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The freshwater leaf fish

The freshwater leaf fish Pristolepis rubripinnis belongs to the family Pristolepididae, restricted to Pamba and Chalakudy rivers of Kerala, India. In the present study, we sequenced the complete mitogenome of P. rubripinnis and analysed its phylogeny in the order Anabantiformes. The 16622-bp long genome comprised of 13 protein-coding genes, two rRNA genes, 22 transfer RNAs (tRNAs) genes and had a noncoding control region. All the protein-coding genes, tRNA and rRNA were located on the heavy strand, except nad6 and eight tRNAs (glutamine, alanine, asparagine, cysteine, tyrosine, serine, glutamic acid and proline) transcribed from L strand. The genome exhibited an overlapping between atp8 and atp6 (2 bp), nad4 and nad4l (2 bp), tRNAIle and tRNAGln. (1 bp), tRNAThr and tRNAPro (1 bp). Around 157 bp, an intergenic spacer was identified. The overall GC-skews and AT-skews of the H-strand mitogenome were -0.35 and 0.079, respectively, revealing that the H-strand consisted of equal amounts of A and T and that the overall nucleotide composition was C skewed. All tRNA genes exhibited cloverleaf secondary structures, while the secondary structure of tRNASer lacked a discernible dihydrouridine stem. The phylogenetic analysis of available mitogenomes of Anabantiformes revealed a sister group relationship between Pristolepididae and Channidae. The whole mitogenome of Pristolepis rubripinnis will form a molecular resource for further taxonomic and conservation studies on this endemic freshwater fish.

In the zero-water exchange shrimp culture pond maintained with the application of indigenous bioaugmentor, low levels of total ammonia–nitrogen were reported, indicating the relevance of indigenous microbial communities. Sediments (0–5 cm layer) were sampled from the pond (85th day) and the bacterial and archaeal communities; specifically, the ammonia oxidizers (ammonia-oxidizing bacteria, ammonia-oxidizing archaea, and anaerobic ammonia-oxidizing bacteria) in the sediment metagenome of the pond were analysed using the 16S rRNA and functional genes. Bacterial and archaeal 16S rRNA genes showed the relative abundance of Delta-Proteobacteria and Bacteroidetes groups performing sulphur respiration and organic matter degradation, archaeal groups of anaerobic sulphur respiring Crenarchaeotae, and chemolithoautotrophic ammonia oxidizers belonging to Thaumarchaeota. The presence of these diverse bacterial and archaeal communities denotes their significant roles in the cycling the carbon, nitrogen, and sulphur thereby bringing out efficient bioremediation in the bioaugmented zero-water exchange shrimp culture pond. Similarly, the functional gene-specific study showed the predominance of Nitrosomonas sp. (ammonia-oxidizing bacteria), Nitrosopumilus maritimus (ammonia-oxidizing archaea), and Candidatus Kuenenia (anaerobic ammonia-oxidizing bacteria) in the system, which points to their importance in the removal of accumulated ammonia. Thus, this study paves the way for understanding the microbial communities, specifically the ammonia oxidizers responsible for maintaining healthy and optimal environmental conditions in the bioaugmented zero-water exchange shrimp culture pond.

To mitigate the toxicity of ammonia in aquaculture systems, marine and brackish water ammonia-oxidizing bacterial consortia have been developed and are used for activation of nitrifying bioreactors integrated to recirculating aquaculture systems. To shed more light on to these biological entities, diversity of both the consortia were analyzed based on random cloning of 16S rRNA gene and ammonia-oxidizing bacterial specific amoA gene sequences. The dendrograms of representative clones on the basis of amplified ribosomal DNA restriction analysis generated 22 and 19 clusters for marine and brackish water nitrifying consortia, respectively. Phylogenetic analysis demonstrated the presence of various autotrophic nitrifiers belonging to α-, β- and γ-Proteobacteria, anaerobic ammonia oxidizers, heterotrophic denitrifiers, Bacteroidetes, and Actinobacteria. Distribution patterns of the organisms within the two consortia were determined using the software Geneious and diversity indices were investigated using Mega 5.0, VITCOMIC and Primer 7. The abundance of ammonia oxidizers was found in the order of 2.21 ± 0.25 × 109 copies/g wet weight of marine consortium and 6.20 ± 0.23 × 107 copies/g of brackish water consortium. Besides, marine ammonia-oxidizing consortium exhibited higher mean population diversity and Shannon Wiener diversity than the brackish water counterparts.

