• Journal of fish pathology
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• v.34 no.1
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• pp.71-80
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• 2021

Cephalexin, a semi-synthetic cephalosporin antibiotic, has long been used in fish aquaculture in various countries under legal authorization. The drug is thus widely available for use in other aquatic species except fishes like the crustacean whiteleg shrimp. This study aims to develop a sensitive method for laboratory residue studies to adopt in withdrawal period determinations. Through repeated trials from the existing methods developed for other food animal tissues, it was possible to achieve a sensitive high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method. The results showed that at a concentration of 0.1 mg/kg, the recovery rate was 81.79%, and C.V. value was 8.2%, which meet the recovery rate and C.V. recommended by Codex guideline. After satisfactory validation of analytical procedures, applicability to the shrimp tissue was confirmed in experimentally cephalexin-treated whiteleg shrimp. As a result, most muscle samples were detected below the limit of quantification (0.05 mg/kg) after day 3, and most hepatopancreas samples were detected below the limit of quantification after day 14. In particular, the limit of quantification 0.05 ppm with the presently developed method suggests sufficient sensitive over the current legal maximum residue limit of 0.2 mg/kg set for fishes.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.10
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• pp.1434-1437
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• 2004

The study was conducted to evaluate the effects of montmorillonite anocomposite (MNC) on mercury residues in growing/finishing pigs. A total of 96 cross bred pigs ($Duroc{\times}Landrace{\times}large$ white, 48 barrows and gilts respectively), with similar initial weight (27.87${\pm}$1.15 kg), were used in this study. The animals were randomly assigned to two concentrations of mercury (0.1 and 0.3 ppm from $HgCl_2$) and two levels (0 and 0.3%) of MNC in a $2{\times}2$factorial arrangement of treatments. Each group has 3 pens (replications), and each pen has 8 pigs (4 barrows and 4 gilts). The experiment lasted for 90 days. The results showed that pig growth performances were not affected significantly by inclusion of Hg and addition of MNC (p$\geq$0.05). It indicated that the extent of intoxication in these pigs were not severe enough to impair growth performances. Both on the bases of 0.1 ppm and 0.3 ppm mercury supplementations, addition of 0.3% MNC markedly decreased mercury levels of blood, muscle, kidney and liver tissue (p<0.05). These results implied that the addition of non-nutritive sorptive material, MNC, could effectively reduce the gastrointestinal absorption of mercury via its specific adsorption, with a consequent reduction of mercury residues in body tissues. MNC had offered an encouraging solution to produce safe animal products with mercury contaminated feed.

• Toxicological Research
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• v.17
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• pp.281-287
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• 2001

The Artichoke extract at 10% was used to add in drinking water to understand its effect on Aflatoxicosis of chickens. The Artichoke extract at the dose of 6 ml per liter of drinking water was given (experiment group) or not (control group) and to Hybro chickens (150 birds), during the first 49 days of life. Also, the chickens were fed with foodstuff containing 200 ppb or 500 ppb Aflatoxin $B_1$. Results showed that, the chickens having Artichoke extract: (1) Had overcome the growth retardation caused by the toxin at concentration of 200 ppb and 500 ppb of Aflatoxin $B_1$ (an addittonal weight gain of about 200-400 g/bird). (2) The feed conversion was improved (a reduction of 200-400 g of feed per kg of bird living weight). (3) Aflatoxicosis lesions were mild in the chickens, which fed 500 ppb of Aflatoxin $B_1$ or not found in those having the toxin 200 ppb. The blood examinations at 28th and 49th days of the trial gave the following results: (1) The Artichoke extract had an effect of suppressing the changes of blood cell numbers, hemoglobin amount. packed cell volume. leukocyte formula that were caused by Aflatoxin $B_1$. (2) The Artichoke extract had an effect of suppressing the diminution oj sugar, protein levels and the increase of the levels of GOT, GPT, alkaline phosphatase and bilirubin in the blood of intoxicated chickens. There was not or very Jew residue of Aflatoxin $B_1$ contained in the liver and muscle of chickens intoxicated by Aflatoxin $B_1$ having Artichoke, that was much lower than the allowed level in animal products.

