• Applied Biological Chemistry
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• v.34 no.2
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• pp.174-179
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• 1991

Protoplast fusion was carried out between Brevibacterium flavum and Corynebacterium glutamicum. For the protoplast fusion, various mutants were isolated from Brevibacterium flavum ATCC 21493 and Corynebacteriurn glutamicum ATCC 21831. The optimum conditions for protoplast fusion of these mutants were examined. In the present work, the authors obtained a fusant, MWF 9031, by the intergeneric protoplast fusion between Brevibacterium flavum 108-125 and Corynebacterium glutamicum 41-214A, which was excellent in L-arginine fermentation. Fusant MWF 9031 was found to accumulate a large amount of L-arginine reached 32.5 mg/ml with a medium containing 10% glucose. The fusant possessed intermediate characteristics between the parental strains and the stability was found to retain for 60 days.

• Journal of Ecology and Environment
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• v.48 no.1
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• pp.17-23
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• 2024

Background: Honey bees play a crucial role in pollination and ecological balance. Apis mellifera L. colonies, especially those located in specific geographic regions, such as the palm garden in Eastern Thailand, are susceptible to potential threats from microbial contaminants. Understanding and detecting microbial organisms in these beehives is essential for the preservation of bee health, honey production, and the broader ecosystem. However, the problem of microbial infection and antibiotic-resistant bacteria is more severe and continuously increasing, resulting in a health, economic, and social crisis. The purpose of this study is to determine the prevalence of microorganisms in A. mellifera beehives in palm gardens in Rayong province, Eastern Thailand. Results: Ten swabs in transport media were swabbed and obtained from different parts of each beehive (1 swab per beehive), for a total of 10 hives. Traditional microbial culture-based methods, biochemical tests, and antimicrobial susceptibility (disc-diffusion) tests were used to detect microbial organisms and antibiotic resistance in bacteria. The swab tests from nine beehives resulted in the detection of Gram-positive bacteria (63.64%), Gram-negative bacteria (27.27%), and fungi/yeast (9.09%). These microorganisms are classified as a group of coagulase-negative Staphylococcus spp. and made up 40.91% of the bacteria discovered. Other bacteria found were Coryneform bacteria (13.64%), Pantoea spp. (13.64%), Bacillus spp. (9.09%), yeast (9.09%), glucose non-fermentative Gram-negative bacilli (9.09%), and Pseudomonas spp. (4.55%). However, due to the traditional culture-based and 0biochemical tests usually used to identify the microbial organisms in clinical specimens and the limitation of identifying some environmental microbial species, the results of the antimicrobial susceptibility test cannot reveal if the organism is resistant or susceptible to the drug. Nevertheless, drug-sensitive inhibition zones were formed with each antibiotic agent. Conclusions: Overall, the study supports prevention, healthcare, and public health systems. The contamination of microorganisms in the beehives may affect the quality of honey and other bee products or even the health of the beekeeper. To avoid this kind of contamination, it is therefore necessary to wear personal protective equipment while harvesting honey and other bee products.

• Microbiology and Biotechnology Letters
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• v.23 no.2
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• pp.164-169
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• 1995

An endogenous cryptic plasmid, pBL1, which has been used to construct plasmid vectors for coryneform bacteria producing amino acids, was eliminated from Brevibacterium lactofermentum. The pBL1 was partially digested with Sau3AI and the resulting DNA fragments were subcloned into a suicide vector pEM1 which contains a kanamycin-resistant (km$^{r}$) gene. KM$^{r}$ B. lactofermentum transconjugants were obtained by conjugal transfer of the pEM1 derivatives containing pBL1 DNA fragments from Escherichia coli into B. lactofermentum. A km$^{r}$ transconjugant was analyzed to contain a plasmid pEB14, which occurred in vivo by homologous recombination between pBL1 and the conjugal-transferred plasmid. The pEB14 including the pEM1-derived km$^{r}$ gene was found to be lost concomitantly with km$^{r}$ phenotype, resulting in the construction of a pBL1-free strain of B lactofermentum. Based on transformation efficiencies and plasmid stability, the resultant pBL1- free strain is more useful than wild strain as a host cell for genetic manipulation. It could be concluded that foreign plasmid DNAs are efficiently isolated and analyzed from the pBL1-free strain because of the absence of endogenous pBL1 plasmid.

