Hamadi I. Boga

Hydrogen-Dependent Oxygen Reduction by Homoacetogenic Bacteria Isolated from Termite Guts

ABSTRACT Although homoacetogenic bacteria are generally considered to be obligate anaerobes, they colonize the intestinal tracts of termites and other environments that are not entirely anoxic in space or time. In this study, we investigated how homoacetogenic bacteria isolated from the hindguts of various termites respond to the presence of molecular oxygen. All strains investigated formed growth bands in oxygen gradient agar tubes under a headspace of H2-CO2. The position of the bands coincided with the oxic-anoxic interface and depended on the O2 partial pressure in the headspace; the position of the bands relative to the meniscus remained stable for more than 1 month. Experiments with dense cell suspensions, performed with Clark-type O2 and H2 electrodes, revealed a large capacity for H2-dependent oxygen reduction in Sporomusa termitida and Sporomusa sp. strain TmAO3 (149 and 826 nmol min−1 mg of protein−1, respectively). Both strains also reduced O2 with endogenous reductants, albeit at lower rates. Only in Acetonema longum did the basal rates exceed the H2-dependent rates considerably (181 versus 28 nmol min−1 mg of protein)−1). Addition of organic substrates did not stimulate O2 consumption in any of the strains. Nevertheless, reductive acetogenesis by cell suspensions of strain TmAO3 was inhibited even at the lowest O2 fluxes, and growth in nonreduced medium occurred only after the bacteria had rendered the medium anoxic. Similar results were obtained with Acetobacterium woodii, suggesting that the results are not unique to the strains isolated from termites. We concluded that because of their tolerance to temporary exposure to O2 at low partial pressures (up to 1.5 kPa in the case of strain TmAO3) and because of their large capacity for O2 reduction, homoacetogens can reestablish conditions favorable for growth by actively removing oxygen from their environment.

Influence of temperature on virulence of fungal isolates of Metarhizium anisopliae and Beauveria bassiana to the two-spotted spider mite Tetranychus urticae.

Twenty-three isolates of Metarhizium anisopliae (Metschnikoff) Sokorin and three isolates of Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales: Clavicipitaceae) were assessed for their virulence against the two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae). Based on the screening results, nine isolates of M. anisopliae and two isolates of B. bassiana were tested for their virulence against young adult (1- to 2-day-old) female T. urticae at constant temperatures of 20, 25, 30 and 35°C. At all temperatures tested, all the fungal isolates were pathogenic to T. urticae but mortality varied with isolates and temperatures. Fungal isolates were more virulent at 25, 30 and 35°C than at 20°C. The lethal time to 50% mortality (LT50) and lethal time to 90% mortality (LT90) values decreased with increased temperature. There were no significant differences in virulence between fungal isolates at 30 and 35°C; however, significant differences were observed at 20 and 25°C.

Isolation and characterisation of bacteria from the haloalkaline Lake Elmenteita, Kenya

Culture-independent studies show that soda lake environments harbour diverse groups of bacteria and archaea. In this study different enrichment and isolation media were used in an attempt to isolate novel groups of bacteria from Lake Elmenteita. Different media were prepared using filter-sterilised water from the lake. The isolates recovered were purified on tryptic soy agar supplemented with 1% sodium carbonate and 4% sodium chloride. Phylogenetic analysis of 181 partial 16S rRNA gene sequences with excellent quality showed that the majority of the isolates were affiliated to the class Gammaproteobacteria and to the genus Bacillus. Isolates from the genus Halomonas and Bacillus constituted 37 and 31% of the total sequenced isolates, respectively. Other groups recovered were related to Marinospirillum, Idiomarina, Vibrio, Enterococcus, Alkalimonas, Alkalibacterium, Amphibacillus, Marinilactibacillus and the actinobacteria Nocardiopsis and Streptomyces. Fifty-one different genera were represented with 31 and 15 cultures scoring with their nearest neighbour similarities below 98 and 97%, respectively. Some novel taxa were identified which had not been isolated previously from the soda environment. The results show that the use of different media with varying compositions can help retrieve novel bacterial diversity from the soda lake environment.

Molecular epidemiology of geographically dispersed Vibrio cholerae, Kenya, January 2009-May 2010.

Isolates represent multiple genetic lineages, a finding consistent with multiple emergences from endemic reservoirs.

Factors Affecting Uptake of Cervical Cancer Early Detection Measures Among Women in Thika, Kenya

In this study, 50 in-depth interviews were carried out with women from the general population in Thika, Kenya. We explored awareness, attitudes, and behavior toward cervical cancer and screening measures among the women. The concept of the health belief model (HBM) was used to develop the topics for the in-depth interviews. Our findings highlight the lack of awareness of cervical cancer and the benefits of early detection measures as critical barriers that affect womens participation in screening programs. We provide a basis for designing programs that will be acceptable and accessible to a larger population, resulting in a reduced cervical cancer burden.

Archaeal Diversity in the Haloalkaline Lake Elmenteita in Kenya

A non-culture approach was used to study the archaeal diversity in Lake Elmenteita, Kenya. Five different sampling points were selected randomly within the lake. Wet sediments and water samples were collected from each sampling point. In addition, dry mud cake was collected from three points where the lake had dried. DNA was extracted from these samples and the 16S rRNA genes were amplified using primers described to be Domain-specific for Archaea. Eleven clone libraries were constructed using PCR-amplified 16S rRNA genes. A total of 1,399 clones were picked and analysed via ARDRA. 170 ARDRA patterns were unique and the respective clones were selected for sequencing. 149 clones gave analysable sequences. BLAST analysis showed that 49 belong to the Domain Archaea while the others were either chimera or affiliated to eukaryotic taxa. Comparative sequence analysis of archaeal clones affiliated them to a wide range of genera. The order Halobacteriales was represented by members of the genera Natronococcus, Halovivax, Halobiforma,Halorubrum, and Halalkalicoccus. The highest percentage (46%) of the clones, however, belonged to uncultured members of the Domain Archaea in the order Halobacteriales. The results show that the archaeal diversity in the lake could be higher than previously reported.

