Isolation and Characterization of Cellulolytic Bacteria Diversity in Peatland Ecosystem and Their Cellulolytic Activities

Peatlands are terrestrial wetland ecosystems formed from piles of organic matter that decompose into organic deposits. Peat soil has a high potential to produce cellulose which, can be reused by cellulolytic bacteria. This study aims to find out the potential strain of cellulolytic bacteria isolated from peatland ecosystems. The method used was experimental, sequentially, the stages are isolation and screening for cellulolytic bacteria, quantitative testing of cellulolytic activity, characterizing the morphology and physiology of bacteria, and the identification of bacteria based on Bergey’s Manual of Determinative Bacteriology. The screening results obtained seven isolates of cellulolytic bacteria capable of hydrolysed cellulose on 1% Carboxy Methyl Cellulose (CMC) Agar Medium, namely SPS1, SPS2, SPS 3, SDG1, SDG 2, SPW1, and SPW4. Three of seven isolates obtained the highest cellulolytic index sequentially, namely SPS2 of 2.82, SPS3 of 2.65, and SDG1 of 2.47. The cellulolytic activity was indicated by the value of a halo zone around the colonies on 1 % CMC medium after being dripped with Congo red. The halo zone is an early indication to determine the ability of bacteria to decompose cellulose. Based on Bergey’s Manual of Determinative Bacteriology showed that the three isolates had the same characteristics as the genus Bacillus, Lactobacillus and Corynebacterium.

Screening and Characterization of Calcite Solubilizing Bacteria isolated from Marble Slurry dumped in Sukher, Udaipur, Rajasthan, India

Rajasthan is known as mineral majestic state as more than 50 types of minerals are produced including marble. Various types of marbles are spread over in 16 belts in 33 districts of the State. Large amount of marble slurry is discharged as a waste generated by the quarries which is dumped in empty pits in the forest area; thereby creating huge dumping area. The present study was undertaken for screening and characterization of calcite solubilizing bacteria from marble slurry collected from dumping yard in Sukher, Udaipur, India. Screening of calcite solubilizing bacteria was done on calcite agar medium. Calcite solubilizing efficiency (CSE) and calcite solubilizing activity index (CSAI) of the isolates were determined using 0.1% calcite agar medium at 37C for 10 days of incubation. Characterization of the isolates was done on the basis of morphological, biochemical and molecular methods. A total of 27 isolates appeared on calcite agar medium after an incubation of 24 hours at 37°C. Among them only 3 isolates namely isolate CS6, CS13 and CS25 showed clear halo zone of 20 mm, 29 mm and 2mm diameters respectively after 10 days of incubation indicating demonstrable calcite solubilizing activity. The maximum CSE and CSAI 362% and 4.62 respectively were observed for isolate CS13. Isolates CS6, CS13 and CS25 were identified as Exiguobacterium aquaticum CS6 (accession no. MK353511), Staphylococcous aureus CS13 (accession no. KY694446) and Bacillus endophyticus CS25 (accession no. MK353513) respectively. Two isolates that showed remarkable calcite solubilizing activity can be further analyzed for their possible use in restoration of marble slurry contaminated soil.

Isolation and Characterization of Cellulase Producing Bacteria for the Purpose of Converting Spent Mushroom Edible Canna-Waste into Organic Fertilizer

Converting spent mushroom substrates into organic fertilizer helps to tackle the problem of pollution in edible canna starch processing villages and adds new value to the production chain of edible canna. To successfully turn the spent substrates into compost, there is certainly an indispensable role for cellulolytic microorganisms, in which Bacillus strains are always important. Several bacterial strains have been isolated from spent edible canna substrate after cultivation of monkey head mushroom in this study. Among isolated strains, the strain NDK5 has been selected exhibiting the highest cellulolytic activities with solubilization indexes of 6.14 and 18.3 mm for the ratio between the halo zone diameters and the colony diameters in the point cultivation method (SIratio) and the offset between the halo zone diameters and the agar hole diameters (SIoffset), respectively. The highest CMCase activity was 4.29 ± 0.071 U/ml. Morphological, physiological, biochemical, and 16S rRNA sequence analyses (100% homology with B. amyloliquefaciens sp. plantarum FZB42) were further carried out for the selected strain, leading to the identification of the strain as B. amyloliquefaciens sp. plantarum NDK5 strain. In addition, NDK5 was proved to have a capacity for synthesizing indole-3-acetic acid, a plant growth hormone, on an L-tryptophan-containing medium. Trial incubation of spent mushroom edible canna-substrate with the strain NDK5 showed increases in several quality criteria of the waste after 20 days of incubation, that meet the standard criteria for bio-organic fertilizer according to TCVN 7185:2002.

