Microbial natural biocides have attracted much more attention in recent years in order to avoid the unrestricted use of chemical biocides in the environment. The aim of this study is to analyze the antibacterial and antifungal activities of secondary metabolites and growth promoting, nematicidal, and soil enzyme activity mediated by Trichoderma hamatum FB10. The bactericidal and fungicidal activities were performed using cell-free extract. Results revealed that the selected strain exert antibacterial activity against Acidovorax avenae, Erutimacarafavora, and Xanthomonas campestris. The selected fungal strain FB10 showed antagonistic activity against fungal pathogens such as, S. sclerotiorum, Rhizoctonia solani, Alternaria radicina, Alternaria citri, and Alternaria dauci. Among the bacterial pathogens, A. avenae showed least MIC (30 ± 2.5 µg/mL) and MBC (70 ± 1.25 µg/mL) values. T. hamatum FB10 strain synthesized bioactive volatile secondary metabolite, which effectively inhibited the growth of bacteria and fungi and indicated the presence of 6-pentyl-alpha-pyrone as the major compound (67.05%). The secondary metabolite synthesized by T. hamatum FB10 showed nematicidal activity against M. incognita eggs. Egg hatch inhibition was 78 ± 2.6% and juvenile stage mortality rate was 89 ± 2.5% when the strain FB10 was treated with nematode. The cell free extract of T. hamatum FB10 showed protease, amylase, cellulase, chitinase, glucanase activities. T. hamatum FB10 inoculated with green gram increased 11% plant height, compared to the control. The fresh weight of the experimental group inoculated with T. hamatum FB10 increased 33.6% more compared to the control group. The green gram seedlings inoculated with T. hamatum FB10 increased 18% more dry weight than control group. Soil enzymes such as, urease, phosphatase, catalase and saccharase were improved in the soil inoculated with T. hamatum FB10. These biochemical components play potent role in soil fertility, energy conversion, and in soil organic matter conversion.

In recent years, studies have focused on the therapeutic properties of probiotics to eliminate pathogenic microorganisms associated with various diseases. Lactobacilli are important probiotics groups that have been found to possess many health-promoting activities. This study was carried out to isolate LactobacillusreuteriLR12 and L. lactisLL10 from pineapple puree. The invitro analysis to evaluate probiotic characteristics of the isolated bacteria included survival in bile and acid tolerance. The cell-free supernatant of L. reuteri LR12 was effective against various pathogenic bacteria and fungi compared with L. lactisLL10. These two bacterial strains have strong anti-biofilm activity (100%) against Enterococcus faecalis, Staphylococcus aureus, and Bacillus cereus. The bacterial strains exhibited adhesion properties to HT-29 cells (human colorectal adenocarcinoma). These bacteria showed DPPH- (2,2-diphenyl-1-picryl-hydrazyl-hydrate) free radical scavenging activity, scavenging of hydroxyl radical activity, superoxide radical scavenging activity, and reducing power activity in the range of 72% ± 3%to 89.3% ± 1.7%, 64% ± 2.7%to 66.8% ± 1.5%, 59.8% ± 4.1% to 63.8% ± 2.1%, and 60.4% ± 1.8%to 66.1% ± 3.3%, respectively. Pineapple puree was used as the starter culture with milk for 2 days for yogurt preparation. Pineapple puree increased flavor and showed the physicochemical properties of yogurt. The finding of the sensory evaluation revealed no significant change compared with the control, except the appearance of yogurt. These findings show that Lactobacilli and pineapple puree have potential use in various probiotic preparations for the fermentation industry.

