The Euryarchaeota comprise both methanogenic and nonmethanogenic orders and many lineages of uncultivated archaea with unknown properties. One of these deep-branching lineages, distantly related to the Thermoplasmatales, has been discovered in various environments, including marine habitats, soil, and also the intestinal tracts of termites and mammals. By comparative phylogenetic analysis, we connected this lineage of 16S rRNA genes to a large clade of unknown mcrA gene sequences, a functional marker for methanogenesis, obtained from the same habitats. The identical topologies of 16S rRNA and mcrA gene trees and the perfect congruence of all branches, including several novel groups that we obtained from the guts of termites and cockroaches, strongly suggested that they stem from the same microorganisms. This was further corroborated by two highly enriched cultures of closely related methanogens from the guts of a higher termite (Cubitermes ugandensis) and a millipede (Anadenobolus sp.), which represented one of the arthropod-specific clusters in the respective trees. Numerous other pairs of habitat-specific sequence clusters were obtained from the guts of other termites and cockroaches but were also found in previously published data sets from the intestinal tracts of mammals (e.g., rumen cluster C) and other environments. Together with the recently described Methanomassiliicoccus luminyensis isolated from human feces, which falls into rice cluster III, the results of our study strongly support the idea that the entire clade of "uncultured Thermoplasmatales" in fact represents the seventh order of methanogenic archaea, for which the provisional name "Methanoplasmatales" is proposed. Methanogenesis is an important process in the carbon cycle, with a significant impact on global warming. Methane is produced exclusively by methanogenic archaea-strictly anaerobic microorganisms that occur in almost all anoxic habitats on earth, from the marine environment to freshwater sediments to soils, including hot springs and the deep subsurface, in sewage sludge, and in the digestive tracts of animals and humans (33).All methanogens belong to the phylum Euryarchaeota. They presently comprise members of six orders. The basal groups are Methanopyrales, Methanococcales, and Methanobacteriales (class I); Methanomicrobiales (class II) (3); and Methanosarcinales (class III) (2), with the recently recognized sister group Methanocellales (50). It has been hypothesized that the genes for hydrogenotrophic methanogenesis were already present in a common ancestor and were vertically inherited in a broader monophyletic unit encompassing all methanogens (3). Consequently, it has to be postulated that methanogenesis was lost in the Archaeoglobales (which fall among class I methanogens), the Thermoplasmatales, and the Halobacteriales (which fall between class I and class II) (3).In addition, there are many deep-branching lineages of archaea that are exclusively represented by their 16S rRNA genes (19,53,60) and whose properties cannot be safe...

BackgroundTermites are important contributors to carbon and nitrogen cycling in tropical ecosystems. Higher termites digest lignocellulose in various stages of humification with the help of an entirely prokaryotic microbiota housed in their compartmented intestinal tract. Previous studies revealed fundamental differences in community structure between compartments, but the functional roles of individual lineages in symbiotic digestion are mostly unknown.ResultsHere, we conducted a highly resolved analysis of the gut microbiota in six species of higher termites that feed on plant material at different levels of humification. Combining amplicon sequencing and metagenomics, we assessed similarities in community structure and functional potential between the major hindgut compartments (P1, P3, and P4). Cluster analysis of the relative abundances of orthologous gene clusters (COGs) revealed high similarities among wood- and litter-feeding termites and strong differences to humivorous species. However, abundance estimates of bacterial phyla based on 16S rRNA genes greatly differed from those based on protein-coding genes.ConclusionCommunity structure and functional potential of the microbiota in individual gut compartments are clearly driven by the digestive strategy of the host. The metagenomics libraries obtained in this study provide the basis for future studies that elucidate the fundamental differences in the symbiont-mediated breakdown of lignocellulose and humus by termites of different feeding groups. The high proportion of uncultured bacterial lineages in all samples calls for a reference-independent approach for the correct taxonomic assignment of protein-coding genes.Electronic supplementary materialThe online version of this article (doi:10.1186/s40168-015-0118-1) contains supplementary material, which is available to authorized users.

