• The Plant Pathology Journal
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• 제37권3호
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• pp.215-231
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• 2021

Brown leaf spot disease caused by Nigrospora guilinensis on Phellodendron chinense occurs in a large area in Dayi County, Chengdu City, Sichuan Province, China each year. This outbreak has severely reduced the production of Chinese medicinal plants P. chinense and caused substantial economic losses. The bacterial isolate JKB05 was isolated from the healthy leaves of P. chinense, exhibited antagonistic effects against N. guilinensis and was identified as Bacillus megaterium. The following fermentation medium and conditions improved the inhibitory effect of B. megaterium JKB05 on N. guilinensis: 2% glucose, 0.1% soybean powder, 0.1% KCl, and 0.05% MgSO₄; initial concentration 6 × 10⁶ cfu/ml, and a 42-h optimal fermentation time. A composite of 0.1% nano-SiO₂ JKB05 improved the thermal stability, acid-base stability and ultraviolet resistance by 16%, 12%, and 38.9%, respectively, and nano-SiO₂ was added to the fermentation process. The best formula for the wettable powder was 35% kaolin, 4% polyethylene glycol, 8% Tween, and 2% humic acid. The following quality test results for the wettable powder were obtained: wetting time 87.0 s, suspension rate 80.33%, frequency of microbial contamination 0.08%, pH 7.2, fineness 95.8%, drying loss 1.47%, and storage stability ≥83.5%. A pot experiment revealed that the ability of JKB05 to prevent fungal infections on P. chinense increased considerably and achieved levels of control as high as 94%. The use of nanomaterials significantly improved the ability of biocontrol bacteria to control this disease.

• Journal of Veterinary Science
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• 제21권4호
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• pp.57.1-57.11
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• 2020

Background: Streptococcus dysgalactiae subspecies equisimilis (SDSE) acts as an etiological agent for lameness, neurological signs, and high mortality in pigs. Despite its importance in pig industries and zoonotic potential, little is known about the effects of this pathogen. Objectives: This study aimed to determine the molecular characteristics and antimicrobial resistance of SDSE strains isolated from diseased pigs. Methods: A total 11 SDSE isolates were obtained from diseased pigs. Bacterial identification, PCR for virulence genes, emm typing, and antimicrobial resistance genes, multilocus sequence typing, and antimicrobial susceptibility test were performed. Results: Nine isolates were from piglets, and 8 showed lameness, sudden death, or neurological signs. The isolates were PCR-positive for sla (100%), sagA (100%), and scpA (45.5%), and only 1 isolate amplified the emm gene (stL2764). Eight different sequence types were detected, categorized into 2 clonal complexes and 4 singletons. All the isolates in this study were included in a small cluster, which also contained other strains derived from humans and horses. The minimum inhibitory concentrations for the tested beta-lactams were low, while those for macrolides, tetracyclines, and fluoroquinolones were relatively high. PCR analysis of the macrolide and tetracycline resistance genes demonstrated that the isolates carried erm(B) (18.2%, n = 2), mef(A/E) (9.1%, n = 1), tet(M) (18.2%, n = 2), and tet(O) (90.2%, n = 10). Two isolates presented a mutation in parC, which is associated with fluoroquinolone resistance. Conclusion: This study provided insight into swine-derived SDSE, as it is related to veterinary medicine, and elucidated its zoonotic potential, in the context of molecular epidemiology and antimicrobial resistance in public health.

• Journal of Animal Science and Technology
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• 제64권6호
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• pp.1144-1172
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• 2022

Salmonella enterica serovar Typhimurium isolate HJL777 is a virulent bacterial strain in pigs. The high rate of salmonella infection are at high risk of non-typhoidal salmonella gastroenteritis development. Salmonellosis is most common in young pigs. We investigated changes in gut microbiota and biological function in piglets infected with salmonella via analysis of rectal fecal metagenome and intestinal transcriptome using 16S rRNA and RNA sequencing. We identified a decrease in Bacteroides and increase in harmful bacteria such as Spirochaetes and Proteobacteria by microbial community analysis. We predicted that reduction of Bacteroides by salmonella infection causes proliferation of salmonella and harmful bacteria that can cause an intestinal inflammatory response. Functional profiling of microbial communities in piglets with salmonella infection showed increasing lipid metabolism associated with proliferation of harmful bacteria and inflammatory responses. Transcriptome analysis identified 31 differentially expressed genes. Using gene ontology and Innate Immune Database analysis, we identified that BGN, DCN, ZFPM2 and BPI genes were involved in extracellular and immune mechanisms, specifically salmonella adhesion to host cells and inflammatory responses during infection. We confirmed alterations in gut microbiota and biological function during salmonella infection in piglets. Our findings will help prevent disease and improve productivity in the swine industry.

