Lysinibacillus Fusiformis Research Articles

Selection of probiotic bacteria for use in shrimp larviculture

Three candidate probiotics, Bacillus foraminis, Bacillus cereus biovar toyoi and Bacillus fusiformis, were isolated from hydrogen-producing fermented solution and identified using 16S rRNA sequence analysis. Bacillus foraminis and B. cereus biovar toyoi exhibited strong antagonism against Streptococcus iniae and Photobacterium damselae subsp. piscicida in in vitro co-culture for competitive exclusion assay and then were conducted in the larviculture system of Penaeus monodon reared from zoea 1 to postlarva 1. The daily addition of B. cereus biovar toyoi resulted in significantly deleterious effects on survival (P<0.01) whereas the daily addition of B. fusiformis showed highest survival rate (88.7±0.7%) but no statistically significant difference from control (73.3±12.1%). Bacillus fusiformis was continuously applied in the larviculture system of Litopenaeus vannamei. Administration of B. fusiformis significantly increased survival (P<0.01) in both treatments added daily (87.9±1.7%) and every other day (54.7±1.2%), respectively, at a concentration of 105 CFU mL−1 over control (41.2±1.3%).

Metal adsorption capabilities of clinoptilolite and selected strains of bacteria from mine water

Small-scale mining has socio-economic advantages such as the reduction of unemployment and the general improvement of the economy. However, these operations if not properly managed or controlled have a potential to cause environmental damage, particularly with respect to the contamination of groundwater and water supplies that are not distant from where these mining activities take place. This paper focuses on metal removal from water contaminated by heavy metals emanating from small-scale mining operations using clinoptilolite and bacteria. Removal of As, Ni, Mn, Au, Co, Cu and Fe was carried out on mine water samples using original and HCl-activated (in 0.02 M and 0.04 M) natural clinoptilolite and bacterial strains (a mixed consortia of Bacillus strains ( Bacillus subtilis, Bacillus cereus, Bacillus firmus, Bacillus fusiformis, Bacillus macroides and Bacillus licheniformis), Pseudomonas spp., Shewanella spp. and a mixed consortia of Acidithiobcillus caldus, Leptospirillum spp., Ferroplasma spp. and Sulphobacillus spp.). The purpose of the study was to compare the removal efficiencies of the bacterial strains versus natural clinoptilolite adsorbents for metal cations. The Bacillus consortia removed most of the metals up to 98% metal removal efficiency with the exception of nickel where clinoptilolite showed good removal efficiency. The 0.02 M HCl-activated clinoptilolite also demonstrated excellent removal capabilities with Cu, Co and Fe removal efficiency of up to 98%. Both clinoptilolite and bacteria demonstrated capabilities of removing Cu 2+, Co 2+, Fe 2+, Mn 2+, As 3+ and Au from solution which augurs well for metal recovery from mining and mineral processing solutions, as well as in water decontamination.

Decolorization and degradation of Disperse Blue 79 and Acid Orange 10, by Bacillus fusiformis KMK5 isolated from the textile dye contaminated soil

The release of azo dyes into the environment is a concern due to coloration of natural waters and due to the toxicity, mutagenicity and carcinogenicity of the dyes and their biotransformation products. The dye degrading bacterial strain KMK 5 was isolated from the textile dyes contaminated soil of Ichalkaranji, Maharashtra, India. It was identified as Bacillus fusiformis based on the biochemical and morphological characterization as well as 16S rDNA sequencing. KMK 5 could tolerate and degrade azo dyes, Disperse Blue 79 (DB79) and Acid Orange 10 (AO10) under anoxic conditions. Complete mineralization of DB79 and AO10 at the concentration of 1.5 g/l was observed within 48 h. This degradation potential increased the applicability of this microorganism for the dye removal.

Evaluation of In vitro Cr(VI) reduction potential in cytosolic extracts of three indigenous Bacillus sp. isolated from Cr(VI) polluted industrial landfill

