Standardization of Various Parameters for Mycoherbicidal metabolites production from Fusarium sp. FGCCW#16 for Parthenium hysterophorus Management

Influence of nutrition and culturing conditions for optimum growth and antimicrobial metabolite production by Streptomyces sp. 201

Antibiotic resistance is one of the biggest global problem of our times. The leading producers of antibiotics that can be used to curb this problem are actinomycetes. This study was conceived to isolate antibiotic producing actinomycetes from the soils of Menengai crater and identify the cultural and physical factors that favoured production of antibiotics by selected actinomycetes. Soil samples were collected from 32 randomly selected sampling points within Menengai crater. The actinomycetes were isolated using serial dilution technique. The actinomycetes were tested for antagonism against selected bacterial and fungal pathogens using primary and secondary screening bioassays. Based on broad spectrum of activity and the size of zone of inhibition, four potent actinomycetes were selected for further studies. The effect of growth media, pH, temperature, incubation period, aeration, inoculum concentration, carbon source, nitrogen source and salt concentration on growth and production of antibiotic metabolites was determined. The actinomycetes isolated presented varying morphological characteristics. There was a significant difference in the diameters of zones of inhibition produced by the test pathogens when subjected to the antibiotic metabolites from the selected actinomycetes (F = 6.6046 P = 0.001338). The growth and production of antibiotics by the selected actinomycetes was favoured by use of Luria Bertani as the culture medium, a pH of 6, incubation temperature of 28oC, incubation period of 7d, aeration rate of 200rpm, inoculum concentration of 1%, glycerol as carbon source, oat meal as nitrogen source and a salt concentration of 1.5%. The growth and production of antibiotics by the selected actinomycetes is affected by culture medium, pH, incubation temperature, incubation period, aeration rate, inoculum concentration, carbon source, nitrogen source and salt concentration. There is need to curry out structure elucidation of the antibiotics from the selected actinomycetes.

Fluoranthene and pyrene are polycyclic aromatic hydrocarbons of high molecular weight that are recalcitrant and toxic to humans; therefore, their removal from the environment is crucial. From hydrocarbon-contaminated soil, 25 bacteria and 12 filamentous fungi capable of growth on pyrene and fluoranthene as the sole carbon and energy source were isolated. From these isolates, Ochrobactrum anthropi BPyF3 and Fusarium sp. FPyF1 were selected and identified because they grew quickly and abundantly in both hydrocarbons. Furthermore, O. anthropi BPyF3 and Fusarium sp. FPyF1 were most efficient at removing pyrene (50.39 and 51.32 %, respectively) and fluoranthene (49.85 and 49.36 %, respectively) from an initial concentration of 50 mg L(-1) after 7 days of incubation. Based on this and on the fact that there was no antagonism between the two microorganisms, a coculture composed of O. anthropi BPyF3 and Fusarium sp. FPyF1 was formed to remove fluoranthene and pyrene at an initial concentration of 100 mg L(-1) in a removal kinetic assay during 21 days. Fluoranthene removal by the coculture was higher (87.95 %) compared with removal from the individual cultures (68.95 % for Fusarium sp. FPyF1 and 64.59 % for O. anthropi BPyF3). In contrast, pyrene removal by the coculture (99.68 %) was similar to that obtained by the pure culture of Fusarium sp. FPyF1 (99.75 %). The kinetics of removal for both compounds was adjusted to a first-order model. This work demonstrates that the coculture formed by Fusarium sp. FPyF1 and O. anthropi BPyF3 has greater potential to remove fluoranthene than individual cultures; however, pyrene can be removed efficiently by Fusarium sp. FPyF1 alone.

