Enrofloxacin is a fluoroquinolone antibiotic approved for the treatment of infections in animals. Because of the side effects to consumers of animal products, the maximum residue limits (MRLs) of enrofloxacin in animal tissues for consumption are regulated. In this study, a monoclonal antibody (mAb) against enrofloxacin was prepared and characterized for the development of a direct competitive enzyme-linked immunosorbent assay (ELISA). The obtained mAb, Enro44, was highly specific for enrofloxacin and had a 50% inhibition concentration ($IC_{50}$) of 1.99 ng/ml in a competitive ELISA, and the limit of detection (LOD) was 0.50 ng/ml. The cross-reactivity of the mAb with other quinolones and fluoroquinolones was lower than 0.01%. The subclass of the mAb Enro44 was identified as IgG1. The antigen (Ag)-captured direct competitive ELISA using the mAb Enro44 was tested on different spiked samples, including chicken muscle, cattle milk, and cattle urine, and the assay demonstrated recoveries of 82-112%, 80-125%, and 78-124%, respectively. Furthermore, the quantitation of enrofloxacin obtained from the ELISA and from high-performance liquid chromatography (HPLC) was in good agreement, with the linear regression coefficient between 0.933 and 1.056. The cDNAs encoding a heavy-chain Fd fragment (VH and CH1) and a light chain of the mAb Enro44 were cloned and sequenced. Taken together, the results obtained reveal a potential use of this mAb in an ELISA for the detection of enrofloxacin in food samples. The information of amino acid sequence of this mAb will be useful for further modification and production of the mAb in a bioreactor.
Jung, Yujung;Jeon, Youngsic;Kim, Hyung Ja;Kang, Ki Sung;Kim, Yong Kee;Kim, Su-Nam
YAKHAK HOEJI
/
v.58
no.5
/
pp.294-299
/
2014
In this study, we investigate anti-allergic and anti-inflammatory effects of Leonurus sibiricus seed (LSS) extract in basophilic leukemia RBL-2H3 cells. To identify anti-allergic actions of LSS, the degranulation was evaluated in IgE and DNP-BSA stimulated RBL-2H3 cells. At the concentration of $100{\mu}g/ml$ of methanol (MeOH) extract and Methylene chloride (MC) and Ethyl acetate (EtOAc) fractions, the degranulation was significantly inhibited 16.7%, 16.7% and 27.9% respectively. And then, to assess anti-inflammatory effects of LSS, IL-4 and IL-13 mRNA level were detected in PMA/ionomycin (PI)-induced RBL-2H3 cells and cell proliferation and IL-4 mRNA level in isolated splenocytes from Balb/c mice. LSS MeOH extract and MC and EtOAc fractions significantly decreased the level of IL-4 and IL-13 mRNA in PI-induced RBL-2H3 cells and showed inhibitory effects on cell proliferation and expression of IL-4 mRNA level in mouse splenocytes. Taken together, these results suggest that LSS has potential anti-allergic and anti-inflammatory effects and EtOAc fraction is the most effective in regulating immune responses.
Objectives : In order to investigate the efficacy of BJBB on atopic dermatitis, various anti-inflammatory factors were studied. Methods : In-vitro, inflammatory mediators, such as MTT and nitric oxide were detected after the addition of LPS with or without BJBB in Raw 264.7 cells. In-vivo, in order to verify the effectiveness of BJBB in atopic dermatitis animal model, its role in inflammation factors and histological changes were observed in NC/Nga mice. Results : BJBB showed cell viability of 100% or higher in all concentration in Raw 264.7 cells. BJBB inhibited LPS-induced productions of inflammatory mediators nitric oxide in RAW 264.7cells. BJBB treated group showed significant decrease in the expression of IL-1b, IL-6 and TNF-a by 40%, 80% and 44% respectively. Also the group showed decrease in the transcription of IL-1b, IL-6 and TNF-a mRNA in spleen by 41%, 93% and 39% respectively. BJBB treated group showed significant decrease in WBC, neutrophil, lympocyte and monocytes immune cell ratio in blood by 54%, 63%, 57% and 86% respectively. BJBB treated group showed decrease in the expression of IgG by 39% respectively. Also, infiltration of adipocytes into skin was suppressed and the thickness of epidermis and dermis were relatively decreased in the BJBB treated group. Conclusion : BJBB has an anti-inflammatory effects in NC/Nga mouse. Thus, these results suggested a beneficial effect of BJBB in treatment with Atopic dermatitis and inflammatory.
