This experiment was carried out to develop an in vitro culture system for rabbit embryos. The zygotes or 2-cell embryos were collected from the oviducts of the superovulated and mated does with D-PBS/10% FCS at 24 hours after hCG injection. The in vitro developmental rate of blastocyst formation and the number of nuclei in the embryos were examined under the following treatments; 1) TCM-199 with 10% FCS, 2) EBSS with 10% FCS, 3) rabbit vitreous humor(VH), 4) TCM-199 with 10% FCS+BOEC, 5) TCM-199 with 10% FCS+ROEC, 6) EBSS with 10% FCS+BOEC and 7) EBSS with 10% FCS+ROEC. For a comparative study of in vivo and in vitro development, the fresh blastocysts, which were developed in vivo for 96 hours after hCG injection, were collected from the uterus and their numbers of nuclei were counted. 1. The zygotes or 2-cell embryos developed to the blastocyst stage in TCM-199, EBSS and VH at the rates of 93, 92 and 89%, respectively. 2. The higher developmental rates 95~98% of blastocyst formation was achieved when the embryos were co-cultured with a monolayer of bovine or rabbit oviductal epithelial cells in TCM-199 or EBSS. No significant difference in developmental rates was shown between bovine and rabbit oviductal epithelial cells. 3. In a comparative study of in vivo and in vitro development, the total numbers of nuclei were significantly less in the in vitro cultured embryos(104~224) than the in vivo developed embryos(1, 0090 at 96 hours after hCG injectin. 4. The mean cell cycle numbers in the embryos cultured for 72 hours in TCM-199 with 10% FCS, EBSS with 10% FCS, TCM-199 with 10% FCS+BOEC, TCM-199 with 10% FCS+ROEC, EBSS with 10% FCS+BOEC and in vivo was 7.38, 6.63, 7.76, 7.69, 7.01 and 9.92, respectively. From these results, it can be suggested the optimal culture system for in vitro culture of rabbit embryos is a co-culture system with bovine or rabbit oviductal epithelial cells in TCM-199 with 10% FCS. Considering the significant reduction in total numbers of nuclei in the in vitro cultured embryos, the advanced research on development of in vitro culture system for rabbit embryos is expected.
To evaluate effects of Sam so on Pye Tang(SSOPT) Water Extract plasma cortisol concentration and plasma $CO_2$ in the Rabbit. The results obtained were as follows. 1. Intravenous administration of SSOPT water extract at the dose of 0.5ml/kg remarkably increased plasma cortisol concentration on 1 hour. 2. Intravenous adminitration of SSOPT Water extract at the does of 1.0ml/kg significantly decreased plasma $CO_2$ on 1 hour. 3. Intravenous adminitration of SSOPT Water extract at the does of 0.5ml/kg remarkable decreased plasma $Na^+$ from 1 to 4 hour and 1.0ml/kg significantly decreased plasma $Na^+$ on 1 hour. 4. Intravenous administration of SSOPT Water extract at the does of 1.0ml/kg remarkably increased plasma $k^+$ from 1 to 3 hour. 5. Intravenous administration of SSOPT water extract at the does of 0.5ml/kg significantly decreased plasma $CI^-$ on 3, 4 hours. and 1.0ml/kg remarkably increased plasma $CI^-$ on 4 hour. These results suggest that therapeutic action of SSOPT water extract for athma may be reated with the increment of plasma cortisol concentration and the decrease plsma $CO_2.$.
Global warming is a key challenge subjecting animals to heat stress conditions resulting in multiple physiological alterations in tropical climate. Dietary approach seems to be the more friendly approach to curb the adverse effects of heat stress in rabbits. Some herbs have been categorized to have high potential for promotion of immune responses for amelioration of heat stress. Thus, this research aims to evaluate the potential of Mistletoe (Viscum album), Moringa (Moringa oleifera) and Phyllanthus (Phyllanthus amarus) leaf meal as herbal supplements for the alleviation of heat stress in female rabbits by measuring improvement in sex and stress hormonal responses in serum biochemistry. 80 Rabbit does were exposed to 4 dietary groups supplemented with each of Mistletoe, Moringa, Phyllanthus and a control in an 84-day trial at the summit of thermal stress in South west Nigeria. Growth indices were monitored throughout the study, blood samples were compiled at the end of the trial to assess serum biochemistry, stress and sex hormonal responses of the Does using standard protocols. The results revealed that final weight and weight gain of Does fed on Phyllanthus were significantly (p < 0.05) higher (11.46% and 14.25%, respectively) than Does on control. The herbal supplements enhance glucose, protein, albumin and globulin, reduced cholesterol, and creatinine of Does under heat stress conditions. Among the herbal treatment groups, mistletoe, moringa and phyllanthus had 12.42%, 18.39% and 16.90%, respectively, lower corticosterone than control groups which had 39.76ng/ml. Triiodothyronine of Does fed control were significantly (p < 0.05) lower than Does on Moringa oleifera and Phyllanthus amarus supplements. Estradiol and Follicle stimulating hormone of rabbit Does fed on moringa supplement were significantly (p < 0.05) higher than other treatments. In conclusion, the herbal supplements tend to mitigate the detrimental outcome of thermal stress on Does by suppressing stress hormones. Moringa oleifera and Phyllanthus amarus enhanced sex hormones while Phyllanthus amarus confered growth promoting effects on the Does.
