Pigs subjected to heat stress (HS) decrease their feed intake and growth. The objectives of the experiment were to determine the effects of live yeast (LY) supplementation (Saccharomyces cerevisiae var boulardii CNCM I-1079) on feeding behaviour, energy metabolism and faecal microbiota composition of finishing boars (n 10) housed in a respiration chamber at thermoneutrality (7 d at 22°C) or during HS (seven plus six days at 28°C). Dietary LY supplementation increased DM intake (P = 0·01) whatever the ambient temperature, whereas HS decreased feed intake whatever the dietary supplementation (P = 0·01). Dietary LY supplementation increased the number of meals (P = 0·02). Energy retention was higher with dietary LY supplementation (P < 0·01) but decreased during HS (P < 0·01). The skin temperature of the supplemented pigs was lower at thermoneutrality and increased during HS to a lesser extent than that of non-supplemented pigs (P < 0·01). Faecal microbiota composition was determined using 16S rRNA gene sequencing. Treponema, Christensenellaceae R-7, Ruminococcaceae UCG-002, Rikenellaceae RC9, Clostridium sensu stricto 1 and Romboutsia genera and some bacteria belonging to Alloprevotella, Oxalobacter and Anaeroplasma genera were more abundant under HS. LY supplementation attenuated HS effects on Romboutsia abundance, while decreasing the abundance of some bacteria from Ruminoccocus, Coprococcus, Peptococcus and Oxalobacter genera and increasing the abundance of beneficial bacteria from Lactococcus and Subdoligranulum genera. Our results suggest that higher level of the keystone species Ruminococcus bromii at thermoneutrality may be one of the causes for higher energy retention observed under subsequent HS.

The digestive microbiota plays a decisive role in shaping and preserving health throughout life. Rabbit younglings are born with a sterile digestive tract but then it gets progressively colonised by the microbiota of the nursing mother, by entering in contact with or ingesting the maternal droppings present in the nest. Here we posit that (i) offspring survival and (ii) lifespan of female rabbits are linked to how diverse their microbiota are. To test the hypothesis that maternal microbiota evolves in females having had different levels of offspring survival in their lifetime, we obtained 216 hard faecal samples from 75 female rabbits at ages 19.6, 31.6, 62.6 and 77.6 weeks. The annual mean offspring survival (MOS) at 64 days was calculated for each female then crossed against three alpha-diversity indexes (operational taxonomic units (OTUs), inverse Simpson index and Shannon index). Age was also analysed against these three parameters. The alpha-diversity indexes of the female faecal microbiota did not correlate with MOS, but they did decrease with age (e.g. from 712 OTUs at age 19.6 weeks to 444 OTUs at 77.6 weeks; P < 0.05). The age effect was also found in beta-diversity non-metric multidimensional scaling plots using the Bray–Curtis dissimilarity index and the unweighted UniFrac index but not for MOS. The ability of the microbiota composition from the faecal samples of young females (19.6 weeks old) to predict their lifespan was also evaluated. After subdividing the initial population into two classes (females that weaned a maximum of three litters and females living longer), we found no clear distinction between these two classes. To our knowledge, this is the first long-term study to characterise the gut microbiota of adult female rabbits through their reproductive life, thus laying foundations for using the gut microbiota data and its influence in studies on adult rabbits.

The present study investigates the photobiont diversity of the boreal felt lichen, Erioderma pedicellatum. Previously sampled genetic data from Newfoundland were reanalyzed and new sequence data (16S rDNA, rbcLX) of the boreal felt lichen from Alaska (USA), Kamchatka (Russia), and North Trøndelag (Norway) were generated. The highest genetic diversity of the photobiont is found in Alaska and Kamchatka, indicating that these may be the primary sources of the species in the Northern Hemisphere. In Newfoundland, the photobiont of E. pedicellatum was screened on leaves of the symbiotic liverwort Frullania asagrayana and it was found to occur on trees where no other lichens were present, demonstrating that the geographical distribution, and possibly also the ecological requirement of the photobiont of E. pedicellatum, is wider than that of the lichen phenotype. Finally, a postulated association between the occurrence of the vegetatively reproducing Coccocarpia palmicola and the occurrence of the compatible photobiont of E. pedicellatum on the same tree could not be established.

