• Asian-Australasian Journal of Animal Sciences
• /
• 제29권10호
• /
• pp.1490-1499
• /
• 2016

In this study, 69 lactobacilli isolated from Tibetan Qula, a raw yak milk cheese, were screened for their potential use as probiotics. The isolates were tested in terms of: Their ability to survive at pH 2.0, pH 3.0, and in the presence of 0.3% bile salts; tolerance of simulated gastric and intestinal juices; antimicrobial activity; sensitivity against 11 specific antibiotics; and their cell surface hydrophobicity. The results show that out of the 69 strains, 29 strains (42%) had survival rates above 90% after 2 h of incubation at pH values of 2.0 or 3.0. Of these 29 strains, 21 strains showed a tolerance for 0.3% bile salt. Incubation of these 21 isolates in simulated gastrointestinal fluid for 3 h revealed survival rates above 90%; the survival rate for 20 of these isolates remained above 90% after 4 h of incubation in simulated intestinal fluid. The viable counts of bacteria after incubation in simulated gastric fluid for 3 h and simulated intestinal fluid for 4 h were both significantly different compared with the counts at 0 h (p<0.001). Further screening performed on the above 20 isolates indicated that all 20 lactobacilli strains exhibited inhibitory activity against Micrococcus luteus ATCC 4698, Bacillus subtilis ATCC 6633, Listeria monocytogenes ATCC 19115, and Salmonella enterica ATCC 43971. Moreover, all of the strains were resistant to vancomycin and streptomycin. Of the 20 strains, three were resistant to all 11 elected antibiotics (ciprofloxacin, erythromycin, tetracycline, penicillin G, ampicillin, streptomycin, polymyxin B, vancomycin, chloramphenicol, rifampicin, and gentamicin) in this study, and five were sensitive to more than half of the antibiotics. Additionally, the cell surface hydrophobicity of seven of the 20 lactobacilli strains was above 70%, including strains Lactobacillus casei 1,133 (92%), Lactobacillus plantarum 1086-1 (82%), Lactobacillus casei 1089 (81%), Lactobacillus casei 1138 (79%), Lactobacillus buchneri 1059 (78%), Lactobacillus plantarum 1141 (75%), and Lactobacillus plantarum 1197 (71%). Together, these results suggest that these seven strains are good probiotic candidates, and that tolerance against bile acid, simulated gastric and intestinal juices, antimicrobial activity, antibiotic resistance, and cell surface hydrophobicity could be adopted for preliminary screening of potentially probiotic lactobacilli.

• 생명과학회지
• /
• 제10권3호
• /
• pp.262-272
• /
• 2000

설사환자에 대한 기초 역학자료의 일환으로 부산지역의 설사환자로부터 분리한 대장균의 혈청형 분 포와 항생제 내성유형을 확인한 결과 다음과 같은 성적을 얻었다. 1. 부산시 관내의 설사환자 1,239명의 분변으로부터 대장균을 분리한 결과 1,239명중 621명의 환자분변으로부터 721주의 대장균을 분리 동정 하였다. 2. 설사 환자의 월별 발생 분포는 8월에 131명으로 가장 높은 발생율을 나타낸 반면, 1월에 59 명으로 가장 낮은 발생율을 보였다. 그러나 1월에 발생한 설사환자를 제외하고는 계절에 크게 관계없이 연중 지속적으로 고른 발생 분포를 나타내었다. 연령별 발생분포는 1세 이하의 유아와 10세 이하의 어 린이가 633명으로 전체 설사환자의 50% 이상을 나타내었고, 50대의 경우 123명, 40대의 경우 114명, 60 대의 경우 99명, 30대의 경우 79명, 10대의 경우 74명, 20대의 경우 63명의 순으로 높은 발생율을 나타 내었으며 70세 이상의 노인들은 54명으로 가장 갖은 발생율을 보였다. 3. 혈청형별 분포는 혈청형 O44 가 121주(16.8%)로서 가장 높은 분포도를 나타내었으며, O153 (8.6%), O1(7.5%), 등에는 분리균 모두 내 성을 나타내었다. 공시약제에 대한 분리주의 내성유형은 14제 내성형(26.1%), 13제 내성형(22.3%), 12제 내성형(17.3%), 11제 내성형(14.8%), 10제 내성형(8.5%), 9제 내성형(6.0%) 등의 순으로 높은 분포를 보 였으며, 본 실험에 공시된 16종의 항생제에 대한 설사환자유래 대장균의 내성유형은 총 125종류로 분류 되었다. 특히 Am Cb Cl Co Cp Em Gm Mx Nb Pg St Tb Tc Tm 의 14제 내성형이 전체 분리균주의 24.3%로 가장 높은 내성유형을 나타내었다.

