Kim, Tae-Shin;Yang, Cao;Cheong, Hee-Tae;Yang, Boo-Keun;Lee, Sang-Young;Park, Choon-Keun
Reproductive and Developmental Biology
/
v.30
no.3
/
pp.189-194
/
2006
Sperm-mediated gene transfer(SMGT) can be used to transfer exogenous DNA into the oocyte at fertilization. The main objective of this study was to assess efficiency of transferring mitochondrial DNA(mtDNA) fragment into boar spermatozoa in either presence or absence of liposome and quality of transfected spermatozoa. The mtDNA of chicken liver was isolated and purified by phenol and alkaline lysis extraction, and it was inserted to plasmid. The genome of transfected spermatozoa treated with DNase I was purified by alkaline lysis, and then amplified by the PCR analysis. After electrophoresis, DNA quantitation of each well was calculated by comparison of the band intensity with standard. As a result, exogenous DNA was composed of mtDNA fragment(1.2 kb) and plasmid(2.7 kb). On the other hand, efficiency of transfection by liposome($9.0{\pm}0.34ng/{\mu}l$) in SMGT was higher than that by DNA solution($6.9{\pm}0.53ng/{\mu}l$). However, there was no significant difference. Transfering exogenous DNA into spermatozoa was completed within 90 min of incubation. In another experiment, there were significant (p<0.05) differences between transfected spermatozoa using both DNA solution and DNA/liposome completes with unheated spermatozoa for viability ($70.8{\pm}1.80$ and $68.0{\pm}2.16%$ vs. $83.3{\pm}1.69%$, respectively) and motility($78.7{\pm}1.59$ and $79.3{\pm}2.14%$ vs. $86.7{\pm}1.59%$, respectively). This study indicates that exogenous mtDNA can be efficiently transferred into boar spermatozoa regardless of the presence of liposome, and transfected spermatozoa can also use insemination and in vitro fertilization to generate transgenic pig.
Park, Sun-Ae;Kim, Lee-Kyung;Park, Chang-Min;Kim, Seung-Chang;Lee, Seung-Hwan;Lee, Ji-Woong;Choi, Bong-Hwan
Reproductive and Developmental Biology
/
v.37
no.4
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pp.199-203
/
2013
This experiment was conducted to investigate the genetic effects of candidate genes on the growth of spleen and liver tissues using dietary Curcuma longa (C. longa) supplementation. Expression analyses of candidate genes regarding animal growth was performed in order to determine the factors affecting the growth related to immune components of Curucumin, Turmerone, and Zingiberene as the bile secretion Paratolyl methyl carbinol (PTMC). The animals were divided into four groups of five chicks supplied with experimental diets of C. longa at 0.25, 0.5 and 1% and controls. The 19 growth-related genes were known to cell maturation, differentiation significant expression patterns in this analysis. Expression of growth response-related genes in chicks supplemented with 1% of C. longa showed better growth performance than chicks with 0.25 and 0.5% in spleen (p<0.05). The IGF1, MSTN, POU1F1, ADCYAP1 gene were known to central roles in mediating gonadotropin function, regulating steroidogenesis and promoting oocyte growth and maturation. Sex steroids, androgen and estrogen can affect sex differentiation and also can affect muscle development. On the other hand, GHSR and FABP3 gene showed significant expression patterns in this analysis. The results would be used as basic information for the variation of growth-related genes expression on the cell growth, sex cell growth, and sex hormones according to dietary supplementation with C. longa in chickens.
Protective efficacy of trivalent Salmonella inactivated vaccine containing Chlorhexidine-inactivated S. Enterltidis (SE), S. Typhimurium (ST), and S. Gallinarum (SG) strains, was evaluated in this study. A total of 70 brown nick layers were divided into 7 groups, A to G, containing 10 hens per group. All hens in groups B to D were intramuscularly immunized with approximately 7×108 cells (3×108 cells of SE+1×108 SE+1×108 cells of ST+3×108 cells of SG) of the trivalent vaccine in 0.5 mL of PBS. All chickens in groups E to G were injected with sterile PBS. All hens of groups B and E, groups C and F, and groups D and G were orally challenged with approximately 2 ×109 CFU of wild-type SE, ST, and SG, respectively. Serum IgG titers and CD3+CD4+ T-cells, and CD3+CD8+ T-cells levels of groups B to D significantly higher than those of group A. In addition, all animals in groups A to C, E and F showed no clinical symptoms and survived after the virulent challenges, whereas one chicken in group D died and all chickens in group G died following the challenge. The protection against wild-type SE and ST in liver, spleen, cecum, and cloaca of groups B and C chickens was significant effective as compared with those in groups E and F. These indicate that the trivalent inactivated vaccine can be an effective tool for prevention of Salmonella infections by inducing robustly protective immune responses and cellular immune response in chickens.
