본 시험은 무항생제 사육방식이 한우육내 코티졸과 지방산 함량에 미치는 영향을 구명하고자 실시하였다. 무항생제 및 관행 축산으로 생산된 한우육 1등급 등심(M. longissimus)부위를 3가지의 브랜드별로 각각 3회씩 구매하여 시료로 공시하였고, 분석한 결과는 다음과 같다. 한우육 등심내 코티졸 함량은 무항생제 처리구에서 관행사육 처리구 대비 유의적(p=0.0176)으로 낮게 나타났으며, 포화지방산 및 불포화지방산 함량에서는 유의적(p>0.05)인 차이를 나타내지는 않았으나, 관행사육 처리구에서는 포화지방산이, 무항생제 처리구에서는 불포화지방산이 각각 높은 경향으로 분석되었다. 또한, 불포화지방산 중 n-3과 n-9 지방산을 제외한 n-6 지방산(linoleic acid, γ-lino-lenic acid)에서 유의적(p<0.01)인 차이를 보였다. 또한, 한우육내 코티졸 농도와 n-6 지방산 농도간의 상관관계를 분석한 결과, 특히 linoleic acid와 유의적(p=0.0140)으로 부(—)의 상관관계가 성립되었다.결과적으로, 무항생제 인증으로 사육된 한우육은 관행적으로 사육된 한우육에서 보다 상대적으로 코티졸 함량이 낮았으며, 건강에 유익한 불포화지방산 함량이 다소 높은 경향으로 분석됨으로써 육질을 향상시키는 것으로 나타났다.

Minimal inhibitory concentration (MIC) is the lowest concentration of antibiotics that inhibits the visible growth of a microorganism. It has been reported that sub-MIC of antibiotics may result in morphological alterations along with biochemical and physiological changes in bacteria. The purpose of this study was to examine morphological changes of periodontal pathogens after treatment with sub-MIC antibiotics. Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Porphyromonas gingivalis were used in this study. The MIC for amoxicillin, doxycycline, metronidazole, penicillin, and tetracycline were determined by broth dilution method. The bacterial morphology was observed with bright field microscope after incubating with sub-MIC antibiotics. The length of A. actinomycetemcomitans and F. nucleatum were increased after incubation with metronidazole; penicillin and amoxicillin. P. gingivalis were increased after incubating with metronidazole and penicillin. However, F. nucleatum showed decreased length after incubation with doxycycline and tetracycline. In this study, we observed that sub-MIC antibiotics can affect the morphology of periodontal pathogens.

The objective of this study was to develop a simultaneous method of 8 penicillin antibiotics including amoxicillin, ampicillin, cloxacillin, dicloxacillin, nafcillin, oxacillin, penicillin G and penicillin V in meat using LC-MS/MS. The procedure involves solid phase extraction with HLB cartridge and subsequent analysis by LC-MS/ MS. To optimize MS analytical condition of 8 compounds, each parameter was established by multiple reaction monitoring in positive ion mode. The chromatographic separation was achieved on a C18 column with a mobile phase of 0.05% formic acid and 0.05% formic acid in acetonitrile at a flow rate of 0.2 mL/min for 20 min with a gradient elution. The developed method was validated for specificity, linearity, accuracy and precision in beef, pork and chicken. The recoveries were 71.0~106%, and relative standard deviations (RSD) were 4.0~11.2%. The limit of detection (LOD) and the limit of quantification (LOQ) were 0.003~0.008 mg/kg and 0.01~0.03 mg/kg, respectively, that are below maximum residue limit (MRL) of the penicillins. This study also performed survey of residual penicillin antibiotics for 193 samples of beef, pork and chicken collected from 9 cities in Korea. Penicillins were not found in all the samples except a sample of pork which contained cloxacillin (concentration of 0.08 mg/kg) below the MRL (0.3 mg/kg).

A simultaneous determination was developed for 9 aminoglycoside antibiotics (amikacin, apramycin, dihydrostreptomycin, gentamicin, hygromycin B, kanamycin, neomycin, spectinomycin, and streptomycin) in meat by liquid chromatography tandem mass spectrometry (LC-MS/MS). Each parameter was established by multiple reaction monitoring in positive ion mode. The developed method was validated for specificity, linearity, accuracy, and precision based on CODEX validation guideline. Linearity was over 0.98 with calibration curves of the mixed standards. Recovery of 9 aminoglycosides ranged on 60.5~114% for beef, 60.1~112% for pork and 63.8~131% for chicken. The limit of detection (LOD) and limit of quantification (LOQ) were 0.001~0.009 mg/kg and 0.006~ 0.03 mg/kg, respectively in livestock products including beef, pork and chicken. This study also performed survey of residual aminoglycoside antibiotics for 193 samples of beef, pork and chicken collected from 9 cities in Korea. Aminoglycosides were not found in any of the samples.

