Animal gastrointestinal tracts are populated by highly diverse and complex microbiotas. The gut microbiota influences the bioavailability of dietary components and is closely associated with physiological processes in the host. Clostridium butyricum reportedly improves growth performance and affects the gut microbiota and immune functions in post-weaning piglets. However, the effects of C. butyricum on finishing pigs remain unclear. Therefore, we herein investigated the effects of C. butyricum MIYAIRI 588 (CBM588) on the gut microbiota of finishing pigs. 16S rRNA gene sequencing was performed using fecal samples and ileal, cecal, and colonic contents collected after slaughtering. The α-diversity of the small intestinal microbiota was lower than that of the large intestinal microbiota, whereas β-diversity showed different patterns depending on sample collection sites. The administration of CBM588 did not significantly affect the α- or β-diversity of the microbiotas of fecal and intestinal content samples regardless of the collection site. However, a linear discriminant ana­lysis Effect Size revealed that the relative abundance of Lactobacillaceae at the family level, Bifidobacterium at the order level, and Lactobacillus ruminis and Bifidobacterium pseudolongum at the species level were higher in the fecal samples and cecal and colonic contents of the treatment group than in those of the control group. Therefore, the administration of CBM588 to finishing pigs affected the composition of the gut microbiota and increased the abundance of bacteria that are beneficial to the host. These results provide important insights into the effects of probiotic administration on relatively stable gut microbial ecosystems.

Excess nitrate (NO₃^–) and nitrite (NO₂^–) in surface waters adversely affect human and environmental health. Bacteria with the ability to remove nitrogen (N) have been isolated to reduce water pollution caused by the excessive use of N fertilizer. To obtain plant growth-promoting rhizobacteria (PGPR) with salt tolerance and NO₃^–-N removal abilities, bacterial strains were isolated from plant rhizosphere soils, their plant growth-promoting effects were evaluated using tomato in plate assays, and their NO₃^–-N removal abilities were tested under different salinity, initial pH, carbon source, and agriculture wastewater conditions. The results obtained showed that among the seven strains examined, five significantly increased the dry weight of tomato plants. Two strains, Pseudomonas stutzeri NRCB010 and Bacillus velezensis NRCB026, showed good plant growth-promoting effects, salinity resistance, and NO₃^–-N removal abilities. The maximum NO₃^–-N removal rates from denitrifying medium were recorded by NRCB010 (90.6%) and NRCB026 (92.0%) at pH 7.0. Higher NO₃^–-N removal rates were achieved using glucose or glycerin as the sole carbon source. The total N (TN) removal rates of NRCB010 and NRCB026 were 90.6 and 66.7% in farmland effluents, respectively, and 79.9 and 81.6% in aquaculture water, respectively. These results demonstrate the potential of NRCB010 and NRCB026 in the development of novel biofertilizers and their use in reducing N pollution in water.

The present study investigated the effects of the endophytic fungus, Penicillium pinophilum EU0013 on fruit phytochemical indices and essential trace elements in five tomato cultivars. In a completely randomized design, inoculated and uninoculated seedlings of tomato cultivars (Momotaro, Rodeo, Anaya, Reika, and Cherry) were raised for sixteen weeks in a greenhouse. Fruit fresh weights and root colonization by P. pinophilum were significantly higher in the Rodeo cultivar than in the other cultivars tested. Significant effects of the cultivar, inoculation, and interaction on fruit dry weights were‍ ‍observed with higher values in Anaya inoculated with P. pinophilum. Cultivar and inoculation effects were significant for ascorbic acid and soluble sugars in four cultivars, with increases being observed due to the P. pinophilum inoculation. Lycopene levels increased in Rodeo and decreased in Anaya, while β-carotene levels increased in four cultivars due to the inoculation. Manganese concentrations were significantly increased in Cherry, while iron concentrations were increased in Reika and Cherry. Increases due to the inoculation were observed for gibberellic acids (GA₁ and GA₄) in Reika and Anaya, whereas decreases were detected in Cherry and Rodeo. Similar results were obtained for abscisic acids (ABA) with increases in Reika and Anaya due to the inoculation. P. pinophilum EU0013 demonstrated the ability to improve the nutritive value of tomato fruits via modulations to phytochemicals in addition to increases in Mn and Fe concentrations, particularly in Cherry and Rodeo. Cultivar responses to the P. pinophilum inoculation are a factor that need to be considered for its use in increasing fruit quality indices in tomato.