Azospirillum species were collected from the rhizosphere and free soil of different plants in Al-Jabal El- Akhdar region. The isolates were identified according to biochemical activities. Isolate (H3) which showed the higher solubilization efficiency (SE) on Pikovskaya medium (PVK) containing insoluble phosphate (inorganic phosphate) Ca3(po4)2 Isolate (H3) was identified as Azospirillum lipoferum (H3), which used as an inoculum as free cell suspension or as alginate formulation. Phosphate solubilization was measured by A.lipoferum (H3) as free or alginate immobilized cells in (PVK) liquid medium and recording the pH of the medium at the same time. The results showed in the phosphorus content by immobilized bacteria in liquid (PVK) broth medium from 2nd day of incubation (2.48 µg/ ml) to 10th day (3.70 µg/ ml) and the free bacteria from 2nd day (8.81 µg/ ml) to 10th day (4.49 µg/ ml). pH of the liquid (PVK) broth medium was recorded from 2nd day to 10th day after incubation which decreased by immobilized A.lipoferum (H3) from 7.00 pH to 4.47 on the 10th day and by the free A.lipoferum (H3) to 6.00, which improved the production of organic acids from sugars which was response of decreasing the pH of the medium. This present study contribute to make agriculture more productive with less harm to the environment and for developing countries where the use of fertilizers is costly, and to encourage of using the biofertilizers ‘plant growth promoting rhizobacteria’ (PGPR) instead of the chemical fertilizers to enhance the plant growth which is the main goal to increase the food production in a healthy way.

Hottle, G. A. (Naval Biological Laboratory, University of California, Berkeley), and D. N. Wright. Growth and survival of Mycoplasma neurolyticum in liquid media. J. Bacteriol. 91:1834-1839. 1966.-Maximal growth of Mycoplasma neurolyticum (between 10(8) and 10(9) colony-forming units per ml) was obtained after 3 days of incubation at 36 C in broth media containing 10% agamma horse serum. When whole horse serum was used in the medium, a complement-mediated inhibition was observed. This inhibition could only be detected when growth was followed by daily plate counts. Maximal growth was delayed for about 24 hr by the horse serum, and the inhibition was spontaneously reversed at the temperature of incubation. Penicillin G was also found to have a temporary inhibitory effect. This was detected with as little as 40 units per ml. Maximal growth was delayed until the 6th day of incubation, when 200 units per ml was present, and until the 16th day, when 1,000 units per ml was present. The survival of M. neurolyticum at undetectable levels in cultures during the incubation period presented an "eclipse" phenomenon which has not been explained. The recrudescence of growth in such cultures late in the incubation period illustrates the events which may occur when mycoplasmas are isolated from clinical material by prolonged incubation in the presence of inhibitors. Survival data showed that M. neurolyticum had greatest stability at pH 8.0, with reduced viability at pH 9.0, 7.0, 10.0, and 6.0, in that order The data on growth and stability suggest a close relationship between the species. of Mycoplasma studied and bacteria.

Grobogan district is the center for maize cultivation in Central Java province with production contribution of 21.8% in 2016. The types of agricultural land in this district is moor and forest land (pesanggem), thus it is interesting to study the sustainability of maize farming in this location. Farmers are taking consideration of sustainability in terms of land, social, economy and agronomy for the next generation. This study aims to (1) describe maize farming implementation, (2) analyze maize farming sustainability and (3) formulate the efforts to improve maize cultivation sustainability in Grobogan district. Descriptive analysis Majewski Index and SWOT were applied to analyze the data. The results showed that (1) farmers tried to increase production by applying “siti methuk” cultivation technique, where farmers began planting again even though the maize had not been harvested (2) the order of priority for sustainability by farmers is the aspect of land - social - economic - agronomy. The average score of sustainability analysis with The Synthetic Farm Sustainability Index of Majewski is 51.15, (3) the alternative sustainable improvement of maize farming such as subsidies for the price of inputs, facilitation of land conservation infrastructure with deep wells, and seed price subsidies.

