Abstract: Five highly efficient phosphate solubilizing bacteria, viz., Pantoea sp. A3, Pantoea sp. A34, Kosakonia sp. A37, Kosakonia sp. B7 and Bacillus sp. AH9 were isolated from termitorial soils of Sanjivani island of southern Maharashtra, India. These isolates were characterized and explored for phosphate solubilization and plant growth promotion. Among these, Bacillus sp. AH9 showed highest phosphate solubilization index (3.5) and solubilization efficiency (250%) on Pikovskaya agar. Interestingly, Pantoea sp. A34 displayed maximum mineral phosphate solubilization (1072.35 mg/L) in liquid medium and during this period the pH dropped to 3.13. All five isolates had highest P solubilization at 48 h after inoculation. During mineral phosphate solubilization, both gluconic acid and 2-keto gluconic acid were produced by Kosakonia and Bacillus isolates, while only 2-keto gluconic acid was detected in Pantoea isolates. Highest organic acid (39.07 ± 0.04 g/L) production was envisaged in Bacillus sp. AH9, while Pantoea sp. A34 produced the least amount (13.00 ± 0.01 g/L) of organic acid. Seed bacterization with Pantoea sp. A3 and Kosakonia sp. A37 resulted in ~ 37% and ~ 53% increase in root length of tomato seedlings, respectively, while Pantoea sp. A34 and Kosakonia sp. B7 had deleterious effects on root length as well as overall growth of the seedlings. To our knowledge, this is the first report of plant growth promoting potential of microorganisms isolated from termitorial soil of Sanjivani island, which is a drought-prone area. Therefore, such efficient growth promoting P solubilizers can offer an effective solution for sustainable agriculture in arid, dryland farming and drought-prone regions.

Abstract: Plant growth promoting rhizobacteria (PGPR) enhance the plant growth directly by assisting in nutrient acquisition and modulating plant hormone levels, or indirectly by decreasing the inhibitory effects of various pathogens. The aim of this study was to select effective PGPR from a series of indigenous bacterial isolates by plant growth promotion and antifungal activity assays. This study confirmed that most of the isolates from maize rhizosphere were positive for PGPR properties by in vitro tests. Azotobacter and Bacillus isolates were better phosphate solubilizers and producers of lytic enzymes, hydrocyanic acid (HCN), and siderophores than Pseudomonas. Production of indole-3-acetic acid (IAA) and antifungal activity were the highest in Azotobacter, followed by Bacillus and Pseudomonas. The most effective Azotobacter isolates (Azt₃, Azt₆, Azt₁₂) and Bacillus isolates (Bac₁₀, Bac₁₆) could be used as PGPR agents for improving maize productivity. Further selection of isolates will be necessary to determine their efficiency in different soils.

Abstract: Probiotic microorganisms are generally considered to promote the balance of intestinal microbiota and generate health benefits to the host. However, selection of probiotics for incorporation into specific diets requires a scrutiny in the form of both in vitro and in vivo tests. In this work, in vitro probiotic properties of Bacillus subtilis strains available in probiotic formulations were investigated. The isolates CHR01 and FTC01 showed broad antimicrobial spectra, inhibiting Gram-positive bacteria and Aspergillus species, but only Aeromonas hydrophila was inhibited among Gram-negative bacteria. Highest antioxidant activity was measured for isolates KM01 and FTC01. A marked difference in adhesion to hydrocarbons and auto-aggregation properties, ranging from 2.2 to 56.4% and 4.4 to 52.9%, respectively, was observed among the tested bacteria. The Bacillus isolates were mostly tolerant to bile salts and acid pH, although the strains FPR02 and CHR01 were sensitive to ox bile. Among tested cultures, FTC01 and CP01 showed more than 90% survival at pH 2. None Bacillus tested showed positive hemolytic reaction. Four strains were evaluated for surfactin production and higher production was observed by Bacillus FPR01, independently of the substrate.