Streptomyces sp. strain MBCN152-1, isolated from cabbage, has potential as a biocontrol agent for Alternaria brassicicola on cabbage seedlings. The present study examined its mode of action. Light microscopy showed that appressorium formation by A. brassicicola was significantly suppressed on cabbage seedlings bacterized with MBCN152-1. Furthermore, scanning electron microscopy revealed that the mycelia of MBCN152-1, which were epiphytically growing on the cotyledon leaves of cabbage seedlings, intensively coiled around the germinating conidia of A. brassicicola. In vitro co-culture experiments demonstrated that MBCN152-1 is an aggressive mycoparasite of A. brassicicola, but not of A. brassicae or Colletotrichum higginsianum. Biocontrol experiments indicated that MBCN152-1 did not control diseases caused by A. brassicae or C. higginsianum. These results suggest that mycoparasitism is the primary mode of action for MBCN152-1. This is the first study to clearly demonstrate the significance of mycoparasitism in the biocontrol efficacy of endophytic Streptomyces.

Integrative and conjugative elements (ICEs) play a role in the horizontal transfer of antibiotic resistance genes (ARGs). We herein report an ICE from Shewanella halifaxensis isolated from fish intestine with a similar structure to both a clinical bacterial ICE and marine bacterial plasmid. The ICE was designated ICEShaJpn1, a member of the SXT/R391 family of ICEs (SRIs). ICEShaJpn1 has a common core structure with SRIs of clinical and fish origins and an ARG cassette with the pAQU1 plasmid of Photobacterium damselae subsp. damselae, suggesting that the common core of SRIs is widely distributed and ARG cassettes are collected from regional bacteria.

RNA viruses are distributed throughout various environments, and most have recently been identified by metatranscriptome sequencing. However, due to the high nucleotide diversity of RNA viruses, it is still challenging to identify novel RNA viruses from metatranscriptome data. To overcome this issue, we created a dataset of RNA-dependent RNA polymerase (RdRp) domains that are essential for all RNA viruses belonging to Orthornavirae. Genes with RdRp domains from various RNA viruses were clustered based on amino acid sequence similarities. A multiple sequence alignment was generated for each cluster, and a hidden Markov model (HMM) profile was created when the number of sequences was greater than three. We further refined 426 HMM profiles by detecting RefSeq RNA virus sequences and subsequently combined the hit sequences with the RdRp domains. As a result, 1,182 HMM profiles were generated from 12,502 RdRp domain sequences, and the dataset was named NeoRdRp. The majority of NeoRdRp HMM profiles successfully detected RdRp domains, specifically in the UniProt dataset. Furthermore, we compared the NeoRdRp dataset with two previously reported methods for RNA virus detection using metatranscriptome sequencing data. Our methods successfully identified the majority of RNA viruses in the datasets; however, some RNA viruses were not detected, similar to the other two methods. NeoRdRp may be repeatedly improved by the addition of new RdRp sequences and is applicable as a system for detecting various RNA viruses from diverse metatranscriptome data.

Many stinkbugs in the superfamily Coreoidea (Hemiptera: Heteroptera) develop crypts in the posterior midgut, harboring Caballeronia (Burkholderia) symbionts. These symbionts form a monophyletic group in Burkholderia sensu lato, called the “stinkbug-associated beneficial and environmental (SBE)” group, recently reclassified as the new genus Caballeronia. SBE symbionts are separated into the subclades SBE-α and SBE-β. Previous studies suggested a regional effect on the symbiont infection pattern; Japanese and American bug species are more likely to be associated with SBE-α, while European bug species are almost exclusively associated with SBE-β. However, since only a few insect species have been investigated, it remains unclear whether region-specific infection is general. We herein investigated Caballeronia gut symbionts in diverse Japanese, European, and North American populations of a cosmopolitan species, the Western conifer seed bug Leptoglossus occidentalis (Coreoidea: Coreidae). A mole­cular phylogenetic ana­lysis of the 16S rRNA gene demonstrated that SBE-β was the most dominant in all populations. Notably, SBE-α was rarely detected in any region, while a third clade, the “Coreoidea clade” occupied one fourth of the tested populations. Although aposymbiotic bugs showed high mortality, SBE-α- and SBE-β-inoculated insects both showed high survival rates; however, a competition assay demonstrated that SBE-β outcompeted SBE-α in the midgut crypts of L. occidentalis. These results strongly suggest that symbiont specificity in the Leptoglossus-Caballeronia symbiotic association is influenced by the host rather than geography, while the geographic distribution of symbionts may be more important in other bugs.

Ammonia-oxidizing archaea (AOA), key players in agricultural upland soil nitrification, convert soil ammonium to nitrite. The microbial oxidation of ammonia to nitrite is an important part of the global biogeochemical nitrogen cycle. In the present study, we recovered six novel AOA metagenome-assembled genomes (MAGs) containing genes for carbon (C) fixation and nitrogen (N) metabolism by using a deep shotgun metagenomic sequencing strategy. We also found that these AOA MAGs possessed cobalamin synthesis genes, suggesting that AOA are vitamin suppliers in agricultural upland soil. Collectively, the present results deepen our understanding of the metabolic potential and phylogeny of AOA in agroecosystems.