Benzoic acid-degrading bacteria from the intestinal tract of Macrotermes michaelseni Sjöstedt

The intestinal tracts of termites host a wide variety of microbial symbionts, which have been implicated in degradative processes. In this study, a fungus‐cultivating termite, Macrotermes michaelseni was found to harbor 2.2 × 106 bacterial cells per ml of gut homogenates capable of degrading benzoic acid. Two benzoic acid degrading bacteria were isolated from the highest dilution of gut homogenates in oxic media with benzoic acid as the sole carbon source. Isolate CBC was related to Stenotrophomonas maltophila LMG 958T, Xanthomonas campestris DSM 3586T and Stenotrophomonas acidaminophila DSM 13117T with a sequence similarity of 98.3%, 94.7% and 94.2%, respectively. Isolate CBW was related to Enterobacter aerogenes JCM 1235T and Raoultella ornithinolytica ATCC 31898T with sequence similarity of 98.4% and 97.8%, respectively. In addition to growing on benzoic acid (up to 9 mM) aerobically, isolate CBW also degraded benzoic acid under anoxic conditions with nitrate as electron acceptor. Isolate CBC did not degrade bezoic acid with nitrate but could degraded resorcinol under oxic conditions. (© 2007 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

Attraction response of adult Rhipicephalus appendiculatus and Rhipicephalus pulchellus (Acari: Ixodidae) ticks to extracts from Calpurnia aurea (Fabaceae).

Experiments were carried out to investigate the response of two tick species Rhipicephalus pulchellus Gerstaker, 1873 and Rhipicephalus appendiculatus Neumann, 1901 to three different extracts (acetone, aqueous and oil) of the dried leaves of Calpurnia aurea (Aiton) Benth in both an inverted glass tube and a dual choice T-olfactometer. The oil extract at 50 and 100mg/ml attracted 46.7% and 65.9% of R. appendiculatus, respectively, in the inverted glass tube assay, which was comparable to 47.8% of the attraction-aggregation-attachment pheromone (AAAP) used as positive control. At a dose of 100mg/ml the oil extract attracted 52.4% of R. pulchellus in the T-olfactometer bioassay. The relative attraction of both tick species to plant extract was also tested in semi-field plot experiments using a trap baited with different concentrations of emulsifiable extract of C. aurea. A dose of 100mg/ml attracted 52.2% of R. pulchellus and 44.4% of R. appendiculatus from a distance of 1m while 14.4% of R. pulchellus and 12.2% of R. appendiculatus were attracted from 5m distance at the same dose. Addition of CO(2) to the plant extract-baited-trap at the dose of 100mg/ml increased the range of attraction of adult R. pulchellus (44.4% from 5m distance) and up to 33.3% of adult R. appendiculatus tick from a distance of 4m. The results of this study suggest that extracts from C. aurea can potentially be used as baits in a trap for the control of ticks in the field.

Rhodococcus opacus strain RW, a resorcinol- degrading bacterium from the gut of Macrotermes michaelseni

The population of resorcinol degrading bacteria in the intestinal tract of fungi-cultivating termite, Macrotermes michaelseni , was estimated to be 6.8 × 10 2 cells/ml. A gram-positive bacterium designated RW, capable of degrading resorcinol, was isolated from the highest positive dilution. Isolate RW could also degrade phenol and benzoic acid aerobically, and anaerobically using nitrate as an electron acceptor. The isolate is a rod-shaped bacterium that exhibited evolutionary relatedness with the genus Rhodococcus, as determined by phenotypic traits and physiological tests, and a 16S rRNA gene sequence similarity value of 99.6% to the closest Rhodococcus opacus strain. On the basis of these results isolate RW is proposed as a new strain in the species R. opacus . The ability of the isolate to degrade resorcinol, phenol and benzoic acid makes it a potential candidate for use in bioremediation of environments contaminated by such or related compounds. African Journal of Biotechnology Vol. 4 (7), pp. 639-645, 2005

Efficacy of Metarhizium anisopliae in controlling the two-spotted spider mite Tetranychus urticae on common bean in screenhouse and field experiments.

The efficacy of aqueous and emulsifiable formulations of the fungus Metarhizium anisopliae isolate ICIPE78 was evaluated on the population density of Tetranychus urticae infesting common bean plants under screenhouse and field conditions. Synthetic acaricide abamectin was included as a check. Bean plants were artificially infested with T. urticae and allowed to multiply. Three treatments were applied in the screenhouse and 1 treatment in field trials. Mite density was recorded 2 d before spraying and weekly postspraying. The number of pods per plant, number of seeds per pod, and the dry weight of seeds per plant were recorded only in the screenhouse trials. In both screenhouse and field trials, fungal formulations applied at the concentration of 108 conidia/mL and the acaricide reduced the population density of mites as compared to the controls. There were significant differences in T. urticae population densities between the treatments at the various post‐spraying sampling dates. In the screenhouse, the mite densities were near zero from 3‐week postspraying in the treated leaves. At 4‐week postspraying, there were no more leaves in the untreated control (T1) and in the control water + Silwet‐L77 (T2). Fungal formulations were as effective as abamectin in reducing mite densities in both screenhouse and field experiments. There were significant differences in the production parameters during the 2 screenhouse trials, with fungal and abamectin treatments generally having the highest yield. Results of this study underline the potential of the M. anisopliae isolate ICIPE78 as an alternative to acaricides for T. urticae management.