Abstract: Aim The aim of this study was to evaluate the physicochemical properties of the crude pectinase activity from three Bacillus isolates of ruminant dung origin and study their synergism with crude xylanases from the same Bacillus spp. and a commercial cellulase to evaluate their accessory role in improved biomass saccharification. Methods and results Pectinolytic crude culture filtrate obtained from three ruminant dung isolates, Bacillus safensis M35, Bacillus altitudinis R31 and Bacillus altitudinis J208, on crude pectin containing medium possessed polygalacturonate hydrolase, pectate lyase and pectin lyase activities. Studies regarding their stability under various temperature and pH conditions revealed their mild acidic to alkaline and mesophilic nature with enzyme activity falling within the pH range 6·0-9·0 and temperature range 30-60°C. The pectinase activity was categorized as endolytic as it brought about ~50% reduction in relative viscosity of pectic polymer within initial 10 min of incubation. Synergism of pectinase activity with crude xylanase activities and/or commercial cellulase was clearly demonstrated as ~1·6 to ~1·9-fold increase in agrowaste biomass saccharification was obtained confirming the role of pectinases as accessory enzymes. Conclusion Synergism of the broad-spectrum endopectinase activity obtained from three Bacillus isolates with accessory crude xylanases from the same isolates and commercial cellulase enhanced the agrowaste saccharification and confirmed the accessory role of crude pectinase as they formed an efficient enzyme cocktail functioning in a contributive manner for improvement of agrowaste biomass saccharification. Significance and impact of the study Mesophilic crude endopectinases obtained from Bacillus spp. isolated from ruminant dung possessed activity in broad pH and temperature ranges as well as broad substrate specificity. Moreover, their synergism with crude xylanase and Primfast® 200 cellulase demonstrated the potential to form efficient enzyme cocktail for application in plant biomass saccharification process.

Abstract: Achieving metabolomic data with satisfactory coverage is a formidable challenge in metabolomics because metabolites are a chemically highly diverse group of compounds. The knowledge concerning the behavior of these Bacilli as antagonists and metabolite analysis is essential for their effective use and the commercialization. The present study was focused on selection of best biocontrol antifungal Bacillus strain against aflatoxin producing Aspergillus by antagonism on PDA medium. About 16 different strains of bacteria were isolated from healthy and infested rhizosphere of groundnut using N-agar medium. The isolates were identified based on morphological and microscopic characters. Bacterial isolate JND-KHGn- 29-A and JND-KSGn-30-L were recorded to be a best antagonist as of its ability to inhibit most toxic fungus A. flavus JAM-JKB-BHA-GG20 (58.20%) after screening with 16 Bacillus isolates. GCMS analysis of best and least bacterial antagonist Bacillus subtilis (JND-KHGn-29-A, Accession KU984480) inoculated onto N-agar medium identified total 55 and 42 compounds respectively. Whereas GCMS analysis from best bacterial antagonist Pseudomonas isolate no. 14 (JNDKSGn- 30-L) inoculated onto N-agar identified total 60 compounds.

Abstract: The present disclosure provides isolated Bacillus strains, as well as compositions comprising one or more of the isolated strains and and methods of using the isolated strains and compositions to enhance crop growth/yield.

Abstract: Yanti Y, Warnita, Reflin, Nasution CR. 2018. Characterizations of endophytic Bacillus strains from tomato roots as growth promoter and biocontrol of Ralstonia solanacearum. Biodiversitas 19: 906-911. Bacterial wilt caused by Ralstonia solanacearum is the most damaging vascular pathogens in tomato and many other crops in tropical, subtropical and warm temperate areas of the world limiting its production. Biological agents such as Plant growth Promoting Rhizobacteria (PGPR) is considered as a potential biological control agent for the suppression of plant diseases such as bacterial wilt. Bacillus spp. are one of the most potential genera of PGPR group used for controlling pathogens and promoting plant growth because of their spore-forming ability which increases their adaptation to the environment. The aims of the research were to isolate Endophytic Bacillus isolates, to characterize its ability as plant growth promoter and pathogen controller, and to identify its molecular genetic using 16S rRNA. Bacillus strains were isolated from healthy tomato roots. All Bacillus spp. strains acquired from isolation were then screened directly on plants in completely randomized design experiments with 3 replications. All potential strains were screened and identified using 16S rRNA with 27F and 1492R primers. Results showed that out of 15 obtained isolates, 6 of them showed a good ability to both promote growth and control R. solanacearum. All isolates were identified as B. Pseudomycoides strain NBRC 101232, B. cereus strain CCM 2010, B. toyonensis strain BCT-7112, B. anthracis strain ATCC 14578, B. cereus strain JCM 2152 and B. cereus ATCC 14579.

