• Title/Summary/Keyword: Gram positive

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Analysis of Aminoglycoside Antibiotics in Meat and Cell Culture Medium Coupled with Direct Injection of an Ion-pairing Reagent (이온쌍 시약 직접 주입법을 활용한 육류 및 세포배양액 내 아미노글리코사이드계 항생제 분석)

  • Kyung-Ho Park;Song-Yi Gu;Geon-Woo Park;Jong-Jib Kim;Jong-soo Lee;Sang-Gu Kim;Sang-Yun Lee;Hyang Sook Chun
    • Journal of Food Hygiene and Safety
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    • v.38 no.5
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    • pp.319-331
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    • 2023
  • Aminoglycoside antibiotics, also known as aminoglycosides (AGs), are veterinary drugs effective against a wide range of gram-negative and gram-positive bacteria. Owing to their recent use in cultured meats, it has become essential to establish an analytical method for safety management. AGs are highly polar compounds, and ion-pair reagents (IPRs) are used to ensure component separation. Owing to the high possibility of potential mechanical problems resulting from IPR addition to the mobile phase, an analytical method in which IPRs are added directly to the vial was explored. In this study, methods for analyzing 10 AGs via liquid chromatography-tandem mass spectrometry (LC-MS/MS) with the addition of two IPRs were validated for selectivity, detection limit, quantitation limit, recovery, and precision. The detection limit was 0.0001-0.0038 mg/kg, the quantification limit was 0.004-0.011 mg/kg, and the linearity (R2) within the concentration range of 0.01-0.5 mg/kg was over 0.99. Recovery and precision (expressed as relative standard deviation) evaluated in the two matrices (beef and cell culture media) ranged from 70.7% to 120.6% and 0.2% to 24.7%, respectively. The validated AG analytical method was then applied to 15 meats prepared from chicken, beef, and pork, and 6 culture media and additives used in cultured meat. No AGs were detected in any of the 15 meats distributed in Korea; however, streptomycin and dihydrostreptomycin were detected at levels ranging from 695.85 to 1152.71 mg/kg and 6.35 to 11.11 mg/kg, respectively, in the culture media additives. The LC-MS/MS method coupled with direct addition of IPRs to the vial can provide useful basic data for AG analysis and safety evaluation of meats as well as culture media and additives for cultured meats.

Epidemiological Studios on Clonorchis sinensis Infection along the Nam-river in Gyeongnam Province, Korea (남강유역 간흡충감염의 역학적 조사연구)

