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The insulin-like growth factor signalling system comprises insulin-like growth factors, insulin-like growth factor receptors and insulin-like growth factor-binding proteins. Along with the growth hormones, insulin-like growth factor signalling is very pivotal in the growth and development of all vertebrates. In fishes, insulin-like growth factors play an important role in osmoregulation, besides the neuroendocrine regulation of growth. Insulin-like growth factor concentration in plasma can assess the growth in fishes and shellfishes and therefore widely applied in nutritional research as an indicator to evaluate the performance of selected nutrients. The present review summarizes the role of insulin-like growth factor signalling in fishes and shellfishes, its significance in aquaculture and in evaluating growth, reproduction and development, and discusses the utility of this system as biomarkers for early indication of growth in aquaculture.

One of the major problems to be addressed in aquaculture is the prominence of antimicrobial resistance (AMR). The occurrence of bacterial infections in cultured fishes promotes the continuous use of antibiotics in aquaculture, which results in the selection of proliferated antibiotic-resistant bacteria and increases the possibility of transfer to the whole environment through horizontal gene transfer. Hence, the accurate cultivation-dependent and cultivation-independent detection methods are very much crucial for the immediate and proper management of this menace. Antimicrobial resistance determinants carrying mobile genetic transfer elements such as transposons, plasmids, integrons and gene cassettes need to be specifically analysed through molecular detection techniques. The susceptibility of microbes to antibiotics should be tested at regular intervals along with various biochemical assays and conjugation studies so as to determine the extent of spread of AMR. Advanced omic-based and bioinformatic tools can also be incorporated for understanding of genetic diversity. The present review focuses on different detection methods to unearth the complexity of AMR in aquaculture. This monitoring helps the authorities to curb the use of antibiotics, commencement of appropriate management measures and adequate substitute strategies in aquaculture. The long battle of AMR could be overcome by the sincere implementation of One Health approach. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of antibiotics and increased antimicrobial resistance (AMR) are of major concerns in aquaculture industry. This could result in global health risks through direct consumption of cultured fishes and dissemination of AMR to natural environment through horizontal gene transfer. Hence, timely detection of the antimicrobial-resistant pathogens and continuous monitoring programmes are inevitable. Advanced microbiological, molecular biological and omic-based tools can unravel the menace to a great extent. This will help the authorities to curb the use of antibiotics and implement appropriate management measures to overcome the threat.

The whole mitogenome of the freshwater fish Channa diplogramma (Malabar snakehead), a vulnerable species of snakehead from the Western Ghats, India was sequenced for the first time. The genome is 16,571 bp in length, comprised of 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes, and a non-coding control region. All the protein-coding genes, tRNA and rRNA were located in the heavy strand except nad6 and 8 tRNAs (Glutamine, Alanine, Asparagine, Cysteine, Tyrosine, Serine, Glutamic acid and Proline) transcribed from L strand. The genome exhibited an overlapping of 7 bp in 2 different locations from 8100 to 10,342 bp. These overlaps were detected between protein-coding genes (atp8 – atp6, nad4l – nad4). In addition, intergenic spacers of 167 bp were also present. The overall GC- and AT-skews of the H-strand mitogenome were 0.1077 and − 0.2439, respectively revealing that the H-strand had equal amounts of A and T and that the overall nucleotide composition was C skewed. All tRNA genes had cloverleaf secondary structures, while the secondary structure of tRNASer lacked a discernible dihydrouridine stem. The phylogenetic analysis of C. diplogramma shows its close similarity to C. micropeltes, native to Southeast Asia, Malay Peninsula and Indonesia.

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.

The insulin-like growth factor signalling system comprises insulin-like growth factors, insulin-like growth factor receptors and insulin-like growth factor-binding proteins. Along with the growth hormones, insulin-like growth factor signalling is very pivotal in the growth and development of all vertebrates. In fishes, insulin-like growth factors play an important role in osmoregulation, besides the neuroendocrine regulation of growth. Insulin-like growth factor concentration in plasma can assess the growth in fishes and shellfishes and therefore widely applied in nutritional research as an indicator to evaluate the performance of selected nutrients. The present review summarizes the role of insulin-like growth factor signalling in fishes and shellfishes, its significance in aquaculture and in evaluating growth, reproduction and development, and discusses the utility of this system as biomarkers for early indication of growth in aquaculture.