A microsporidian infecting the skeletal muscle of hybrid jundiara (Leiarius marmoratus × Pseudoplatystoma reticulatum) in a commercial aquaculture facility in Brazil is described. Affected fish exhibited massive infections in the skeletal muscle that were characterized by large opaque foci throughout the affected fillets. Histologically, skeletal muscle was replaced by inflammatory cells and masses of microsporidial developmental stages. Generally pyriform spores had a wrinkled bi-layer spore wall and measured 4·0 × 6·0 µm. Multinucleate meronts surrounded by a simple plasma membrane were observed. The polar filament had an external membrane and a central electron dense mass. The development of sporoblasts within a sporophorous vesicle appeared synchronized. Ultrastructural observations and molecular analysis of 16S rDNA sequences revealed that the microsporidian was Pleistophora hyphessobryconis. This study is the first report of a P. hyphessobryconis infection in a non-ornamental fish.

The emergence of metagenomics-based approaches in biology has overcome historical culture-based biases in microbiological studies. This has also enabled a more comprehensive assessment of the microbial ecology of environmental samples. The subsequent development of next-generation sequencing technologies, able to produce hundreds of millions of sequences at improved cost and speed, necessitated a computational shift from user-supervised alignment and analysis pipelines, that were used previously for vector-based metagenomic studies that relied on Sanger sequencing. Current computational advances have expanded the scope of microbial biogeography studies and offered novel insights into microbial responses to environmental variation and anthropogenic inputs into ecosystems. However, new biostatistical and computational approaches are required to handle the large volume and complexity of these new multivariate datasets. While this has allowed more complete characterization of taxonomic, phylogenetic and functional microbial diversity, these tools are still limited by methodological biases, incomplete databases, and the high cost of fully characterizing environmental biodiversity. This review addresses the evolution of methods to monitor surface waters and characterize environmental samples through the recent computational advances in metagenomics, with an emphasis on the study of surface waters. These new methods have provided an abundance of opportunities to expand our understanding of the interaction between microbial communities and public health. Specifically, they have allowed for comprehensive monitoring of bacterial communities in surface waters for changes in community structure associated with faecal contamination and the presence of human pathogens, rather than relying on only a few indicator bacteria to direct public health concerns.

Zoantharians are one of the least studied orders of benthic cnidarians of the Persian Gulf. A survey and molecular analysis was conducted to determine zoantharian species diversity in the Persian Gulf. For this purpose, 63 colonies of zoantharians were collected from Hengam and Larak Islands in the Strait of Hormuz and some morphological characteristics of each specimen were recorded, i.e. sand encrustation, polyp shape and colour, oral groove, oral zone and oral disc colours as well as tentacle number and colour. After DNA extraction, mitochondrial 16S ribosomal DNA (16S rDNA) and cytochrome oxidase subunit I (COI) were amplified by polymerase chain reaction (PCR). Based on obtained 16S rDNA and COI gene sequences, five putative species-level clades were identified: Zoanthus sansibaricus (N = 30), Palythoa tuberculosa (N = 12), Palythoa mutuki (N = 2), Palythoa aff. mutuki (N = 18) and Neozoanthus sp. Iran (N = 1). While the first three are known species, the last two were potentially novel undescribed species. Palythoa aff. mutuki has an external appearance similar to Palythoa mutuki. However, mitochondrial DNA sequences obtained from these specimens placed them in a previously undescribed species group. The Neozoanthus specimen was morphologically and molecularly different from other described Neozoanthus species. This is the first record of this genus from the Persian Gulf and neighbouring areas. Since there is not much work on zoantharians identification in the Persian Gulf, further sampling and investigation is needed to speculate on the accuracy of these potentially new species and to complete the knowledge of zoantharian diversity in this area.

Artificial neural networks (ANN) methodology, molecular analyses and comparative morphology of the male postabdomen were used successfully in parallel for species identification and resolution of some taxonomic problems concerning West Palaearctic species of the genus Tachina Meigen, 1803. Supervised feed-forward ANN with back-propagation of errors was applied on morphometric and qualitative characters to solve known taxonomic discrepancies. Background molecular analyses based on mitochondrial markers CO I, Cyt b, 12S and 16S rDNA and study of male postabdominal structures were published separately. All three approaches resolved taxonomic doubts with identical results in the following five cases: case 1, the four presently recognized subgenera of the genus Tachina were confirmed and the description of a new subgenus was recommended; case 2, the validity of a new boreo-alpine species (sp.n.) was confirmed; case 3, the previously supposed presence of T. casta (Rondani, 1859) in central Europe was not supported; case 4, West Palaearctic T. nupta (Rondani, 1859) was contrasted with East Palaearctic specimens from Japan, which seem to represent a valid species not conspecific with central European specimens; T. nupta needs detailed further study; case 5, T. nigrohirta (Stein, 1924) resurrected recently from synonymy with T. ursina Meigen, 1824 was confirmed as a valid species. This parallel application of three alternative methods has enabled the principle of ‘polyphasic taxonomy’ to be tested and verified using these separate results. For the first time, the value of using the ANN approach in taxonomy was justified by two non-mathematical methods (molecular and morphological).

