The API 20B microtube system to aid in the identification of N2-fixing Bacillaceae

1987 ◽  
Vol 33 (6) ◽  
pp. 504-509 ◽  
Author(s):  
R. J. Rennie
Keyword(s):  
Heterotrophic Bacteria ◽  
General Procedure ◽  
Bacillus Species ◽  
Computer Assisted ◽  
Growth Media ◽  
Natural Environments ◽  
Bacterial Isolates ◽  
Biochemical Tests ◽  
Isolation And Identification ◽  
Carbohydrate Utilization

N2-fixing bacterial isolates from soil have been identified successfully using the API 20E and 50E microtubes in conjunction with a computer-assisted biochemical profile search. The 20E and 50E result in few positive tests with N2-fixing Bacillaceae. Recently, a new battery of 20 microtube tests, the 20B, has been introduced for identifying aerobic, heterotrophic bacteria isolated from natural environments. This system was compared with the 20E and evaluated for N2-fixing bacillus species from soil. It was then used in conjunction with guanine-plus-cytosine analysis to identify the C-11-25 (5DN+) bacillus strain that fixes N2 in association with Canadian wheat cultivars. The API 20E and 20B gave similar results for many biochemical tests, but the 20E resulted in negative tests for carbohydrate utilization by bacillus strains. The use of different growth media or 0.85% NaCl to suspend these bacteria prior to inoculating the cupules did not alter the inability of the 20E in this manner. Carbohydrate utilization in the 20B system agreed well with that in the traditional utilization tests. Hence, I concluded that the API 20E is not suitable to evaluate carbohydrate utilization in N2-fixing Bacillaceae. The spore-forming, acetylene-reducing C-11-25 strain of bacillus was biochemically similar to the ATCC type culture of Bacillus polymyxa (strain 842). Its guanine-plus-cytosine ratio of 48.0 mol% was similar to that of B. polymyxa (43–46 mol%). A general procedure for the isolation and identification of N2-fixing Bacillus from soil is proposed.

scholarly journals SUSCEPTIBILITY PROFILES TO ANTIBIOTICS IN STRAINS OF THE GENUS BACILLUS ISOLATED FROM EXTREME AQUATIC ENVIRONMENTS OF ECUADOR

2021 ◽  
pp. 27-34
Author(s):  
Marco González ◽  
Francisco Viteri ◽  
Luis Villacís ◽  
Jessica Escobar ◽  
Liliana Araujo ◽  
...  
Keyword(s):  
Membrane Filtration ◽  
Sample Volume ◽  
Diffusion Method ◽  
Bacillus Species ◽  
Natural Environments ◽  
Biochemical Tests ◽  
Volcanic Lake ◽  
Antibiotic Resistance Profile ◽  
High Concentrations ◽  
Susceptibility Profiles

Introduction. The water of aquatic ecosystems considered extreme, given the values of its physicochemical and chemical parameters, such as high concentrations of salts, oligotrophic environments, extreme pH, high radiation and extreme temperatures, there is a bacterial population that has adapted to these conditions and that they can be an important reservoir of natural resistomes. Objective. The objective of the present work was to know the profiles of susceptibility to various antibiotics in strains of the Bacillus genus isolated from mineromedicinal water spas and water from a volcanic crater lake in Ecuador. Materials and methods. A total of 16 mineromedicinal water samples and 32 samples of crater volcanic lake water were analyzed. The isolation of the Bacillus colonies was carried out by the membrane filtration technique, using Millipore filters of 0.45 μm pore, a sample volume of 100 mL and R2A agar. The isolated strains were identified following the schemes of MacFaddin (2004), complemented with the biochemical tests of the Microgen galleries for Bacillus. The antibiotic resistance profile was determined by the plate diffusion method of Kirby and Bauer (1966), interpreted according to the CLSI (2019). Results. 19 Bacillus strains were identified. Most of the strains were resistant and multi-resistant to the antibiotic clindamycin, erythromycin, gentamicin, oxacillin, and penicillin. Conclusions. The results indicate the presence of Bacillus species and resistomes associated with this genus in the water of extreme natural environments in Ecuador, which suggests that these environments may be an important reservoir of bacteria resistant to antibiotics.

