• Title/Summary/Keyword: Antibiotic quantification

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Novel Approaches to Monitoring and Remediation of Veterinary Antibiotics in Soil and Water: A Review

  • Awad, Yasser M.;Lee, Sang-Soo;Kim, Sung-Chul;Yang, Jae-E.;Ok, Yong-Sik
    • Korean Journal of Environmental Agriculture
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    • v.29 no.4
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    • pp.315-327
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    • 2010
  • A vast increase of antibiotics usage in concentrated animal feeding operations (CAFOs) over the last few decades has led to an environmental risk due to the presence of antibiotic residuals in different environmental compartments. Especially in Korea, the use of antibiotics in CAFOs is much greater than in other developed countries. One of the primary adverse impacts of antibiotic residuals in the environment is that they readily produce antibiotic resistant bacteria (ARB), which exert detrimental effects on the ecosystem as well as human health. In this article, the impacts of veterinary antibiotic residuals with regard to their quantification and management, and desirable remediation technologies have been widely reviewed. This review article concluded that the continuous monitoring should be required to ensure the safety of antibiotic residuals in the surrounding environments. Furthermore, the management guidelines of antibiotic residuals need to be developed in the future.

Protective effect of bacteriophages against Salmonella Typhimurium infection in weaned piglets (이유자돈에서 Salmonella Typhimurium 감염에 대한 박테리오파지의 방어 효능)

  • Kim, Sung-Jae;Kim, Jae-Hoon;Jun, Soo-Yeon;Paik, Hyoung Rok;Han, Jeong-Hee
    • Korean Journal of Veterinary Service
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    • v.37 no.1
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    • pp.35-43
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    • 2014
  • Salmonellosis has caused heavy losses in swine industry and implications for public health. Recently, the urgent problem of antibiotic resistance due to multidrug-resistant Salmonella spp. has been on the rise. The use of host-specific bateriophages as a biocontrol is one possible alternative. In this study, clinical signs, growth performance, quantification and detection of antigen, histopathological changes of gastrointestinal tracts were analyzed comparatively in weaned piglets according to administration of bacteriophages and challenge with Salmonella (S.) Typhimurium. Piglets challenged with S. Typhimurium after administered with bacteriophages showed reduced clinical signs, higher growth performance, lower bacterial shedding, lower quantificational value of antigens in intestines, higher V/C ratio and higher the number of goblet cells in intestines than piglets administered without bacteriophage and challenged with S. Typhimurium. These results indicate that feeding contained with bacteriophages has effect to prevent infection of S. Typhimurium in weaned piglets and suggest that a use of bacteriophage can be considered a valid antibiotic alternative.

Determination of Residual Erythromycin Antibiotic in Fishery Products by Liquid Chromatography-electrospray Ionization Mass Spectrometry (LC-MS/MS를 이용한 어류 및 갑각류의 잔류 Erythromycin 항생제 분석)

  • Jo, Mi-Ra;Mok, Jong-Soo;Lee, Doo-Seog;Kim, Min-Jung;Kim, Poong-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.42 no.1
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    • pp.15-19
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    • 2009
  • A simple and sensitive method for erythromycin quantification by liquid chromatography electrospray mass spectrometry (LC-MS/MS) in fishery products was developed. Samples were extracted by liquid-liquid extraction using 70% acetonitrile. Lipids were removed by acetonitrile saturated hexane. LC separation was performed on a Shiseido UG C-18 column ($150\;mm{\times}2.0\;mm$ internal diameter.) with a gradient system of 0.2% acetic acid-acetonitrile containing 0.2% acetic acid as a mobile phase at flow rate of 0.2 mL/min. The mass spectrometer was operated in selected reaction monitoring with positive electro-spray interface. Transitions were monitored a m/z $734{\to}577$ and $734{\to}158$, with m/z $734{\to}577$ chosen for quantification. Recovery of erythromycin from fish and shrimp fortified at the 10 ng/mL, 50 ng/mL and 100 ng/mL were 91.6-109.4%, 84.4-111.2% and 98.8-109.6% with high precision, respectively. Limits of quantification and limits of detection of erythromycin in both fish and shrimp were 10.0 ng/mL and 1.0 ng/mL, respectively. This analysis method for erythromycin has been proposed for registration in the Korean Official Methods of Food Analysis and has been utilized for fishery products analysis by the Korea Food and Drug Adminstration and the National Fisheries Products Quality Inspection Service.