Abstract: The objective of this work was to evaluate the commercial formulations of Bacillus subtilis QST-713 and Bacillus pumilus QST-2808 on mycelial growth inhibition of Thielaviopsis paradoxa , for the control of the pineapple disease and growth promotion in sugarcane . The inhibition of mycelial growth was evaluated in paired culture tests. Bacillus isolates were transferred to Petri dishes 48 hours before the pathogen, and the zone of inhibition was determined. The direct effect of the autoclaved commercial formulations at 0, 0.001, 0.01, 0.1, 1.0, and 10.0% on the inhibition of pathogen mycelial growth was evaluated. Bacillus spp. potential in controlling the disease and promoting plant growth was evaluated in assays on plant development and disease severity, both under controlled conditions and in the field. Bacillus isolates inhibited pathogen mycelial growth; however, the isolates did not control the disease effectively in the growth chamber and in the field. In the field assay without infestation by the pathogen, all treatments with biocontrol agents and fungicide increased the yield of cane stalks and sugar per hectare, compared with the control. In the assay with infestation by the pathogen, the B. pumilus -based product (2.0 L ha -1 ) and fungicide differed from the control only for the variable number of tillers.

Abstract: Objectives : Screening of antibacterial activity, identification, and L-lactic acid production of spore forming Bacillus isolates. Materials and methods : Fourteen Bacillus isolates were characterized based on their phenotypic characteristics and 16SrRNA gene sequence analyses. The antibacterial activity was determined by agar diffusion method and lactic acid production was analyzed using a high-performance liquid chromatography. Four isolates that produced above 85.0 g/L of lactic acid were selected for lactic acid fermentation using mid log seed culture. Results : All isolates were identified as Bacillus coagulans based on the phenotypic characteristics and 16SrRNA gene sequence similarity (99.02-99.78 %). Strain JC-15 and JC-16 exhibited antibacterial activity against Kocuria rhizophila ATCC 9341 T . All isolates produced L-lactic acid ranged from 56.7± 0.54 to 92.05±0.23 g/L at 37 o C under microaerobic conditions with 99.12±0.1 to 100±0.00 % optical purity. Bacillus coagulans PP-16 showed highest L-lactic acid concentration at 92.05±0.23 g/L, yield of 0.77± 0.00 g/g and productivity of 0.96±0.00 g/L.h. After controlled seed preparation conditions, the four selected strains produced L-lactic acid in the ranged of 93.25±0.07-103.25±0.21 g/L and 102.85±0.49-118.85±0.21 g/L at 37 o C with 98.75±0.00 - 99.70±0.01 and 88.65±0.02 -99.60±0.01 % optical purity under microaerophilic and anaerobic conditions respectively. Yield and productivity were also increased. Fermentation under anaerobic conditions showed higher L-lactic acid titers and yield whereas microaerobic conditions gave higher productivity. Strain PP-16 gave the highest optical purity in both conditions.

