• Asian-Australasian Journal of Animal Sciences
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• 제32권10호
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• pp.1580-1590
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• 2019

Objective: The aims of this study were to select one strain of Lactobacillus plantarum (L. plantarum) for a potential indigenous safe starter culture with low level antibiotic resistant and low biogenic amine production and evaluate its effect on biogenic amines reduction in Moo som. Methods: Three strains of indigenous L. plantarum starter culture (KL101, KL102, and KL103) were selected based on their safety including antibiotic resistance and decarboxylase activity, and fermentation property as compared with a commercial starter culture (L. plantarum TISIR543). Subsequently, the effect of the selected indigenous safe starter culture on biogenic amines formation during Moo som fermentation was studied. Results: KL102 and TISIR 543 were susceptible to penicillin G, tetracycline, chloramphenicol, erythromycin, gentamycin, streptomycin, vancomycin, ciprofloxacin and trimethoprim (MIC90 ranging from 0.25 to $4{\mu}g/mL$). All strains were negative amino acid-decarboxylase for lysis of biogenic amines in screening medium. For fermentation in Moo som broth, a relatively high maximum growth rate of KL102 and TISIR543 resulted in a generation time than in the other strains (p<0.05). These strain counts were constant during the end of fermentation. Similarly, KL102 or TISIR543 addition supported increases of lactic acid bacterial count and total acidity in Moo som fermentation. For biogenic amine reduction, tyramine, putrescine, histamine and spermine contents in Moo som decreased significantly by the addition KL102 during 1 d of fermentation (p<0.05). In final product, histamine, spermine and tryptamine contents in Moo som inoculated with KL102 were lower amount those with TISIR543 (p<0.05). Conclusion: KL102 was a suitable starter culture to reduce the biogenic amine formation in Moo som.

• Childhood Kidney Diseases
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• 제23권1호
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• pp.22-28
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• 2019

Purpose: We studied the pathogens and trends in antibiotic sensitivity pattern in children with urinary tract infection (UTI) over 8 years in order to evaluate adequate treatment. Methods: We performed a retrospective review of medical records of children with UTI from January 2009 to December 2016 in Daegu Fatima Hospital. Uropathogens and antibiotic sensitivity patterns were selected. Only 1 bacterial species with a colony count of ${\geq}105CFU/mL$ was considered a positive result. We compared 2 periods group (A: 2009~2012, B: 2013~2016) to investigate trends of antibiotic sensitivity pattern. Results: During the 8 year period, 589 cases are identified (E. coli was cultured in 509 cases, 86.4%). Among all patients, this study investigated the antibiotic sensitivity of E. coli. Antimicrobial susceptibility to ampicillin was steadily low for both periods (A: 32.6%, B: 40.1%, P=0.125), and to amikacin was consistently high for both periods (A: 99.4%, B: 99.3%, P=1.000). Antibiotic sensitivity to third-generation cephalosporin decreased from period A to B (A: 91.7%, B: 75.5%, P=0.000). Antibiotic sensitivity to quinolone significantly decreased from A to B (A: 88.4%, B: 78.2%, P=0.003). The prevalence of extended-spectrum ${\beta}$-lactamase-producing E. coli increased from period A to B (A: 6.1%, B: 17.1%, P=0.000). Conclusion: This study showed that conventional antibiotic therapy for the treatment of pediatric UTI needs to be reevaluated. A careful choice of antibiotic is required due to the change in antibiotic sensitivity and the emergence of antibiotic-resistant bacteria.