Swidden agriculture, commercial logging and plantation development have been considered to be the primary common causes of degradation and loss of tropical rain forests in Southeast Asia. In this paper, I chose a part of northeastern Sarawak, East Malaysia as my case study area to analyze the changes in its land‐use characteristics. In the study area, as well as primeval forests, we see that land use began about 100 years ago by a native group called the Iban; commercial logging began in the 1960s, and the development of oil palm plantations began recently. I describe the changes in land use as well as their social and economic causes by referring to aerial photographs, literature surveys, interviews with government officers and the Iban, and observation of land use. My analysis of land use demonstrates that on “state land”, where commercial logging and oil palm plantation development are occurring, large areas of forest have been disturbed in a short period of time. The objective is to benefit economically in response to the social and economic conditions surrounding the study area. On the other hand, in the “Iban territory,” where the Iban practice their land use, land conversion has not occurred on a large scale and in a short period of time, even though the forest has been cut and agricultural fields have been created in response to social and economic conditions as well. They disperse small agricultural fields throughout their forest land. Therefore, the landscape of the “Iban territory” is based on secondary forest, composed of patches of forest in various stages and with several types of agricultural land. Today in Sarawak, monocrop plantations are rapidly expanding and little primeval forest remains. Given these conditions, the land‐use practices of natives such as the Iban will be evaluated from the viewpoint of ecosystem and biodiversity conservation. It could play an important role in providing habitats for natural wildlife.

The objective of this study was to optimize the nutritional and cultural conditions of Streptomyces strain ERI-1, ERI-3 and ERI-26 for the production of antimicrobial metabolites under shake-flask conditions. Effect of eight fermentation medium, different temperature, pH, incubation time, different carbon and nitrogen sources and different concentration of sodium chloride on production of antimicrobial metabolites were studied. Antimicrobial activity of the fermentation medium was evaluated by cup plate method by measuring the zone of inhibition. Nutritional and cultural conditions for the production of antimicrobial metabolites by Streptomyces strain ERI-1, ERI-3 and ERI-26 under shake-flask conditions have been optimized. Modified nutrient medium was found to be good base for fermentation. Glucose and ammonium nitrate were identified as best carbon and nitrogen sources, respectively for growth and production of more antimicrobial compounds. Similarly, initial production medium pH of 7.0, incubation temperature of 30°C and incubation time of 96 h was found to be optimal. Optimization of medium and cultural conditions resulted in better antibacterial and antifungal activity. The zone of inhibition of ERI-26 against Aspergillus niger was 25 and 20 mm for Curvularia lunata, respectively. It is clear that novel Stretomyces strains ERI-1, ERI-3 and ERI-26 produced extra cellular antimicrobial metabolites effective against pathogenic bacteria and fungi, moreover the medium and cultural conditions for better antimicrobial metabolites production have been optimized. Key words: Streptomyces, antimicrobial activity, nutritional requirements, cultural conditions, optimized media.

Cellulases were produced from fungus Humicola fuscoatra MTCC 1409 by solid state fermentation under different cultural conditions viz. pH, incubation temperature, inoculum size and days of incubation in order to optimize the conditions for maximum enzyme production. The potential of cellulase pretreatment to increase the digestibility of paddy straw was also ascertained. Maximum enzyme production was achieved at pH 6.0 of Mandel media and at temperature 45°C. Inoculum size of 1×107 spores/ml was found to be optimum for maximum enzyme production. Enzyme production increased with the increase in days of incubation from 2 to 6 days and then declined thereafter. Cellulase units at the concentration of 1, 1.5 and 2 µmole/g were exogenously added to paddy straw and change in chemical composition of paddy straw was determined after 18, 24, 30 and 36 h of treatment. With increase in enzyme concentration and incubation period, the content of neutral detergent fibre (NDF), acid detergent fibre (ADF), cellulose and hemicellulose reduced gradually with simultaneous increase in lignin and silica content. The concentrations of NDF, ADF, cellulose and hemicellulose decreased by 9.2, 5.9, 10.8 and 23.4% respectively, however lignin and silica content increased by 9.7 and 6.4% respectively as compared to control (with no cellulase added) at 2 µmole enzyme concentration after 36 h of pretreatment. These results show that the enzyme produced from cellulolytic fungus H. fuscoatra is capable of increasing paddy straw digestibility and thus enhancing the utilization of paddy straw for different purposes. Key words: Cellulase production, Humicola fuscoatra, paddy straw, paddy straw digestibility.