Corals live in close association with bacterial communities, but the nature of the relationship is still poorly understood. In this study, three scleractinian coral species, Galaxea fascicularis, Pavona cactus and Turbinaria reniformis were incubated under different laboratory conditions, and the composition of the bacterial community associated with their tissue or skeleton was compared between species or between species and seawater using denaturing gradient gel electrophoresis (DGGE). The amount of dissolved organic carbon (DOC) excreted and the mucus glycoconjugate composition were also determined for each species. The aim of the study was to assess if the bacterial community composition was species-specific or linked either to the seawater composition, or to the quality and quantity of carbon released by each coral. Results obtained showed that DOC release was significantly different (P < 0.0001) for the three species, with the highest excretion rate for G. fascicularis. Also, the mucus of G. fascicularis and P. cactus mainly contained galactose and glucose whereas the mucus of T. reniformis contained more glucose and xylose. Cluster analyses of microbial community composition showed that the bacterial community was species-specific in the coral tissue but not in the skeleton, in all conditions. It remained specific when corals were incubated in the same or in different aquaria, and under different seawater renewal rates. Since DOC release rates and bacterial composition were both different according to the coral species considered, a link might be suggested between the two parameters. Sequencing of DGGE bands indicated that some bacterial phylotypes were consistently retrieved in all samples of a given species.

With the rapid development of nanotechnology, the security of nanomaterials has been an increasing cause for concern. In this study, the impact of titanium dioxide nanoparticles (nano-TiO2) on the phyllosphere bacterial community were analysed by both a culturable-dependent method and a polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) method. The quantity of culturable phyllosphere bacteria was significantly reduced with an increased concentration of nano-TiO2. With increasing concentrations from 0.002 to 20 mg/ml of nano-TiO2, the quantity of culturable phyllosphere bacteria decreased from 1.8×106 to 3.1×105 cfu/g. The phyllosphere bacteria community was analysed by PCR-DGGE, and when the concentrations of nano-TiO2 were higher than 0.02 mg/ml, the DGGE bands were significantly lower than in the control. Sequencing results of the bands from the DGGE gel showed that there were at least seven genera in the phyllosphere bacteria. Only one uncultured bacterium was unaffected by the concentration of nano-TiO2.

Enterococci represent an important part of bacterial microbiota in different types of artisanal cheeses, made from either raw or pasteurized milk. Polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) of ribosomal DNA is currently one of the most frequently used fingerprinting method to study diversity and dynamics of microbial communities and also a tool for microbial identification. Among several primer pairs for DGGE analysis published so far, six primer pairs amplifying different variable regions of 16S rDNA were selected and applied in our DGGE analysis of 12 species belonging to genus Enterococcus and eight other bacterial species often found in cheeses (seven lactobacilli and one Lactoccocus lactis). When DGGE procedures were optimized, the same set of primers was used for DGGE analysis of five cheese samples. Our study demonstrates that the use of different primer pairs generate significant differences in DGGE analysis of enterococcal population, consequently, appropriate primers regarding the purpose of analysis can be selected. For differentiation and identification of pure enterococcal isolates, primer pair P1V1/P2V1 showed the most promising results since all 12 enterococcal isolates gave distinctive DGGE fingerprints, but with multiple bands patterns; therefore, these primers do not seem to be appropriate for identification of enterococcal species in mixed cultures. Use of primer pairs HDA1/HDA2 and V3f/V3r amplifying V3 region showed better potential for detection and identification of enterococci in mixed communities, but since some bacterial species showed the same fingerprint, for clear identification combination of DGGE and some other method (e.g. species specific PCR) or combined DGGE analysis using two primer pairs generating distinctive results should be used.