Analysis of antibiotic residues in milk from healthy dairy cows treated with bovine mastitis ointment using ultra-performance liquid chromatography coupled with electrospray tandem mass spectrometry

  • Park, Eun-Kee;Ryu, Yong-Jae;Cha, Chun-Nam;Yoo, Chang-Yeul;Kim, Suk;Lee, Hu-Jang
    • Korean Journal of Veterinary Research
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    • v.56 no.4
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    • pp.233-239
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    • 2016
  • This study was conducted to analyze penicillin G (PEG), streptomycin (STR) and neomycin (NEO) residues in milk of healthy lactating cows. Milk samples were collected from all four quarters of 12 dairy cows 2−7 days after intramammary infusions of an ointment containing PEG, STR and NEO once (n = 4; group I) or twice (n = 4, group II) daily. Ultra-performance liquid chromatography coupled with electrospray tandem mass spectrometry was used to determine the antibiotic residues in the samples. The correlation coefficient ($r^2$) of the calibration curves for all antibiotics was > 0.999 and the limits of detection and quantification were $0.002-0.005{\mu}g/mL$ and $0.007-0.02{\mu}g/mL$, respectively. Recovery rates were ranged from 75.5 to 92.3%. In group I, PEG, STR and NEO residues were detected in milk at 2, 3 and 2 days post-treatment, respectively, which were below the maximum residue limit (MRL). In group II, PEG, STR and NEO residues were detected in milk at 2, 3 and 3 days post-treatment, respectively, which were bellow the MRL. These results suggest that a 3-day for milk withdrawal period after the ointment treatment might be sufficient for reduction of the antibiotic residues below the MRL.

Inhibitory effects of tuberostemonine on Staphylococcus aureus biofilm (Tuberostemonine에 의한 Staphylococcus aureus의 생물막 억제 효과)

  • Yum, Su Jin;Kim, Seung Min;Kwon, Jun Hyeok;Jeong, Hee Gon
    • Korean Journal of Food Science and Technology
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    • v.54 no.2
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    • pp.241-246
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    • 2022
  • Antibiotic resistance is a serious problem to food safety as well as human healthcare. To avoid this, there are several approaches for a new class of antibiotic agents that target only production of virulence factors such as biofilm without bacterial growth defect. The objective of this study was to investigate the antibiofilm activity of tuberostemonine in Staphylococcus aureus. Tuberostemonine significantly reduced the biofilm formation (26.07-47.02%) in the crystal violet assay whereas there were no effect on S. aureus growth. The dispersion in preformed biofilm was also observed by confocal laser scanning microscopy (CLSM). Quantification real-time PCR revealed that the icaA and agrA expression having an important role in biofilm production of S. aureus were strongly affected with tuberostemonine. These results suggest that tuberostemonine has potential for controlling biofilm formation and dispersion by effect on virulence regulation of S. aureus.

Ambient Air Waste Sorting Facilities Could Be a Source of Antibiotic Resistant Bacteria

  • Calheiros, Ana;Santos, Joana;Ramos, Carla;Vasconcelos, Marta;Fernandes, Paulo
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.367-373
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    • 2021
  • The antimicrobial resistance of Staphylococcus spp. and Gram negative strains present in air samples from waste sorting facilities was assessed. Phenotypic studies have revealed a high percentage of strains of Staphylococcus spp. resistant to methicillin. Genotypically and by RT-PCR, it was found that the mecA gene usually associated with methicillin resistance was present in 8% of the Staphylococcus strains isolated. About 30% of the Gram negative strains from the same samples also displayed resistance to meropenem and 79% of these were resistant to multiple antibiotics from different classes, namely cephalosporins and β-lactams. The results suggest that in professional activities with high levels of exposure to biological agents, the quantification and identification of the microbial flora in the work environment, with the determination of the presence of potential agents displaying multi-resistances is of relevance to the risk assessment. The personal protection of workers is particularly important relevance in these cases, since many of the strains that exhibit multi-resistance are potential opportunistic agents.

A new cell-direct quantitative PCR based method to monitor viable genetically modified Escherichia coli

  • Yang Qin;Bo Qu;Bumkyu Lee
    • Korean Journal of Agricultural Science
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    • v.49 no.4
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    • pp.847-859
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    • 2022
  • The development and commercialization of industrial genetically modified (GM) organisms is actively progressing worldwide, highlighting an increased need for improved safety management protocols. We sought to establish an environmental monitoring method, using real-time polymerase chain reaction (PCR) and propidium monoazide (PMA) treatment to develop a quantitative detection protocol for living GM microorganisms. We developed a duplex TaqMan quantitative PCR (qPCR) assay to simultaneously detect the selectable antibiotic gene, ampicillin (AmpR), and the single-copy Escherichia coli taxon-specific gene, D-1-deoxyxylulose 5-phosphate synthase (dxs), using a direct cell suspension culture. We identified viable engineered E. coli cells by performing qPCR on PMA-treated cells. The theoretical cell density (true copy numbers) calculated from mean quantification cycle (Cq) values of PMA-qPCR showed a bias of 7.71% from the colony-forming unit (CFU), which was within ±25% of the acceptance criteria of the European Network of GMO Laboratories (ENGL). PMA-qPCR to detect AmpR and dxs was highly sensitive and was able to detect target genes from a 10,000-fold (10-4) diluted cell suspension, with a limit of detection at 95% confidence (LOD95%) of 134 viable E. coli cells. Compared to DNA-based qPCR methods, the cell suspension direct PMA-qPCR analysis provides reliable results and is a quick and accurate method to monitor living GM E. coli cells that can potentially be released into the environment.