Screening and Isolation of Effective Microbes for Decomposition Process among Different Organic Waste

Organic waste is material that is biodegradable and comes from either plant or animal. It can be decomposed over time by microbes into methane, carbon dioxide, water, and other simple organic molecules. Microorganism especially bacteria and fungus are able to decompose cellulose by an enzyme known as cellulase. Since plants are made up from cellulose, this study is focused on isolating and identifying the microbes that are responsible in decomposing the organic waste by screening for the microorganisms that are able to produce cellulose enzyme. In this experiment, the bacteria were isolated from three different samples, which are; compost product, decayed wood and empty fruit bunch mix with oil palm frond. The samples were collected from the fresh decomposing organic waste. The samples undergo serial dilution, screening the ability of the microorganism to solubilize phosphate by using National Botanical Research Institute’s Phosphate media (NBRIP) and screening the ability of the microorganism to produce cellulose enzyme by using Carboxymethyl Cellulose Media (CMC) treated with Congo Red and NaCl for confirmation of the targeted effective microbes. The result shows that the targeted effective microbes will produce the halo zone or clear zone formation after being treated with Red Congo and NaCl solution on CMC media and producing a halo zone on NBRIP media. In this experiment, we manage to isolate and identify the bacteria that is responsible in producing cellulase enzyme and solubilizing phosphate that is known as Streptomyces sanglieri. Application of effective microbes will promote the plant uptake due to its ability in converting the nutrient into available form to the plant.

Seleksi Aktivitas Antibakteri dari Beberapa Gorgonia dari Perairan Manado Terhadap Bakteri Staphylococcus aureus, Escherichia coli, dan Edwarsiella tarda

Telah dilakukan penelitian dengan tujuan menguji aktivitas antibakteri dari beberapa Gorgonia yang berasal dari Perairan Pulau Bunaken Manado. Tahapan penelitian yaitu pengambilan sampel gorgonia pada lokasi Perairan Bunaken Manado dengan metode purposive sampling, identifikasi, ekstraksi dengan metode maserasi, peremajaan bakteri uji dengan metode goresan, pembuatan larutan Mc. Farland, pengujian aktivitas antibakteri dengan metode Kirby-Bauer. Bakteri uji yang digunakan adalah Staphylococcus aureus, Escherichia coli, dan Edwarsiella tarda. Hasil penelitian dari ke 9 sampel Gorgonia teridentifikasi 7 sampel Gorgonia yaitu Annella sp, Melithaea sp, Astrogorgia sp, Mopsella sp *cf, Siphonogorgia geodeffry *cf, Gorgonia sp, dan Iciligorgia sp *cf serta 2 sampel yang belum teridentifikasi. Pengujian antibakteri didapatkan untuk bakteri uji Staphylococcus aureus, ada 4 sampel yang memiliki zona hambat dengan kategori sedang, 1 sampel dengan kategori kuat dan 4 sampel tidak memiliki zona hambat tetapi memiliki zona halo. Bakteri uji Escherichia coli, ada 5 sampel memiliki zona hambat dengan kategori sedang, 1 sampel dengan kategori kuat dan 3 sampel tidak memiliki zona hambat tetapi memiliki zona halo. Bakteri uji Edwarsiella tarda, ada 2 sampel memiliki zona hambat dengan kategori sedang, dan 7 sampel tidak memiliki zona hambat tetapi memiliki zona halo. Berdasarkan hasil penelitian dapat disimpulkan bahwa ke 9 sampel Gorgonian memiliki sifat bakterisida (membunuh bakteri) dan bakteriostatik (menghambat pertumbuhan bakteri)The aim of the research to examine the antibacterial activity of some Gorgonian from Bunaken Island Waters. The stages of the research are Gorgonian sampling, identification, extraction by maceration method, making Mc. Farland solution, and testing the antibacterial activity by the Kirby-Bauer method. The test bacteria used Staphylococcus aureus, Escherichia coli, and Edwarsiella tarda. The results of the nine Gorgonian samples were identified namely Annella sp, Melithaea sp, Astrogorgia sp, Mopsella sp *cf, Siphonogorgia geodeffry *cf, Gorgonia sp, and Iciligorgia sp and three unidentified samples. Antibacterial testing was obtained for the Staphylococcus aureus bacteria, there were four samples that had medium inhibitory categories, one sample with strong category and four samples did not have inhibitory zones but had a halo zone. Escherichia coli bacteria, there are five samples having inhibition zone with medium categories, one sample with strong category and three samples do not have inhibitory zone but have a halo zone. Edwarsiella tarda bacteria, there are two samples having inhibition zone with medium categories, and seven samples do not have inhibitory zone but have a halo zone. Based on the results of the study it can be concluded that the nine Gorgonian samples have bactericidal and bacteriostatic properties