• Journal of Microbiology and Biotechnology
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• v.21 no.8
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• pp.802-807
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• 2011

Cationic liposomes have been actively used as gene delivery vehicle because of their minimal toxicity, but their relatively low efficiency of gene delivery is the major disadvantage of these vectors. Recently, cysteine residue incorporation to HIV-1 Tat peptide increased liposomemediated transfection compared with unmodified Tat peptide. Therefore, we designed a novel modified Tat peptide having a homodimeric (Tat-CTHD, Tat-NTHD) and closed structure (cyclic Tat) simply by using the disulfide bond between cysteines to develop a more efficient and safe nonviral gene delivery system. The mixing of Tat-CTHD and Tat-NTHD with DNA before mixing with lipofectamine increased the transfection efficiency compared with unmodified Tat peptide and lipofectamine only in MCF-7 breast cancer cells and rat vascular smooth muscle cells. However, cyclic Tat did not show any improvement in the transfection efficiency. In the gel retardation assay, Tat-CTHD and Tat-NTHD showed more strong binding with DNA than unmodified Tat and cyclic Tat peptide. This enhancement was only shown when Tat-CTHD and Tat-NTHD were mixed with DNA before mixing with lipofectamine. The effects of Tat- CTHD and Tat-NTHD were also valid in the experiment using DOTAP and DMRIE instead of lipofectamine. We could not find any significant cytotoxicity in the working concentration and more usage of these peptides. In conclusion, we have designed a novel transfection-enhancing peptide by easy homodimerization of Tat peptide, and the simple mix of these novel peptides with DNA increased the gene transfer of cationic lipids more efficiently with no additional cytotoxicity.

• Journal of Microbiology and Biotechnology
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• v.23 no.1
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• pp.69-75
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• 2013

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.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.2
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• pp.187-191
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• 2004

Actin is a ubiquitous and highly conserved protein found in eukaryotic organisms. In this study, we describe the cDNA cloning and mRNA expression of an actin gene from the mulberry longicorn beetle, Apriona germari. The A. germari actin cDNA is 1524 bp containing a complete 1128 bp open reading frame that encodes a polypeptide of 376 amino acid residues with a predicted molecular weight of about 41.5 kDa. The deduced amino acid sequence of the A.germari actin cDNA showed 99% protein sequence identity to Homalodisca coagulata actin, differing at only two amino acid positions, and 92-98% protein sequence identity to known insect species actins. The predicted three-dimensional structure of A. germari actin revealed the four residue hydrophobic pulg loop characteristic of the actin family. Northern blot analysis showed that A. germari actin is highly expressed in epidermis and muscle, and less strongly in midgut, but not in the fat body of A. germari larva.

• Food Science of Animal Resources
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• v.44 no.4
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• pp.873-884
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• 2024

Flunixin is a veterinary nonsteroidal anti-inflammatory agent whose residues have been investigated in their original form within tissues such as muscle and liver. However, flunixin remains in milk as a metabolite, and 5-hydroxy flunixin has been used as the primary marker for its surveillance. This study aimed to develop a quantitative method for detecting flunixin and 5-hydroxy flunixin in milk and to strengthen the monitoring system by applying to other livestock and fishery products. Two different methods were compared, and the target compounds were extracted from milk using an organic solvent, purified with C₁₈, concentrated, and reconstituted using a methanol-based solvent. Following filtering, the final sample was analyzed using liquid chromatography-tandem mass spectrometry. Method 1 is environmentally friendly due to the low use of reagents and is based on a multi-residue, multi-class analysis method approved by the Ministry of Food and Drug Safety. The accuracy and precision of both methods were 84.6%-115% and 0.7%-9.3%, respectively. Owing to the low matrix effect in milk and its convenience, Method 1 was evaluated for other matrices (beef, chicken, egg, flatfish, and shrimp) and its recovery and coefficient of variation are sufficient according to the Codex criteria (CAC/GL 71-2009). The limits of detection and quantification were 2-8 and 5-27 ㎍/kg for flunixin and 2-10 and 6-33 ㎍/kg for 5-hydroxy flunixin, respectively. This study can be used as a monitoring method for a positive list system that regulates veterinary drug residues for all livestock and fisheries products.