Three efficient Cr(VI) reducing bacterial strains were isolated from Cr(VI) polluted landfill and characterized for in vitro Cr(VI) reduction. Phylogenetic analysis using 16S rRNA gene sequencing revealed that the newly isolated strains G1DM20, G1DM22 and G1DM64 were closely related to Bacillus cereus, Bacillus fusiformis and Bacillus sphaericus, respectively. The suspended cultures of all Bacillus sp. exhibited more than 85% reduction of 1000 μM Cr(VI) within 30 h. The suspended culture of Bacillus sp. G1DM22 exhibited an ability for continuous reduction of 100 μM Cr(VI) up to seven consecutive inputs. Assays with the permeabilized cells and cell-free extracts from each of Bacillus sp. demonstrated that the hexavalent chromate reductase activity was mainly associated with the soluble fraction of cells and expressed constitutively. The Cr(VI) reduction by the cell-free extracts of Bacillus sp. G1DM20 and G1DM22 was maximum at 30 °C and pH 7 whereas, Bacillus sp. G1DM64 exhibited maximum Cr(VI) reduction at pH 6. Addition of 1 mM NADH enhanced the Cr(VI) reductase activity in the cell-free extracts of all three isolates. Amongst all three isolates tested, crude cell-free extracts of Bacillus sp. G1DM22 exhibited the fastest Cr(VI) reduction rate with complete reduction of 100 μM Cr(VI) within 100 min. The apparent K m and V max of the chromate reductase activity in Bacillus sp. G1DM22 were determined to be 200 μM Cr(VI) and 5.5 μmol/min/mg protein, respectively. The Cr(VI) reductase activity in cell-free extracts of all the isolates was stable in presence of different metal ions tested except Hg 2+ and Ag +.

Microbial combinatorics: a simplified approach for isolating insecticidal bacteria

Bacteria can control pest insects that damage food crops, vector diseases and defoliate trees. Conventionally, isolation of these bacteria has been from soil and sporadically from dead insects. A simplified approach for isolating insecticidal bacteria from soil using the target insect as the selective agent was employed in this study. Instead of isolating single strains of bacteria from soil and testing each individual strain for insect toxicity, mixtures of bacteria present in each soil sample were tested together directly for toxicity using Manduca sexta (Linnaeus) (Lepidoptera: Sphingidae) as a model insect. Thirty-five soil suspensions or bacterial suspensions of the 40 suspensions tested killed at least one M. sexta larva. All but one bacterial culture isolated from dead larvae and retested for toxicity, killed at least one M. sexta larva. Nineteen bacterial strains isolated from larvae killed in the first test, were identical to the bacteria fed to the retested larvae. Of the 19 strains isolated, 14 were identified by 16S rDNA sequencing as belonging to the Bacillus cereus group including three strains that formed crystals that were identified as B. thuringiensis. Of the three other spore-forming strains, two were identified as psychrotrophic B. weihenstephanensis and the third as Lysinibacillus fusiformis. Two others were identified as Enterococcus faecalis. This approach, microbial combinatorics, reduces the number of insects necessary for toxicity screening and associated time and resources compared to conventional methods that first isolate bacteria and then individually test for toxicity as well as a means of discovery of new pathogens using the insect as the selective agent.

Analysis of rhizosphere bacteria of rice cultivated in Andosol lowland and upland fields using molecular biological methods

Bacteria in the rhizosphere influence plant growth and interact with plant roots. Microscopy and culture method have been used for studies of microorganisms of the rhizosphere, but these methods are insufficient for evaluation because most rhizosphere bacteria are viable but non-culturable (VBNC). Bacteria in the rhizosphere of rice cultivated in Andosol lowland and upland fields were analyzed in this study using PCR-DGGE and FISH, in combination with modified pretreatments. Results show that the two methods with the pretreatments, more than conventional methods, provided a rapid and simple analysis of rhizosphere bacteria. The 16S rDNA band pattern of bacteria in the rhizosphere obtained using PCR-DGGE indicated different species composition of bacterial community in the two ecosystems and greater diversity of bacteria in the rhizosphere in upland field. Sequencing of major 16S rDNA bands identified Bacterium A35 and Clostridium bifermentans as dominant bacteria in the rhizosphere of rice in lowland fields and Klebsiella planticola and Bacillus fusiformis in upland fields. Furthermore, FISH observation indicated the predominance of gram-positive low GC bacteria in both rhizospheres and a higher proportion of Clostridium spp. in lowland fields, which is consistent with results of PCR-DGGE analysis. The results suggest that the bacteria in the rice rhizosphere can be changed depending on aerobic and anaerobic conditions of fields. It is expected to apply the PCR-DGGE and FISH to agricultural field experiments as reliable methods to evaluate the rhizosphere bacteria.