ISOLATION AND IDENTIFICATION OF AZOTOBACTER OF SOME TYPE OF LAND USE IN JEGU VILLAGES

Conventional farming systems have a negative impact on the life of Azotobacter. Through the results of this study, we want to provide information on the existence of Azotobacter in several rhizosphere of cultivated plants, and looking for isolates that have advantages as biofertilizers and decomposers. This research is an exploration of bacteria Azotobacter spp. from several plant rhizosphere namely: organic and inorganic rice paddy, cassava, coconut, and chocolate. Three samples was taken from each type of land use, so that the number of isolate sources were 15 samples. All analyzes were conducted at Soil Biology Laboratory, Faculty of Agriculture Udayana University. This study aims to find isolates of Azotobacter spp. which is superior to be utilized as biofertilizer and decomposer. Parameters used to support isolation and identification results are total population of bacteria Azotobacter spp., soil respiration, gram staining, halo zone and optical density. The results showed that organic rice rhizosphere was the best isolate source compared with inorganic rice rhizosphere, coconut, cassava and chocolate. Isolate from this organic rice rhizosphere has the highest total population (40.10 cfu x107g-1 soil), on positive catalase test yield bubbles and optical density (average 1.217ABS at 550 nm wavelength). While the superior isolates of Azotobacter spp. As biofertilizer and decomposer candidates are TSO2 isolates (samples from organic rice plant rhizosphere) with soil respiration rate (8.057mgC-CO2 kg-1 soil/day), high optical density (1.147 ABS on spectrophotometer with 550 nm wavelength) and highest halo zone diameter (10 mm).

Isolation and characterization of insoluble inorganic phosphate solubilizer rice rhizosphere strain Enterobacter cloacae BAU3

The objective of the present study was to isolate and characterize most efficient phosphate solubilizing bacteria (PSB) from rice rhizosphere. The study was carried out during the Kharif season’2018 at Department of Soil Science and Agricultural Chemistry, Bihar Agricultural University, Sabour, Bhagalpur, Bihar. The availability of phosphorous to plants for uptake and utilization is limited in soil due to fixation in the form of Fe-P, Al-P and Ca-P. The use of phosphate solubilizing bacteria can prove to be helpful measure to supply phosphorous to the crops to increase the productivity. In the present investigation, a total of 10 isolates were obtained from rice rhizosphere soil samples. All ten isolated isolates were shown phosphorus solubilization. Out of ten isolates BAU3 was found to be most potent phosphate solubilizers showing clear halo zone around its colony. The isolate BAU3 showed 20.00 mm phosphate solubilizing halo zone around its colony. The solubilization index (SI) of the isolate BAU3 was also calculated at the end of the incubation period and observed phosphate solubilization index (SI) of 3.22. The isolate BAU3 showed maximum insoluble phosphate solubilization of 450.24 ?g ml-1 and isolates BAU3 was selected for subsequent studies. The bacterial isolates BAU3 was gram negative, non-spore forming rods shaped. On the basis of the 16SrDNA sequencing, isolate BAU3 was identified as Enterobacter cloacae strain BAU3 (Genebank Accession No. MK033472). The isolated strain of bacterial has potential to solubilize insoluble phosphorus and it can be utilized for preparation of microbial inoculants or biofertilizers.

Phosphate Solubilizing Bacteria in Certain Agricultural Crop Soils of Delhi

Phosphate solubilizing bacteria (PSB) helps in the solubilization of insoluble phosphates and thus lead to increase in crop yields. A study was conducted to isolate and characterize biochemically PSB from different agricultural crop soils of Delhi such as Garlic, Radish, Chilli, Onion and Cabbage. PSB were isolated in Pikovskaya solid medium and formation of solubilization (halo) zone was measured. 16 PSB were isolated and identified. The selected PSB differed in phosphate solubilizing efficiency, production of organic acids and phosphatases. Citrobacter sp. and Pseudomonas sp. were dominant in all the crop plants. Among all the isolates, Pseudomonas sp. proved to be an efficient phosphate solubilizer.

Isolation of Ethidium Bromide Degrading Bacteria from Jharkhand

Ethidium bromide (EtBr) is a carcinogenic and mutagenic agent which is widely used in research laboratories to probe nucleic acids by gel electrophoresis. It is generally buried underground (for solid waste) or disposed of pouring it down the sink (in case of liquid waste). Soil or drain microbial community may be able to take care of such substance else it will lead to contamination of our underground resources or others through defined and undefined routes. In view of the above assumption and literature reports the present study was undertaken to isolate and evaluate bacteria for removal, by bioaccumulation and /or biotransformation, of EtBr from contaminated sources and wastes, before their disposal to the environment. Two distinct bacteria both motile BR3 and BR4 could be identified from agarose-gel-waste containing 0.5-1.0 µg/ml ethidium bromide. Both bacteria were found to grow on EtBr-NA plate (Nutrient-Agar supplemented with EtBr at a concentration of 30 µg/ml) however only BR3 isolate showed large non-fluorescent-halo zone (characteristic to degradation of EtBr) when exposed to trans-UV light. Other isolate BR4 could accumulate EtBr within the colony biomass but did not showed clear (non-fluorescent) hallow zone around it. However the bacterium is not able to utilize the EtBr as a sole carbon source.Int. J. Appl. Sci. Biotechnol. Vol 5(3): 293-301