Hae Won Shin;Xing Hao Jin;Min Jin Gim;Yoo Yong Kim
Animal Bioscience
/
v.36
no.5
/
pp.776-784
/
2023
Objective: This experiment was conducted to evaluate the inclusion of dietary nontoxic sulfur (NTS) on growth performance, immune response, sulfur amino acid composition and meat characteristics in growing-finishing pigs. Methods: A total of 140 crossbred pigs ([Yorkshire×Landrace]×Duroc) with an average body weight of 34.73±0.66 kg were used for the 12-week feeding trial. Experimental pigs were allotted to one of 5 treatments in 4 replicates of 7 pigs per pen in a randomized complete block (RCB) design. The experimental treatments were as follows (0%, 0.1%, 0.2%, and 0.4% NTS levels): i) Control, corn soybean meal (SBM)-based diet; ii) NTS 0.1, basal diet + NTS 0.1%; iii) NTS 0.2, basal diet + NTS 0.2%; iv) NTS 0.4, basal diet + NTS 0.4%. Results: Body weight increased linearly as dietary NTS levels increased up to 0.2% (linear; p = 0.04) in the early finishing phase (9 weeks). During the whole experimental period, body weight and average daily gain linearly increased as the dietary NTS level increased in the diet (linear; both p = 0.01), but quadratic responses in body weight and average daily gain were observed with the addition of NTS 0.4% (quadratic, both p = 0.01). In the late finishing period, the IgG concentration increased linearly (linear; p = 0.01) as the dietary NTS level increased up to 4%. In the finishing period, a linear response was observed as a dietary NTS level was added (linear; p = 0.03), and supplementation with 0.2% NTS resulted in a higher methionine content than the other treatments (quadratic; p = 0.01). NST 0.2% had a lower value of thiobarbituric acid reactive substances (quadratic; p = 0.01). Conclusion: Consequently, supplementation with dietary NTS up to 0.2% could improve growth performance, amino acid composition in hair and meat antioxidation capacity.
Ki Whan Kim;Seok Han Ra;Gereltuya Renchinkhand;Woo Jin Ki;Myoung Soo Nam;Woan Sub Kim
Korean Journal of Agricultural Science
/
v.50
no.2
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pp.281-294
/
2023
As a byproduct obtained from cheese manufacture, whey protein was developed as a functional food that contains multi-functional proteins. In this study, the biochemical activity of fermented milk prepared by fortifying whey protein with excellent physiological activity was investigated. Immunoglobulin (IgG) content was higher in 10% fortified whey protein fermented milk than in the control. The viable cell counts were 20% higher in the fermented milk with 10% fortified whey protein than in the control group. The antibacterial effect of 10% fortified whey protein fermented milk compared to the control group was shown to be effective against four pathogenic microorganisms, Escherichia coli (KCTC1039), Pseudomonas aeruginosa 530, Salmonela Typhimurium (KCTC3216), and Staphylococcus aureus (KCTC1621). The antioxidant effect by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities wasincreased two-fold in 10% fortified whey protein fermented milk compared to the control. The 10% fortified whey protein fermented milk inhibited the expression of the inflammatory cytokines (interleukin [IL]-6, tumor necrosis factor [TNF]-α, and induced nitric oxide synthase [iNOS]) in a concentration-dependent manner. In a piglets feeding test, the weight gain with 10% fortified whey protein fermented milk was increased by 18% compared to the control group, and no diarrhea symptoms appeared. Our results clearly demonstrated that 10% fortified whey protein fermented milk could be a useful functional ingredient for improving health.