• 한국식품과학회지
• /
• 제37권3호
• /
• pp.456-464
• /
• 2005

본 연구에서는 최근 전세계적으로 문제가 되고 있는 항생제 내성균에 대해 그 항균효과를 천연의 약용식물을 이용하여 분자생물학적 수준에서 비교해보고 천연항균제의 개발에 우선하여 직접 식품에 첨가하여 그 효과를 검색하고자 하였다. 병원내감염의 주된 원인균으로 알려진 황색포도상구균(Staphylococcusaureus)은 여러 항균제에 대하여 내성을 획득함으로써 질병의 치료를 어렵게 한다는 점에서 매우 심각한 문제이다. 이러한 내성균은 다양한 방법으로 동 식물에 빠르게 전파되고 있으므로 소비자가 내성균을 지닌 식품을 섭취해 잠재적인 내성균을 보유하게 되는 결과를 낳게 된다. 이러한 항생제 내성 황색포도상구균(Methicillin Resistant S. aureus, MRSA)에 대해 천연 약용 식물 중 우수한 항균력을 가지는 감초를 순차 분획하여 각 분획물을 이용한 MRSA에 대한 항균 활성을 알아본 결과 hexane, chloroform 분획물에서 항균활성을 나타냈으며, 동일농도의 생육저해비교 실험에서 대부분의 내성균주에 대해 대조군과 유사한 효과를 나타낸 penicillin에 비해 감초분획물이 대수기를 넘는 억제효과를 나타내었으며 이 중 chloroform 분획물이 가장 높은 저해활성을 나타내었다. 또한 RT-PCR을 통하여 표준균주와 내성균주의 내성 정도와 내성 유전자를 관찰해본 결과 KCCM 40510은 내성획득유전자인 mecA, mecRI, mecI, femA에 발현이 높게 나타나 고도내성균임을 알 수 있었으며, 고도내성을 나타내는 균주를 이용하여 감초의 chloroform 분획물을 농도별로($50,\;100,\;250\;{\mu}/mL$) 처리한 결과 농도 의존적으로 유전자 발현이 억제됨을 볼 수 있었다. 특히 고도내성균주인 KCCM 40510은 메티실린 내성을 상승시키는 mecA 유전자의 발현을 억제시키는 mecRI 유전자의 pathway와 상응하는 것으로 보여지므로 감초가 이러한 고도내성균에 효과가 있음을 알 수 있었다. 또한 SEM을 통한 형태학적 관찰을 통해 분획물 처리 시 세포벽이 파괴되는 것을 확인할 수 있었다. 한편 이러한 감초추출물을 직접 식품에 첨가하여 내성 균주에 대한 억제 효과도 확인한 결과 추출물 200mg/100mL 농도에서 포도상구균에 대해 항균효과를 나타내는 것을 볼 수 있었다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• 제32권5호
• /
• pp.436-446
• /
• 2006