• 한국미생물·생명공학회지
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• 제51권1호
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• pp.10-17
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• 2023

Dairy products are extensively used as carriers of probiotic strains that have potential health benefits. Assessment of the viability of probiotic strains during manufacturing is important to ensure that products meet recommended levels. Hence, the method for accurately quantifying lactic acid bacteria (LAB) in probiotic or dairy products is required. The present study aims to examine the performance of de-Man Rogosa Sharpe (MRS), plate count agar with bromocresol purple (PCA with BCP), and glucose blood liver (BL) agars recommended in the Korea Food Code guidelines for counting LAB. Analysis of the performance of culture media containing 19 lactic acid bacterial species commonly encountered in probiotic and dairy products showed no statistically significant difference between 18 reference strains and three culture media (p > 0.01). Furthermore, the suitability of three culture media was verified for the quantitative assessment of LAB in 25 probiotic and dairy products. The number of LAB in three culture media was determined to be more than 10⁷ colony-forming unit (CFU)/ml for fermented milk products and 10⁸ CFU/ml for condensed fermented milk and probiotic products, indicating that they all satisfied the Korea Food Code guidelines. Moreover, there was no statistically significant difference in the amount of LAB counted in all three culture media, suggesting that they can be used to isolate or enumerate LAB in commercial products. Finally, three culture media will be useful for isolating and enumerating LAB from fermented foods as well as gut microflora.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.67.2-68
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• 2003

A rice cultivar, Taebaegbyeo, is highly resistant to rice blast and moderately resistant to bacterial leaf blight (BLB) caused by Magnaporthe grisea and Xanthomonas oryzae pv. oryzae, respectively. To study the rice disease resistance mechanism, we generated rice deletion M3 mutants by gamma-ray irradiation. Blast and BLB responses of 16,000 M3 mutants were screened by inoculating mixtures of 4 races (KJ-201, H-1113a, KI-313, KI-409) of M. grisea and 3 Korean races of X. oryzae pv. oryzae. We selected so far 21 M3 mutants of Taebaegbyeo showing high susceptibility to the diseases. One of the mutants, KCT-6417, was susceptible to KI-1113a race of M. grisea, suggesting the deletion of a race-specific blast resistance gene in the mutant. To isolate rice genes involved in blast resistance and defense response, we take a PCR-based suppression subtractive hybridization approach using cDNAs of blast-inoculated wild type and the KCT-6417 as a tester and a driver, respectively. Genes specifically expressed in the wild type will be presented. The selected genes would give us a clue to understand mechanism for the race specific resistance and defense responses against M. grisea H-1113a in Taebaegbyeo.

• Journal of Microbiology and Biotechnology
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• 제33권1호
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• pp.61-74
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• 2023

The global increase in multidrug-resistant (MDR) bacteria has inspired researchers to develop new strategies to overcome this problem. In this study, 23 morphologically different, soil-isolated actinomycete cultures were screened for their antibacterial ability against MDR isolates of ESKAPE pathogens. Among them, isolate BOGE18 exhibited a broad antibacterial spectrum, so it was selected and identified based on cultural, morphological, physiological, and biochemical characteristics. Chemotaxonomic analysis was also performed together with nucleotide sequencing of the 16S rRNA gene, which showed this strain to have identity with Streptomyces lienomycini. The ethyl acetate extract of the cell-free filtrate (CFF) of strain BOGE18 was evaluated for its antibacterial spectrum, and the minimum inhibitory concentration (MIC) ranged from 62.5 to 250 ㎍/ml. The recorded results from the in vitro anti-biofilm microtiter assay and confocal laser scanning microscopy (CLSM) of sub-MIC concentrations revealed a significant reduction in biofilm formation in a concentration-dependent manner. The extract also displayed significant scavenging activity, reaching 91.61 ± 4.1% and 85.06 ± 3.14% of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), respectively. A promising cytotoxic ability against breast (MCF-7) and hepatocellular (HePG2) cancer cell lines was obtained from the extract with IC₅₀ values of 47.15 ± 13.10 and 122.69 ± 9.12 ㎍/ml, respectively. Moreover, based on gas chromatography-mass spectrometry (GC-MS) analysis, nine known compounds were detected in the BOGE18 extract, suggesting their contribution to the multitude of biological activities recorded in this study. Overall, Streptomyces lienomycini BOGE18-derived extract is a good candidate for use in a natural combating strategy to prevent bacterial infection, especially by MDR pathogens.

• 한국환경과학회지
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• 제31권3호
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• pp.265-274
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• 2022

Lipoxygenase is an enzyme, mainly produced by plants, capable of converting unsaturated fatty acids to fatty acids. It has vast application potential in the food, pharmaceutical and agricultural industries. The aim of this study was to isolate novel lipoxygenase-producing bacteria from the environment and to investigate the lipoxygenase enzymatic properties for industrial production. The strain, NC1, isolated from cultivation soils, was identified as Bacillus subtilis based on the phenotypic characteristics and 16S rRNA gene sequencing. This strain formed a pink color around the colony when cultured on indamine dye formation plates. The production of lipoxygenase by B. subtilis NC1 was influenced by the composition of the medium and linoleic acid concentrations. The optimum temperature and pH for lipoxygenase activity was determined to be 40 ℃ and pH 6, respectively. The enzyme showed relatively high stability at temperatures ranging from 20-50 ℃ and acid-neutral regions. In addition, the lipoxygenase produced by B. subtilis NC1 was able to degrade commercially available oils including sunflower seed oil and Perilla oil. In this study, a useful indigenous bacterium was isolated, and the fundamental physicochemical data of bacterial lipoxygenase giving it industrial potential are presented.