Isolation and algae-lysing characteristics of the algicidal bacterium B5

Water blooms have become a worldwide environmental problem. Recently, algicidal bacteria have attracted wide attention as possible agents for inhibiting algal water blooms. In this study, one strain of algicidal bacterium B5 was isolated from activated sludge. On the basis of analysis of its physiological characteristics and 16S rDNA gene sequence, it was identified as Bacillus fusiformis. Its algaelysing characteristics on Microcystis aeruginosa, Chlorella and Scenedesmus were tested. The results showed that: (1) the algicidal bacterium B5 is a Gram-negative bacterium. The 16S rDNA nucleotide sequence homology of strain B5 with 2 strains of B. fusiformis reached 99.86%, so B5 was identified as B. fusiformis; (2) the algal-lysing effects of the algicidal bacterium B5 on M. aeruginosa, Chlorella and Scenedesmus were pronounced. The initial bacterial and algal cell densities strongly influence the removal rates of chlorophyll- a. The greater the initial bacterial cell density, the faster the degradation of chlorophyll- a. The greater the initial algal cell density, the slower the degradation of chlorophyll- a. When the bacterial cell density was 3.6 fi 10 7 cells/ml, nearly 90% of chlorophyll- a was removed. When the chlorophyll- a concentration was less than 550 μg/L, about 70% was removed; (3) the strain B5 lysed algae by secreting metabolites and these metabolites could bear heat treatment.

Biotransformation of isoeugenol to vanillin by Bacillus fusiformis CGMCC1347 with the addition of resin HD-8

A novel strain, Bacillus fusiformis CGMCC1347, with high vanillin yield from isoeugenol, was isolated from soil. This strain was utilized successfully to transform isoeugenol to vanillin and the product inhibition could be well avoided by addition of resin HD-8. After 72 h reaction, the vanillin concentration reached 8.10 g l −1 vanillin from 50 g l −1 isoeugenol under the condition of adding 12.5 g resin HD-8 in 20 ml reaction solution at pH 7.0, 37 °C and 180 rpm. Isoeugenol and vanillin in HD-8 could be well separated. Vanillin separation yield and isoeugenol recovery were 78.4% and 69.9%, respectively.

Alkane Utilization by Rhodococcus Strain NTU‐1 Alone and in Its Natural Association with Bacillus fusiformis L‐1 and Ochrobactrum sp.

Linear (n-hexadecane) and branched (pristane) alkanes were degraded by a mixed culture isolated from an oil-contaminated field. The degradation was accompanied by formation of biofloccules. The culture was composed of Rhodococcus strain NTU-1, Bacillus fusiformis L-1, and Ochrobactrum sp. Rhodococcus strain NTU-1 carried out the degradation of the alkane via a hydroxylase. Bacillus fusiformis L-1 and Ochrobactrum sp. did not degrade the alkanes but aided the flocculation by forming more rigid bacterial aggregates that enhanced the trapping of alkanes. In batch cultures, transformation and removal of the linear and branched alkanes was achieved within 66 h with more than 95% efficiency.

Inhibition of the American foulbrood bacterium, Paenibacillus larvae larvae, by bacteria isolated from honey bees

SUMMARYWhile honey bees (Apis mellifera) support a diverse microbial community, the impacts of most of their associated microbes on honey bee health remain unresolved. Here, pairwise inhibition assays were used to identify honey bee bacterial symbionts that inhibit a primary pathogen, the Gram-positive bacterium Paenibacillus larvae larvae. Four bacterial taxa isolated from bee larvae appeared especially promising with respect to inhibition of P. l. larvae and, in fact, completely inhibited P. l. larvae growth in pairwise plate assays. These isolates were identified by 16S ribosomal RNA sequencing as Stenotrophomonas maltophilia, Acinetobacter sp., Brevibacillus formosus and Bacillus fusiformis. A PCR-based survey confirmed that these bacterial isolates are present in bee larvae, at frequencies ranging from 2% (1/48) for B. formosus to 79% (39/48) for Acinetobacter sp. An understanding of the distributions of these co-occurring bacteria could elucidate variation across colonies in susceptibility to American foulbrood disease. In addition, supplementation of colonies with these naturally occurring bacteria or their antagonistic products can provide a novel way of controlling foulbrood disease.

Isolation and characterization of diazotrophic growth promoting bacteria from rhizosphere of agricultural crops of Korea

Free-living nitrogen fixing bacteria were isolated from rhizosphere of seven different plant namely sesame, maize, wheat, soybean, lettuce, pepper and rice grown in Chungbuk Province, Korea. Five isolates with nitrogenase activity above 150nmol(-1) mg(-1) protein were identified based on, phenotypic and 16S rDNA sequences analysis. The strains were identified as Stenotrophomonas maltophilia (PM-1, PM-26), Bacillus fusiformis (PM-5, PM-24) and Pseudomonas fluorescens (PM-13), respectively. All the isolates produced indole-3-acetic acid (IAA), in the presence of tryptophan, ranging from 100.4 microg ml(-1) (PM-13) to 255 microg ml(-1) (PM-24). The isolate PM-24 (Bacillus fusiformis) exhibiting highest nitrogenase activity (3677.81 nmol h(-1) mg(-1) protein) and IAA production (255microg ml(-1)) has a promising potential for developing as a plant growth promoting rhizobacteria.