Kang, Mun Hee;Park, Sol;Lee, Sang-Woo;Kim, Hyun-A;Lee, Byung-Tae;Eom, Ig-Chun;Kim, Soon-Oh
Journal of Korean Society of Environmental Engineers
/
v.37
no.4
/
pp.218-227
/
2015
A prerequisite for precise quantification of nanomaterials contained in environmental samples is to prepare suitable preservation conditions of samples. This study was initiated to suggest preservation conditions of aqueous samples for analyses of metal nanomaterials. Variation in the size of silver nanomaterial (cit-AgNP) was observed according to change in various conditions, such as pH, electrolyte concentration, temperature, nanomaterial concentration, and time. Aggregation of AgNP was characterized for each environmental condition, and finally proper preservation conditions of samples were proposed based on experimental results on AgNP aggregation. In addition, the preservation period of sample was computed by the doublet time of AgNP. The results indicate that the aggregation rate of cit-AgNP was close to 0 at the conditions of pH of ${\geq}7$, electrolyte ($Ca(NO_3)_2$) concentration of ${\leq}3mM$, temperature of $4^{\circ}C$, and cit-AgNP concentration of ${\leq}2mg/L$. Furthermore, the experimental results on doublet time of cit-AgNP suggest that maximum preservation period was evaluated to be 15.79~17.53 days when the concentration of 100 nm cit-AgNP is assumed to be $1{\mu}g/L$ which is considered as an environmentally-relevant concentration of engineered nanomaterials. Our results suggest that samples should be preserved at $4^{\circ}C$ and analyzed within 2 weeks.
A total of 140 weaning pigs ((Landrace${\times}$Yorkshire)${\times}$Duroc, BW = $6.47{\pm}0.86$ kg) were used in a 5-wk growth trail to determine the effects of phytoncide supplementation on growth performance, nutrient apparent total tract digestibility (ATTD), blood profiles, diarrhea scores and fecal microflora shedding. Pigs were assigned randomly by BW into 5 treatments, dietary treatments were: i) NC, basal diet; ii) PC, NC+0.05% tylosin; iii) EO, NC+0.1% essential oil; iv) PP, NC+0.2% PP (phytoncide with 2% citric acid), and v) PA, NC+0.2% PA (phytoncide). Each treatment had 7 replicate pens with 4 pigs per pen. All pigs were housed in pens with a self-feeder and nipple drinker to allow ad libitum access to feed and water throughout the experimental period. During 0 to 2 wks, supplementation with essential oil and PA decreased (p<0.05) G/F compared with the other treatments. During 2 to 5 wks, supplementation with PA led to a higher (p<0.05) G/F than the other treatments. At 2 wk, ATTD of dry matter (DM) and gross energy (GE) in EO treatment were decreased (p<0.05) compared with NC treatment. Dietary PC treatment improved (p<0.05) ATTD of DM and E compared with the CON group, and PA and PP treatments showed a higher (p<0.05) ATTD of E than that in NC treatment. Pigs fed phytoncide (PA and PP) had a greater (p<0.05) ATTD of DM than those of NC and EO treatments at 5 wk. Moreover, supplementation with phytoncide elevated (p<0.05) the concentration of immunoglobulin (IgG) in blood at 2 wk. The inclusion of EO, PP and PA treatments showed a greater (p<0.05) amount of fecal Lactobacillus compared with CON group. However, no difference (p>0.05) was observed in diarrhea scores among treatments. In conclusion, phytoncide can elevate feed efficiency, nutrient digestibility, and improve the fecal Lactobacillus counts in weaning pigs. Our results indicated that the phytoncide could be used as a good antibiotics alternative in weaning pigs.