scholarly journals Isolation and Identification of Elite Phosphate Solubilizing Bacteria from Soil under Paddy Cultivation

2014 ◽  
Vol 16 ◽  
pp. 62-68 ◽  
Author(s):  
A. Gandhi ◽  
G. Muralidharan ◽  
E. Sudhakar
Keyword(s):  
Bacterial Species ◽  
Essential Element ◽  
Broth Culture ◽  
Bacillus Species ◽  
Available Phosphorus ◽  
Growth Media ◽  
Screening Methods ◽  
Phosphate Solubilizing Bacteria ◽  
Isolation And Identification ◽  
Phosphate Solubilizing

A considerable number of bacterial species are able to exert a beneficial effect upon plant growth. Mostly they are associated with the plant rhizosphere, so they are called as rhizobacteria. Phosphorus is an essential element for plant development and growth making up about 0.2 % of plant dry weight. Several scientists have reported the ability of different bacterial species to solubilize insoluble inorganic phosphate compounds, such as tricalcium phosphate, dicalcium phosphate, hydroxyapatite, and rock phosphate. Detection and estimation of the phosphate solublization ability of microorganisms have been possible using plate screening methods. Phosphate solubilizers produce clearing zones around the microbial colonies in growth media. In the present investigation a total number of fifteen phosphate solubilizing bacterial colonies isolated from different paddy soils in Cuddalore district of Tamilnadu, India. The isolated PSB were identified and characterized for effective use in the field. All the PSB isolates were identified as Bacillus species and designated as P with serial number from 1 to 15. Among the fifteen isolates, the PSB isolate P6 showed highest amount of phosphate solubilization. The quantity of available phosphorus estimated in the P6 grown Sperber broth culture medium on 7th day was maximum of 321.7 μg/ml which was the highest value compared to other PSB isolates.

Isolation and identification of nitrogen-fixing Enterobacter cloacae and Klebsiella planticola associated with rice plants

1983 ◽  
Vol 29 (10) ◽  
pp. 1301-1308 ◽  
Author(s):  
J. K. Ladha ◽  
W. L. Barraquio ◽  
I. Watanabe
Keyword(s):  
Heterotrophic Bacteria ◽  
Enterobacter Cloacae ◽  
Dry Weight ◽  
Nitrogen Fixing ◽  
Biochemical Tests ◽  
Isolation And Identification ◽  
Cross Reactions ◽  
Rice Plants ◽  
Klebsiella Planticola ◽  
Percentage Incidence

Acid- and gas-producing nitrogen-fixing bacteria associated with rice roots and leaf sheaths were isolated. These isolates along with reference enterobacteria strains were characterized biochemically and serologically. Using selected cultural and biochemical tests, all isolates from rice were identified as Enterobacter cloacae, except two strains which were found similar to Klebsiella pneumoniae. However, further biochemical tests showed that the Klebsiella isolates were K. planticola. Antisera and fluorescent antibodies (FA) were prepared against K. pneumoniae M5a1, K. planticola DWUL2, and E. cloacae EnSs. The results of FA cross-reactions of 28 strains isolated from rice plants and of other type cultures showed a separation into two different serogroups: E. cloacae and K. planticola. However, the cross-reactions in gel immunodiffusion were found to be extensive and widespread. All strains of Enterobacteriaceae tested produced at least one immunodiffusible precipitin band with any one or all antisera. The population of N2-fixing enterobacteria associated with the root and stem of rice fell within the range of 103 and 105 per gram dry weight. The percentage incidence of N2-fixing enterobacteria among the aerobic heterotrophic bacteria in the root and stem was found to be less than 1%.

scholarly journals Occurrence of hydrocarbon degrading bacteria in soil in Kukawa, Borno State

