• Title/Summary/Keyword: Pathogenic bacterial

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Disruption of the metC Gene Affects Methionine Biosynthesis in Pectobacterium carotovorum subsp. carotovorum Pcc21 and Reduces Soft-Rot Disease

  • Seonmi, Yu;Jihee, Kang;Eui-Hwan, Chung;Yunho, Lee
    • The Plant Pathology Journal
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    • v.39 no.1
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    • pp.62-74
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    • 2023
  • Plant pathogenic Pectobacterium species cause severe soft rot/blackleg diseases in many economically important crops worldwide. Pectobacterium utilizes plant cell wall degrading enzymes (PCWDEs) as the main virulence determinants for its pathogenicity. In this study, we screened a random mutant, M29 is a transposon insertion mutation in the metC gene encoding cystathionine β-lyase that catalyzes cystathionine to homocysteine at the penultimate step in methionine biosynthesis. M29 became a methionine auxotroph and resulted in growth defects in methionine-limited conditions. Impaired growth was restored with exogenous methionine or homocysteine rather than cystathionine. The mutant exhibited reduced soft rot symptoms in Chinese cabbages and potato tubers, maintaining activities of PCWDEs and swimming motility. The mutant was unable to proliferate in both Chinese cabbages and potato tubers. The reduced virulence was partially restored by a complemented strain or 100 µM of methionine, whereas it was fully restored by the extremely high concentration (1 mM). Our transcriptomic analysis showed that genes involved in methionine biosynthesis or transporter were downregulated in the mutant. Our results demonstrate that MetC is important for methionine biosynthesis and transporter and influences its virulence through Pcc21 multiplication in plant hosts.

Highlighting the Microbial Community of Kuflu Cheese, an Artisanal Turkish Mold-Ripened Variety, by High-Throughput Sequencing

  • Talha Demirci
    • Food Science of Animal Resources
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    • v.44 no.2
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    • pp.390-407
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    • 2024
  • Kuflu cheese, a popular variety of traditional Turkish mold-ripened cheeses, is characterized by its semi-hard texture and blue-green color. It is important to elucidate the microbiota of Kuflu cheese produced from raw milk to standardize and sustain its sensory properties. This study aimed to examine the bacteria, yeasts, and filamentous mold communities in Kuflu cheese using high-throughput amplicon sequencing based on 16S and ITS2 regions. Lactococcus, Streptococcus, and Staphylococcus were the most dominant bacterial genera while Bifidobacterium genus was found to be remarkably high in some Kuflu cheese samples. Penicillium genus dominated the filamentous mold biota while the yeasts with the highest relative abundances were detected as Debaryomyces, Pichia, and Candida. The genera Virgibacillus and Paraliobacillus, which were not previously reported for mold-ripened cheeses, were detected at high relative abundances in some Kuflu cheese samples. None of the genera that include important food pathogens like Salmonella, Campylobacter, Listeria were detected in the samples. This is the first experiment in which the microbiota of Kuflu cheeses were evaluated with a metagenomic approach. This study provided an opportunity to evaluate Kuflu cheese, which was previously examined for fungal composition, in terms of both pathogenic and beneficial bacteria.

Investigation of supplementation with a combination of fermented bean dregs and wheat bran for improving the growth performance of the sow

  • Junze Liu;Kai Wang;Liangyu Zhao;Yansen Li;Zhaojian Li;Chunmei Li
    • Journal of Animal Science and Technology
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    • v.66 no.2
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    • pp.295-309
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    • 2024
  • To investigate the effect of dietary supplementation with a fermented mixture of bean dregs and wheat bran (FBW) on sow performance. FBW was given to sows during late gestation and lactation; in total, 24 sows were randomly assigned to 4 groups (control diet; 3% FBW diet; 6% FBW diet; 9% FBW diet, n = 6). The weight ratio of bean dregs (wet) to wheat bran was 4:6. Sows were fed different diets from 85 d of gestation until weaning. The results showed that supplementation with FBW increased average daily feed intake (ADFI) during lactation (p < 0.05). FBW supplementation also increased litter weight and milk yield (p < 0.05). The contents of Escherichia coli in the feces of the treatment groups were significantly reduced by FBW supplementation (p < 0.01). FBW supplementation significantly improved the fecal morphology (p < 0.05), alleviating sows' constipation. In conclusion, FBW could increase the ADFI, improve lactation and piglet litter weight in sows and reduce the pathogenic bacterial content in sow feces and constipation.

