The effect of adding casein hydrolysate as a protein source in the culture of the bacteria paenibacillus polymyxa at wilker office of food and horticultural plants protection (PTPH) Bojonegoro

agencia(Biological Control AgentsBiological Control Agents) are currently widely used for the purpose of controlling pests and diseases or plant-disturbing organisms. One of the biological agents that has many benefits for controlling several types of diseases in both food crops and horticulture is Paenibacillus polymyxa. These bacteria are beneficial in nitrogen fixation, promotion of plant growth, solubilization of soil phosphorus, and production of exopolysaccharides, hydrolytic enzymes, antibiotics, and cytokinins. Paenibacillus polymyxa also produces polymyxin antibiotics and it is known that these bacteria contain the hormone gibberellins. Casein hydrolyzate is one of the growth media that can be used for the growth of microorganisms. The media is a complex mixture of 18 amino acids, vitamins, calcium, phosphate, and several microelements which results in their high price. Casein hydrolyzate is known to be more effective for plant tissue culture. Therefore, the purpose of this study was to determine whether the amino acid content in casein hydrolyzate could affect the growth ofbacteria Paenibacillus polymyxa. The simple medium used in this study was soybean boiled water. Soybean as an alternative medium for protein sources to substitute beef extract, beef extract and bacto peptone for the growth ofbacteria Paenibacillus polymyxa. Based on TPC calculation with graded dilution in Paenibacillus polymyxa, it was found that there was no difference in the number of bacteria to the addition of casein. This shows that bacteria can grow optimally even though casein hydrolyzate is not given during the growth process.

Method Development for Enteric Virus Recovery from Primary Sludge

Enteric viruses, such as poliovirus, are a leading cause of gastroenteritis, which causes 2–3 million deaths annually. Environmental surveillance of wastewater supplements clinical surveillance for monitoring enteric virus circulation. However, while many environmental surveillance methods require liquid samples, some at-risk locations utilize pit latrines with waste characterized by high solids content. This study’s objective was to develop and evaluate enteric virus concentration protocols for high solids content samples. Two existing protocols were modified and tested using poliovirus type 1 (PV1) seeded into primary sludge. Method 1 (M1) utilized acid adsorption, followed by 2 or 3 elutions (glycine/sodium chloride and/or threonine/sodium chloride), and skimmed milk flocculation. Method 2 (M2) began with centrifugation. The liquid fraction was filtered through a ViroCap filter and eluted (beef extract/glycine). The solid fraction was eluted (beef extract/disodium hydrogen phosphate/citric acid) and concentrated by skimmed milk flocculation. Recovery was enumerated by plaque assay. M1 yielded higher PV1 recovery than M2, though this result was not statistically significant (26.1% and 15.9%, respectively). M1 was further optimized, resulting in significantly greater PV1 recovery when compared to the original protocol (p < 0.05). This method can be used to improve understanding of enteric virus presence in communities without liquid waste streams.

In-Vitro Screening and Biosynthesis of Secondary Metabolites from a New Streptomyces sp. SA1 from a Marine Environment

Background: Streptomyces sp. produces various antibiotic agents and the number of lead molecules from the genus Streptomyces increased rapidly in recent years. Drug resistance against various commercially available antibiotics is one of the important problems throughout the world. Streptomyces spp. produce various antimicrobials with potent activity against drug-resistant bacteria. Methods: Streptomyces sp. SA1 was isolated from the marine environment for the biosynthesis of antibiotics. The important variables influencing secondary metabolite biosynthesis were optimized to increase the biosynthesis of antimicrobial agents using the traditional method and statistical approach. Results: Streptomyces sp. SA1 produced novel antibiotics and the process variables were optimized by the traditional method (One-variable-at-a-time approach). Maltose showed maximum antimicrobial activity (220 U/mL). Analysis of the nitrogen, the effect of nitrogen sources revealed that beef extract incorporated culture medium showed rich antibacterial activity (188/mL). Among the ionic sources, KCl significantly influenced antibiotic production. Maltose, beef extract and KCl were considered as the most influencing medium components. Antimicrobial agent biosynthesis was achieved with maltose 1.22 g/L, beef extract 0.93 g/L and KCl 0.27 g/L in response surface methodology. Conclusion: Actinomycetes, especially Streptomyces, play an important role as a source for bioactive compounds that are used to treat infections, and many other diseases. The isolated Streptomyces sp. was a good producer of antibacterial agent, which required various nutritional supplements in the culture medium. The optimized medium components investigated in this study will be useful for future studies with the mass production of secondary metabolites.

Evaluation of Alternative Components in Growth Media of Lactobacillus brevis for Halal Probiotic Preparation

Probiotic has been widely used in functional food because of numerous advantages for health. MRS broth is commonly used as standard medium in studying lactobacilli. However, in some communities - like muslim and vegetarian society, components in MRS broth/medium become an issue. Beef extract and peptone – animal derived substances as nitrogen sources in the MRS medium should be avoided for the vegetarian. Meanwhile, for the muslim society, all components must be halal-certified including those animal derived ingredients. Therefore, several alternative sources for beef extract and peptone substitution were studied. Combination of alternative nitrogen sources was applied. In order to increase the effect of the alternative nitrogen sources, alternative carbon sources were also included. This is the first report about effects of L. brevis media components on cells growth to expression level of surface layer protein (Slp). Whey, lactose, sucrose, and galactose showed high contribution to L. brevis growth. However, the tested concentration of those substances were not sufficient to obtain equal bacterial growth and Slp expression than that of MRS broth. In addition, yeast extract appeared necessary to maintain cell wall and Slp expression.

Promising entomopathogenic strain of Bacillus thuringiensis 0428 effective against the Colorado beetle

Most of the world produced biopesticides are made by entomopathogenic bacteria B. thuringiensis. So, searching for new strains of it is always necessary. In 2006, the strain B. thuringiensis 0428 was isolated from the caterpillar of the ringed silkworm. The strain 0428 is entomopathogenic against Colorado beetle larvae. The effectiveness of the strain for 5 days was 100%. On beef-extract agar this Gram-positive bacterium formed round or irregular colonies with an average diameter of 6-10 mm. The relief of the colonies is flat; the surface is matte. Colonies of B. thuringiensis 0428 are fast-growing, appearing on the surface of the beef-extract agar on the second or third day at 26-30ºC. The average cell size is 6.48±0.16 (large diameter) and 2.62±0.06 (small diameter) microns. The study of the physiological and biochemical properties of the isolated strain shown that B. thuringiensis 0428 is able to form acetyl-methyl-carbinol and lecithinase. B. thuringiensis 0428 is not able to form ureases or pigments, as well as to use citrates and galactose. But it is able to use sucrose, glucose, mannose, and salicin as a source of carbon. The strain 0428 has proteolytic activity. The strain is capable of synthesizing an insecticidal crystalline protein Cry1A and β-exotoxin. All these characteristics allow us to identify the isolated entomopathogenic strain 0428 as B. thuringiensis var. thuringiensis.