Phylogeny of Bacillus sphaericus-like organisms.

The mesophilic round-spored bacteria embrace four species, namely Bacillus sphaericus, Bacillus fusiformis, Bacillus silvestris and Bacillus pasteurii. Although not displayed by all strains, mosquito pathogenicity is a noteworthy characteristic of B. sphaericus sensu lato. Phylogenetic analysis based on 16S rDNA sequences from 58 strains identified as B. sphaericus was used to examine the genetic heterogeneity of the taxon. Results from sequence analysis were compared with whole-cell fatty acid profiles and other phenotypic determinations. The B. sphaericus-like strains segregated into seven distinct clusters in a phylogenetic tree generated from 16S sequences. One cluster represented B. sphaericus and another B. fusiformis. A third cluster containing all of the pathogenic strains was closely related to, or was possibly part of, the B. fusiformis group. The remaining four groups were distinct and represented unnamed taxa that were more closely related to B. sphaericus and B. fusiformis than to the psychrophilic round-spored species, Bacillus globisporus and Bacillus psychrophilus. Groups based on phenotypic analysis corresponded to the 16S rDNA phylogenetic clusters. Data showed that B. sphaericus was genetically and phenotypically a highly heterogeneous taxon including at least seven genetically distinct taxa. The pathogenic strains were members of a distinct group and not of the species B. sphaericus sensu stricto. This heterogeneity partially accounts for the apparent variability of mosquito pathogenicity among B. sphaericus strains.

A numerical classification of the genus Bacillus.

Three hundred and sixty-eight strains of aerobic, endospore-forming bacteria which included type and reference cultures of Bacillus and environmental isolates were studied. Overall similarities of these strains for 118 unit characters were determined by the SSM, SJ and DP coefficients and clustering achieved using the UPGMA algorithm. Test error was within acceptable limits. Six cluster-groups were defined at 70% SSM, which corresponded to 69% SP and 48-57% SJ. Groupings obtained with the three coefficients were generally similar but there were some changes in the definition and membership of cluster-groups and clusters, particularly with the SJ coefficient. The Bacillus strains were distributed among 31 major (4 or more strains), 18 minor (2 or 3 strains) and 30 single-member clusters at the 83% SSM level. Most of these clusters can be regarded as taxospecies. The heterogeneity of several species, including Bacillus brevis, B. circulans, B. coagulans, B. megateriun, B. sphaericus and B. stearothermophilus, has been indicated and the species status of several taxa of hitherto uncertain validity confirmed. Thus on the basis of the numerical phenetic and appropriate (published) molecular genetic data, it is proposed that the following names be recognized; Bacillus flexus (Batchelor) nom. rev., Bacillus fusiformis (Smith et al.) comb. nov., Bacillus kaustophilus (Prickett) nom. rev., Bacillus psychrosaccharolyticus (Larkin & Stokes) nom. rev. and Bacillus simplex (Gottheil) nom. rev. Other phenetically well-defined taxospecies included 'B. aneurinolyticus', 'B. apiarius', 'B. cascainensis', 'B. thiaminolyticus' and three clusters of environmental isolates related to B. firmus and previously described as 'B. firmus-B. lentus intermediates'. Future developments in the light of the numerical phenetic data are discussed.