In Vitro Screening of Tannic Acid-based Eco-friendly Farming Material (notice no. 2-4-064) against Plant Pathogenic Bacteria (탄닌산을 함유한 친환경농자재(공시번호 2-4-064)의 식물병원세균 기내 억제효과)

  • Han, Kyu Suk;Ju, Ho-Jong;Hong, Jin Sung;Chung, Jong-Sang;Park, Duck Hwan
    • Korean Journal of Organic Agriculture
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    • v.24 no.4
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    • pp.945-955
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    • 2016
  • To date, chemical managements of plant bacterial diseases are complicated by limitations on use of antibiotics in agriculture, antibiotic resistance development, and limited efficacy of alternative control agents. In this study, thus, we performed screening of eco-friendly farming material (notice no. 2-4-064) containing tannic acid as new antibacterial-activity against 7 plant bacterial pathogens: Pectobacterium carotovorum subsp. carotovorum (Pcc), Ralstonia solanacearum (Rs), Acidovorax avenae subsp. citrulli (Aac), Xanthomonas cirti pv. citri (Xcc), Erwinia pyrifoliae (Ep), Clavibacter michiganensis subsp. michiganensis (Cmm), and Streptomyces scabies (Sc), Initial screening of antibacterial effects of eco-friendly farming material was performed using the paper disk diffusion method and co-cultivation in broth culture. Tannic acid based eco-friendly farming material showed inhibitory effect against Pcc, Rs, Aac, Xcc, Cmm, and Ss, whereas it did not show inhibition zone against Ep. However, when it tested by co-cultivation in broth culture, it showed strong inhibition effect against all pathogens that declined growth curve compared to bacterial pathogen only. Interestingly, when we observed morphological changes on those pathogens by SEM, cell morphologies of 7 pathogens were slightly changed that seem to be malfunction in their cell envelope.

Molecular Biological Identification of Bacteria in Middle Ear Effusion Using 16S rDNA Multiplex PCR (중이 삼출액 미생물의 16S rDNA 복합중합효소연쇄반응을 이용한 분자생물학적인 진단)

  • 이정구;이인숙;박지연;정상운;오충훈
    • Korean Journal of Microbiology
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    • v.39 no.1
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    • pp.36-39
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    • 2003
  • The rapid and reliable 16S rDNA multiplex polymerase chain reaction (PCR) assay was established to characterize bacterial etiologies of middle ear effusion. These etiologies included Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumonia, which were detected in middle-ear effusion (MEE) samples taken from patient with otitis media. A total of 39 MEE samples were aspirated from 26 patients. DNA was extracted from MEE samples, and PCR was done with DNA extracts by using the common primers, which is localized at C4 region in the 16S rDNA gene of all bacterial species, and species-specific primers: (i) Haemophilus-specific primer, (ii) Moraxella- specific primer, and (iii) Streptococcus-specific primer. Among 39 samples tested, 24 (61.5%) were positive for H. influenzae, 10 (25.6%) were positive for M. catarrhalis, 3(7.7%) were positive for S. pneumonia, and 11 (28%) were negative for 165 rDNA multiplex PCR reaction. Nine samples (28.6%) exhibited a mixed infection and were positive for both H. infuenzae and M. catarrhalis. We suggested that 16S rDNA multiplex PCR is a useful method to identify rapidly for rapid identification of the pathogenic bacteria and characterization of bacterial etiologies of middle ear effusion.