Abstract: The objective of the present work was to determine the effect of Bacillus isolates against the phytopatogenic fungi Fusarium oxysporum and Fusarium moniliforme . The antagonistic at seven and fifteen days effect of 14 stains of Bacillus against two Fusarium species was determine. In the antagonistic assay against F. oxysporum , nine bacterial isolates kept the fungal inhibition over 80 % for seven and fifteen days. In the experiment against F. moniliforme , only six isolates showed inhibition percent of over 50 % after 15 days of incubation. In this work the high potential of new isolates of Bacillus as antagonists of Fusarium , phytopatogens of vegetables crops, was demonstrated. RCGr32, RCQ7, and RCGr33 were selected as the best promising isolates for the biological control of these fungi. Key words: biological control, Bacillus , Fusarium oxysporum, Fusarium moniliforme, vegetables.

Abstract: Papaya target-spot, caused by fungus Corynespora cassiicola presents a wide geographic distribution, causing leaf spots in several hosts. The aim of this research was to evaluate the behavior of papaya varieties and/or selections to the natural occurrence of target spot, and test the fungi toxic effect of neem, citronella and eucalyptus extracts, and fungi static effects of Bacillus isolates on the fungus C. cassiicola in vitro. To evaluate the natural occurrence of the disease in field, symptoms on leaves of 11 papaya varieties and/or selections have been assessed. The pathogenicity of isolates was verified by inoculation of plants in greenhouse. Neem, citronella and eucalyptus extracts were prepared at concentrations of 10, 15 and 20% and poured into Petri dishes containing C. cassiicola mycelium discs. For , Bacillusnine isolates were used, and the antagonistic potential was evaluated by the pairing of pathogen and isolate colonies by the circle method. Papaya varieties and/or selections presented all leaves with injury, and Canaan Sunrise Solo and Canaan Golden selections showed the greatest number of injuries per leaf, showing that papaya varieties and/or selections respond differently to the phytopathogen. All plant extracts tested showed inhibitory effect on the mycelial growth of C. cassiicola in vitro. Bacillus isolates showed an antagonistic potential on the mycelial growth of the fungus, with B. methylotrophicus (Iso 31 and Iso 41) being the most promising as a biological control agent. The results indicate that forms of alternative control of diseases are promising in the management of papaya target-spot.

Abstract: Fifty bacterial cultures were isolated from 156 soil samples In order to identify six bacteria in both systems, a fatty acid methyl ester (FAMEs) analysis was conducted, and carbon utilization profiles were assessed, using microbial identification, Biolog Microplac Systems, and the VITEK bacterial identification systems (bioMerieux, Prod No 21341 and 21342) Results showed that four species of Bacillus ssp, isolated from soil, were safe and efficient biological control agent for plant pests in Ordu, Turkey These bacteria were Bacillus mycoides, B cereus, B thuringiensis, Paenibacillus validus, B atrophaeus, and Arthrobacter globiformis Laboratory tests were conducted to assess the potential of the isolates against the great spruce bark beetle, Dendroctonus micans (Kugelann) (Coleoptera: Curculionidae, Scolytinae) (Curculionidae, Scolytinae) Mortality rates of larvae and adults were 60 and 50%, 40 and 30%, and 80 and 70% when using B mycoides, B cereus, and B thuringiensis, respectively P validus, B atrophaeus, and A globiformis showed insufficient rates of mortality, 30 and 20% on the larvae and adults of D micans, respectively Also, these isolates had no antimicrobial effect on pathogen microorganisms They have, however, a lethal effect on some insect groups that are agriculture and forest pests The results indicated that the Bacillus isolates in question can be used as one of the biological control agent

Abstract: The study investigated the genetic basis of the bioremediation activities of two bacterial isolates. Waste engine oil polluted soil was collected from a mechanic workshop and five heterotrophic bacteria were isolated from the soil sample. These were subjected to a preliminary bioremediation experiment by inoculating each of the isolates into a mineral salt medium (MSM) broth, contaminated with waste engine oil. At the end of the experiment, two bacterial isolates, Corynebacterium and Bacillus were selected for genetic study. Plasmid curing was carried out on the selected isolates; plasmid DNA was isolated from them and the extracts were fractionated on 0.8 % agarose gel electrophoresis. Another round of hydrocarbon degradation experiment was set up using varying concentrations (5, 10, 15 and 20 % contamination levels) of waste engine oil as the sole source of carbon source in MSM broth. Data for pH, electrical conductivity (EC) and total hydrocarbon content (THC) were collected at 5 days’ interval for 20 days and statistical analysis was done. The results obtained showed that pH and THC values were decreasing with increase in incubation period, while EC values were increasing with increase in incubation period, suggesting a biodegradation activities going on in cultures of both isolates. There was no significant difference between the values obtained for pH, THC and EC for both the wildtype and plasmid-cured Corynebacterium or Bacillus isolates, which suggested equal rate of bioremediation activities with or without the presence of the plasmid. The results obtained from this study indicate that the gene(s) associated with bioremediation activity may not reside on the plasmid, hence may be chromosomal-based.

