fold (6–9-fold). Based on the weight of 1 g of the insect host infected with Mattesia sp., small stored grain insect hosts (e.g. L. turcicus, S. zeamais, and R. dominica) seemed to achieve Mattesia spore yields more than the larger ones (e.g. P. interpunctella). The increase in spore yields over that used for the inoculum, based on an average of 25 P. interpunctella larvae per bioassay container, was ca. 2 to 31-fold. These results revealed that the Indianmeal moth, P. interpunctella, could serve as a potential host for mass propagating the isolated entomopathogenic protozoan, Mattesia sp. Besides Mattesia larval mortality, survivors of Mattesia infection suffered deformities and noticeable undersized pupae or adults than the control ones. Also, many copulated moths (ca.46%) were unable to become separated after copulation until they had died. Bioassay of siftings, obtained from L. turcicus-protozoan-infected stock cultures, was carried out in order to emphasize the suppressive potent role of such protozoan entomopathogens in long-term storage. With the highest tested concentration of the studied siftings (10%), mortality responses due to Mattesia infection ranged from 13 to 68% at 14–169 days post-treatment. The corresponding figures for Adelina infection were 7–42%.

Effect of oil cakes and garlic aqueousbased formulations of trichoderma viride on management of meloidogyne incognita in chilli

Root-knot nematode, Meloidogyne incognita (Kofoid and White) is a major threat to chilli (Capsicum annum) cultivation, by forming root galls and subsequent wilting. These nematodes live in soil, roots debris and reported surviving in other Solanaceae crops. Chemical application of nematicides cause impact on environment, therefore biocontrol using antagonistic fungi is desired to tackle this problem. This research is aimed to evaluate the effect of Trichoderma viride and its formulations on management of M. incognita. Oil cakes such as neem (Azadiracta indica), gingelly (Sesamum indicum), Mahua (Madhuca longifolia) and garlic (Allium sativum) extracts were used to evaluate the growth and sporulation of T. viride. Chilli variety MI-2 was used. Among the four preparations, neem oil cake recorded high spore yield of 1.75 ×107 spores/ml. Similarly gingelly oil cake and garlic produced the spore yields of 1.57 ×107 spores/ml and 1.368 ×107 spores/ml, respectively. Comparatively Mahua oil cake obtained low spore yield of 7.87×106 spores/ml. Plant growth was significant (P<0.05) in the application of neem oil cake formulation (30.42 cm). Extent of galling was significantly low with garlic (9.00) and neem (9.25). These results confirmed that the formulations of neem, gingelly oil cake extracts and garlic aqueous extract stimulated the chilli plant growth, productivity and reduced the nematode infestation. Above all, neem oil cake and garlic are the best formulations that can be used to manage M. incognita