본 연구는 하수 및 가축분뇨 처리장 방류수, 하천수를 이용하여 반코마이신 내성 장구균(Vancomycin Resistant Enterococci, VRE)의 검출여부를 확인하고 검출된 장구 균의 표현형을 분석함으로써 하천에 대한 공공처리시설 의 방류수에 포함되어있는 엔테로코커스의 영향을 파악 하여 항생제 내성균이 환경에 미치는 영향 및 항생제 내 성 연구에 기초 자료가 되고자 하였다. 엔테로코커스의 동정시험 결과 모두 32균주가 검출되었고 이는 모두 E. faecium으로 동정되었다. Multiplex PCR 시행한 결과 32 균주 모두 Van A 표현형에 해당하는 반코마이신 내성 장 구균으로 확인되었다. 조사지점별 세균의 E. faecium 집 락수의 측정 결과 하수처리장 지역에서 평균적으로 가장 높게 나타났고, 하천 지역의 경우 상류지점에서 하류 지 점으로 갈수록 세균의 colony 숫자도 늘어나는 것으로 조 사되었다. 하수처리장 방류수 시료 26균주에 대한 최소억 제농도 검사 결과, 19개의 항생제 중 내성, 감수성을 보인항생제 수는 각각 14, 5종으로 내성을 보이는 항생제의 수가 약 70% 이상으로 대부분을 차지하였다. 하천 시료 6균주에 대한 최소억제농도 검사 결과, 19개의 항생제 중 내성, 감수성을 보인 항생제 수는 각각 14, 5종으로 내성을 보이는 항생제의 수가 약 70% 이상으로 대부분 을 차지하여 하수처리장 방류수 시료의 검사 결과와 동 일하였다.

본 논문에서는 축산물 중 닭고기 내 마크로라이드계(MLs) 항생물질(spiramycin, josamycin, tilmicosin, 그리고tylosin) 4종을 분석하기 위하여, 액-액 추출 과정을 거쳐서 PDA 검출기가 장착된 액체크로마토그래피를 이용하여MLs를 효율적으로 동시분석하는 방법을 확립하였다. 컬럼은 Unison UK-C18 (150 mm × 3 mm id, 3 μm), 이동상 용매는 0.1% TFA와 0.1% TFA in ACN로 기울기 용리를사용하였으며, 유속은 0.7 mL/min, 그리고, 주입량은 10 μL로 설정하여 분석하였다. 확립된 분석조건으로, 표준검정곡선은 50-1000 μg/kg의 농도범위에서 상관계수가 0.9975이상의 양호한 직선성을 나타내었으며, 회수율은 저, 중,고농도로 첨가하여 분석한 결과, 80.4-99.1%를 나타내었다. 검출한계는 8.8-19.6 μg/kg이었고, 정량한계는 26.6-59.4μg/kg이었으며, 일내(intra-day)와 일간(inter-day) 정밀도(CV%)는 0.9-13.2%, 2.4-13.1%이었다. 따라서, 확립된 분석방법은 축산물 중 닭고기 시료 내 MLs를 효과적으로분석하는데 이용될 수 있을 것으로 사료된다.

A investigation of electrochemical analysis of antibiotics Neomycin (C23H46N6O13) was searched using electrochemical square wave (SW) stripping and cyclic voltammetry (CV) using working sensor of the modified carbon nanotube combination electrodes, optimum diagnostic parameters were searched by anodic stripping, final conditions were attained to working range of 1.0-14.0 ng/L, detection limit (S/N) was found to be 0.6 ng/L. The developed method was discovered to be fitting in quality control in the food, pharmaceutical and other manufacturing sectors.

A rapid, simple and reliable LC-MS/MS method, which can be used on a routine basis, was developed for the simultaneous detection of 8 penicillin antibiotics (amoxicillin, ampicillin, penicillin-G, penicillin-V, oxacillin, cloxacillin, nafcillin and dicloxacillin) in swine muscle and kidney. The antibiotics were extracted from samples with water and methanol. The extract was centrifuged, filtered and analyzed by liquid chromatography coupled with a tandem mass spectrometer (LC-MS/MS), using a C18 reversed phase column with water/acetonitrile gradient containing 0.05 % formic acid. Mass spectral acquisition was achieved in an electrospray positive ion mode by applying multiple reaction monitoring (MRM) of 2 fragment ion transitions to provide a high degree of sensitivity and specificity. The ion rations were consistent and could be used for confirmation of identity of the penicillin antibiotics. Recoveries of eight penicillins at three fortification levels (10, 20 and 40㎍/㎏) ranged from 79.8 to 102.4% and 72.8 to 103.4% in swine muscle and kidney, respectively. The coefficient of variation was than 9% in all samples. The estimated limits of quantification (LOQs) ranged from 1.0 to 3.2㎍/㎏ in swine muscle and kidney, respectively. The LOQs of this method are below the MRLs of penicillin antibiotics in animal tissues established in Korea and other countries.