Abstract: The genus Bacillus is frequently found in soil, water and food. Bacillus cereus and Bacillus anthracis are the main pathogens causing foodborne diseases and serious infections in humans. A total of 52 Bacillus spp. from meat samples was tested for determination of biofilm production, antimicrobial resistance pattern and beta-lactamase activity. The 24 (46.1%) Bacillus isolates were found to be for biofilm production. Of the 24 (46.1%) biofilm producer Bacillus isolates, 13 (25%), 6 (11.5%) and 5 (9.6%) were considered as strong, moderate and weak biofilm producer, respectively. The most common species for the production of biofilm was Bacillus thuringiensis (80%). Antimicrobial disk susceptibility tests of Bacillus spp. revealed high resistance to ampicillin (84.6%) followed by penicillin (75%), cefepime (34.6%), and cefoxitin (26.9%). A multidrug resistance to at least 3 or more antimicrobials was observed in the 25 isolates (48.1%). All Bacillus spp. were sensitive to vancomycin, gentamicin, amikacin, ciprofloxacin, and imipenem. The susceptibility rate to streptomycin, chloramphenicol, and trimethoprim-sulphamethoxazole was 94.2%. Among the isolates, the 6 (11.5%) isolates were found to be sensitive to all antimicrobial agents tested. Besides, only one isolate from meat was found to be positive for beta-lactamase test. The existence of biofilm production as a virulence factor and of multidrug resistance in bacteria isolated from food should not be underestimated in terms of food safety, public health and economic concerns.

Abstract: Plant growth promoting rhizobacteria (PGPR) are naturally occurring soil bacteria that aggressively colonize plant roots and can enhance plant growth by a wide variety of mechanisms. Bacteria are the most abundant among different rhizospheric microbes. Among Plant growth promoting bacteria, Bacillus is one of the most potential genera due to their spore forming ability, thereby increasing the adaptation of Bacillus strain to commercial formulation and field application In the present study, an attempt was made to isolate, screen and select different isolates of Bacillus and subjected to different test to identify their morpho-physiological, biochemical and In vitro characterization to study their plant growth promoting activities. The Rhizospheric soil samples were collected from different plants i.e. Triticumaestivum (wheat), Cynodondactylon (doobghas), Cicerarietinum (chana), Trifoliumamoenum (bersin) and from different locations of Dehradun and processed to isolate different isolates of Bacillus on nutrient agar medium. A total of 50 Bacillus isolates were isolated from the 22 soil samples. Interestingly, in the present study 17 isolates out of 50 were formed endospores, 5 isolates recorded as capsulated, 35 isolates were reduced the nitrate and all were recorded positive for IAA production. 6 isolates were found to be positive for siderophore production Biochemical characterization revealed that 12 isolates belonged to genus Bacillus belonging to five different species. Isolates NAB-1 seems to be B. alvei. Isolates NAB-8, NAB-11 and NAB-22 seem to be B. laterosporus. Isolates NAB-18, NAB-48, seems to be B. subtillis. Isolates, NAB-20, NAB-21 and NAB-25 seem to be B. larvae. Isolates NAB-23, NAB-43, NAB-44 seem to be B. alcalophilus.