• The Korean Journal of Malacology
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• v.24 no.3
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• pp.253-260
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• 2008

A total of 15 different residues of polycyclic aromatic hydrocarbons (PAHs) in each 20 samples of Pacific oysters, dried laver and rockfish obtained from seafood markets were analyzed. The prevalence of samples in which more than one PAH residues were found was 75% in oyster, 35% in rock fish hepatopancreas, 0% in rockfish muscle and laver, respectively. To estimate factors contributing to this residue level difference among organisms, tissue concentrations were analyzed after exposing three organisms to phenanthrene, a representative PAH, with concentration of 0.01 or $0.1{\mu}g/mL$ for 2 weeks. Phenanthrene levels after exposure were higher in the oyster digestive gland, laver and rockfish hepatopancreas, but were lower in the oyster whole meat or rockfish muscle. This finding disproved that any close relationship between the residue difference of market samples and concentrating properties of PAHs. The second possible factor analyzed was total lipid contents in the three organisms. Although higher lipid level in hepatopancreas of rockfish may contribute accumulation of PAH residues in the rockfish, lipid factor did not affect to PAH levels in other organism samples. Activity of 7-ethyoxyresorufin-O-deethylase (EROD), a kind of cytochrome $P_{450}$ enzyme, was measured to evaluate the eliminated amount of PAHs through metabolism. The higher EROD activity in rockfish, compared to that in oyster, was likely to contribute to the lower PAH residues in the rockfish. More factors, such as different exposure history, organisms' ability to escape, ingestion through prey organisms, and post-harvest loss, should be studied in the future.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.11
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• pp.1655-1661
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• 2007

Prolyl 4-hydroxylase (P4H) plays a central role in collagen synthesis by catalyzing the hydroxylation of the proline residue in the X-Pro-Gly amino acid sequence, and controls the biosynthesis of collagen that influences overall meat quality. In order to verify expression level of the catalytic ${\alpha}$ subunit of P4H, a 2.7 kb clone of the ${\alpha}$ subunit gene for P4H was selected from a cDNA library prepared from the muscular tissue of Sancheong berkshire pigs, and the whole gene sequence was determined. As expression level of the ${\alpha}$ subunit of P4H differed between tissues of pigs, we intended to assess more precisely the level of ${\alpha}$-subunit expression between tissues of Sancheong Berkshire pigs by using RT-PCR. Muscular and adipose tissues were taken from each pig grouped by growth stage (weighing 60, 80, and 110 kg) of Yorkshire and Sancheong Berkshire pigs, and the expression levels of the ${\alpha}$-subunit of P4H were examined. Since there were significant differences in the expression level with respect to variation in growth stage (p<0.01), an attempt was made to identify any influences of pig species and tissue variation. The muscular and adipose tissues of pigs weighing 110 kg showed higher expression levels than pigs weighing 60 kg and 80 kg. In general, significantly higher expression levels were found in muscular than in adipose tissue. The expression levels in Sancheong Berkshire were significantly higher than in Yorkshire pigs (p<0.01 or p<0.05). Since expression level of the ${\alpha}$-subunit of P4H affects the activity of P4H and is connected to the biosynthesis of collagen and increased collagen can improve meat texture, this finding may explain why meat quality of the Sancheong Berkshire pig is acclaimed in Korea. Given the higher expression levels of the ${\alpha}$-subunit gene in adipose than in muscular tissue, and also in the heavier pigs, more intensive studies are required to assess the correlation between expression level of the ${\alpha}$ subunit gene and overall meat quality.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.10
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• pp.1668-1675
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• 2004

The effect of pH on surface hydrophobicity, sulfhydryl group, infrared spectrum, SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) pattern and enthalpy was investigated in recovered protein from mackerel and frozen blackspotted croaker by alkaline processing. Hydrophobic residue in myofibrillar protein exposed to the surface of protein, and hydrophobic interaction were the highest around 6$0^{\circ}C$. The surface hydrophobicity was different between myofibrillar protein and myofibrillar protein including sarcoplasmic protein (recovered protein). The peak at 1636 c $m^{-l}$ was increased with pH, and the recovered protein was unfolded in alkali pH. Difference of surface and total sulfhydryl group at pH 7.0 and 10 was comparative high, and decrease of surface sulfhydryl group indicated formation of S-S bonds. Mackerel and frozen blackspotted croaker in alkaline pH showed bands of polymerized myosin heavy chain on SDS-PAGE pattern. The transition temperatures of recovered protein were 33.1, 44.3 and 65.5$^{\circ}C$. Gelation of recovered protein from alkali processing was estimated by increase of $\beta$-sheet structure by pH treatment, S-S bonds by oxidation of surface sulfhydryl group in heating, polymerization of myosin heavy chain in order.r.