  • 배경훈;안영겸
    • Parasites, Hosts and Diseases
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    • v.21 no.2
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    • pp.116-186
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    • 1983
  • An epidemiological study on Clonorchis sinensis infection along the Nam-river (total length ; 186km) flowing in Gyeongsang-nam-do, southern part of Korea, was carried out. Formalin-ether concentration technique and Stoll's egg counting method were employed to figure out the prevalence of C. sinensis infection. For the detection of cercariae from Parafossarulus sp., the snail host of C. sinensis, each snail was placed in aerated tap water, and examined for expelled cercariae. For observing the metacercariae the fresh water fish favorably eaten in raw conditions wore prepared by means of pressing the muscles between two slide glasses and/or by digesting them with artificial gastric juice. The fresh water fish were fed to the rabbits to get the worms and to identify the morphology of adult C. sinensis. The results are summarized as follows: 1. Overall Prevalence of C. sinensis infection was 38.75 from 5,291 examinees; 44.15 (1,408 out of 3,196) in male and 30.4% (637 out of 2,095) in female. 2. The prevalence rates were 42.0% at the upper stream, 41.2% around the vicinity of Jinyanglake, 34.2% at Jinju city, 34.2% at middle stream and 40.3% at down stream regions, respectively. 3. By age, the highest positive rate (53.4∼54.3%) was observed in 30 to 59 years of age. In this age group, the rate in males was 59.7∼62.2%, and in females 42.2∼44.4%, In the age group of less than 19 years it was 7.5∼20.9%. 4. By social strata, the positive rate was 16.5% in the primary school children, 22.6% in school students, 46.2% in teachers and local officers and 49.6% in the general inhabitants. 5. The quantitative examinations with the stool collected from clonorchiasis cases revealed that the light infection (less than 4,000/EPG) was 53.6%, moderate infection (4,001∼10,000 /EPG) 30.3% and heavy infection (more than 10,001/EPG) 16.1%, respectively. More than half of total cases examined were light infection, and 73.2% of female examinees were lightly infected with this cuke. 6. The average value of EPG was 4,963 (male, 6,057; female, 2,557 and the highest value was obtained from the age group of 30 to 59 years (5,240∼6,454). 7. The prevalence of Metagonimus yokogawai infection in humans was 5.5%, and 89.8% of metagonimiasis cases wore double-infected arith C. sinensis. The highest prevalence rate was observed in Jinju city (11.2%). 8. Total of 5,005 Parafossarulus sp., the snail intermediate host of C. sinensis, were examined for the detection of cercariae. The cercarial expulsion rate was 0.34%, and the snails collected in Jin-yang-lake side and in the down stream expelled mainly the cercariae of C, sinensis. 9. About 788 cercariaejday (range: 127∼1,503) were expelled daily from a snail naturally infected with C. sinensis. The snails which released more than 1,000 cercariae/day were 30.8% out of total collected. A snail uniquely released 5,840 cercariae/day in this study. 10. The other trematode cercariae besides C. sinensis were also detected, and the rates out of total snails were the cercariae of Losogenes liberum 6.71%, Cyathocotyle orientalis 0.26%, Notocotylus attenuatus 2.52% and Mucobucaris 0.54%, 11. Ten out of 18 species of fresh water fish caught along the river harbored the metacercariae of C, sinensis. The highest rate of metacercarial infection in fish was detected in Pseudorasbora larva (85.9%). The fish mainly eaten by the inhabitants along the Nam-river, and the metacercarial infection rates were: Zacco platypus 8.0%, Hemibarbus sp. 18.2∼26.7%, Gnathepcgcn sp. 37.5%, Ischikauia steenackeri 42.9% and Pseudogobio esocinus 16.7%. 12. Out of 36 P. larva, the number of metacercariae were about 109 (ranEe; 18∼446) per fish and 27 per gram of flesh. The fish caught in spring harbored the highest number of metacercariae. As indicated above, the prevalence of C. sinensis infection in the inhabitants residing around the Nam-river was relatively high. The farther toward the upper stream areas, the higher was the positive rate of C. sinensis infection, and most of clonorchiasis cases were lightly infected. The snail hosts of C. sinensis distributed all around the Nam-river. Several species of freshwater fish were infected with the metacercariae of C. sinensis and the infection rates were relatively high. To prevent C. sinensis infection in the endemic areas, the effective health education system is suggested as a control measure, although mass treatment is also expected to be useful, using chemotherapeutic agents such as "praziquantel", a recently developed anthelminthic for C. sinensis inftraction.

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Effect of Graded Levels of Mustard Oil Cake Supplementation on Intake, Nutrient Digestibility, Microbial N Yield of Adult Cannulated Native (Bos Indicus) Bulls Fed Rice Straw

  • Chowdhury, S.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.5
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    • pp.715-722
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    • 1999
  • On a urea-molasses-straw (3:15:82, UMS) based diet, effect of graded levels of mustard oil cake (MOC) supplementation on the performances of native bulls has been studied. Four cannulated adult Bos indicus bulls of $415({\pm}44.6)kg$ live weight and 80 months old, were given daily either of 0, 200, 400 or 800 g of MOC in four periods in a $4{\times}4$ latin square design. Besides, each animal also received 200 g of each of molasses and wheat bran and a mineral mixture. For unit (1 g) increase in MOC intake, total DM intake increased by 0.8 g/d ($r^2=0.88$) but no change in the straw DM intake. With the increasing levels of MOC, crude protein (CP) digestibility increased exponentially with an asymptotic value of 72%. However, MOC level had no effect on the digestibilities of DM, OM and ADF. Similarly, rumen degradability of rice straw was also not affected by the level of dietary MOC, and mean straw DM degradabilities were 15, 21. 28, 37, 47 and 51% at 8, 16, 24, 48, 72 and 96 hours of incubation respectively. Microbial N yield per kg digestible organic matter apparently fermented in the rumen were 7.46, 8.77, 6.88 and 5.96 g respectively for 0, 200, 400 or 800 g of dietary MOC. For each gram increase in dietary MOC, N intake and N balance increased by 0.054 g/d ($r^2=0.998$) and $0.59mg\;N/kg\;W^{0.75}/d$ ($r^2=0.99$) respectively. Nitrogen balance was estimated to be attained at the N intakes of $246mg\;N/kg\;W^{0.75}/d$. Thus, on a UMS-based diet supplementation of MOC up to 800 g (10% of total intake) of the dietary intake had little or no effect on intake, digestibility, rumen parameters, and microbial N yield but slightly increased the N balance. However marginal response to MOC supplementation is probably due to the high degradability of MOC protein in the rumen. Thus, any substantial positive response of MOC supplementation on a UMS-based diet can probably be achieved by reducing its protein degradability in the rumen.