The purpose of this study was to isolate and identify the bacteria in chronic maxillary sinusitis (CMS) lesions from 3 patients and to determine the antimicrobial susceptibility of them against 10 antibiotics. One of them was odontogenic origin and the others were non-odontogenic origin. Pus samples were collected by needle aspiration from the lesions and examined by culture method. Bacterial culture was performed in three culture systems (anaerobic, CO2, and aerobic incubator). Identification of the bacteria was performed by 16S rRNA gene (16S rDNA) nucleotide sequencing method. To test the sensitivity of the bacteria isolated from the maxillary sinusitis lesions against seven antibiotics, penicillin G, amoxicillin, tetracycline, ciprofloxacin, cefuroxime, erythromycin, clindamycin, and vancomycin, minimum inhibitory concentration (MIC) was performed using broth dilution assay. Our data showed that enterobacteria such as Enterobacter aerogenes (30%), Klebsiella pneumoniae (25%), and Serratia marcescens (15%) were predominately isolated from the lesion of non-odontogenic CMS of senile patient (70 year old). Streptococcus spp. (40.3%), Actinomyces spp. (27.4%), P. nigrescens, M. micros, and P. anaerobius strains were isolated in the lesion of odontogenic CMS. In the lesion of non-odontogenic CMS, Streptococcus spp. (68.4%), Rothia spp. (13.2%), and Actinomyces sp. (10.5%) were isolated. The susceptibility pattern of 10 antibiotics was determined according to the host of the bacteria strains ratter than the kinds of bacterial species. Even though the number of CMS was limited as three, these results indicate that antibiotic susceptibility test must be accompanied with treatment of CMS. The combined treatment of two or more antibiotics is better than single antibiotic treatment in the presence of multidrug-resistant bacteria in the CMS lesions.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• 제34권5호
• /
• pp.562-570
• /
• 2008

Most purulent maxillofacial infections are of odontogenic origin. Treatment of infection includes the surgical intervention, such as incision and drainage, and adjunctive treatment. The use of high-dose antibiotics is also indicated. The choice of an antibiotics should be based on the knowledge of the usual causative microbes and the results of antibacterial sensitivity test. We have undertaken clinical studies on 119 patients in Dept. of Oral and Maxillofacial Surgery, Inha University Hospital from January 2000 to December 2007. Many anaerobic microbes are killed quickly when exposed to oxygen. Thus the needle aspiration techniques and the transfer under inert gas were used when culturing. The aim of this study was to obtain informations for the bacteriologic features and the effective antimicrobial therapy against maxillofaical odontogenic infections. The obtained results were as follows: 1. The most frequent causes of infections were odontogenic (88.3%), and in odontogenic cause, pulpal infections were the most common causes(53.8%). 2. The buccal and submandibular spaces (respectively 23.5%) were the most frequent involved fascial spaces, followed by masticator spaces (14.3%). 3. The most common underlying medical problems were diabetes (17.6%), however the relation with prognosis was not discovered. 4. The complications were the expiry, mediastinitis, necrotizing fasciitis, orbital abscess, and osteomyelitis. 5. The most common admission periods were 1-2 weeks, and the most patients were discharged within 3 weeks. However, patients who admitted over 5 weeks were about 10%. 6. A total of 99 bacterial strains (1.1 strains per abscess) was isolated from 93 patients (78.2%). The most common bacterium isolated was Streptococcus viridans (46.2%), followed by $\beta$-hemolytic group streptococcus (10.1%). 7. Penicillins (penicillin G 58.3%, oxacillin 80.0%, ampicillin 80.0%) have slightly lower sensitivity. Thus we recommend the antibiotics, such as glycopeptides (teicoplanin 100%, vancomycin 100%) and quinolones (ciprofloxacin 90.0%) which have high susceptibility in cases in which peni cillin therapy failed or severe infections.