2014 ◽  
Vol 3 (2) ◽  
pp. 36-47 ◽  
Author(s):  
IA Allamin ◽  
UJJ Ijah ◽  
HY Ismail ◽  
ML Riskuwa
Keyword(s):  
Crude Oil ◽  
Heterotrophic Bacteria ◽  
Bacterial Species ◽  
Petroleum Exploration ◽  
Bacterial Isolates ◽  
Biochemical Tests ◽  
As Species ◽  
Degrading Bacteria ◽  
Different Depths ◽  
Hydrocarbon Utilizing Bacteria

Soil samples were collected from five sites covering petroleum exploration station in Kukawa, Kukawa Local Government Area of Borno State, Nigeria between October, 2012 and February, 2013 at two different depths (0-10cm and 10-20cm) to enumerate and identify hydrocarbon degrading bacteria in the soil. Total aerobic heterotrophic bacteria (TAHB) were enumerated on Nutrient agar (NA), and Hydrocarbon utilizing bacteria (HUB) enumerated on Oil agar (OA). The bacterial isolates were identified using morphological and biochemical tests. It was observed that the microorganisms (TAHB, and HUB) were more densely populated at 10cm depth. (TAHB: 5.3×108 - 11.4×108cfu/g, and HUB: 2.4×105 - 5.3×105 cfu/g, than at 20 cm depth (TAHB: 3.0×108 - 5.7×108 cfu/g, and HUB: 2.1×105 - 4.8×105 cfu/g). The HUB was identified as species of Bacillus, Pseudomonas, Klebsiella, Lactobacillus, Micrococcus, Corynebacterium, and Actinomyces. Bacillus, and Pseudomonas species were more constantly isolated than other isolates and they constitute 100% of total bacterial isolates. The potential of hydrocarbon utilizing bacteria isolated to degrade hydrocarbon was studied. Nineteen (19) bacterial species was screened, Bacillus subtilis, Pseudomonas aeruginosa, Bacillus cereus, Klebsiella pneumoniae, Micrococcus leteus,and Lactobacillus casei, utilized and degrade crude oil at considerably high rates after 21 days of incubation. The degradation efficiency was confirmed by GC-MS analysis, which indicated that the bacterial isolates utilized most of the crude oil components particularly straight chain alkanes and cycloalkanes DOI: http://dx.doi.org/10.3126/ije.v3i2.10503 International Journal of the Environment Vol.3(2) 2014: 36-47

Biochemical Methods for Automated Bacterial Identification and Testing Metabolic Activities in Water and Wastewater

1988 ◽  
Vol 20 (11-12) ◽  
pp. 221-227 ◽  
Author(s):  
W. Dott ◽  
P. Kämpfer
Keyword(s):  
Heterotrophic Bacteria ◽  
Carbon Substrate ◽  
Computer Assisted ◽  
Identification System ◽  
Bacterial Identification ◽  
Test Results ◽  
Biochemical Tests ◽  
Conventional Tests ◽  
Water And Wastewater ◽  
Metabolic Activities

The improvements of computer-assisted identification were used to develop a new microplate system for characterization and identification of various gram-negative and gram-positive heterotrophic bacteria from the environment. In standard microtitration plates about 90 biochemical tests, some of them conventional tests, carbon substrate assimilation tests and enzyme tests using chromogenic substrates can be performed. Reading of the test results is done automatically by a photometer coupled to a computer. The applications of this identification system are shown by two waste water samples.

scholarly journals ISOLATION AND IDENTIFICATION OF MICROORGANISMS FROM SOME SPOILED FRUITS AND VEGETABLES COLLECTED FROM NAGAON, ASSAM

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Dipanjali Devi ◽  
Rupjyoti Gogoi
Keyword(s):  
Fruits And Vegetables ◽  
Sufficient Condition ◽  
Bacterial Isolates ◽  
Biochemical Tests ◽  
Isolation And Identification ◽  
Great Loss ◽  
Local Markets