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.
Sohn, Sea Hwan;Jang, In Surk;An, Young Sook;Moon, Yang Soo
Korean Journal of Poultry Science
/
v.42
no.1
/
pp.51-59
/
2015
Chickens are exposed to the external and internal stressors such as low and high temperature, high stocking density, feed restriction and disease. There have been a few studies on gene expressions through the investigation of chickens under direct exposure to the stress of high stocking density. The objective of the present study was to determine the expressions of genes associated with stress, endoplasmic reticulum (ER)-stress, lipid and glucose metabolism in two strains of chickens, Korean Native Chicken (KNC) and White Leghorn (WL), raised in high stocking density. A total of 164 chickens aged 40 weeks were randomly allotted to a $540cm^2/bird$ stocking density (control), whereas the chickens in a high density group were assigned in a $311cm^2/bird$ stocking density with feeding ad libitum for 10 weeks. Total RNA was extracted from the live for qRT-PCR. The expression levels of hsp70 and $hsp90{\alpha}$ were higher in WL subjected to stress with high stocking density compared with those genes in control (P<0.05), while the expressions of genes were not affected in KNC. ER stress marker gene XBP1 was also highly expressed in WL with stress (P<0.05), but the stress of high stocking density did not influence to ER stress marker genes in KNC. Lipid metabolism associated genes including FABP4, FATP1 and ACSL1 were highly expressed in WL compared with KNC when subjected to high stocking density stress (P<0.05). The expression of glucose transport gene GLUT2 and GLUT8 were increased in chickens exposured to the stress of high stocking density (P<0.05). The data indicate that WL is more sensitive to the stress of high stocking density compared with KNC and the stress may influence the modulation of lipid and glucose metabolism in the liver of chickens.
The Siberian ginseng and Eucommia are a kind of medicinal plant with powerful anti-oxidant activity. An experiment was conducted to investigate the effect of Siberian ginseng leaf and Eucommia leaf at level of 0.5% and 1% per feed in Ross commercial broiler for 4 to 35 days of age on performance, organ weight, blood biochemical profiles and telomere quantity. Chickens consuming diets containing 1% Siberian ginseng had higher feed conversion ratio than the other treated chicken during experimental period whereas no significant differences were detected in body weight, weight gain and feed intake. The weight of bursa of fabricius was significantly increased in chickens with dietary supplementation compared with chickens fed control but this was not seen in liver, spleen and thymus. In blood biochemical profiles, chickens with dietary supplementation had higher concentration than chickens fed control in triglyceride, cholesterol and glucose. The concentration of aspartate aminotransferase, alanine aminotransferase, albumin and total protein, however, was not significantly different between dietary supplemented chickens and control chickens. The relative amount of telomeric DNA of lymphocytes in chickens with dietary supplementation was significantly higher than that of control chickens but the difference was not found in liver, heart and testis tissues. In conclusion, dietary supplementation of Siberian ginseng and Eucommia in broiler improved immune activity and telomere length without decreasing chicken growth performance.
Kim, Jimin;Yoon, Hyung-sook;Hwangbo, Jong;Kim, Sang-Ho;Choi, Yang-Ho
Journal of Animal Environmental Science
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v.18
no.3
/
pp.183-190
/
2012
The objective of this study was to evaluate the effects of an unexpected change in housing environment on stress in poultry. Laying hens (Hy-line Brown), adapted to a free-range housing system for more than a month, were randomly divided into two groups to be subjected to an unexpected change of their housing environment: one half of them were individually housed into conventional battery cages and the other continually left in the same housing system throughout the entire period of the study. The sudden change resulted in an increase in egg production and albumen height (P<0.05), but decrease in eggshell thickness (P<0.05). The change had tendency to increase Haugh unit ($P{\leq}0.061$) and to decrease eggshell color ($P{\leq}0.074$), but did not affect body weight for 5 days thereafter. No significant changes were detected in liver color. Plasma concentrations of corticosterone was acutely increased one day after the change (P<0.05), then returned to similar control levels. The results of the present study indicate that unexpected changes in housing environment cause acute increase in stress hormone concentrations, but interpretation of the results should be cautious due to the experimental conditions.