Relationship assessment of the residual antibiotics and the amount of N component by different production stages of liquid fertilizer from livestock manure (가축분뇨 유래 액비 생산단계별 항생제 잔류 농도와 질소 성분 함량과의 상관성 평가)

  • Song-Hee Ryu;Jin-Wook Kim;Young-Kyu Hong;Sung-Chul Kim;Jun-Hyeong Lee;Eun-A Jeong;Chang-Gyu Kim;Young-Man Yoon;Oh-Kyung Kwon
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.258-265
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    • 2023
  • After application of veterinary antibiotics, they may be partially metabolized before they are excreted by feces or urine either as unaltered form or as metabolites. Residual antibiotics enter the adjacent agricultural environments by spraying manure-based compost and liquid fertilizer on farmlands and lead to secondary pollution. The objective of this study was to compare the residual concentrations of 6 veterinary antibiotics by different production stages of liquid fertilizer from livestock manure recycling facilities. The relationship between concentration change of the residual antibiotics and the amount of liquid fertilizer component was also assessed. Pre-treatment showed the recovery of 63.4-106.7% at ppb level and the limit of quantification of 0.009-0.037 ㎍/L.As the result of analyzing the relationship between the residual concentrations of antibiotics and the amount of N component in liquid fertilizer by different production stages, the residual concentrations of antibiotics and N tended to decrease as the stabilization period elapsed during the liquid fertilization process. Average concentrations of sulfamethazine in raw materials, middle and final products of liquid fertilizer were 40.85, 26.17, 3.54 ㎍/L, respectively. Those of chlortetracycline decreased from 2.32 to 1.25 ㎍/L. The other 4 antibiotics also showed a decreasing trend by different production stages of liquid fertilizer. The amount of liquid fertilizer component N decreased from 0.21 to 0.096% by production stages of liquid fertilizer. It is considered that the correlation between residual antibiotic concentrations and N content can be applied as basic data for setting antibiotic reduction indicators.

Determination of Veterinary Antibiotic Residues: IV. Comparable Analytical Methods with EPA Methods 1694_A Review (시료 중 잔류 항생제 분석 방법: IV. EPA method 1694와 비교 가능한 기기 분석 방법)

  • Kim, Chansik;Ryu, Hong-Duck;Chung, Eu Gene;Kim, Yongseok;Rhew, Doug Hee
    • Journal of Korean Society on Water Environment
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    • v.32 no.6
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    • pp.670-699
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    • 2016
  • In this study, 16 antibiotics were selected from among the top 30 veterinary antibiotics sold in South Korea in 2014, as well as from among the pharmaceuticals targeted by EPA method 1694, in order to review analytical methods for the detection of trace levels of antibiotics in environmental samples: surface water, soils, animal origin foods, and manures. LC-MS/MS was heavily used. In the chromatography for the detection of the selected antibiotics, the $C_{18}$ column was mostly used at the temperature of $30{\sim}40^{\circ}C$. Water and methanol/acetonitrile were commonly chosen as a nonpolar and a polar mobile phase, respectively. Gradient elution was applied to separate multiclass antibiotics. Volatile additives, such as formic acid, acetic acid, and ammonium acetate were mixed with the mobile phase to improve the ionization efficiency of analytes and the sensitivity in MS detection. Electrospray ionization (ESI) was widely used in the LC-MS/MS and positive ionization was preferred to determine the selected antibiotics. A protonated $[M+H]^+$ molecule was selected as a precursor ion, and its two transitions were analyzed, one for quantitative measurement and the other for confirmation. This study reviewed linearity of the calibration curve, recovery, repeatability, method detection limits (MDLs), and method quantification limits (MQLs) for each target compound used to validate the developed analytical methods.

Determination of Veterinary Antibiotic Residues: III. Analytical Methods_A Review (시료 중 잔류 항생제 분석 방법: III. 기기 분석 방법)

  • Kim, Chansik;Ryu, Hong-Duck;Chung, Eu Gene;Kim, Yongseok;Rhew, Doug Hee
    • Journal of Korean Society on Water Environment
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    • v.32 no.6
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    • pp.649-669
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    • 2016
  • This study explored the analytical conditions for 21 veterinary antibiotics which have been popularly sold in South Korea in 2014 but have not yet been targeted in EPA method 1694. Most of the selected antibiotics were separated by a reverse-phase C18 column with a combination of (buffered) water and organic polar solvent, which was commonly methanol and acetonitrile in the gradient elution mode. Volatile additives such as formic acid, ammonium acetate and ammonium formate were usually added to the mobile phases to minimize asymmetrical and tailing of antibiotics' peaks and to increase their ionization in mass spectrometry. The analytical methods of aminoglycoside antibiotics were distinct from those of the other antibiotics in terms of adoption of ion-pair chromatography (IPC) and hydrophilic interaction liquid chromatography (HILIC) capable of retaining and separating extremely polar compounds due to their hydrophilicity. Trifluoroacetic acid or heptafluorobutyric acid was frequently added to the mobile phase as an ion-pair reagent for the IPC. Tandem mass spectrometry was numerously applied to the detection of antibiotics using positive electrospray ionization (ESI) and the selected reaction monitoring (SRM) mode. All reviewed analytical methods had been/were validated by evaluating recovery, limits of detection and quantification, decision limit or detection capability of the methods.