• Asian-Australasian Journal of Animal Sciences
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• v.15 no.10
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• pp.1412-1421
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• 2002

To examine the feasibility of using a sperm vector system for gene transfer, we have investigated the binding and the uptaking of foreign DNA into the sperm nucleus by PCR, in situ hybridization and LSC. We have also examined the transportation of exogenous DNA into oocytes by immunofluorescene via PCR. Sperm cells were incubated with DNA/liposome complexes (1:4 ratio) in fertilization medium with BSA or without BSA. In situ hybridization demonstrated that the transfection rate of sperm cells with and without BSA was 41 and 68% respectively, when the cells were treated with liposome/DNA complexes and 13% for DNA alone. LSC analysis showed that the binding of exogenous DNA was greatly reduced by DNase I treatment which digests DNA bound onto spermatozoa, suggesting that some of the DNA was internalized into the sperm membrane. To find out whether transfected DNA was internalized into sperm intracytomembrane, sperm DNA was amplified by inverse PCR. No PCR products were detected from sperm cells, indicating that the foreign DNA was simply bound onto the sperm membrane. To investigate transfer rates of exogenous DNA into oocytes via sperm cells, we used immunofluorescene method to follow the distribution of foreign DNA via spermatozoa: a few exogenous DNA was located in the cytoplasm of early embryos (13/60, 21.7% for DNA+/liposome+/BSA) and was not located in the pronucleus and/or nucleus. These results suggest that most of the transfected sperm cells could carry the foreign DNA into the egg by in vitro fertilization, but that the transferred DNA is degraded in the developing embryos without stable integration into the zygote genome. Therefore, we have directly injected with transfected sperm cell into oocyte cytoplasm and observed that some of the exogenous DNA was detected in preimplantation embryonic cytoplasm and expressed at preimplantation stages, suggesting that exogenous DNA in early zygote has their integrity. In this study, we have not identified a noble mechanism that interfering transportation of foreign DNA into zygote genome via spermatozoa. Our data, however, demonstrated that inverse PCR and immunofluorescene methods would be used as a new tool for follow-up of gene distribution in oocyte via sperm cells.

• Korean Journal of Plant Tissue Culture
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• v.22 no.6
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• pp.317-322
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• 1995

As a basic research for breeding new varieties, reciprocal -intergeneric crosses between Citrus junos and P.trifoliata were made. F$_1$ hybrid production using in vitro ovule culture, gametogenesis, and fertilization phenomena were investigated. Frequency of fruit set resulting from crossing of Citrus junos and Poncirus Trifoliata was 16.6% while that of Poncirus Trifoliata and Citrus junos was 11.7%. Callus formation occurred well when ovules at the 6th week after pollination were cultured on MT (Murashige and Tucker) medium supplemented with zeatin 0.5 mg/L and NAA 1.0 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4-D 0.1 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4 D 0.1 or 0.5 mg/L. The invitro germination rates of 20-week-old ovules set C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 54.5% and 48.6%, respectively. The emergence ratios of trifoliate hybrids obtained by C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 56.7% and 100%, respectively. The chromosome number of C. junos and P. Trifoliata was n = 9 or 2n = 18, and the sizes of their pollen grain were 33.75 $\mu$ and 25.0 $\mu$. The length and width of embryo sac in C. junos and P. Trifoliata were 69.38~79.23 $\mu$ and 27.50~38.56 $\mu$, and those of egg cells were 17.50~41.50 $\mu$ and 6.25~8.12$\mu$. Fertilization of C. junos and P. trifoliata terminated 72 h after pollination.

• Korean Journal of Poultry Science
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• v.42 no.3
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• pp.247-256
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• 2015

There are approximately 1,500 broiler farms in Korea, each raising 55,000 birds. Ninety-five percent of the farms are contracted with Integration Company. According to the Korean broiler performance index, broilers in Korea are marketed at 32 days with 1.52 kg of body weight. In contrast, the market age and body weight of broilers are 47 days/2.8 kg in the United States and 42 days/2.5 kg in Europe. Because of the younger market age of the Korean broiler, the pre-starter feed is important. Chicks exhibit poor absorption of dietary nutrients up to 7 days after hatching due to an immature digestive system and low enzyme secretion rate and activity. At the beginning of hatching, chicks obtain their nutrients from the egg yolk sac. It is highly recommended that chicks, after consuming the nutrients in the egg yolk sac, are given supplemented pre-starter feed to increase early growth rates and improve the performance of broiler production. Pre-starter nutrient requirements are not expressed in NRC, so Korean feeding standards for poultry and commercial breeding companies determine the nutrient requirements of pre-starter broiler chickens. Three approaches are followed to formulate specially designed pre-starter feeds for broiler chicks: (i) selective use of raw materials, (ii) proper standards of nutrient supply, and (iii) application of feed additives such as exogenous enzymes. In the selection of raw materials, those with high digestibility can be used. The absorption rate of carbohydrates in grains can be increased through feed processing at high temperature and high pressure. Soy proteins and fish meal can also be added as protein sources. As an energy source, vegetable oils are preferred over animal fats because of the former's high digestibility. It is suggested that the levels of proteins and amino acids are higher in pre-starter feed than in starter feed. With regard to energy, the sources of energy are more important than the levels of energy in feed. Feed additives such as exogenous enzymes can be used to improve nutrient digestibility. In addition, organic acids and plant extracts can be used as alternatives to animal growth promoters to stimulate immunity and prevent diseases. The growth performance of broilers is affected by various factors, such as management and disease control, in addition to the nutritional strategy; however, nutritional strategies play an important role in improving the productivity of broilers. Therefore, nutritional strategies, along with management and disease control, are required for improving the productivity of broilers in Korea.