BACTERIOLOGICAL STUDIES ON TYMPANOPLASTY

Bacteriological studies were carried out for the purpose of determining the kinds of bactaria detectable before, during and after tympanoplasty as well as to find out their survival rate on 55 cases (61 ears) with chronic otitis media who had been treated in our clinics during the two-year period from July 1958 to June 1960. The following are the results.1. Out of the 98 strains of bacteria detected before the operation, the most numerous were Staphylococcus aureus (16 strains), followed by Pseudomonas aeruginosa (12 strains), Diphtheroid (12 strains), Diplococus pneumoniae (8 strains), Pneumococcus mucosus (7 strains.) and Micrococcus (7 strains) in the order mentioned.2. In connection with the preoperative detection of bacteria, mixed infection was found to be a two-fold simple infection. As for the different kinds of bacteria, simple infection by Staphylococus aureus was twice as frequent as by other bacteria, while in the cases of infection by the other bacteria the majority proved to be of mixed infection.3. Simple infection decreased during and after the operation, whereas mixed infection, although it did decrease during the operation, increased again postoperatively.4. Staph. aureus completely disappeared during or after the operation, while Pseudomonas aeruginosa, Diplococcus pneumoniae, and Diphtheroid proved to be persistent, not decreasing so readily during or after the operation. Therefore, in the cases where these bacteria are detected prior to tympanoplasty, every possible care needs to be taken for the disinfection during the operation.5. Of the detectable bacteria, Gram-positive ones were twice as numerous as the Gram-negative.6. Such saprophytes as Bacillus fusiformis, spore forming bacteria and Serratia marcescens were also detected, although they were very small in number.7. Those that could not be detected at all were Mycobacterium tuberculosa, Haemophilus influenzae and true Fungi such as Candid.

ELECTRON-MICROSCOPICAL STUDIES ON BACILLUS FUSIFORMIS, ESPECIALLY ON ITS FLAGELLA

A kind of “gram negative” fusiform obligate aerobic bacteria was isolated from tooth coating of a normal mouth and was identified as Bacillus fusiformis, for its various biological characters.This bacterium was studied electron-microscopically, by using emulsion specially prepared to avoid the morphological changes of the cultured bacteria.The most interesting result was the discovery of the existence of flagella by some strains of B. fusiformis. (Fig. 3)According to the observation, several flagella exist irregulary around the bacterial body, so apparently this bacterium does not belong to Monotricha, Amphytricha or Lophotricha, but Peritricha.(This is the briefest synopsis of the publication made at the 5th Annual Meeting of the Japanese Stomatological Society in April 1951)

Decontaminating solution

Effects associated with aflatoxins (AFs), principally aflatoxin B1 (AFB1) have necessitated strategies to eliminate their occurrence in commodities along the food chain. This study therefore, investigated the AFB1 biodegradation ability of Staphylococcus warneri, Sporosarcina sp. and Lysinibacillus fusiformis liquid cultures and cell lysates (disrupted in the presence or absence of protease inhibitors to obtain lysates). These were incubated with AFB1 (2.5 μg/mL) for 3, 6, 12, 24 and 48 h. AFB1 degradation was subsequently monitored on high performance liquid chromatography (HPLC) and results indicated that after 48 h, % AFB1 degradation by the liquid cultures of Lysinibacillus fusisormis, S. warneri and Sporosarcina sp. were 61.3, 47.7 and 46.9%, respectively. After 12 h of incubation, a 100% AFB1 degradation was observed for all protease inhibited lysates tested. To establish toxicity of the AFB1 biotransformed products, results from a cytotoxicity study against human lymphocytes demonstrated that the products exhibited significantly (p ≤ 0.05) lower cytotoxic effect compared to the parent AFB1. From this study, it can be deduced that the mechanism of AFB1 degradation was enzymatic and that protease inhibition of cells before disruption, could increase this enzymatic activity. Conclusively, the potential of these lysates as a biotechnological approach towards decontaminating AFB1 is promising.

Morphological Changes in Bacillus fusiformis

IN a communication in Nature1, Webster and Frey have reported changes in the morphology of the Bacillus fusiformis found in cases of ulcerative gingivitis after the application of penicillin. The organisms had developed central round or spindle-shaped swellings. In an investigation of tropical ulcer in Madras, I have, however, noted in smears from untreated cases similar changes in the morphology of the fusiform bacillus. The central swelling is seen both by dark-ground illumination and after staining. It would appear, therefore, that the changes in morphology are due to the influence of other factors besides any specific treatment.

Inoculation Experiments withBacillus fusiformisisolated from Tropical Ulcer with Observations on the Bacillus

The isolation of fusiform bacilli from typical tropical ulcer is described and an account is given of the motility and staining properties of the organisms cultivated.Experimental inoculations, both in man and animals, are recorded in tabular form and the results of the investigation discussed.

Tropical Ulcer: The Distribution of Bacillus Fusiformis and Spirochæta Schaudinni in the Lesion

Tropical Ulcer: The Distribution of Bacillus Fusiformis and Spirochæta Schaudinni in the Lesion

An Unidentified Organism Exhibiting Cultural Characteristics Similar to those of the Bacillus Fusiformis

An Unidentified Organism Exhibiting Cultural Characteristics Similar to those of the Bacillus Fusiformis