Thalassic biogas production from sea wrack biomass using different microbial seeds: cow manure, marine sediment and sea wrack-associated microflora.

Sea wrack (dislodged sea grasses and seaweeds) was used in biogas production. Fresh water scarcity in island communities where sea wrack could accumulate led to seawater utilization as liquid substrate. Three microbial seeds cow manure (CM), marine sediment (MS), and sea wrack-associated microflora (SWA) were explored for biogas production. The average biogas produced were 2172±156 mL (MS), 1223±308 mL (SWA) and 551±126 mL (CM). Though methane potential (396.9 mL(CH4) g(-1) volatile solid) computed from sea wrack proximate values was comparable to other feedstocks, highest methane yield was low (MS=94.33 mL(CH4) g(-1) VS). Among the microbial seeds, MS proved the best microbial source in utilizing sea wrack biomass and seawater. However, salinity (MS=42‰) observed exceeded average seawater salinity (34‰). Hence, methanogenic activity could have been inhibited. This is the first report on sea wrack biomass utilization for thalassic biogas production.

Vibrio sp. causing Porites ulcerative white spot disease.

The causative agent of the Indo-Pacific coral disease, Porites ulcerative white spot syndrome (PUWS), that affects Porites spp. and a few other coral genera has so far remained unidentified. Inoculation of thiosulphate citrate bile sucrose (TCBS) agar with tissue material from Porites cylindrica infected with white spot produced colonies of approximately 3 mm diameter consisting of Gram-negative, motile, non-sucrose-fermenting, slightly curved rods with a minimum NaCl requirement of 0.3%. Three of these putative Vibrio sp. isolates were used for infection trials that included different stages of cell growth. Four modes of inoculation and 3 stages of bacterial cell growth were considered for testing Koch's postulates. Stationary phase cells proved more consistently infectious than did exponentially growing or starved cells using a 1-step immersion technique at cell concentrations of 10(4) cells ml(-1). A 1-step immersion technique proved more reliable in producing signs of white spot than did other techniques, such as injection, smearing and 2-step immersion of the inoculum. At inoculum densities >10(4) cells ml(-1) further signs of disease, such as tissue degradation and bleaching, also became evident. At elevated temperatures (>29 degrees C) bleaching remained absent for at least 2 mo from non-inoculated corals serving as controls, but was observed in artificially infected coral fragments. Of the 9 seawater aquaria containing healthy specimens of P. cylindrica, 6 showed signs of white spot 15 d after infection with an isolate tentatively identified as Vibrio sp. Based on 99% similarity of its 16S rRNA gene sequence and selected phenotypical features, this isolate revealed a close relationship to V. natriegens and V. parahaemolyticus.

Epizoic communities of prokaryotes on healthy and diseased scleractinian corals in Lingayen Gulf, Philippines.

In search for microbiological indicators of coral health and coral diseases, community profiles of coral-associated epizoic prokaryotes were investigated because of their dual potential as a source of coral pathogens and their antagonists. In pairwise samples of visually healthy and diseased coral specimens from Bolinao Bay (Pangasinan, Philippines), mixed biofilm communities of ectoderm- and mucus-colonizing epizoic prokaryotes were compared using fluorescent in situ hybridization (FISH). Oligonucleotide probes targeted 13 phylotypes representing the main taxonomic groups of marine prokaryotes. Coral taxa tended to show specific community profiles. An attempt to separate the profiles of healthy and diseased specimens by applying principal component analysis (PCA) to a (nonselective) collection of corals (affected by various diseases) proved unsuccessful. On the other hand, separate PCA clusters were obtained from healthy and diseased corals belonging to a single species (Pocillopora damicornis) only. This cluster formation was dominated by principal component 1 with the genus Vibrio accounting for 18%. At the same time, reef-site-specific clusters were formed as well. At a reef site exposed to pollution from intensive fish cage (Chanos chanos) farming, healthy P. damicornis were mainly (93%) colonized by unicellular cyanobacteria. The formal calculation of diversity parameters suggested that evenness in particular was driven by both health status and reef site location. Despite the low resolution of taxonomic levels achieved with FISH probes targeting only large phylotype groups, significant differences between healthy and diseased corals and also between polluted and nonpolluted reef sites were observed.