• Journal of Microbiology and Biotechnology
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• 제32권6호
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• pp.749-760
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• 2022

α-Galactosidase is a debranching enzyme widely used in the food, feed, paper, and pharmaceuticals industries and plays an important role in hemicellulose degradation. Here, T26, an aerobic bacterial strain with thermostable α-galactosidase activity, was isolated from laboratory-preserved lignocellulolytic microbial consortium TMC7, and identified as Parageobacillus thermoglucosidasius. The α-galactosidase, called T26GAL and derived from the T26 culture supernatant, exhibited a maximum enzyme activity of 0.4976 IU/ml when cultured at 60℃ and 180 rpm for 2 days. Bioinformatics analysis revealed that the α-galactosidase T26GAL belongs to the GH36 family. Subsequently, the pET-26 vector was used for the heterologous expression of the T26 α-galactosidase gene in Escherichia coli BL21 (DE3). The optimum pH for α-galactosidase T26GAL was determined to be 8.0, while the optimum temperature was 60℃. In addition, T26GAL demonstrated a remarkable thermostability with more than 93% enzyme activity, even at a high temperature of 90℃. Furthermore, Ca²⁺ and Mg²⁺ promoted the activity of T26GAL while Zn²⁺ and Cu²⁺ inhibited it. The substrate specificity studies revealed that T26GAL efficiently degraded raffinose, stachyose, and guar gum, but not locust bean gum. This study thus facilitated the discovery of an effective heat-resistant α-galactosidase with potent industrial application. Meanwhile, as part of our research on lignocellulose degradation by a microbial consortium, the present work provides an important basis for encouraging further investigation into this enzyme complex.

• 대한임상검사과학회지
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• 제54권2호
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• pp.119-124
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• 2022

녹농균(Pseudomonas aeruginosa), 대장균(Escherichia coli) 및 포도알균(Staphylococcus aureus)은 기회감염균이다. 또한 이들 세균은 다제내성(Multiple-Drug Resistance, MDR) 세균의 성질을 가지는 것으로 알려져 있다. 녹농균, 대장균, 포도알균에 대한 다양한 효능을 지닌 6가지 발효액의 항균활성을 분석하였다. 발효액을 섭취했을 때 대장균에서 유전적 발현 변화가 일어나는지 알아보기 위해 annealing control primer를 사용하여 유전자발현 분석을 수행하였다. 디스크확산법을 통해 무화과식초와 대봉감식초가 다른 발효액에 비해 억제대의 크기가 가장 크게 나타났고, 토종약초발효소는 항균효과가 없었다. 대장균에 5% 무화과식초를 처리하여 21일간 매일 계대배양하여 Escherichia coli O157:H7 OmpW gene for outer membrane protein W 유전자 발현이 감소하며, Synthetic construct Lao1 gene for L-amino acid oxidase, complete cds 유전자가 발현이 증가함을 확인하였다. 발효액 중에서 특히 무화과식초를 섭취하는 것은 우리 주변에 항상 존재하는 대장균에 대해 방어능력을 더욱 단단하게 가질 수 있음을 의미하며, 나아가 다제내성균에 대한 천연치료제로 유용하게 쓰일 수 있기를 기대한다.

• Journal of Periodontal and Implant Science
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• 제52권2호
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• pp.141-154
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• 2022

Purpose: C57BL/6 mice, which are among the most common backgrounds for genetically engineered mice, are resistant to the induction of periodontitis by oral infection with periodontal pathogens. This study aimed to develop a periodontitis model in C57BL/6 mice using coaggregation between human pathogens and the mouse oral commensal Streptococcus danieliae (Sd). Methods: The abilities of Porphyromonas gingivalis ATCC 33277 (Pg33277), P. gingivalis ATCC 49417 (Pg49417), P. gingivalis KUMC-P4 (PgP4), Fusobacterium nucleatum subsp. nucleatum ATCC 25586 (Fnn), and F. nucleatum subsp. animalis KCOM 1280 (Fna) to coaggregate with Sd were tested by a sedimentation assay. The Sd-noncoaggregating Pg33277 and 2 Sd-coaggregating strains, PgP4 and Fna, were chosen for animal experiments. Eighty C57BL/6 mice received oral gavage with Sd once and subsequently received vehicle alone (sham), Fna, Pg33277, PgP4, or Fna+PgP4 6 times at 2-day intervals. Mice were evaluated at 5 or 8 weeks after the first gavage of human strains. Results: Fnn, Fna, and PgP4 efficiently coaggregated with Sd, but Pg33277 and Pg49417 did not. Alveolar bone loss was significantly higher in the PgP4 group at both time points (weeks 5 and 8) and in all experimental groups at week 8 compared with the sham group. The PgP4 group presented greater alveolar bone loss than the other experimental groups at both time points. A higher degree of alveolar bone loss accompanied higher bacterial loads in the oral cavity, the invasion of not only PgP4 but also Sd and Fna, and the serum antibody responses to these bacteria. Conclusions: Periodontitis was successfully induced in C57BL/6 mice by oral infection with a P. gingivalis strain that persists in the oral cavity through coaggregation with a mouse oral commensal bacterium. This new model will be useful for studying the role of human oral bacteria-host interactions in periodontitis using genetically engineered mice.