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 complete mitochondrial genome of the endangered fish Tor malabaricus, (Malabar mahseer) was sequenced for the first time. The mitochondrial genome size was determined to be 16580 bp in length, consisting of 13 protein-coding genes, 22 transfer RNAs, 2 ribosomal RNA genes and a control region (D-loop). The overall base composition was A: 31.88%, T: 24.87%, G: 15.68% and C: 27.58%. The phylogenetic analysis of 13 protein-coding genes of available mitogenome of mahseer species showed that the T. malabaricus had closer relationship to the black mahseer T. khudree. The complete mitochondrial genome sequence of T. malabaricus would contribute to the better understanding of molecular systematics, selection, conservation, species identification and evolution of this species.

The Indian major carp, mrigal (Cirrhinus mrigala), is a bottom-dwelling fish that can survive hypoxic episodes in its natural environment. We hypothesise that it can better survive hypoxic conditions by altering metabolic responses through GABA (Gamma-aminobutyric acid) supplementation. In the first experiment, the hypoxia tolerance time of the fishes was evaluated under extreme anoxic conditions after feeding with GABA, which showed that GABA had improved survival time under hypoxia. To study the response of dietary GABA in hypoxia-exposed fish, the branchial HIF-1α expression levels, serum thyroid hormone levels and hepatic metabolic responses were assessed in the subsequent experiment. The treatment groups were fed for 60 days with experimental diets containing 4 levels of GABA (0.00% G, 0.50% G, 0.75% G and 1.0%G) and were subjected to 72-h hypoxia exposure (0.5 ± 0.02 mg L-1 dissolved oxygen (DO)) whereas a control group was maintained under normoxic conditions (6.0 ± 0.21 mg L-1 DO). The five treatment groups with three replicates were C0 (0% G + normoxia), H0 (0% G + hypoxia), H0.5 (0.50% G + hypoxia), H0.75 (0.75% G + hypoxia) and H1.0 (1.00% G + hypoxia). The results indicated that GABA supplementation triggered downregulation of HIF 1 alpha expression. When compared with the control group, decreased thyroxine (T4) and triiodothyronine (T3) levels were observed in the GABA-fed hypoxic groups. However, TSH (thyroid stimulating hormone) level remained unchanged in all the treatments. The LDH (lactate dehydrogenase) level in hypoxia-exposed groups was decreased by GABA supplementation. Our study demonstrated that GABA supplementation restores acute hypoxia-induced HIF-1α expression, thyroid hormone levels and LDH activities. On the other hand, it enhanced the citrate synthase (CS) activities at 0.5-1.00%, which showed a sharp decline in hypoxia. Hypoxia caused increase in the serum metabolites such as glucose, lactate, cholesterol and triglycerides. However, GABA supplementation was partially effective in reducing glucose and lactate level while triglycerides and cholesterol values remained unchanged. Overall, our results suggested a potential role of GABA in suppressing metabolism during hypoxia exposure, which can increase the chances of survival of the species Cirrhinus mrigala during hypoxia.

Hypoxia in aquatic environment is often a recurrent stressor which can affect growth and health of fishes. However, nutritional intervention can help aquatic animals to cope up with environmental stressors so as to alleviate the adverse effects. Here, we examined the impact of L-Arginine (Arg) supplementation on hypoxia-induced changes in immune responses and resistance to Aeromonas hydrophila infection in Indian Major Carp, Cirrhinus mrigala. The fish were fed with graded levels of Arg for 60 days and exposed to 72 h sublethal level of hypoxia (0.50 ± 0.16 mg/L dissolved oxygen [DO]). The six treatment groups with three replicates having different levels of Arginine such as N0.0 (0% Arg + Normoxia), H0.0 (0% Arg + Hypoxia), N0.7 (0.70% Arg + Normoxia), H0.7 (0.70% Arg + Hypoxia), N1.4 (1.40% Arg + Normoxia), and H1.4 (1.40% Arg + Hypoxia) were used in the study. Subsequently, the fish of different treatment groups were challenged with A. hydrophila. The hypoxia exposed fish exhibited significantly higher (P

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.