Nitrification-denitrification dynamics and community structure of ammonia oxidizing bacteria in a high yield irrigated Philippine rice field.

Nitrogen is the single most limiting factor for rice production. Detailed knowledge on nitrogen dynamics in rice fields is therefore of major importance for developing sustainable rice production. A combination of state-of-the-art microsensor, stable isotope tracer, and molecular techniques was used to evaluate coupled nitrification-denitrification potentials and community structure of ammonia-oxidizing bacteria in a high yield irrigated rice cropping system in the Philippines, without the use of microcosm incubations. The multiple approaches showed a high degree of concordance among methods and thereby clarified the investigated processes. Numbers and potential activity of ammonia-oxidizing bacteria in the system reflected the availability of substrate in three defined soil factions with a ranking of: surface soil > rhizosphere > bulk soil. No nitrification activity was measured between spit applications of N fertilizer. However, nitrification was induced upon nitrogen amendment in intact soil cores. Despite induction by nitrogen amendment, the loss of nitrogen through coupled nitrification-denitrification was less than 10% of the plant nitrogen uptake. Denaturant gradient gel electrophoresis of amoA fragments revealed no differences in diversity profiles between the soil fractions, and phylogenetic analysis, based on amoA genes retrieved from the rice paddy soil, identified a set of mutually very similar sequences related to Nitrosomonas nitrosa.

Seasonal variation of phenol/benzoate-degrading iron-reducing bacteria in irrigated tropical paddy soils as affected by some management practices.

The study provides the first evidence of the presence and abundance of bacterial population that coupled ferric iron reduction to aromatic compounds degradation in tropical irrigated paddy soils in the Philippines. Culturable phenol/benzoate degrading iron-reducing bacteria was enumerated by the most probable number (MPN) counts using phenol or benzoate as sole carbon source, and ferric oxide [Fe(OH)(3)] as the sole electron acceptor. Population density of phenol degrading iron-reducing bacteria (P-IRB) in irrigated paddy soil ranged from 10(2) to 10(8)g(-1) dry soil, and increased with the progressive rice growth in rice cropping seasons; the study also revealed a significant rhizosphere effect on population of P-IRB. However, high enumeration of benzoate degrading iron-reducing bacteria (B-IRB) was obtained in all the tested soil samples averaging at 1.2 x 10(6)g(-1) dry soil, and did not fluctuate significantly over the rice cropping seasons. Statistical data showed that less cropping density with aerated fallow and high nitrogen rate favored the population growth of P-IRB. However, results showed that population size of B-IRB was relatively insensitive to the effect of either seasonal or extrinsic factors tested in this study.

Influence of different cultivars on populations of ammonia-oxidizing bacteria in the root environment of rice.

Comparisons of the activities and diversities of ammonia-oxidizing bacteria (AOB) in the root environment of different cultivars of rice (Oryza sativa L.) indicated marked differences despite identical environmental conditions during growth. Gross nitrification rates obtained by the 15N dilution technique were significantly higher in a modern variety, IR63087-1-17, than in two traditional varieties. Phylogenetic analysis based on the ammonium monooxygenase gene (amoA) identified strains related to Nitrosospira multiformis and Nitrosomonas europaea as the predominant AOB in our experimental rice system. A method was developed to determine the abundance of AOB on root biofilm samples using fluorescently tagged oligonucleotide probes targeting 16S rRNA. The levels of abundance detected suggested an enrichment of AOB on rice roots. We identified 40 to 69% of AOB on roots of IR63087-1-17 as Nitrosomonas spp., while this subpopulation constituted 7 to 23% of AOB on roots of the other cultivars. These results were generally supported by denaturing gradient gel electrophoresis of the amoA gene and analysis of libraries of cloned amoA. In hydroponic culture, oxygen concentration profiles around secondary roots differed significantly among the tested rice varieties, of which IR63087-1-17 showed maximum leakage of oxygen. The results suggest that varietal differences in the composition and activity of root-associated AOB populations may result from microscale differences in O2 availability.