Fruits and vegetables provide sufficient condition for the growth of several fungi and bacteria Microorganisms spoil the fruits and vegetables and changes the texture and makes them uneatable. There is a heavy loss of fruits and vegetables due to spoilage by microorganisms; however during handling and transportation also, there is a greater risk to get damaged .Thus farmers suffered a great loss due to spoilage. In the present study a total of about 3 spoiled fruits and 3spoiled vegetables were taken from some local markets of Nagaon, from which fungal and bacterial isolates were isolated and identified. Biochemical tests for the isolated bacteria were also done.

scholarly journals Biofilm production as a virulence factor in Uropathogenic bacteria and yeasts

2013 ◽  
Vol 7 (1) ◽  
pp. 29-34
Author(s):  
Raghada S. Makia ◽  
Ayad M.A. Fadhil ◽  
Munera Ch. Ismail
Keyword(s):  
Test Tube ◽  
Bacterial Isolates ◽  
Biochemical Tests ◽  
Candida Spp ◽  
Isolation And Identification ◽  
Pseudomonas Spp ◽  
Biofilm Production ◽  
Tract Infections ◽  
Uropathogenic Bacteria

This study includes isolation and identification of different uropathogenes (bacteria- yeasts) collected from catheterized patients suffering from complicated urinary tract infections. Three hundred and fifty urine samples obtained by swabs from catheterized patients were identified for the presence of uropathogenes (bacteria and yeast). 221(63.13%) samples were obtained from females and 118 (33.71%) samples from male, 339(96.85%) sample were identified by culturing as a positive result, while 11(3.14%) sample were negative result. The 339 positive isolates include 303 (89.38%) bacterial isolates and 36 (10.61%) yeast isolates. Results of biochemical tests and Api 20 system for bacterial and yeast isolates reveals E.coli, Proteus spp, Klebseilla spp, Pseudomonas spp and Candida spp represented the main causative uropathogen infect urinary system and causing a complicated type of infection. The determination of bacterial and yeasts ability to form biofilm was carried out using test tube method , 306 isolates which represented (90.26%) were capable to form biofilm with differ in the thickness of formed layer. Pseudomonas spp formed the thicker biofilm followed by E.coli, Candida spp, Proteus spp, and Klebseilla spp.

scholarly journals Isolation and Identification of Major Pathogenic Bacteria from Clinical Mastitic Cows in Asella Town, Ethiopia

2020 ◽  
Vol 2020 ◽  
pp. 1-6
Author(s):  
Gezehagn Kasa ◽  
Betelihem Tegegne ◽  
Belege Tadesse
Keyword(s):  
Bacterial Infections ◽  
Pathogenic Bacteria ◽  
Complex Disease ◽  
Bacterial Species ◽  
Clinical Mastitis ◽  
Species Level ◽  
Bacillus Species ◽  
Small Scale ◽  
Biochemical Tests ◽  
Isolation And Identification

Mastitis is a multietiological and complex disease causing inflammation of the parenchyma of mammary glands and is a problem in many dairy cows. The objective of this study was to isolate and identify the pathogenic bacteria that cause bovine clinical mastitis. A cross-sectional study was undertaken between November 2018 to April 2019 on a small scale and government dairy farms in Asella town. Cow’s udder and teats were physically examined to detect clinical mastitis. A total of 83 milk samples were collected from 46 cows that show clinical sign of mastitis from a total of 12 farms. Isolation and identification of major bacterial species were carried out by culturing different media and using primary and secondary biochemical tests. Out of the 83 samples collected and examined, all (100%) were positive for the cultural isolation of bacterial species. The bacteria were identified to genus and species level. Among the 83 isolates, 32 (38.6%), 24 (28.9%), and 6 (7.2%) were Staphylococcus aureus, Staphylococcus intermedius, and Staphylococcus hyicus, respectively. Other bacteria like Escherichia coli 12 (14.5%) and Streptococcus species 2 (2.4%) were also isolated. Bacillus species 2 (2.4%), Proteus species 2 (2.4%), and 3 (3.6%) of them were mixed bacterial infections. The present study revealed that both contagious and environmental bacterial pathogens were responsible for the occurrence of clinical mastitis. Proper milking practices and farm husbandry practices and future detailed studies up to the species level and on antibiotic profiles of the pathogens are needed.