Abstract: Clove rot caused by Fusarium spp. is a very important disease of common garlic (Allium sativum L.) occuring in many areas of the world. However, there is a lack of data about biocontrol of these pathogens. Bacillus species are attractive for research due to their potential use in the biological control of fungal diseases. The aim of this study was to select effective biocontrol agents from a series of indigenous Bacillus spp. isolated from soil. Bacterial isolates positive for hydrolytic enzymes production were screened for antifungal activity against Fusarium spp. isolated from infected garlic cloves. Polymerase chain reaction (PCR) analyses were used for molecular identification of bacterial (16S rDNA gene) and fungal (EF-1α gene) isolates, and detection of biosynthetic genes for antimicrobial lipopeptides (surfactin, iturin, bacillomycin D and fengycin) in Bacillus spp. The obtained results confirmed the presence of Fusarium tricinctum, F. oxysporum f. sp. cepae, F. proliferatum, F. acuminatum and F. verticillioides as the causal agents of garlic clove rot. Four bacterial isolates identified as Bacillus subtilis exhibited the highest antagonistic effect during in vitro testing of antifungal activity (up to 71% reduction in fungal growth), and caused a significant suppression of garlic clove infection (up to 58% reduction in rot symptoms) in situ. Analysis of the antifungal compounds involved in the antagonistic activity of the examined isolates revealed their ability to produce the antibiotic lipopeptide surfactin. The most effective isolates of B. subtilis could be used as potential biocontrol agents of garlic clove rot.

Abstract: Three different biomaterials viz., boiled cow milk, compost manure and tomato rhizospheric soil were found as habitats of the thermophilic antagonistic bacteria. The isolated bacteria were able to grow satisfactorily at thermophilic temperature range (>55i‚°C). Based on morphological, biochemical and physiological characters, the bacterial isolates were identified as Bacillus licheniformis (boiled cow milk), and Bacillus stearothermophilus (compost manure and tomato rhizospheric soil). All the three thermophilic bacterial isolates exhibited strong antagonism against tested soil-borne fungal plant pathogens in order of B. lechaniformis (inhibition zone of 67.67 mm against R. bataticola) > B. stearothermophilus from compost manure (51.67 mm against R. solani) > B. stearothermophilus from tomato rhizospheric soil (38.33 mm against P. aphanidermatum). The ability to tolerate high temperature (>55i‚°C), pH (6-8) and salt concentrations (up to 8%), and antibiotic resistance properties of the antagonistic thermophilic Bacillus isolates may hold them as potential biocontrol candidates, especially under stressed rhizosphere environments where other biocontrol agents fail. However, the results need further confirmation under field conditions where these bioagents will be applied in a formulated form. Key words: Antagonism, antibiotic sensitivity, biocontrol activity, soil borne pathogens, thermophilic bacteria.

Abstract: Although Bacillus cereus is of particular concern in food safety and public health, the role of other Bacillus species was overlooked. Therefore, we investigated the presence of eight enterotoxigenic genes, a hemolytic gene and phenotypic antibiotic resistance profiles of Bacillus species in retail meat samples. From 255 samples, 124 Bacillus isolates were recovered, 27 belonged to B. cereus and 97 were non-B. cereus species. Interestingly, the non-B. cereus isolates carried the virulence genes and exhibited phenotypic virulence characteristics as the B. cereus. However, correlation matrix analysis revealed the B. cereus group positively correlates with the presence of the genes hblA, hblC, and plc, and the detection of hemolysis (p < 0.05), while the other Bacillus sp. groups are negatively correlated. Tests for antimicrobial resistance against ten antibiotics revealed extensive drug and multi-drug resistant isolates. Statistical analyses didn’t support a correlation of antibiotic resistance to tested virulence factors suggesting independence of these phenotypic markers and virulence genes. Of special interest was the isolation of Paenibacillus alvei and Geobacillus stearothermophilus from the imported meat samples being the first recorded. The isolation of non-B. cereus species carrying enterotoxigenic genes in meat within Egypt, suggests their impact on food safety and public health and should therefore not be minimised, posing an area that requires further research.