BackgroundTuberculosis (TB), an ancient scourge of humanity known for several thousands of years, is still a significant public health challenge in many countries today even though some progress has been made in recent years in controlling the disease. The study’s aim was to determine the prevalence of mutations responsible for drug resistance in Mycobacterium tuberculosis among patients visiting selected health centers in Nairobi, Kenya.MethodsThe cross-sectional study involved 132 TB positive patients visiting Mbagathi and Chandaria hospitals between September 2015 and August 2016. Sputum samples were collected from the participants and handled in a biosafety level 3 laboratory at the Kenya Medical Research Institute (KEMRI). Samples were decontaminated using N-Acetyl-L-Cysteine (NALC) – Sodium Hydroxide (NALC-NaOH), stained using Zeihl–Neelsen (ZN), and cultured in Mycobacterium Growth Indicator Tube (MGIT). DNA extracted from cultured isolates using Genolyse™ technique was subjected to Multiplex PCR amplification and reverse hybridization for detection of drug resistance mutations on rpoB, katG, inhA, gyrA, gyrB, rrs and eis genes using Hain Genotype MTBDRplus and MTBDRsl.ResultsAll 132 (100%) patients included in the study were culture positive for M. tuberculosis. Among them, 72 (54%) were male while the remaining 60 (46%) were female. The mean age of the patients was 26.4 ± 19.4 (SD) with a range of 18 to 60 years. Overall, the prevalence of the resistance to first and second-line TB drugs was 1.5% (2/132). Resistance to isoniazid (INH) was observed in 1 of 132 patients (0.8%), as was multi-drug resistant tuberculosis (MDR-TB), also at 0.8%. No resistance to fluoroquinolones (FQ) or kanamycin (KAN) was observed. The INH resistant strain had the katG mutations S315 T, while mutations detected for the MDR-TB were katG S513 T for INH, rpoB S531 L for rifampicin (RIF) and rrs G1484 T for cross-resistance to aminoglycosides/capreomycin (AG/CP).ConclusionsMolecular analysis confirms transmission of the drug-resistant M. tuberculosis strains. The data suggested that there is homogeneity when it comes to the type of drug resistance and mutation that occurs in the region. This calls for intensified drug resistance surveillance and drug adherence among patients infected with TB.

Introduction:Microorganisms are a preferred source of enzyme production due to their high production capability and low cost of production. Bacterial endophytes occupy unexplored sites hence they represent a new source of enzymes with diverse applications. Mangrove plants in Kenya have traditionally been used as medicinal plants due to their bioactive metabolites. However the enzymatic activity of mangrove plants associated endophytes has not been studied.Aims & Objectives:The study is aimed at bioprospecting for enzymes with potential biotechnological applications from mangrove ecosystems.Methods & Materials:Forty-two bacterial isolates were cultured and isolated from the leaves and roots of six mangrove plants sampled from Mida Creek and Gazi Bay in the coastal region of Kenya. The isolates were screened for pectinases, chitinases, cellulases, proteases, and amylases. The isolates were identified based on morphology and 16S rRNA gene sequences analysis.Results:The study showed bacterial isolates had enzymatic activity as follows; pectinases activity (69% of the isolates), Proteases (95% of the isolates), amylases activity (88% of the isolates), cellulases and chitinases (92% of the isolates each). Bacterial endophytes from leaves showed a higher enzymatic index of cellulases suggesting a potential role in degrading cellulose in the leaves of plants. The enzymes amylases and proteases were mostly exhibited by endophytes in roots suggesting a potential role in metabolizing sugar and amino acids in the roots. Isolates from the mangrove plantSonneratia albashowed highest enzymatic indices. The study also observed that isolates from mangrove plants sampled from Gazi bay had high means of enzymatic indices. Molecular identification showed the isolates were closely related toBacillus, Streptomyces, Myroides, andStaphylococcusspecies. Their respective enzymatic activities have been provided in this study.Conclusion:The study showed that Kenyan Mangrove plant-associated bacterial endophytes provide a good reservoir of enzymes with potential industrial applications.

Several types of odours are involved in the location of host animals by tsetse (Diptera: Glossinidae), a vector of animal African trypanosomiasis. Host animals’ ageing urine has been shown to be the source of a phenolic blend attractive to the tsetse. Nevertheless, limited research has been performed on the microbial communities’ role in the production of phenols. This study aimed at profiling bacterial communities mediating the production of tsetse attractive phenols in mammalian urine. Urine samples were collected from African buffalo ( Syncerus caffer ), cattle ( Bos taurus ) and eland ( Taurotragus oryx ) at Kongoni Game Valley Ranch and Kenyatta University in Kenya. Urine samples, of each animal species, were pooled and left open to age in ambient conditions. Bacteriological and phenols analyses were then carried out, at 4 days ageing intervals, for 24 days. Phenols analysis revealed nine volatile phenols: 4-cresol, ortho-cresol, 3-cresol, phenol, 3-ethylphenol, 3-propylphenol, 2-methyloxyphenol, 4-ethylphenol and 4-propylphenol. Eight out of 19 bacterial isolates from the ageing urine revealed the potential to mediate production of phenols. 16S rRNA gene characterisation of the isolates closely resembled Enterococcus faecalis KUB3006, Psychrobacter alimentarius PAMC 27887, Streptococcus agalactiae 2603V, Morganella morganii sub.sp. morganii KT, Micrococcus luteus NCTC2665, Planococcus massiliensis strain ES2, Ochrobactrum pituitosum AA2 and Enterococcus faecalis OGIRF. This study established that some of the phenols emitted from mammalian urine, which influence the tsetse‘s host-seeking behaviour, are well characterised by certain bacteria. These results may allow the development of biotechnological models in vector control that combines the use of these bacteria in the controlled release of semiochemicals.