• Korean Journal of Microbiology
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• v.23 no.4
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• pp.265-270
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• 1985

Overall procedure of cell fusion between Brevibacterium flavum and Corynebacterium glutamicum was morphologically observed by transmission electron microscopy. Protoplasts formed by treatment of cells with penicillin G and lysozyme in order were released through the pores generated on a certain region of cell walls to be spherical form. When two different protoplasts were met, cell wall and membrane in the contact zone was disappeared and followed by the mutual exchange of cytoplasmic and/or chromosomal materials. Cell xall regeneration speed of the protoplasts fused was slower than that of the non-fused, whereas the size of the former was confirmed as bigger than that of the latter.

• Korean Journal of Soil Science and Fertilizer
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• v.29 no.3
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• pp.303-311
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• 1996

The microflora changes of 10 water-controled treatments combined with livestock manures(pig, chicken) and bulking agents(sawdust, paper sludge) were investigated. The B/F values of the P-1 and C-1(65%, $H_2O$) treatments were 3571 and 5400 respectively, but those of the P-4 and C-4(50%, $H_2O$) treatments showed very low values, 667 and 334, respectively. The B/F values tended to increase with higher water content of the treatments. In the composting processes, the successions of microflora, adapting the compost environments, took place via fluctuating temperature. In the high temperature period, the numbers of mesophilic bacteria and fungi decreased, but that of the spore forming bacteria increased. However, the number of mesophilic bacteria inereased during the cold period. The B/F values of compost ranged 25-300, which indicates a decrease in the quantity of bacteria. The time required for the temperature of compost to reach $60^{\circ}C$ showed different patterns. There was no pathogenic microorganism in the treatments which reached a high temperature in a short period of time, but, in the treatments which reached a high temperature over a Long period of time, the pathgenic microorganism was not still alive.

• Microbiology and Biotechnology Letters
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• v.18 no.3
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• pp.296-300
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• 1990

For interspecific portoplast fusion, Brevibacterium flauum lOAHR (Rifr axg his) and Corynebacterium glutamicum 11TS ($Sm-r$ trp) were induced by UV and NTG treatment. The protoplast fusion frequency between E. flavum XOAHR and C. glutamicum llTS was $3.7\times 10^{-6}$ with the lysozyme treatment (300 P $\mu g$ml) for 18 hrs. Genotypes of recombinants were analized as FMM ($Rif^r\; Sm^r$), FA (Rift $Sm^r$ arg), FH ($Rif^r\; Sm^r$ his), FT ($Rif^r\; Sm^r$ trp), FAH ($Rif^r\; Sm^r$ arg trp), FAT ($Rif^r\; Sm^r$ arg trp), and FAHT ($Rif^r\; Sm^r$ arg his trp). FAH 1 produced 12 fold of glutamate production compared to parental type, E. flauum 10AHR. In glutamine productivity, it produced 2.6 fold to parental type, C. glutamicum 11TS. Production of glutamate or glutamine by recombinants was involved in the specific activities of glutamate dehydrogenase (GDH) and glutamine synthetase (GS), respectively.

• Journal of Veterinary Clinics
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• v.23 no.2
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• pp.169-174
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• 2006

This work described 3 infection cases caused by Rhodococcus equi in foals between 3 and 5 months of age. The disease histories were not fully taken from local veterinarians. At least 1 sick foal has been treated with cephalothins followed by penicillins during approximately 1 week, but died without effectiveness and other foals rearing with the animal have been suffering from severe pneumonia which show high fever, laboring respiration, cough and/or nasal discharge. There were many abscessations into lungs of 2 foals in postmortem examination and another 1 sample was pus collected from abscess around the shoulder, indicating the osteomyelitis. Those bacteria were grampositive coryneform and were identified as a R. equi by a polymerase chain reaction (PCR) using primers for R. equi-specific vapA gene. The pathogens were usually resistant to penicillin, ampicillin, amoxycillin/clavulanic acid, cefazolin, clindamycin, sulfamethoxazol/trimethoprim, kanamycin, and tetracycline, while were sensitive to ciprofloxacin, norfloxacin, orfloxacin, gentamicin, erythromycin, neomycin, and vancomycin. Some more foals with respiratory symptoms in 1 horse farm were treated by orally administration with erythromycin during 2 weeks. Because the combination of erythromycin and rifampin has recommended as the treatment for R. equi infections in foals, the local equine veterinarian can choose those antibiotics for the treatment of this disease in future. However, another antimicrobial agent may be necessary if R. equi resistant to both agents is isolated.