FBs, secondary metabolites of several species of Fusaria, especially Fusarium moniliforme and F proliferatum, are commonly contaminated in com and other food grains throughout the world. Only recently identified, these mycotoxins have been associated field outbreaks of ELEM in horses and PPE in pigs. Currently, naturally or experimentally induced FB toxicosis has been studied in poultry, ruminants and rabbits. Poultry fed FB showed decreased growth rate, performance, and immune competence, as well as embryopathic, and embryocidal effects, and ricktes. Ruminants seem to be relatively less susceptible to FBs than other doestic animal. FB toxicosis reveals that liver is a target organ in all species, although other organs are affected in a species specific manner. Recently, the main target organs for $FB_1$ toxicity in rabbits was shown to be the kidney. Even low concentrations of FBs are likely to be a problem for animal health. A current study being conducted showed that feed containing low level of $FB_1$ reduces the ability of pulmonary intravascular macrophages in pig to clear blood-borne particles which would increase the susceptibility of animals to bacterial disease. The mechanism of FB toxicity remains unknown, but may be related to altered sphingolipid biosynthesis by inhibiting sphinganine N-acyltransferase. Elevations of serum and tissue SA:SO ratio have been observed in horse, pig, chicken, turkey, and rabbit, which could could serve as in effective biomarker for consumption of FB-containing feeds. There is limited information detailing dose-effect relationships either from field cases or in the laboratory. More research on the factors, including the prevalence and tolerance levels of FBs in feedstuffs that cause domestic animal disease associated with FBs, is urgently needed.
Different herbs and spices have been used as feed additives for various purposes in poultry production. This study was conducted to assess the effect of feed supplemented with black pepper (Piper nigrum), turmeric powder (Curcuma longa), coriander seeds (Coriandrum sativum) and their combinations on the performance of broilers. A total of 210 (Cobb) one-d-old chicks were divided into seven groups of 30 birds each. The treatments were: a control group received no supplement, 0.5% black pepper (T1), 0.5% turmeric powder (T2), 2% coriander seeds (T3), a mixture of 0.5% black pepper and 0.5% turmeric powder (T4), a mixture of 0.5% black pepper and 2% coriander seed (T5), and a mixture of 0.5% black pepper, 0.5% turmeric powder and 2% coriander seeds (T6). Higher significant values of body weight gain during the whole period of 5 weeks (p<0.001) were observed in broilers on T1, T3, T5, and T6 compared to control. Dietary supplements with T1, T2, T3, and T6 improved the cumulative G:F of broilers during the whole period of 5 weeks (p<0.001) compared with control. The dressing percentage and edible giblets were not influenced by dietary supplements, while higher values of relative weight of the liver (p<0.05) were obtained in T5 and T6 compared to control. The addition of feed supplements in T5 and T6 significantly increased serum total protein and decreased serum glucose, triglycerides and alkaline phosphatase concentrations compared with the control group (p<0.05). Broilers on T6 showed significant decrease in the serum glutamate pyruvate transaminase concentration (p<0.05) compared to control. The broilers having T5 and T6 supplemented feed had relatively greater antibody titre (p<0.001) at 35 d of age than control. It is concluded that dietary supplements with black pepper or coriander seeds or their combinations enhanced the performance and health status of broiler chickens.
This experiment was conducted to investigate the effects of dietary garlic powder (GP) on growth performance and mRNA expression of hepatic 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoA reductase) in broiler chickens. Day-old male chicks (Ross, n=270) were fed diets containing 0% [positive control (PC) with antibiotics or negative control (NC) without antibiotics], 1%, 3% and 5% GP for 6 wks. There were no significant differences in feed intake among the treatments throughout the experimental period. Body weight gains in groups fed dietary GP at 3% and 5% were significantly higher than that of NC group (P<0.05). Feeding GP up to 5% did not exert any adverse effect on weight gain and feed intake. There were no significant differences in the relative weights of liver, spleen, cecum and breast muscle. The content of meat cholesterol in GP containing dietary groups tended to be reduced as compared to NC group. Average infectious bronchitis antibody titers in chicks fed GP containing diets were significantly higher than that of NC group (P<0.05). The mRNA expression of hepatic HMG-CoA reductase was reduced by dietary GP. These results indicate that dietary GP has growth promoting effects and tended to alter cholesterol metabolism in broiler chickens.
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