Diversity and Antimicrobial Activity of Actinomycetes from Fecal Sample of Rhinoceros Beetle Larvae (장수풍뎅이 유충의 분변에 존재하는 방선균의 다양성 및 항균활성)

  • Lee, Hye-Won;Ahn, Jae-Hyung;Kim, Minwook;Weon, Hang-Yeon;Song, Jaekyeong;Lee, Sung-Jae;Kim, Byung-Yong
    • Korean Journal of Microbiology
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    • v.49 no.2
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    • pp.156-164
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    • 2013
  • Actinomycetes produce diverse secondary metabolites which have the primary importance in medicine, agriculture and food production, and key to this is their ability to interact with other organisms in natural habitats. In this study, we have investigated the taxonomical and functional diversity of actinomycetes in fecal sample of rhinoceros beetle larvae (Allomyrina dichotoma L.) by using culture-dependent and -independent approaches. For the culture-independent approach, the community DNA was extracted from the sample and 16S rRNA genes of actinomycetes were amplified using actinomycetes-specific PCR primers. Thirty-seven clones were classified into 15 genera and 24 species of actinomycetes. For the culture-dependent approach, 53 strains were isolated from larval feces, of which 27 isolates were selected based on morphological characteristics. The isolates were classified into 4 genera and 14 species, and 24 isolates (89%) were identified as the genus Streptomyces. Many of the representative isolates had antimicrobial activities against plant pathogenic fungi and Gram-positive bacteria. In addition, most of the isolates (78%) showed biochemical properties to hydrolyze cellulose and casein. The results demonstrated that diverse and valuable actinomycetes could be isolated from insect fecal samples, indicating that insect guts can be rich sources for novel bioactive compounds.

Screening for Biological Activity of Crude Extracts from Medicinal plants (생약추출물로부터 생리활성의 검색)

  • Kwag, Jung-Sook;Oh, Hyun-Ju;Lee, Hyun-Ok;Perry, Nigel B.;Baek, Seung-Hwa
    • Journal of dental hygiene science
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    • v.3 no.2
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    • pp.67-70
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    • 2003
  • The biological effects of the crude extracts from medicinal plants, Brachyglottis monroi and Trichocolea hatcheri were investigated. The crude ethanol extract inhibited the growth of the Gram positive bacterium Bacillus subtilis (ATCC 19659, 1 mm inhibition zone at $150{\mu}g/disc$) and the dermatophyte Trichophyton mentagrophytes (ATCC 28185, 6 mm inhibition zone at $150{\mu}g/disc$), and was toxic to P388 murine leukaemia cells ($IC_{50}$ $23.96{\mu}g/mL$ at $75{\mu}g/disc$). B. monroi ethanol extract showed stronger antiviral activity than that of T. hatcheri against Herpes simplex Type I virus (ATCC VR 733) and Polio Type I virus (Pfizer vaccine strain) (50% activity, @ 5 mg/ml at $150{\mu}g/disc$). The crude ethanol extract of T. hatcheri showed stronger antimicrobial activity than that of B. monroi. However, this extract was inactive against P388 murine leukaemia cells.