Generation of antibodies against N-terminus fragment of AgI/II protein from Streptococcus mutans GS-5 (연쇄상구균(Streptococcus mutans GS-5)의 항원단백질 AgI/II의 N-terminus절편에 대한 항체형성)

  • Han, Ji-Hye;Baik, Byeong-Ju;Yang, Yeon-Mi;Park, Jeong-Yeol;Kim, Jae-Gon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.33 no.3
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    • pp.401-410
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    • 2006
  • Dental caries results from localized demineralization of tooth enamel by acids of bacterial origin produced from the fermentation of dietary sugars. A group of related oral bacteria, collectively known as mutans streptococci, are implicated as the primary etiological agents of human caries. Within this group, Streptococcus mutans has been known as a causative agent for dental caries. As well as acid production yielding the demineralization of tooth enamel, adherence and colonization of S. mutans to the teeth are also important for their virulence Cell-surface fibrillar proteins, which mediate adherence to the salivary pellicle are virulence components of mutans streptococci, and primary candidates for a human caries vaccine. Here we report that the AgI/II gene from S. mutans GS-5 were cloned by PCR amplification of the bacterial chromosomal DNA and the integrity of cloned genes were confirmed by nucleotide sequencing. Sequence analyses showed the sequence alignment of 280 nucleotides between the cloned AgI/II and the reported sequence of S. mutans GS-5 showed the perfect match The cloned genes which signal nucleotide was truncated, were transferred into bacterial expression vector and the recombinant proteins were purified as His-tag fusion proteins In order to generate polyclonal antibodies against the recombinant proteins, AgI/II mr, some $100{\mu}g$ of the proteins was injected into mice three times. It can be used for an effective vaccine production to prevent dental caries caused by pathogenic S. mutans.

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Antifungal Activity of Bacillus sp. GJ-1 Against Phytophthora capsici (Bacillus sp. GJ-1의 Phytophthora capsici에 대한 항진균활성)

  • Lee, Gun-Joo;Han, Joon-Hee;Shin, Jong-Hwan;Kim, Heung Tae;Kim, Kyoung Su
    • The Korean Journal of Mycology
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    • v.41 no.2
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    • pp.112-117
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    • 2013
  • Phytophthora capsici is one of major limiting factors in production of pepper and other important crops worldwide by causing foliage blight and rot on fruit and root. Increased demand for the replacement of fungicides has led to searching a promising strategy to control the fungal diseases. To meet eco-friendly agriculture practice, we isolated microorganisms and assessed their beneficial effects on plant health and disease control efficacy. A total of 360 bacterial strains were isolated from rhizosphere soil of healthy pepper plants, and categorized to 5 representative isolates based on colony morphology. Among the 5 bacterial strains (GJ-1, GJ-4, GJ-5, GJ-11, GJ-12), three bacterial strains (GJ-1, GJ-11, GJ-12) presented antifungal activity against P. capsici in an fungal inhibition assay. In phosphate solubilization and siderophore production, the strain GJ-1 was more effective than others. The strain GJ-1 was identified as Bacillus sp. using 16S rDNA analysis. Bacillus sp. GJ-1 was also found to be effective in inhibiting other plant pathogenic fungi, including Rhizoctonia solani, Pythium ultimum and Fusarium solani. Therefore, the Bacillus sp. GJ-1 can serve as a biological control agent against fungal plant pathogens.

Inhibition of in Vitro Growth of Three Soil-borne Turfgrass Diseases by Antagonistic Bacteria from Composted Liquid Manure (가축분뇨액비의 길항미생물에 의한 토양전염성 병원균의 생육억제 효과)

  • Ryu, Ju Hyun;Shim, Gyu Yul;Kim, Ki Sun
    • Horticultural Science & Technology
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    • v.32 no.6
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    • pp.879-886
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    • 2014
  • This study was conducted to test in vitro the antagonistic effect of composted liquid manure (CLM) against soil-borne turfgrass pathogenic fungi, Rhizoctonia solani AG-2-2 (IIIB) (brown patch), R. solani AG-2-2 (IV) (large patch), and Sclerotinia homoeocarpa (dollar spot) for environmentally friendly turfgrass management. CLMs were collected from 9 livestock excretion treatment facilities around the country including Gunwi (GW), Hapcheon (HC), Hoengseong (HS), Icheon (IC), Iksan (IS), Muan (MA), Nonsan (NS), and Yeoju (YJ). CLMs of IC, GW, and IS showed s ignificant (p < 0.05) mycelium growth inhibition that was 17.8%, 20.4%, and 48.0% against R. solani AG-2-2 (IIIB), R. solani AG-2-2 (IV), and S. homoeocarpa, respectively. A t otal of 110 bacterial isolates were obtained from the CLMs that showed antagonistic effects. Among them, 5, 4, and 10 microbe isolates showed promising antifungal activity against mycelium growth of R. solani AG-2-2 (IIIB), R. solani AG-2-2 (IV), and S. homoeocarpa, respectively. The bacterial isolates ICIIIB60, GWIV70, and ISSH20 effectively inhibited the mycelial growth of three soil-borne turfgrass pathogens. Selected bacterial isolates were identified as Alcaligenes sp., Bacillus licheniformis Ab2, and B. subtilis C7-3 through 16s rDNA gene sequence analysis. Among 5 fungicides, the most compatible fungicide with ICIIIB60, GWIV70, and ISSH20 was tebuconazol, toclofos-methyl and toclofos-methyl, respectively. These findings suggested that CLMs could be effectively used not only as organic liquid fertilizer sources but also as biological control agents for soil-borne turfgrass diseases such as brown patch, large patch, and dollar spot.

