The effects of a marine photosynthetic bacteria Rhodovulum sulfidophilum on the growth and survival rate of Marsupenaeus japonicus (kuruma shrimp)

The sustainability of the shrimp aquaculture depends largely on disease control and the health status of shrimp. Probiotics, which make shrimps healthier and more resistant to pathogens, are promising countermeasure for shrimp diseases. In this study, the effects of the marine purple non-sulfur photosynthetic bacterium (PNSB) Rhodovulum sulfidophilum on Marsupenaeus japonicus (kuruma shrimp) growth and survival were examined in 177 m2 aquaria (140 tons of water) for 70 days. The shrimp received feed containing 0.01 % fresh weight (106 colony forming unit/g) of R. sulfidophilum cells. The survival rate significantly improved (P < 0.001) (R. sulfidophilum-fed = 81.9 %; control = 71.5 %), the feed conversion rate improved (R. sulfidophilum-fed = 1.83; control = 2.11), and there was no difference in the shrimp average body weight. The approximate bacterial cell cost was $0.003 to $0.005 per 1 kg feed, indicating that the R. sulfidophilum approach is economically feasible and a promising candidate for probiotic bacteria in shrimp aquaculture. Keywords: photosynthetic bacteria, Rhodovulum sulfidophilum, Marsupenaeus japonicus, shrimp, probiotics

Peptide-Mediated Gene Transfer into Marine Purple Photosynthetic Bacteria

Use of photosynthetic organisms is one of the sustainable ways to produce high-value products. Marine purple photosynthetic bacteria are one of the research focuses as microbial production hosts. Genetic transformation is indispensable as a biotechnology technique. However, only conjugation has been determined to be an applicable method for the transformation of marine purple photosynthetic bacteria so far. In this study, for the first time, a dual peptide-based transformation method combining cell penetrating peptide (CPP), cationic peptide and Tat-derived peptide (dTat-Sar-EED) (containing D-amino acids of Tat and endosomal escape domain (EED) connected by sarcosine linkers) successfully delivered plasmid DNA into Rhodovulum sulfidophilum, a marine purple photosynthetic bacterium. The plasmid delivery efficiency was greatly improved by dTat-Sar-EED. The concentrations of dTat-Sar-EED, cell growth stage and recovery duration affected the efficiency of plasmid DNA delivery. The delivery was inhibited at 4 °C and by A22, which is an inhibitor of the actin homolog MreB. This suggests that the plasmid DNA delivery occurred via MreB-mediated energy dependent process. Additionally, this peptide-mediated delivery method was also applicable for E. coli cells. Thus, a wide range of bacteria could be genetically transformed by using this novel peptide-based transformation method.

A marine photosynthetic microbial cell factory as a platform for spider silk production

Photosynthetic microorganisms such as cyanobacteria, purple bacteria and microalgae have attracted great interest as promising platforms for economical and sustainable production of bioenergy, biochemicals, and biopolymers. Here, we demonstrate heterotrophic production of spider dragline silk proteins, major ampullate spidroins (MaSp), in a marine photosynthetic purple bacterium, Rhodovulum sulfidophilum, under both photoheterotrophic and photoautotrophic growth conditions. Spider silk is a biodegradable and biocompatible material with remarkable mechanical properties. R. sulfidophilum grow by utilizing abundant and renewable nonfood bioresources such as seawater, sunlight, and gaseous CO2 and N2, thus making this photosynthetic microbial cell factory a promising green and sustainable production platform for proteins and biopolymers, including spider silks.

IDENTIFICATION AND CHARACTERIZATION OF A PURPLE NONSULFUR BACTERIUM ISOLATED FROM COASTAL AREA OF HAI PHONG FOR USING IN PRODUCTION OF UNSATURATED FATTY ACID (OMEGA 6, 7, 9)

Purple nonsulfur bacteria are a group that has so much biotechnological applications, particularly in producing of functional food rich with unsaturated fatty acids. A purple nonsulfur bacterium (named HPB.6) was chosen based on its strong growth, high lipid and synthesis of unsaturated fatty acid (omega 6,7,9). Studying on basic biological characteristics showed that the cells of HPB.6 were observed as ovoid-rod shape, none motility, Gram negative staining. The diameter of single bacterium was about 0.8-1.0 µm. The cells divide by binary fission and had bacteriochlorophyll a (Bchl a). This bacterium grew well on medium with carbon and nitrogen sources such as acetate, succinate, pyruvate, butyrate, glutamate, arginine, leucine, tyrosine, alanine, methionine, threonine, glutamine, yeast extract and NH4Cl. This selected strain grew well on medium with salt concentrations from 1.5 - 6.0% (optimum 3%), pH from 5.0 to 8.0 (optimum at pH 6.5) and could withstand Na2S at 4.0 - 5.2 mM. Based on morphological, physiological properties and 16S rRNA analysis received demonstrated that HPB.6 strain belongs to the species Rhodovulum sulfidophilum.

Investigation of a Farm-scale Multitrophic Recirculating Aquaculture System with the Addition of Rhodovulum sulfidophilum for Milkfish (Chanos chanos) Coastal Aquaculture

Globally, coastal aquaculture is growing due to the large demand for marine products. Specific impacts caused by coastal aquaculture on the environment include the discharge of culture farm effluents, stress on ground water (the absence of recycling), nutrient pollution, and diseases of cultured animals. Three methods, integrated multitrophic aquaculture (IMTA), recirculating aquaculture system (RAS), and beneficial bacteria for aquaculture, have been developed to solve these problems. In this study, the advantages of IMTA and RAS were integrated to develop a novel multitrophic recirculating aquaculture system (MRAS) to adapt to the farm-scale culturing of milkfish (Chanos chanos). The photosynthetic bacteria Rhodovulum sulfidophilum was added to enhance the performance of the farm-scale milkfish MRAS. This setting could promote growth of beneficial bacteria, such as the nitrogen cycle-associated microbial community and the anoxygenic phototrophic Acidobacteria community. The ammonia level was reduced, and the total phosphorous level was stable in the water recycled in the MRAS. The cyanobacteria, algae, Vibrio, Escherichia, and other potential pathogenic bacteria communities were inhibited in the MRAS. This study provides an effective design of a water recycling aquaculture system. Milkfish, Asian tiger shrimp (Penaeus monodon), Asian hard clam (Meretrix lusoria), and seaweed (Gracilaria sp.) can be cultured and simultaneously produced in the system.