Agro-industrial residues and cow dung were used as the substrate for the production of alkaline protease by Bacillus cereus strain AT. The bacterial strain Bacillus cereus strain AT produced a high level of protease using cow dung substrate (4813 ± 62 U g(-1)). Physiological fermentation factors such as the incubation time (72 h), the pH (9), the moisture content (120%), and the inoculum level (6%) played a vital role in the enzyme bioprocess. The enzyme production improved with the supplementation of maltose and yeast extract as carbon and nitrogen sources, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and zymogram analysis of the purified protease indicated an estimated molecular mass of 46 kDa. The protease enzyme was stable over a temperature range of 40-50 °C and pH 6-9, with maximum activity at 50 °C and pH 8. Among the divalent ions tested, Ca(2+), Na(+) and Mg(2+) showed activities of 107 ± 0.7%, 103.5 ± 1.3%, and 104.6 ± 0.9, respectively. The enzyme showed stability in the presence of surfactants such as sodium dodecyl sulfate and on various commercially available detergents. The crude enzyme effectively de-haired goat hides within 18 h of incubation at 30 °C. The enzymatic properties of this protease suggest its suitable application as an additive in detergent formulation and also in leather processing. Based on the laboratory results, the use of cow dung for producing and extracting enzyme is not cumbersome and is easy to scale up. Considering its cheap cost and availability, cow dung is an ideal substrate for enzyme bioprocess in an industrial point of view.

The shrimp Penaeus monodon was used for the isolation of digestive enzyme producing host-associated probiotic bacteria. Gut was isolated from a healthy animal completely and morphologically different bacterial isolates were screened for the production of hydrolytic enzymes, such as, protease, amylase, lipase and cellulases. Based on their ability to produce enzymes, the potent probiotic bacteria were identified as Bacillus subtilis and B. licheniformis and these two were used for the preparation of probiotic diet for experimental trials. Probiotic diet was prepared by mixing the shrimp feed with 2 g probiotic/100 g artificial diet (F1), 4 g/100 g (F2), 6 g/100 g (F3), 8 g/100 g (F4) and 10 g/100 g (F5). Juvenile shrimp was fed with probiotic and control diet for a period of 7 weeks at 5 and 8% body weight for the first 3 and 7 weeks, respectively. After seven weeks, whole gut was dissected out and protease activity was estimated as 145 ± 12.3 U/g in control animal and increased as 710 ± 15.2 U/ g in F5 feed groups. Amylase activity was 139 ± 10.4 U/g in control and increased as 209 ± 13. 3 U/g in F5 group. Cellulase activities were 171 ± 9.3 in F5 groups and the control group showed only 102 ± 12.4 U/g. Lipase activity was 78 ± 3 U/g in F1 groups and it increased as 85 ± 5 U/g in F3 groups. These findings indicate the potential of host-associated bacteria to enhance the production of enzymes in the gut of juvenile P. monodon.

Cow dung, a cheap and easily available source of energy, was used as the substrate for the production of alkaline protease by solid-state fermentation using the Bacillus subtilis strain VV. In order to achieve the maximum yield of this enzyme, the following optimum process parameters are needed: fermentation period (72 h), pH (10.0), moisture content (140%), inoculum (25%), temperature (30–40°C), carbon source (2% (w/w) maltose) and nitrogen source (1% (w/w) urea). The protease was stable over a broad temperature range (30–50°C) and pH (8.0-10.0), with maximum activity at 50°C and pH 10.0. Among the divalent ions tested, Ca2+ (0.01 M) increased enzyme activity. The purified protease, after being subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis, was found to have a molecular mass of 38.5 kDa. The enzyme was solvent-and surfactant-stable and showed activity even after 24 h incubation along with various commercially available detergents. This enzyme possessed dehairing properties for animal hide after 16 h of incubation at room temperature. From these results it is evident that cow dung is a potential substrate for the production of a detergent-stable, dehairing protease by B. subtilis. This enzyme has a lot of potential applications in the detergent and leather-processing industries.

Amylase production by Bacillus cereus IND4 was investigated by solid state fermentation (SSF) using cow dung substrate. The SSF conditions were optimized by using one-variable-at-a-time approach and two level full factorial design. Two level full factorial design demonstrated that moisture, pH, fructose, yeast extract and ammonium sulphate have significantly influenced enzyme production (p < 0.05). A central composite design was employed to investigate the optimum concentration of these variables affecting amylase production. Maximal amylase production of 464 units/ml of enzyme was observed in the presence of 100% moisture, 0.1% fructose and 0.01% ammonium sulphate. The enzyme production increased three fold compared to the original medium. The optimum pH and temperature for the activity of amylase were found to be 8.0 and 50 °C, respectively. This enzyme was highly stable at wide pH range (7.0-9.0) and showed 32% enzyme activity after initial denaturation at 50 °C for 1 h. This is the first detailed report on the production of amylase by microorganisms using cow dung as the low cost medium.