The Regenerative Enclosed Life Support Module Simulator (REMS) was designed to simulate the conditions aboard the International Space Station (ISS). This unique terrestrial, encapsulated environment for humans and their associated organisms allowed investigations into the microbial communities within an enclosed habitat system, primarily with respect to diversity, phylogeny and the possible impact on human health. To assess time- and/or condition-dependent changes in microbial diversity within REMS, a total of 27 air samples were collected during three consecutive months. The microbial burden and diversity were elucidated using culture-dependent and culture-independent molecular methods. The results indicate that during controlled conditions the total microbial burden detected by culture-dependent techniques (below a detectable level to 102 cells m−3 of air) and intracellular ATP assay was significantly low (102–103 cells m−3 of air), but increased during the uncontrolled post-operation phase (∼104 cells m−3 of air). Only Gram-positive and α-proteobacteria grew under tested culture conditions, with a predominant occurrence of Methylobacterium radiotolerans, and Sphingomonas yanoikuyae. Direct DNA extraction and 16S rDNA sequencing methodology revealed a broader diversity of microbes present in the REMS air (51 species). Unlike culture-dependent analysis, both Gram-positive and proteobacteria were equally represented, while members of a few proteobaterial groups dominated (Rhodopseudomonas, Sphingomonas, Acidovorax, Ralstonia, Acinetobacter, Pseudomonas, and Psychrobacter). Although the presence of several opportunistic pathogens warrants further investigation, the results demonstrated that routine maintenance such as controlling the humidity, crew’s daily cleaning, and air filtration were effective in reducing the microbial burden in the REMS.

Acanthaspid isopods are well known from the deep sea regions of all oceans. Many species have been found also on the continental shelf around Antarctica. Phylogenetic relationships within the Acanthaspidiidae and the genetic differentiation of populations are poorly understood. In this study we analysed 16S rRNA gene sequences of 36 specimens of Acanthaspidiidae, including 17 specimens of Acanthaspidia drygalskii Vanhöffen, 1914. This species is known from several locations along the Antarctic shelf, supporting the idea of a circum-Antarctic distribution of this taxon. Our molecular data support the monophyly of all six species analysed, but there is only limited evidence for the interspecific relationships between the species. However, we were able to identify three distinct groups of haplotypes within Acanthaspidia drygaskii. Our results indicate the evidence of cryptic, reproductively isolated species. Further data are needed to understand mechanisms underlying speciation in deep sea isopods.

PCR amplification of 16S rDNA was used to determine the diversity of bacteria associated with 3 strains of sheep scab mite, Psoroptes ovis. Eight species of bacteria were identified by phylogenetic analysis of the PCR product sequences. Seven of these species are previously unreported in association with sheep scab mites. Five species were matched to Serratia marcesens, Proponibacteium acnes, Phyllobacterium rubiacearum, Pantoea agglomerans, Curacaobacter baltica, whereas the remaining 3 sequences matched unclassified sequences belonging to the gamma proteobacteria, pseudomonads and streptococci. Bacterial diversity of the in vivo cultured strain was very low and did not match the diversity of the 2 wild collected isolates. The diversity of the bacteria in relation to the disease of sheep scab and the possible importance of these bacteria in the diet of the mites are discussed.

Infection with the endosymbiotic bacteria Wolbachia is widespread in filarial nematodes. Previous studies have suggested concordance between the phylogeny of Wolbachia with that of their nematode hosts. However, there is only one published molecular phylogenetic study of filarial species, based on the 5S rRNA gene spacer. The phylogeny proposed by this study is partially incongruent with previous classifications of filarial nematodes, based on morphological characters. Furthermore, both traditional classifications and molecular phylogenies are, in part, inconsistent with the phylogeny of Wolbachia. Here we report mitochondrial cytochrome oxidase I (COI) gene sequences for 11 species of filaria and for another spirurid nematode which was included as an outgroup. In addition, 16S rRNA, wsp and ftsZ gene sequences were generated for the Wolbachia of several filarial species, in order to complete the available data sets and further resolve the phylogeny of Wolbachia in nematodes. We used these data to evaluate whether nematode and Wolbachia phylogenies are concordant. Some of the possible phylogenetic reconstructions based on COI gene were congruent with the phylogeny of Wolbachia and supported the grouping of the rodent filaria Litomosoides sigmodontis with the lymphatic filariae (i.e. Brugia spp. and Wuchereria spp.) and the sister group relationship of Dirofilaria spp. and Onchocerca spp. However, the placement of the Wolbachia-free filaria Acanthocheilonema viteae is ambiguous and dependent on the phylogenetic methods used.