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Molecular Cloning and Expression of the Novel Attacin-Like Antibacterial Protein Gene Isolated from the Bombyx mori (집누에로부터 새로운 attactin 유산 항세균성 펩타이드 유전자의 분리 및 발현)

  • 윤은영;김상현;강석우;진병래;김근영;김호락;한명세;강석권
    • Korean journal of applied entomology
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    • v.36 no.4
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    • pp.331-340
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    • 1997
  • Hyalophora cecropia attacin-like antibacterial gene was isolated from Bombyx mori induced with nonpathogenic bacteria. It was expressed in Spodopfera frugiperda 9 (Sf9 cells using baculovirus expression vector system (BEVS), and examined its antibacterial activity. With a cDNA library constructed from fifthinstar B. mori injected with Escherichia coli(4 X IOhcellsllarva), differential screening was performed using naive and induced mRNA probes. BmInc6 clone was screened by partial nucleotide sequence and GenBank database analysis. A complete nucleotide sequence of Bmlnc6 cDNA was determined (GenBank, AF005384). Its insert size was 852 bp and had open reading frame that started translation at position 35 and stopped at 679. And its putative polyadenylational signal existed at 812 bp. The number of amino acid deduced from Bmlnc6 cDNA was 214 and hydropathy analysis showed that this peptide was hydrophilic. This peptide deduced by BmInc6 was named nuecin. When the nuecin gene was expressed in Sf9 cells using BEVS, about 950 bp of the transcripts was detected. In addition, SDS-PAGE analysis showed that the molecular weights of intracellular expressed protein and the mature protein secreted to culture media were approximately 23 and 20 kDa, respectively. The antibacterial activity of nuecin against E. coli and Bacillus subtilis was significantly high, demonstrating that nuecin had a wider antibacterial spectrum with gram negative and positive bacteria than attacin.

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Crop Rotation in Paddy Soil Exhibiting Crop Failure Following Replanting: Effect on Soil Chemical Properties, Soil Microbial Community and Growth Characteristics of 2-Year-Old Ginseng (인삼 논재배 연작지에서 윤작물 재배가 토양화학성, 토양 미생물상 및 2년생 인삼의 생육에 미치는 영향)

  • Lee, Sung Woo;Park, Kyung Hoon;Lee, Seung Ho;Jang, In Bok;Jin, Mei Lan
    • Korean Journal of Medicinal Crop Science
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    • v.24 no.4
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    • pp.294-302
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    • 2016
  • Background: Crop rotation plays an important role in improving soil chemical properties, minimizing the presence of disease pathogens, and assists in neutralizing autotoxic effects associated with allelochemicals. Methods and Results: Five rotation crops of sudan grass, soybean, peanut, sweet potato, and perilla were cultivated for one year with an aim to reduce yield losses caused by repeated cropping of ginseng. In 2-year-old ginseng grown in the same soil as a previous ginseng crop, stem length and leaf area were reduced by 30%, and root weight per plant was reduced by 56%. Crop rotation resulted in a significant decrease in electrical conductivity, $NO_3$, and $P_2O_5$ content of the soil, whereas organic matter, Ca, Mg, Fe, Cu, and Zn content remained-unchanged. Soil K content was increased following crop rotation with sudan grass and peanut only. Rotation with all alternate crops increased subsequent ginseng aerial plant biomass, whereas root weight per plant significantly increased following crop rotation with perilla only. A significant positive correlation was observed between root rot ration and soil K content, and a significant negative correlation was observed between ginseng root yield and the abundance of actinomycetes. Crop rotation affected the soil microbial community by increasing gram negative microbes, the ratio of aerobic microbes, and total microbial biomass whereas decreases were observed in actinomycetes and the ration of saturated fatty acids. Conclusions: In soil exhibiting crop failure following replanting, crop rotation for one year promoted both soil microbial activity and subsequent ginseng aerial plant biomass, but did not ameliorate the occurrence of root rot disease.