• 현장농수산연구지
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• 제25권3호
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• pp.14-18
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• 2023

2022년 7월, 국내의 저염분 해수 흰다리새우 양식장에서 양식 중이던 흰다리새우가 이상유영과 식욕부진을 보이며 지속적으로 폐사하였다. 일일 평균 폐사율이 0.1%로 완만한 폐사를 보였으나, 2달 넘게 폐사가 지속되었고 성장 지체로 인한 경제적인 피해가 가중되었다. 질병 진단 과정 중, 폐사 새우의 간췌장에서 세균(KNUAF-SHP3)이 분리되었다. 본 세균의 16S rRNA 유전자의 시퀀스 분석 결과, 분리주 KNUAF-SHP3은 Shewanella에 속하는 균으로 동정이 되었고, S. algae MARS 14, S. chilikensis JC5T와 근연 관계를 보이며 같은 그룹으로 분류되었다. 실험 동물을 활용한 인위 감염 실험에서는 상대적으로 고농도(2.1×108CFU/ml와 2.1×109CFU/ml)를 적용한 군에서는 모든 새우들이 감염 증상을 보이며 감염 후 7일 내에 100% 폐사율을 나타내었다. 본 연구는 Shewanella 세균이 저염분 해수 양식 흰다리새우에서의 잠재적인 기회 감염균이 될 수 있음을 기술하고 있다.

• 한국임상수의학회지
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• 제40권6호
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• pp.393-398
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• 2023

We identified mastitis-causing pathogens using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) in an organic dairy farm and evaluated the effects of antimicrobial restriction on antimicrobial susceptibility. A total of 43 Holstein cows without any clinical sign of mastitis were used in this study, and 172 quarter milk samples were cultured on blood agar plates for 24 hours at 37℃. Subsequently, bacterial species were identified and antimicrobial susceptibility tests were performed. The subclinical mastitis infection rates in the cows and quarters were 58.1% (25/43) and 25.6% (44/172), respectively. In the species identification, Staphylococcus aureus (40.9%) was the most prominent isolate, followed by S. chromogenes (22.7%), S. epidermis (18.2%), S. simulans (11.4%), S. haemolyticus (2.3%), S. muscae (2.3%), and S. xylosus (2.3%). In the antimicrobial susceptibility test, all isolates were 100% susceptible to 24 of 28 antibiotics, except for benzylpenicillin, cefalotin, cefpodoxime, and trimethoprim/sulfamethoxazole. The resistance rates of S. aureus, S. chromogenes, and S. muscae isolates to trimethoprim/sulfamethoxazole were 27.8%, 10%, and 100%, respectively, and the resistance rates of S. epidermis and S. xylosus to benzylpenicillin were 50% and 100%, respectively. S. chromogenes, S. epidermis, S. simulans, S. haemolyticus, and S. xylosus were resistant to cefalotin and cefpodoxime. In conclusion, restrictions on antimicrobial use for organic dairy farm certification have resulted in a high Staphylococcus spp. infection rate. Therefore, our study indicates the importance of mastitis management strategies implemented by farmers together with veterinary practitioners, even if mastitis does not appear clinically in organic dairy farms.

• Journal of Microbiology and Biotechnology
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• 제34권5호
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• pp.1040-1050
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• 2024

To isolate and analyze bacteria with Verticillium wilt-resistant properties from the fermentation residue of kitchen wastes, as well as explore their potential for new applications of the residue. A total of six bacterial strains exhibiting Verticillium wilt-resistant capabilities were isolated from the biogas residue of kitchen waste fermentation. Using a polyphasic approach, strain ZL6, which displayed the highest antagonistic activity against cotton Verticillium wilt, was identified as belonging to the Pseudomonas aeruginosa. Bioassay results demonstrated that this strain possessed robust antagonistic abilities, effectively inhibiting V. dahliae spore germination and mycelial growth. Furthermore, P. aeruginosa ZL6 exhibited high temperature resistance (42℃), nitrogen fixation, and phosphorus removal activities. Pot experiments revealed that P. aeruginosa ZL6 fermentation broth treatment achieved a 47.72% biological control effect compared to the control group. Through activity tracking and protein mass spectrometry identification, a neutral metalloproteinase (Nml) was hypothesized as the main virulence factor. The mutant strain ZL6ߡNml exhibited a significant reduction in its ability to inhibit cotton Verticillium wilt compared to the strain P. aeruginosa ZL6. While the inhibitory activities could be partially restored by a complementation of nml gene in the mutant strain ZL6CMߡNml. This research provides a theoretical foundation for the future development and application of biogas residue as biocontrol agents against Verticillium wilt and as biological preservatives for agricultural products. Additionally, this study presents a novel approach for mitigating the substantial amount of biogas residue generated from kitchen waste fermentation.