• Pediatric Infection and Vaccine
• /
• 제21권1호
• /
• pp.65-70
• /
• 2014

선천성 매독은 산전진찰로 예방할 수 있음에도 불구하고, 여전히 문제가 되고 있다. 최근에는 선천성 매독과 관련된 혈뇨나 폐렴이 보고된 증례가 없었으나 저자들은 적절한 산전치료를 받지 못한 산모에게서 태어난 22일된 남아에게서 발생한 선천성 매독에 동반된 신증후군과 폐렴의 증례를 보고한다. 선천 매독의 진단은 혈청학적 검사로 확진하였고, 환아는 페니실린 G 치료를 받고 회복되었다. 신생아에게서 임상증상은 매우 불확실할 수 있어, 질환에 대한 인식이 지연 될 수 있다. 따라서 선천성 매독의 예방에는 철저한 산전관리가 필수적이다. 만약 산모가 적절한 산전진찰을 받지 않았을 경우에는 신속한 혈청학적 진단이 반드시 시행되어야 할 것이다.

• 한국동물번식학회:학술대회논문집
• /
• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
• /
• pp.67-67
• /
• 2003

Early development of porcine oocytes fertilized in vitro was examined in different culture conditions. Porcine ovaries were collected from local slaughter-house. Cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cystein, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for 42~44 hrs. The frozen-thawed spermatozoa were washed by centrifigation 2 times at 1, 500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, 1$\mu\textrm{g}$/ml streptomycin and 1ng/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to 2.5$\times$10$^{6}$ cells/ml motile sperm during fertilization in vitro. At 8hrs h after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine and 4 mg/ml BSA and cultured for 7 days. In first experiment, the mean numbers of oocytes collected from 20 ovaries were 674.4 oocytes, and 4.1(27.6), 12.5(84.0), 25.4(171.6) and 57.9%(390.8) for A, B, C and D grade in morphological classification. In the second experiment, when culture medium was supplemented with various concentrations of EGF, the proportions of oocytes cleaved were 56.9, 55.7, 61.9 and 54.7% for 0, 5, 10 and 20ng/ml EGF. The higher proportions(15.1%) of oocytes developed to morular stage were obtained at concentration of 10ng/ml than 0 and 5ng/ml EGF (P<0.05). However, the proportions of embryos developed to blastocyst stage were not significantly different among concentrations of EGF. In another experiment, when the medium supplemented with catalase was used, the proportions of oocytes cleaved were higher in the concentration of 0 unit (56.5%, 61/108) than 100 and 1, 000 unit/ml of catalase (P<0.05). Although the developmental capacity of embryos was improved by medium with 0 unit/ml compared with 100, 500 and 1, 000 units/ml of catalase in oocytes developed to morula and blastocyst stages, were not significantly different among concentrations of catalase.

• 한국어병학회지
• /
• 제9권2호
• /
• pp.195-201
• /
• 1996

군산근교의 양만장에서 애드와드병에 이환된 뱀장어로부터 총 96균주의 E.tarda균을 분리한 후 이들 분리균의 약제감수성 검사를 실시하고 아울러 내성균의 내성인자 전달 시험을 실시하였다. 공시한 모든 균주는 SM, AK, NF 및 GM등에는 100% 감수성을 보였으나, 대부분의 균주가 SD(86균주), AM(84균주), PM(80균주), NA(67균주), OT(44균주) 및 OA(37균주) 등의 내성을 보였다. 내성유형은 20유형으로서 거의 모든 균주가 2제 이상의 다제내성을 보였으며, 그중 SD-AM-PM-NA-OA(16균주), SD-AM-PM-NA(14균주), SD-AM-PM-NA-OT-OA(12균주), SD-AM-PM-OT(10균주) 및 SD-AM-PM-NA-OT(8균주) 유형등이 고빈도로 출현하였다. 내성균주의 내성인자 전달 빈도는 94균주중에서 78균주가 단계에서 6제까지의 다제내성인자의 전달을 보였다. 그중 AM 및 AM-PM-NA유형(8균주), PM,SD 및 AM-SD유형(6균주), AM-PM, PM-SD 및 AM-SD-OT유형(4균주)등이 고빈도 출현을 보였다. 이러한 결과는 양만장에서 다양한 항균제를 사용하였기 때문에 다제내성현상이 확산되어진 것으로 사료된다.