• Journal of Veterinary Science
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• 제24권6호
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• pp.82.1-82.12
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• 2023

Background: The current conventional serotyping based on antigen-antisera agglutination could not provide a better understanding of the potential pathogenicity of Salmonella enterica subsp. enterica serovar Brancaster. Surveillance data from Malaysian poultry farms indicated an increase in its presence over the years. Objective: This study aims to investigate the virulence determinants and antimicrobial resistance in S. Brancaster isolated from chickens in Malaysia. Methods: One hundred strains of archived S. Brancaster isolated from chicken cloacal swabs and raw chicken meat from 2017 to 2022 were studied. Two sets of multiplex polymerase chain reaction (PCR) were conducted to identify eight virulence genes associated with pathogenicity in Salmonella (invasion protein gene [invA], Salmonella invasion protein gene [sipB], Salmonella-induced filament gene [sifA], cytolethal-distending toxin B gene [cdtB], Salmonella iron transporter gene [sitC], Salmonella pathogenicity islands gene [spiA], Salmonella plasmid virulence gene [spvB], and inositol phosphate phosphatase gene [sopB]). Antimicrobial susceptibility assessment was conducted by disc diffusion method on nine selected antibiotics for the S. Brancaster isolates. S. Brancaster, with the phenotypic ACSSuT-resistance pattern (ampicillin, chloramphenicol, streptomycin, sulphonamides, and tetracycline), was subjected to PCR to detect the corresponding resistance gene(s). Results: Virulence genes detected in S. Brancaster in this study were invA, sitC, spiA, sipB, sopB, sifA, cdtB, and spvB. A total of 36 antibiogram patterns of S. Brancaster with a high level of multidrug resistance were observed, with ampicillin exhibiting the highest resistance. Over a third of the isolates displayed ACSSuT-resistance, and seven resistance genes (β-lactamase temoneira [bla_TEM], florfenicol/chloramphenicol resistance gene [floR], streptomycin resistance gene [strA], aminoglycoside nucleotidyltransferase gene [ant(3")-Ia], sulfonamides resistance gene [sul-1, sul-2], and tetracycline resistance gene [tetA]) were detected. Conclusion: Multidrug-resistant S. Brancaster from chickens harbored an array of virulence-associated genes similar to other clinically significant and invasive non-typhoidal Salmonella serovars, placing it as another significant foodborne zoonosis.

• Animal Bioscience
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• 제36권11호
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• pp.1700-1708
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• 2023

Objective: The objective of this study was to determine the effects of β-mannanase on metabolizable energy (ME) and apparent total tract digestibility (ATTD) of protein in various feedstuffs including barley, copra meal, corn, corn distillers dried grains with solubles (DDGS), palm kernel meal, sorghum, and soybean meal. Methods: A basal diet was formulated with 94.8% corn and 0.77% amino acids, minerals, and vitamins and test diets replacing corn-basal diets with barley, corn DDGS, sorghum, soybean meal, or wheat (50%, respectively) and copra meal or palm kernel meal (30%, respectively). The basal diet and test diets were evaluated by using triplicated or quadruplicated 2×2 Latin square designs consisting of 2 diets and 2 periods with a total of 54 barrows at 20.6±0.6 kg (9 wk of age). Dietary treatments were levels of β-mannanase supplementation (0 or 800 U/kg of feed). Fecal and urine samples were collected for 4 d following a 4-d adaptation period. The ME and ATTD of crude protein (CP) in feedstuffs were calculated by a difference procedure. Data were analyzed using Proc general linear model of SAS. Results: Supplementation of β-mannanase improved (p<0.05) ME of barley (10.4%), palm kernel meal (12.4%), sorghum (6.0%), and soybean meal (2.9%) fed to growing pigs. Supplementation of β-mannanase increased (p<0.05) ATTD of CP in palm kernel meal (8.8%) and tended to increase (p = 0.061) ATTD of CP in copra meal (18.0%) fed to growing pigs. Conclusion: This study indicates that various factors such as the structure and the amount of β-mannans, water binding capacity, and the level of resistant starch vary among feedstuffs and the efficacy of supplemental β-mannanase may be influenced by these factors.