The discovery of antibiotics has helped to save the lives of an uncountable number of people. Antibiotics have been grouped in different classes based on their origin, structure, and mechanism of action. An intrinsic and acquired mechanism of antimicrobial resistance has been identified in many bacterial strains that are of high clinical importance. This has seriously jeopardized the use of antibiotics and has also caused the spread of microbes that are resistant to effective first-choice, or “first-line” drugs. Thus, sensible use of antibiotics and the search for effective alternative measures are of high importance in order to minimize the effect due to existing and emerging antimicrobial resistant microbes.

Postantibiotic effect (PAE) is defined as the length of time that bacterial growth is suppressed following brief exposure to an antibiotic. In this study, the in vitro PAE, postantibiotic sub-MIC effect (PA SME) and sub-MIC effect (SME) of antibiotics on Treponema denticola ATCC 35405 were investigated. The PAE of doxycycline and metronidazole were 20.3 h and 25.0 h, respectively. The PA SMEs examined by addition of 0.1, 0.2 and 0.3X MICs during the postantibiotic phase of the bacteria for metronidazole were longer than those for doxycycline. In contrast, the SMEs for doxycycline were longer than those for metronidazole. The PA-SME and SME values increased as the concentration of antibiotics increased. The present study illustrates the existence of PAE, PA-SME and SME for several antibiotics against T. denticola, thereby extending the pharmacodynamic advantages of these antibiotics.

Antibiotics marker-free with herbicide-resistant tall fescue plants were produced through Agrobacterium: mediated transformation system. Embryogenic calli derived calli were infected with Agrobacterium tumefaciens strain EHA105 harboring the pCAMBIA3300 vector containing the bar and the CP4-EPSPS genes. The PPT-resistant calli and plants were selected with 10 ㎎/L PPT, respectively. Soil-grown plants were obtained about 14-16 weeks after Agrobacterium-mediated transformation. Genetic transformation of the regenerated plants growing under selection was demonstrated by PCR, and Southern blot analysis revealed that one copies of the transgene were integrated into the plant genome of each transgenic plant. Expression of the bar and CP4-EPSPS genes in transgenic plants was confirmed by Northern blot analysis and CP4-EPSPS genes in transgenic plant ELISA uses antibody protein by ELISA Assays. Transgenic plants sprayed of two herbicides with glufosinate ammonium or glyphosate remained green and healthy. We therefore report here a successful and reliable Agrobacterium-mediated transformation of two herbicide-resistances and this method may be useful for routine transformation and has the potential to develop new varieties of tall fescue with several important genes.

Thrombin-induced platelet microbicidal protein (tPMP) is a small cationic peptide that exerts potent in vitro microbicidal activity against a broad spectrum of human pathogens, including Staphylococcus aureus and Streptococcus rattus BHT. Earlier evidence has suggested that tPMP targets and disrupts the bacterial membrane. However, it is not yet clear whether membrane disruption itself is sufficient to kill the bacteria or whether subsequent, presumably intracellular, events are also involved in this process. In this study, we investigated the microbicidal activity of rabbit tPMP toward S. rattus BHT cells in the presence or absence of a pretreatment with antibiotics that differ in their mechanisms of action. The streptocidal effects of tPMP on control cells (no antibiotic pretreatment) were rapid and concentration-dependent. Pretreatment of S. rattus BHT cells with either penicillin or amoxicillin (inhibitors of bacterial cell wall synthesis) significantly enhanced the anti-S. rattus BHT effects of tPMP compared with the effects against the respective control cells over most tPMP concentration ranges tested. On the other hand, pretreatment of S. rattus BHT cells with tetracycline or doxycycline (30S ribosomal subunit inhibitors) significantly decreased the streptocidal effects of tPMP over a wide peptide concentration range. Furthermore, pretreatment with rifampin (an inhibitor of DNA-dependent RNA polymerase) essentially blocked the killing of S. rattus BHT by tPMP at most concentrations compared with the respective control cells. These results suggest that tPMP exerts anti-S. rattus BHT activity through mechanisms involving both the cell membrane and intracellular targets.