Isolation and identification of N2-fixing bacteria associated with sugar cane (Saccharum sp.)

1982 ◽  
Vol 28 (5) ◽  
pp. 462-467 ◽  
Author(s):  
R. J. Rennie ◽  
J. R. de Freitas ◽  
A. P. Ruschel ◽  
P. B. Vose
Keyword(s):  
Sugar Cane ◽  
Carbon Substrate ◽  
Computer Assisted ◽  
Stem Cuttings ◽  
Biochemical Tests ◽  
Isolation And Identification ◽  
Whole Plant ◽  
Reducing Activity ◽  
Acetylene Reducing Activity ◽  
Reducing Bacteria

Acetylene-reducing bacteria isolated from the setts (stem cuttings used as seed pieces) and roots of two sugar cane varieties propagated aseptically from stem cuttings were identified using a computer-assisted scheme based on 75 biochemical tests. Because 106 to 108 acetylene-reducing bacteria per gram (fresh weight basis) were found in the roots, while 10 to 100 times fewer were present in the sett, we suggest that the root is the site of bacterial multiplication. Sterilization of the sett surface before planting or root sterilization at harvest reduced or completely removed acetylene-reducing bacteria and associated whole plant acetylene-reducing activity. This indicates that most of the active bacteria were on the sett and root exteriors. Setts did not exhibit acetylene-reducing activity until after emergence of the roots. Since shoot emergence was not necessary for acetylene-reducing activity, the extensive carbohydrate supply of the sett itself must have provided the carbon substrate for bacterial N2 fixation. The acetylene-reducing bacteria isolated were facultative anaerobes of the families Enterobacteriaceae and Bacillaceae. Klebsiella pneumoniae, Enterobacter cloacae, Erwinia herbicola, and Bacillus polymyxa were present inside the sett and the roots but E. herbicola was the dominant bacterium on the root exterior. No Beijerinckia spp. or Azotobacter spp. were found associated with the sett or the roots.

scholarly journals Heterotrophic Plate Count for Bottled Water Safety Management

Processes ◽  
2020 ◽  
Vol 8 (6) ◽  
pp. 739
Author(s):  
Anna Rygala ◽  
Joanna Berlowska ◽  
Dorota Kregiel
Keyword(s):  
Molecular Analysis ◽  
Safety Management ◽  
Heterotrophic Bacteria ◽  
Industrial Process ◽  
Bottled Water ◽  
Plate Count ◽  
Bacterial Isolates ◽  
Water Safety ◽  
Biochemical Tests ◽  
Adhesion Properties

Heterotrophic bacteria are able to form biofilms in water processing systems, adhering to pipe materials and colonizing surfaces. The aim of our research was to identify the critical points in the process of bottled water production at which controls can be applied to prevent, reduce, or eliminate water safety hazards. Microbiological monitoring was conducted using the plate count method and luminometry. To identify the bacterial isolates, we used polyphasic identification based on biochemical tests and molecular analysis using ribosomal RNA. The heterotrophic plate counts were higher in the water filtration station, ultrafiltration (UV) disinfection station, and holding tank. At these points of the industrial process, the water is stagnant or there is poor flow. Molecular analysis identified the bacterial isolates as belonging to Acinetobacter, Agrobacterium, Aeromonas, Brevundimonas, Citrobacter, Enterobacter, Klebsiella, Pantoea, and Rhizobium genera. Bacterial isolates showed various levels of biofilm formation, and the best adhesion properties were exhibited by the Aeromonas hydrophila and Citrobacter freundii strains.

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