The addition of commercial nitrifying bacterial products has resulted in significant improvement of nitrification efficiency in recirculating aquaculture systems (RAS). We developed two nitrifying bacterial consortia (NBC) from marine and brackish water as start up cultures for immobilizing commercialized nitrifying bioreactors for RAS. In the present study, the community compositions of the NBC were analyzed by universal 16S rRNA gene and bacterial amoA gene sequencing and fluorescence in situ hybridization (FISH). This study demonstrated that both the consortia involved autotrophic nitrifiers, denitrifiers as well as heterotrophs. Abundant taxa of the brackish water heterotrophic bacterial isolates were Paenibacillus and Beijerinckia spp. whereas in the marine consortia they were Flavobacterium, Cytophaga and Gramella species. The bacterial amoA clones were clustered together with high similarity to Nitrosomonas sp. and uncultured beta Proteobacteria. FISH analysis detected ammonia oxidizers belonging to b subclass of proteobacteria and Nitrosospira sp. in both the consortia, and Nitrosococcus mobilis lineage only in the brackish water consortium and the halophilic Nitrosomonas sp. only in the marine consortium. However, nitrite oxidizers, Nitrobacter sp. and phylum Nitrospira were detected in both the consortia. The metabolites from nitrifiers might have been used by heterotrophs as carbon and energy sources making the consortia a stable biofilm.

Since the discovery in 1977 of deep-sea hydrothermal vents, they have been shown to host unique but diverse biological communities, despite the dark, barren ocean-floor settings in which they exist. Recent research has indicated that the production by fault systems of abundant reducing agents such as hydrogen possibly sustains the microbial communities in these chemoautotrophic ecosystems. High-pressure and high-temperature hydrothermal experiments, and friction experiments, have resulted in the development of important new experimental apparatuses. A batch-type (closed) experimental system that creates equilibrium conditions has contributed greatly to our understanding of sub-seafloor hydrothermal reactions. Flow-type experimental systems have allowed investigation of natural systems under non-equilibrium conditions. Friction experiments have recently been developed to better understand generation of the hydrogen that makes fault systems habitable by primary producers. These experiments suggest that microbial ecosystems sustained by chemical energy derived from fault systems might be widely distributed within oceanic crust. Moreover, flow-type systems that can be used to simulate natural hydrothermal environments that include crustal aquifers might provide insights into the ecological significance of microorganisms and their global contribution to biogeochemical cycles in the ocean and crust.

The biological control efficacy of a greenhouse soil bacterial mixture of Lactobacillus farraginis, Bacillus cereus, and Bacillus thuringiensis strains with antinematode activity was evaluated against the root-knot nematode Meloidogyne incognita. Two control groups planted in soil drenched with sterile distilled water or treated with the broadspectrum carbamate pesticide carbofuran were used for comparison. The results suggest that the bacterial mixture is effective as a biocontrol agent against the root-knot nematode.

Recirculation aquaculture systems (RAS) depend on nitrifying biofilters for the maintenance of water quality, increased biosecurity and environmental sustainability. To satisfy these requirements a packed bed bioreactor (PBBR) activated with indigenous nitrifying bacterial consortia has been developed and commercialized for operation under different salinities for instant nitrification in shrimp and prawn hatchery systems. In the present study the nitrification efficiency of the bioreactor was tested in a laboratory level recirculating aquaculture system for the rearing of Penaeus monodon for a period of two months under higher feeding rates and no water exchange. Rapid setting up of nitrification was observed during the operation, as the volumetric total ammonia nitrogen removal rates (VTR) increased with total ammonia nitrogen (TAN) production in the system. The average Volumetric TAN Removal Rates (VTR) at the feeding rate of 160 g/day from 54–60th days of culture was 0.1533 ± 0.0045 kg TAN/m3/day. The regression between VTR and TAN explained 86% variability in VTR (P < 0.001). The laboratory level RAS demonstrated here showed high performance both in terms of shrimp biomass yield and nitrification and environmental quality maintenance. Fluorescent in-situ Hybridization analysis of the reactor biofilm ensured the presence of autotrophic nitrifier groups such as Nitrosococcus mobilis lineage, Nitrobacter spp and phylum Nitrospira, the constituent members present in the original consortia used for activating the reactors. This showed the stability of the consortia on long term operation.

A packed bed bioreactor (PBBR) was devel- oped for rapid establishment of nitrification in brackish water hatchery systems in the tropics. The reactors were activated by immobilizing ammonia-oxidizing (AMON- PCU-1) and nitrite-oxidizing (NIONPCU-1) bacterial consortia on polystyrene and low-density polyethylene beads, respectively. Fluorescence in situ hybridization demonstrated the presence of autotrophic nitrifiers belong to Nitrosococcus mobilis, lineage of b ammonia oxidizers and nitrite oxidizer Nitrobacter sp. in the consortia. The activated reactors upon integration to the hatchery system resulted in significant ammonia removal (P \ 0.01) cul- minating to its undetectable levels. Consequently, a significantly higher percent survival of larvae was observed in the larval production systems. With spent water the reactors could establish nitrification with high percentage removal of ammonia (78%), nitrite (79%) and BOD (56%) within 7 days of initiation of the process. PBBR is con- figured in such a way to minimize the energy requirements for continuous operation by limiting the energy inputs to a single stage pumping of water and aeration to the aeration cells. The PBBR shall enable hatchery systems to operate under closed recirculating mode and pave the way for better water management in the aquaculture industry.