Bacterial Spot Disease of Green Pumpkin by Pseudomonas syringae pv. syringae (Pseudomonas syringae pv. syringae에 의한 애호박 세균점무늬병)

  • Park, Kyoung-Soo;Kim, Young-Tak;Kim, Hye-Seong;Lee, Ji-Hye;Lee, Hyok-In;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.22 no.3
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    • pp.158-167
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    • 2016
  • A pathogen that causes a new disease on green pumpkin in the nursery and the field was characterized and identified. Symptoms of the disease on green pumpkin were water soaking lesions and spots with strong yellow halo on leaf, brown lesion on flower, and yellow spot on fruit. The bacterial isolates from the leaf spot were pathogenic on the 8 curcubitaceae crop plants, green pumpkin, figleaf gourd, wax gourd, young pumpkin, zucchini, cucumber, melon, and oriental melon, whereas they did not cause the disease on sweet pumpkin and watermelon. They were Gram-negative, rod shape with polar flagella, fluorescent on King's B agar and LOPAT group 1a by LOPAT test. Their Biolog substrate utilization patterns were similar to Pseudomonas syringae pv. syringae's in Biolog database. Phylogenetic trees with 16S rRNA gene sequences and multilocus sequence typing (MLST) with nucleotide sequences of 4 housekeeping genes, gapA, gltA, gyrB, rpoD and those of P. syringae complex strains in the Plant Associated and Environmental Microbes Database (PAMDB) showed that the green pumpkin isolates formed in the same clade with P. syringae pv. syringae strains. The clade in MLST tree was in the genomospecies 1 group. The phenotypic and genotypic characteristics suggested that the isolates from green pumpkin lesion were P. syringae pv. syringae.

Microbiological Evaluation for HACCP System Application of Green Vegetable Juice Containing Lactic Acid Bacteria (유산균을 함유한 녹즙의 HACCP 시스템 적용을 위한 미생물학적 위해도 평가)

  • Kwon, Sang-Chul
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.12 no.11
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    • pp.4924-4931
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    • 2011
  • This research performed to evaluate a production processes reporting by the HACCP system of green vegetable juice products, containing lactic acid bacteria, stage of processing raw materials agricultural products and production facilities of general bacteria and pathogenic micro organism. General bacteria are found from four samples of storage of agricultural products at process stage and water was detected 8.67~14.67 CFU/ml. However, all samples were detected less than 105 CFU/ml as a legal standards after the process of UV sterilization. For the outcome of experiment of E.coli, E.coli O157:H7, B.cereus, L.moonocytogenes, Salmonella spp, Staph.aureus as the food poisoning bacterial, E.coli was detected until UV pre-step process in storage process and B.cereus was detected partly till 1st washing. Since all bacterial, Yeast and Mold are detected in main materials, pre-control method is a necessary to establish for decreasing with a number of initial bacteria of main materials and it is considered to establish the effective ways of washing and sterilization such as production facilities for cross contamination prevention of bacteria and Sthaphylococcus. Based on above results, the process of UV sterilization should be managed with CCP as an important process to reduce or eliminate the general and food poisoning bacterial of green vegetable juice products, including lactic acid bacteria. Therefore, it is considered to need an exhaustive HACCP plan such as control manual of UV sterilization, solution method, verification, education and training and record management.