Postantibiotic effects (PAE) refer to suppression of the bacterial growth following limited periods of exposure to an antibiotic and subsequent to the removal of the antibiotic agent. Fusobacterium nucleatum and Porphyromonas gingivalis are Gram-negative anaerobic bacteria associated with several periodontal diseases. In this study, postantibiotic effects (PAE), postantibiotic sub-MIC effect (PA SME) and sub-MIC effect (SME) of antibiotics on F. nucleatum ATCC 25586 and P. gingivalis W50 were investigated. The PAE was induced by 10X the MIC of antibiotic and antibiotic was eliminated by washing. The PA SMEs were studied by addition of 0.1, 0.2 and 0.3X MICs during the postantibiotic phase of the bacteria, and the SMEs were studied by exposition of the bacteria to antibiotic at the sub-MICs only. Amoxicillin, doxycycline and tetracycline induced PAE for F. nucleatum ATCC 25586 and P. gingivalis W50. But metronidazole and penicillin induced PAE for only F. nucleatum ATCC 25586. Metronidazole and doxycycline induced PA SME and SME for both species of anaerobic bacteria used in this study. The PA SME values for both strains were substantially longer than the SME values. The present study showed the existence of PAE, PA SME and SME for various antibiotics against F. nucleatum ATCC 25586 and P. gingivalis W50.

Antibiotics polymer prepared by chemical bonding and simple blending of antibacterial into polymers have attracted much interest because of their long-lasting and antibacterial activity. Antibiotics polymer can significantly reduce losses associated with dissolution, photolytic decomposition and volatillization. Further more, increased efficiency safety and selectivity are additional benefits which may be realized. In this study, Antibiotics polymer was synthesized by chemical reaction of polyacrylic acid with sulfamethazine by N,N'-dicyclohexylcarbodiimide(DCC) method. Antibacterial susceptibility was determined against Streptococcus pyrogenes[gram(+)] and Esherichia coli.[gram(-)] using a standardized disc test. As a result, the synthetic antibiotics polymer exhibited the broad susceptibilty against Streptococcus pyrogenes and Esherichia coli. Especially, the antibiotic effect of antibacterial polymer against Gram negative(Esherichia coli) was much stronger than that against Gram positive(Streptococcus pyrogenes).

Dual-actions are the most recently used delivery system in drug study. Dual-action agents are unique chemical entities comprised of two different type of antibacterial compounds covalently linked together in a single molecule in such a way that both components are able to exert their bactericidal properties. Crosslinked sulfadiazine-sulfanilamide such as antibiotics is synthesized by synthetic handle with glutaraldehyde. As a result, New synthetic antibacterial agent exhibited the broad antibacterial activities against gram(+) and gram(-) of 4 strains and a long durability supposing that the stomach and blood.

Dual-action agents are unique chemical entities comprised of two different types of antibacterial compounds covalently linked together in a single molecule in such a way that both components are able to exert their bactericidal properties. Crosslinked sulfadiazine-sulfacetamide such as antibiotics synthesized by crosslingking reaction such as glutaraldehyde. These structures of the compounds were confirmed by IR, NMR. 4 strains of Gram(+) and Gram(-) revealed effective susceptibility to synthetic crosslinked sulfadiazine-sulfacetamide.

Recently, as concern about the residual antibiotics in milk increase, the detection methods of residual antibiotics used extensevely at the present time were investigated and compared to their properties and the detection limits of variable antibiotics. At first, comparative tests of the detectable sensitivity of 4 teat organisms, B.cereus, B.subtilis, M.luteus, B. stearothermophilus C-953, were performed by disc assay. As a result, B.stearotleermoph.ilus was the most sensitive strain of all other strains and showed the detect limit of 5-50 ppb for penicillins (PCs). And also, B.subtilis was showed the more effective detection limit, 200-400 ppb, for aminoglycosides (AGs) and M.luteus was showed predominant sensitivity , 50-500 ppb for macrolides(MLs) and B.cereus was the most sensitive strain for tetracyclines (TCs) and showed the detection limit of 100-400 ppb. Therefore, each test strains were showed a different sensitivity in the detection of the different antibiotic families. When the detection limit of disc assay and other methods were compared, TTCmethod was less sensitive than other methods showing 5-50 ppb detectable lebel for PCs. Also, for the detection of other antibiotic families TTC method was showed the worst sensitivity and Delvo and Charm Farm tests were similar to the detectable properties of AGs and MLs. Although disc assay was showed the similar detection limit for PCs with Delvo and Charm Farm, it was more widely effective for the detection of kanamycin, erythromycin, chlortetracycline, doxycycline, verginiamycin and so on than Delvo or Charm Farm. CharmII test was showed the best sensitivity for the most of antibiotics except neomycin and gentamycin. But it was necessary that different tests must be performed to each antibiotic family and so it was regarded that the effectiveness of that method was low.