• 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.

• The Plant Pathology Journal
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• 제40권2호
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• pp.106-114
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• 2024

Fusarium head blight (FHB), predominantly caused by Fusarium graminearum and F. asiaticum, is a significant fungal disease impacting small-grain cereals. The absence of highly resistant cultivars underscores the need for vigilant FHB surveillance to mitigate its detrimental effects. In 2023, a notable FHB outbreak occurred in the southern region of Korea. We assessed FHB disease severity by quantifying infected spikelets and grains. Isolating fungal pathogens from infected samples often encounters interference from various microorganisms. We developed a cost-effective, selective medium, named BGT (Burkholderia glumae Toxoflavin) medium, utilizing B. glumae, which is primarily known for causing bacterial panicle blight in rice. This medium exhibited selective growth properties, predominantly supporting Fusarium spp., while substantially inhibiting the growth of other fungi. Using the BGT medium, we isolated F. graminearum and F. asiaticum from infected wheat and barley samples across Korea. To further streamline the process, we used a direct PCR approach to amplify the translation elongation factor 1-α (TEF-1α) region without a separate genomic DNA extraction step. Phylogenetic analysis of the TEF-1α region revealed that the majority of the isolates were identified as F. asiaticum. Our results demonstrate that BGT medium is an effective tool for FHB diagnosis and Fusarium strain isolation.

• 대한소아치과학회지
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• 제36권2호
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• pp.237-244
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• 2009

치근단 감염 시 물리적인 작용으로 증상이 완화되지 않을 때, 근관 내 세균을 제거하기 위해 항균성을 지닌 소독제나 세척제가 필요하다. 천연항균제에 대한 관심이 증가하면서 여러 연구들에 의해 고추냉이 추출물의 구성 성분 중 하나인 allyl isothiocyanate에 대한 항균성이 밝혀졌다. 이에 본 연구에서는 근관내 감염시 자주 발견되는 세균들 중 편성혐기성 세균인 Fusobacterium nucleatum, Prevotella nigrescence, 그리고 항생제에 내성이 커서 항균 작용실험 시 대조균으로 자주 사용되는 Clostidial perfringens에 대한 서양산 고추냉이 뿌리 추출물의 항균효과를 알아보고, 대표적인 항균제인 클로르헥시딘과 비교하여 다음과 같은 결과를 얻었다. 1. 본 연구에 사용된 3종의 편성 혐기성 세균에 대한 서양산 고추냉이 뿌리 추출물의 최소 억제 농도는 평균 87-470 ppm, 최소 살균 농도는 평균 156-625 ppm으로 F. nucleatum에 대해 가장 강한 항균효과를 나타내었으며, 항균제에 저항성 이 큰 C. perfringens에도 항균효과를 나타내었다. 2. 본 연구에 사용된 3종의 편성 혐기성 세균에 대한 클로르헥시딘의 최소억제 농도는 평균 3.12-6.25 ppm, 최소 살균 농도는 평균 10.94 ppm이었다. 3. 서양산 고추냉이 뿌리 추출물은 87-470 ppm의 농도에서 3.12-6.25 ppm의 클로르헥시딘과 비슷한 세균성장 억제 효과를 가지며 156-625 ppm의 서양산 고추냉이 뿌리 추출물은 10.94 ppm의 클로르헥시딘과 비슷한 살균 효과를 보였다.