• Korean Journal of Poultry Science
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• v.43 no.2
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• pp.111-118
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• 2016

Mycotoxins are secondary metabolites produced by molds, such as Aspergillus, Fusarium and Penicillium, that have adverse effects on animals and humans. Aflatoxin, ochratoxin, zearalenone, fumonisin and deoxynivalenol are the mycotoxins of greatest agro-economic importance and cause acute disease called mycotoxicoses. Mycotoxicosis in poultry birds results in decreased meat/egg production, immunosuppressant, and hepatotoxicosis. Some of toxins or their metabolites may be retained in animal or human tissues and induce health problems. This study was designed to develop a sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the simultaneous detection and quantification of mycotoxins, such as aflatoxin $B_1$, aflatoxin $M_1$, ochratoxin A, zearalenone, fumonisin B and deoxynivalenol, in chicken liver and kidney tissues. The mycotoxins were extracted and purified using modified QUECHERS methods, separated by LC and detected by an electrospray ionisation interface (ESI) and tandem MS. Good precision and linearity were observed for most of six mycotoxins. The recovery test for each mycotoxin in liver and kidney tissues mostly indicated good average recovery rates between 80.94% and 98.10% and the coefficient of variation mostly under 13.78%, except for aflatoxin $M_1$ and fumonisin $B_1$. The limit of detection (LOD) for six mycotoxins was $7.6{\sim}145.79{\mu}g/kg$ in liver tissues and $6.07{\sim}197.20{\mu}g/kg$ in kidney tissues. The quantification limits (LOQ) for 6 mycotoxins were in the range $23.04{\sim}441.78{\mu}g/kg$ in liver tissues and $18.40{\sim}597.59{\mu}g/kg$ in kidney tissues, respectively. The developed multi-mycotoxin method in this study permits simultaneous, simple, and rapid determination of several co-existing mycotoxins in chicken liver and kidney tissues.

• Journal of Aquaculture
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• v.9 no.1
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• pp.25-42
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• 1996

The study has been conducted to know an appropriate feeding strategy and effects of the rearing density on larval growth of the rockfish, Sebastes schlegeli. The results obtained are as fellowed ; 1. Thirty-day-old larvae reached at $25.25{\pm}3.76$ mm in total length and $0.23{\pm}0.07$ g in body weight in experiment A, at which rotifer was provided from the beginning to the end of 30-day experiment, Anemia from 3th to 18th day, and artificial feed from 13th to 30th day after hatching. When rotifer was provided for 30 days, Artemia from 6th to 18th day, and artificial feed from 18th to 30th day after hatching (experiment B), these larvae grew up to $27.52{\pm}2.50$ mm in total length and $0.26{\pm}0.06$ g in body weight. On the other hand, when rotifer and artificial feed were supplied with the same time schedule as shown in experiment B, and Artemia was feed from 6th to 30th day after hatching (experiment C), the total length and body weight of those larvae were $23.22{\pm}3.44$ mm and $0.15{\pm}0.05$ g, respectively. The best result for larval growth was obtained from experiment B. The survival rates estimated were $57.6\%$ in experiment A, $66.4\%$ in experiment B, and $44.4\%$ in experiment C. 2. The growth in total length of the larvae according to their rearing days could be represented by the following equations : Experiment A : Y=4.350+0.116X+$1.887X^2$ (r=0.993) Experiment B : Y=4.500+8.931X+$2.221X^2$ (r=0.994) Experiment C : Y=4.478+5.734X+$1.881X^2$(r=0.990) The average number of Artemia nauplius intaken by the larvae was rapidly increased between 15th and 20th day afer hatching, and 9, 212, 242, 750, and 1,171 nauplius were found in the different sizes of larvae, whose total length were 5.65, 6.81, 9.45, 14.96, and 24.52 mm, respectively. 3. Larval growth in total length and body weight reared at four different densites (A: 1.8 $kg/m^3$, B; 4.0 $kg/m^3$, C; 5.0 $kg/m^3$, D; 6.2 $kg/m^3$) indicated that the best growth was found in experiment A, at which the larval were reared at the lower density and the final survival rates extimated were $92.9\%$ in exp. A, $99.5\%$ in exp. C, $89.0\%$ in exp. B, and $88.2\%$ in exp. D. The amount of production per cubic meter turned out to be 30.45 kg in exp. D, 25.89 kg in exp. C, 20.75 kg in exp. B and 10.48 kg in exp. A. therefore, considering both larval growth and survival rate, higher yields seemed to be attainable at the relatively high-rearing density.