BACKGROUND: A packed bed bioreactor (PBBR) activated with an indigenous nitrifying bacterial consortia was developed and commercialized for rapid establishment of nitrification in brackish water and marine hatchery systems in the tropics. The present study evaluated nitrification in PBBR integrated into a Penaeus monodon recirculating maturation system under different substrate concentrations and flow rates. RESULTS: Instant nitrification was observed after integration of PBBR into the maturation system. TAN and NO2-N concentrations were always maintained below 0.5 mg L−1 during operation. The TAN and NO2-N removal was significant (P < 0.001) in all the six reactor compartments of the PBBR having the substrates at initial concentrations of 2, 5 and 10 mg L−1. The average volumetric TAN removal rates increased with flow rates from 43.51 (250 L h−1) to 130.44 (2500 L h−1) gTAN m−3 day−1 (P < 0.05). FISH analysis of the biofilms after 70 days of operation gave positive results with probes NSO 190 ((β ammonia oxidizers), NsV 443 (Nitrosospira spp.) NEU (halophilic Nitrosomonas), Ntspa 712 (Phylum Nitrospira) indicating stability of the consortia. CONCLUSION: The PBBR integrated into the P. monodon maturation system exhibited significant nitrification upon operation for 70 days as well as at different substrate concentrations and flow rates. This system can easily be integrated into marine and brackish water aquaculture systems, to establish instantaneous nitrification. Copyright © 2011 Society of Chemical Industry

Two distinct nitrifying bacterial consortia, namely an ammonia oxidizing non-penaeid culture (AMONPCU-1) and an ammonia oxidizing penaeid culture (AMOPCU-1), have been mass produced in a nitrifying bacterial consortia production unit (NBCPU). The consortia, maintained at 4°C were activated and cultured in a 2 l fermentor initially. At this stage the net biomass (0.105 and 0.112 g/l), maximum specific growth rate (0.112 and 0.105/h) and yield coefficients (1.315 and 2.08) were calculated respectively, for AMONPCU-1 and AMOPCU-1 on attaining stationary growth phase. Subsequently on mass production in a 200 l NBCPU under optimized culture conditions, the total amounts of NH4 +–N removed by AMONPCU-1 and AMOPCU-1 were 1.948 and 1.242 g/l within 160 and 270 days, respectively. Total alkalinity reduction of 11.7–14.4 and 7.5–9.1 g/l were observed which led to the consumption of 78 and 62 g Na2CO3. The yield coefficient and biomass of AMONPCU-1 were 0.67 and 125.3 g/l and those of AMOPCU-1 were 1.23 and 165 g/l. The higher yield coefficient and growth rate of AMOPCU-1 suggest better energy conversion efficiency and higher CO2 fixation potential. Both of the consortia were dominated by Nitrosomonas-like organisms. The consortia may find application in the establishment of nitrification within marine and brackish water culture systems.

For establishing nitrification in prawn (non-penaeid, salinity 10-15 ppt) and shrimp (penaeid, salinity 30-35 ppt) larval production systems, a stringed bed suspended bioreactor (SBSBR) was designed, fabricated, and validated. It was fabricated with 5 mm polystyrene and low density polyethylene beads as the substrata for ammonia and nitrite oxidizing bacterial consortia, respectively, with an overall surface area of 684 cm 2 . The reactors were activated in a prototype activator and were transported in polythene bags to the site of testing. Performance of the reactors activated with the nitrifying bacterial consortia AMONPCU-1 (ammonia oxidizers for non-penaeid culture) and NIONPCU-1 (nitrite oxidizers for non-penaeid culture) was evaluated in a Macrobrachium rosenbergii larval rearing system and those activated with AMOPCU-1 (ammonia oxidizers for pen- aeid culture) and NIOPCU-1 (nitrite oxidizers for penaeid culture) in a Penaeus monodon seed production system. Rapid setting up of nitrification could be observed in both the static systems which resulted in a higher relative per cent survival of larvae.

Surveys for bacteriological analysis of larval samples to isolate the associated vibrios were carried out during 1985^1992, 2001 and 2002 in three diierent hatcheries located on the southwest coast of India. Vibrio isolates were examined for their species diversity, virulence based on haemolysis in prawn blood agar, lipolysis, proteolysis and chitinolysis and antibiotic sensitivity.Vibrio cholerae was the predominant species in the apparently healthy larval samples, whereas V. alginolyticus and V. vulni¢cus dominated during disease and morbidity. No correlation was found between the hydrolytic properties and haemolytic activity of the vibrios associated with the larvae. All isolates were resistant to erythromycin and resistance to oxytetracycline, ampicillin and streptomycin sulphate was prevalent among the larger section of the Vibrio population. This suggested that antibiotic application may not be of much use to protect the larvae from vibriosis. This is the ¢rst report on the diversity of Vibrio species associated with Macrobrachium rosenbergii larvae and their virulence characteristics based on haemolysis in prawn blood agar.

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.