• 한국축산식품학회지
• /
• 제28권5호
• /
• pp.587-595
• /
• 2008

본 연구는 국내 각 지역의 목장에서 수거한 원유에서 분리된 젖산균을 대상으로 10% 환원탈지유에 $37^{\circ}C$에서 pH 4.4에 도달할 때까지 배양한 다음 각각에서 얻어진 유청으로 ACE 저해율을 측정한 결과 88.6%인 우수한 균주를 선발하였다. 선정된 균은 Gram 양성, rod형태의 homo균이며, 당 발효실험과 16S rRNA 분석결과 Lactobacillus zeae로 판명되었고, Lactobacillus zeae RMK354로 명명하였다. 발효유에 적합한 starter인지 확인하기 위해 생리적 특성을 조사하였다. L. zeae RMK354는 배양온도 $40^{\circ}C$에서 빠른 생장을 보였고, pH4.3에 도달하는데 10시간이 소요되었다. 16종의 항생제 중 polymyxin B와 vancomycin에 대해 내성이 높았으며, 효소활성실험에서 esterase와 leucine arylamidase의 활성도가 높았다. 담즙에 대한 내성이 있는 것으로 나타났으며, pH 내성 실험결과 pH 2에서 큰 변화가 없음에 따라 내신성이 있었다. 항균력 시험에서는 Escherichia coli와 Staphylococcus aureus에 대해 억제력이 없었으나 Salmonella typhimurium에 대해 60.0%의 항균력을 보였다. 이러한 결과를 토대로 ACE 억제활성능이 우수한 기능성 발효유 제품의 스타터로 L. zeae RMK354는 적합하다고 할 수 있다.

• Journal of Periodontal and Implant Science
• /
• 제32권4호
• /
• pp.865-873
• /
• 2002

The primary cause of tooth loss after 30 years of age is periodontal disease. Destruction of alveolar bone by periodontal disease is done by bone resorbing activity of osteoclasts. Understanding differentiation and activation mechanism of osteoclasts is essential for controling periodontal disease. The purpose of this study is to identify the possible effects of Vitamin D and cytokines affecting osteoclasts and its precursor cells. Four to six week-old mice were killed and humerus, radius, tibia and femur were removed aseptically and washed two times with Hank's solution containing penicillin-streptomycin and then soft tissue were removed. Bone marrow cells were collected by 22 gauge needle. Cells were cultured in Hank's solution containing 1 mg/ml type II collagenase, 0.05% trypsin, 41mM EDTA. Supernatant solution was removed 5 times after 15 minutes of digestion with above mentioned enzyme solution, and remained bone particles were maintained in alpha-MEM for 15 minutes and $4^{\circ}C$ temperature. Bone particles were agitated for 1 minute and supernatant solution containing osteoclast precursor cells were filtrated with cell stainer. These separated osteoclast precursor cells were dispensed with 100-mm culture dish by $1{\times}10^7$ cells unit and cultured in ${\alpha}$- MEM containing 20 ng/ml recombinant human M-CSF, 30 ng/ml recombinant human soluble osteoclast differentiation factor and 10% fetal calf serum for 2 and 7 days. Total RNA of osteoclast precursor cells were extracted using RNeasy kit. One ${\mu}g$ of total RNA was reverse transcribed in $42^{\circ}C$ for 30 minutes using SuperScriptII reverse transcriptase. Expression of transcribed receptors of each hormone and cytokine were traced with 1 ${\mu}l$ of cDNA solution by PCR amplification. Vitamin D receptor WAS found in cells cultured for 7 days. TNF-${\alpha}$ receptor was found in cells cultured for 2 days and amount of receptors were increased by 7 days. IL-1 type I receptor was not found in cells cultured 2 and 7 days. But, IL-1 receptor type II was found in cells cultured for 2 days. TGF-${\alpha},{\beta}$type I receptor was found in cells cultured 2 and 7 days, and amount of receptors were increased by 7 days of culture. These results implies Vitamin D and cytokines can affect osteoclasts directly, and affecting period in differentiation cycle of osteoclasts is different by Vitamin D and cytokines.