Characteristics and Diagnostic Methods of Streptococcosis Causing Disease in Aquaculture (양식 어류에 질병을 유발하는 연쇄구균증의 특성 및 진단 방법)

  • Kim, Dong-Hwi;Heo, Moon-Soo
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1118-1126
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    • 2018
  • In this study, investigated the general characteristics and diagnostic methods types of streptococcosis among various fish disease pathogens that caused a lot of economic damaged to aquaculture fish based on the previous research paper. Streptococcosis infection of fish is considered a reemerging disease affecting a variety of wild and cultured fish throughout the world. Calssifiacation of Gram positive cocci based on DNA-DNA hybridization coupled with 16S sequencing has shown that at least five different species are considered of significance as fish pathogens: Lactococcus garvieae, L. piscium, Streptococcus iniae, S. agalactiae, S. paruberis, Vagococcus salmoninarum. Symptoms of infection with streptococcosis disease such as body color change, eyeball abnormality, gill discoloration, bleeding, abdominal distension, swelling of the kidney and spleen. In addition, it usually occurs from June to October when the water temperature rise a lot of fish death. Currently, 16S rRNA, 16S-23S rRNA intergenic spacer region (ISR), Random Amplified polymorphic DNA (RAPD), Ribotyion (RT), Loop-mediated isothermal amplification (LAMP) are among the methods for diagnosing streptococcosis. Among them, the LAMP method, which is high applicable to the aquaculture farm has attracted the spotlight, but due to problems such as confirmation of results. This seems to minimize the economic loss of streptococcosis which complements the problem so that it can be easily used from the diagnosis to the results confirmation.

Effect of Factors on the Sporulation of Bacillus cereus and Their Thermal Resistance (여러 조건에 따른 Bacillus cereus 포자 형성과 포자의 열저항성 평가)

  • Kim, Seung-Ju;Jung, Jin-Ho;Tahk, Hong-Min;Baek, Seung-Youb;Lee, Sun-Young
    • Journal of Food Hygiene and Safety
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    • v.24 no.3
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    • pp.256-261
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    • 2009
  • Bacillus cereus is a gram-positive spore-forming bacterium and produces an emetic or diarrheal syndrome induced by an emetic toxin and an enterotoxin, respectively. In this study, the effect of different types of media, temperature, and time on the sporulation of B. cereus, and thermal resistance of B. cereus spores produced in various temperatures were evaluated. The highest levels of spores were detected when they are produced at $25^{\circ}C$. There were no significant differences in levels of spores produced at $25^{\circ}C$ among culture media and times while levels of spores produced at $43^{\circ}C$ were significantly reduced with the increase of time. However, thermal resistance of B. cereus spores could be affected by incubation temperature. In fact, higher D-values (12.0, 10.1, and 5.9 min for 2,4, and 6 weeks, respectively) of spores produced at $43^{\circ}C$ were observed than did in samples produced at other temperatures (25 and $37^{\circ}C$). D-values of spores were 7.7, 8.2, and 12.0 min when they were produced at 25,37, and $43^{\circ}C$ for 2 weeks, respectively. The sporulation of B. cereus at $25^{\circ}C$ could result in high amounts of spores however the sporulation at $43^{\circ}C$ for 2 weeks could be effective to produce thermal resistant spores.

Changes of Antioxidant Activity in Juglans mandshrica Maxim. Leaves by Far Infrared Ray Irradiation (원적외선 조사에 따른 가래나무 잎의 항산화 활성 변화)

  • Eom, Seok-Hyun;Park, Hyoung-Jae;Jin, Cheng-Wu;Park, Seung-Moon;Kim, Myong-Jo;Yu, Chang-Yeon;Cho, Dong-Ha
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.4
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    • pp.266-270
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    • 2007
  • This research was conducted to investigate antioxidant activities of Juglans mandshrica forages by the irradiation of far infrared ray during leaf drying processes before compound extraction, with its potential use for activating antioxidants. The contents of total phenolics and flavonoids in the basis of a gram leaf dry weight were maximized in 20 min at 30 V of far infrared irradiation. Total phenolic content was 25.43 mg/g in the FIR treatment, while it was 21.12 mg/g in the non-FIR treatment control. Total flavonoid content was 39.38 mg/g in the FIR treatment, while it was 32.78 mg/g in the non-FIR treatment control. The contents were decreased when the condition of far infrared ray irradiation was higher voltage and longer treatment time, exhibiting 16.93 mg/g of total phenolics and 23.78 mg/g of total flavonoids in 30 min at 50 V of far infrared irradiation. The antioxidant activities were shown to positive relationships with the contents of total phenolics and total flavonoids. In this study, we concluded that content of bioactive molecules relating antioxidant activity in Juglans mandshrica forages is optimized in 20 min at 30 V of far infrared irradiation for leaf drying process.