Methane metabolism in wetlands involves diverse groups of bacteria and archaea, which are responsible for the biological decomposition of organic matter under certain anoxic conditions. Recent advances in environmental omics revealed the phylogenetic diversity of novel microbial lineages, which have not been previously placed in the traditional tree of life. The present study aimed to verify the key players in methane production, either well-known archaeal members or recently identified lineages, in peat soils collected from wetland areas in Japan. Based on an ana­lysis of microbial communities using 16S rRNA gene sequencing and the mole­cular cloning of the functional gene, mcrA, a marker gene for methanogenesis, methanogenic archaea belonging to Methanomicrobiales, Methanosarcinales, Methanobacteriales, and Methanomassiliicoccales were detected in anoxic peat soils, suggesting the potential of CH₄ production in this natural wetland. “Candidatus Bathyarchaeia”, archaea with vast metabolic capabilities that is widespread in anoxic environments, was abundant in subsurface peat soils (up to 96% of the archaeal community) based on microbial gene quantification by qPCR. These results emphasize the importance of discovering archaea members outside of traditional methanogenic lineages that may have significant functions in the wetland biogeochemical cycle.

Anaerobic ammonium-oxidizing (anammox) bacteria are slow-growing and fastidious bacteria, and limited numbers of enrichment cultures have been established. A metagenomic ana­lysis of our 5 established anammox bacterial enrichment cultures was performed in the present study. Fourteen high-quality metagenome-assembled genomes (MAGs) were obtained, including those of 5 anammox Planctomycetota (Candidatus Brocadia, Ca. Kuenenia, Ca. Jettenia, and Ca. Scalindua), 4 Bacteroidota, and 3 Chloroflexota. Based on the gene sets of metabolic pathways involved in the degradation of polymeric substances found in Chloroflexota and Bacteroidota MAGs, they are expected to be scavengers of extracellular polymeric substances and cell debris.

The gut microbiota influences the phenotype and fitness of a host; however, limited information is currently available on the diversity and functions of the gut microbiota in wild animals. Therefore, we herein examined the diversity, composition, and potential functions of the gut microbiota in three Sceloporus lizards: Sceloporus aeneus, S. bicanthalis, and S. grammicus, inhabiting different habitats in a mountainous ecosystem. The gut bacterial community of S. bicanthalis from alpine grasslands at 4,150‍ ‍m a.s.l. exhibited greater taxonomic, phylogenetic, and functional alpha diversities than its sister species S. aeneus from cornfields and human-induced grasslands at 2,600‍ ‍m‍ ‍a.s.l. Bacteria of the genus Blautia and metabolic functions related to the degradation of aromatic compounds were more abundant in S. bicanthalis than in S. aeneus, whereas Oscillibacter and predicted functions related to amino acid metabolism and fermentation were more abundant in S. aeneus. The structure of the dominant and most prevalent bacteria, i.e., the core microbiota, was similar between the sister species from different habitats, but differed between S. grammicus and S. aeneus cohabiting at 2,600‍ ‍m‍ ‍a.s.l. and between S. grammicus and S. bicanthalis cohabiting at 4,150‍ ‍m a.s.l. These results suggest that phylogenetic relatedness defines the core microbiota, while the transient, i.e., non-core, microbiota is influenced by environmental differences in the habitats. Our comparisons between phylogenetically close species provide further evidence for the specialized and complex associations between hosts and the gut microbiota as well as insights into the roles of phylogeny and ecological factors as drivers of the gut microbiota in wild vertebrates.

Patescibacteria are widely distributed in various environments and often detected in activated sludge. However, limited information is currently available on their phylogeny, morphology, and ecophysiological role in activated sludge or interactions with other microorganisms. In the present study, we identified microorganisms that interacted with Patescibacteria in activated sludge via a correlation ana­lysis using the 16S rRNA gene, and predicted the metabolic potential of Patescibacteria using a metagenomic ana­lysis. The metagenome-assembled genomes of Patescibacteria consisted of three Saccharimonadia, three Parcubacteria, and one Gracilibacteria, and showed a strong positive correlation of relative abundance with Chitinophagales. Metabolic predictions from ten recovered patescibacterial and five Chitinophagales metagenome-assembled genomes supported mutualistic interactions between a member of Saccharimonadia and Chitinophagales via N-acetylglucosamine, between a member of Parcubacteria and Chitinophagales via nitrogen compounds related to denitrification, and between Gracilibacteria and Chitinophagales via phospholipids in activated sludge. The present results indicate that various interactions between Patescibacteria and Chitinophagales are important for the survival of Patescibacteria in activated sludge ecosystems.