One hundred and twenty weanling pigs in experiment 1 (Exp. 1) ($6.91{\pm}0.99kg$; 21 d of age) and Exp. 2 ($10.20{\pm}1.09kg$; 28 d of age) were used in two 42-d and 35-d experiments to evaluate the effect of medium-chain-triglyceride (MCT) on growth performance, apparent total tract digestibility (ATTD) of nutrients and blood profile. In both of Exp. 1 and Exp. 2, the same dietary treatments were utilized as follows : i) negative control (NC), ii) positive control (PC), NC+antibiotics (40 mg/kg Tiamulin, 110 mg/kg Tylosin, and 10 mg/kg Enramycin, iii) MCT3, NC+0.32% (phase 1, 2 and 3) MCT, and iv) MCT5, NC+0.55% (phase 1), 0.32% (phase 2 and 3) MCT. In Exp. 1, the pigs fed MCT5 diets had higher (p<0.05) ADG compared to NC treatment during the first 2 wk. From d 15 to 28, the ATTD of energy was improved (p<0.05) by MCT3 compared to the PC treatment. No effect has been observed on the blood profiles [red blood cell (RBC), white blood cell (WBC), immunoglobulin-G (IgG), lymphocyte concentration] measured in this study. In Exp. 2, the ADG were increased (p<0.05) by the MCT5 treatment than the PC treatment from d 0 to 14. Pigs fed PC treatment diet had lower ADFI (p<0.05) and better FCR (p<0.05) than NC treatment, whereas no differences were shown between MCT treatments and NC or PC treatment from d 15 to 35 and overall phase. The ATTD of DM and nitrogen were improved (p<0.05) by the effect of MCT5 related to the NC and PC treatment at the end of 2nd and 5th wk. The pigs fed MCT3 had higher (p<0.05) energy digestibility than PC treatment. No effects were seen in the blood profiles we measured (WBC, RBC, lymphocyte and immunoglobulin-G). In conclusion, the addition of MCT in the weanling pigs diet can improve the ADG and digestibility during the earlier period (first 2 wks), but had little effect on the blood characteristics.
This study was performed to evaluate the anti-oxidative, anti-inflammatory, anti-allergy, and whitening effects of Zizania latifolia ethanol extracts prepared from 5 different ethanol concentrations (10, 30, 50, 70, and 90%). As the ethanol concentration in the extraction solvent was increased, the radical scavenging activities also increased. The inhibitory activity of Z. latifolia ethanol extracts on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells tended to increase as the content of ethanol increased. The highest inhibitory activity was obtained with 70% ethanol extract. The antiallergy effects of Z. latifolia ethanol extracts were tested by measuring the release of ${\beta}-hexosaminidase$ in IgE-sensitized RBL-2H3 cells. The suppressive effect of Z. latifolia ethanol extracts increased in a dose-dependent manner as the proportion of ethanol increased, except for the 10% ethanol extract. Furthermore, the inhibitory effects of Z. latifolia ethanol extracts against melanin production in ${\alpha}-melanocyte$ stimulated hormone (MSH)-stimulated B16F0 cells increased as the ethanol ratio increased, and 70 and 90% ethanol extracts showed similar inhibitory activities to arbutin, a positive control, at $250{\mu}m$. The present study confirmed the efficacy of Z. latifolia ethanol extracts in various areas, demonstrating antioxidative, anti-inflammation, antiallergy, skin protective, and skin whitening effects, with no cytotoxicity. It could be used as a raw material in functional foods, as well as in cosmetics.
Yang, Seung Hak;Kim, Hyeon Shup;Cho, Won Mo;Kim, Sang Bum;Cho, Sung Back;Park, Kyu Hyun;Choi, Dong Yoon;Hwang, Sung Gu;Yoo, Yong Hee
Journal of Animal Environmental Science
/
v.18
no.sup
/
pp.47-54
/
2012
Effect of commercial Direct Fed Microbials (DFM) or protease treated feed (PTF) supplementation on growth rate and biogenic substances such as BUN, glucose, IgG, GOT, GPT and Vitamin A, C, E from Holstein steers was studied for 7 months. Thirty two steers aged 2~3 months were separated with 4 groups for control, DFM (PS), protease (ES) and their mix (PS + ES) supplementally fed 0, 100, 100 and 50 + 50 g/day respectively. Weight gain was averagely higher in PS than any others, although there were no differences significantly. All treatments enhanced to 3~8% of control in dry matter, crude protein and total digestible nutrient (P>0.05). Metabolic diseases with veterinary cure had not shown in this study. Plasma GOT and GPT were lower in the PS and ES than control. Plasma glucose concentration was also lower in PS than the others. Total cholesterol of ES was higher than the others but that of PS is the lowest. Plasma vitamin C was higher in PS than the others. It was shown that dietary PS affected change from glucose to vitamin C with not overloading liver. Conclusionally, PS and ES were shown to enhance metabolic health of steers during growing period.
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