An LC-MS-based metabolomics approach was used to characterise the variation in secondary metabolite production due to changes in the salt content of the growth media as well as across different growth periods (incubation times). We used metabolomics as a tool to investigate the production of rifamycins (antibiotics) and other secondary metabolites in the obligate marine actinobacterial species Salinispora arenicola, isolated from Great Barrier Reef (GBR) sponges, at two defined salt concentrations and over three different incubation periods. The results indicated that a 14 day incubation period is optimal for the maximum production of rifamycin B, whereas rifamycin S and W achieve their maximum concentration at 29 days. A “chemical profile” link between the days of incubation and the salt concentration of the growth medium was shown to exist and reliably represents a critical point for selection of growth medium and harvest time.

Effect of carbon dioxide during the early stage of duck egg incubation on hatching characteristics and duckling performance

Objective: The purpose of our study was to isolate and identify the bacteriocinogenic strain exhibiting broad range antimicrobial activity and to analyze the effect of different culturing conditions on the production of an antimicrobial metabolites isolated from the soil of Simlipal Biosphere Reserve, India.
 Methods: In the current study, bacterial strains were screened for antimicrobial activity from soil samples of five different regions. The effect of varying culture conditions such as pH, incubation period, and temperature along with carbon and nitrogen sources with and without certain salts was studied. The characterization of the potent strain was studied by morphological, biochemical, and 16S rRNA genetic sequencing. A phylogenetic affiliation of the strain was studied.
 Results: A total of 31 out of 245 strains isolated from soil were screened on the basis of antimicrobial results against the test pathogens. On the basis of bacteriocin-like inhibition studies method, one potential isolate that exhibited the highest inhibition against all the pathogens was selected. The optimization of highest antimicrobial metabolite production by the isolate with the influence of physical parameters was found as the incubation period of 3 days with 37°C temperature at pH 8 and for the chemical parameters dextrose was showed the most effective carbon sources when implemented with salts and yeast extract as the best sources of nitrogen with salts. The crude metabolite showed an absorbance peak value of 1.234 with optimum ƛ-max at 214 nm. The potent isolate showed maximum identity with Bacillus amyloliquefaciens (99% similarity) with highest query coverage on basic local alignment search tool search analysis of the 16S rDNA sequence. Phylogenetic analysis revealed close affiliation of the isolate with B. amyloliquefaciens (KC494392.1) having antimicrobial activity.
 Conclusion: The findings revealed that the incubation period, temperature, pH, and the culture medium have a direct influence on the production of metabolites. These parameters can be modified for the improvement of the fermentation process.

Production, characterization and pesticidal potential of Bacillus species metabolites against sugar ant (Camponotus consobrinus)

The present study aimed to optimize cultural conditions for optimum bioactive metabolite production by endophytic fungus Trichoderma harzianum, isolated by surface sterilization method from the leaf of the eucalyptus plant. The fungus was identified based on morphological characterization. Fungal metabolites were carried out by ethyl acetate solvent. The antibacterial activity was tested against Escherichia coli (ATCC 25922) and Staphylococcus aureus (NCTC 6571). Various carbon, nitrogen sources, pH, temperature, incubation period, and NaCl on the antibacterial metabolite production were studied. Bioactive metabolite production of T. harzianum exhibits a broad spectrum of in vitro antibacterial activity against two strains of bacteria. For the optimum production of bioactive metabolites, Dextrose and Glucose were found to be the best sources of carbon and the best sources of Nitrogen Yeast extract (YE) and (NH4)2SO. The maximum production of bioactive metabolites occurs at pH 7 and 25°C.; the NaCl showed a positive influence on bioactive metabolites.

A combination of physiology, metabolomics, and genetics reveals the two-component system ResS/ResR-mediated Fe and Al release from biotite by Pseudomonas pergaminensis F77