The following results were obtained from the observation on the change of plasma cortisol concentration in the experiment of intravenous administration of Haengsoyeum Water Extract in the rabbit. And the effects of Haengsoyeum extract on the contractile force of the isolated guinea pig trachea smooth muscle. 1. In intravenous administration the plasma cortisol concentration increased significantly about 1 hours after with a does of 0.2 ml/kg. 2. In intravenous administration the plasma cortisol concentration increased significantly about from 1 to 3 hours after with a does of 0.4 ml/kg. 3. The contractile response of the trachea smooth muscle of isolated guinea pig to histamine $10^{-4}\;M$ was significantly inhibited by Haengsoyeum extract. 4. The contractile response of the trachea smooth muscle of isolated guinea pig to acetylcholine $10^{-4}\;M$ was considerably inhibited by Haengsoyeum extract. 5. The contractile response of the trachea smooth muscle of isolated guinea pig to 5-hydroxytryptamine $10^{-4}\;M$ was inhibited by Haengsoyeum extract.
The rabbit pyrogen test and Limulus amoebocyte lysate (LAL) assay have been used to detect endotoxins present in vaccines. Currently, the rabbit pyrogen test is used to detect endotoxins in hepatitis B (HB) vaccines, even though the HB surface protein, which is the active ingredient, is overexpressed in and purified from eukaryotic cells that lack these endotoxins. Although the LAL clot assay is sensitive and reliable and can be used to replace the rabbit pyrogen test, its reaction is limited by the lack of responsiveness to the Gram-positive bacterial components. Furthermore, aluminum hydroxide in the HB vaccine can interfere with the LAL assay. In contrast, macrophages can detect the endotoxin as well as other pyrogens, and secrete $TNF-{\alpha}$. Therefore, this study was undertaken to examine the possibility of replacing the animal tests with a more efficient $TNF-{\alpha}$ secretion assay. With this in mind, we determined if aluminum hydroxide in the HB vaccines affects the $TNF-{\alpha}$ secretion assay. HB vaccines and the HB protein solutions spiked with lipopolysaccharide (LPS) produced the same level of dose-dependent $TNF{\alpha}$ secretion and temperature increase in rabbits, indicating that aluminum hydroxide in the HB vaccine does not interfere with the pyrogenic response in rabbits, nor does it interfere with $TNF-{\alpha}$ secretion. In addition, the $TNF-{\alpha}$ assay was found to be more sensitive than the LAL assay, and correlated well with the pyrogen test and the LAL assay. These results suggest that the $TNF-{\alpha}$ assay in RAW264.7 cells is a good substitute for the current pyrogen assays that are used for detecting LPS in HB vaccines as well as in other vaccines containing aluminum.
This study evaluated the influence of cell stage of donor nucleus on nuclear injection, electrofusion and in vitro development in the rabbit to improve the efficiency of nuclear transplantation in the rabbit. The embryos of 8-, 16- and 32-cell stage were collected from the mated does by flushing viducts with Dulbecco's phosphate buffered saline(D-PBS) containing 10% fetal calf serum(FGS) at 44, 54 and 60 hours after hCG injection. The blastorneres separated from these embryos were used as donor nucleus. The ovulated oocytes collected at 14 hours after hCG injection were used as recipient cytoplasm following removing the nucleus and the first polar body. The separated blastomeres were injected into the enucleated oocytes by micromanipulation and were electrofused in 0.28 M mannitol solution at 1.5 kV /cm, 60 $\mu$sec for three times. The fused oocytes were cocultured with a monolayer of rabbit oviductal epithelial cells in M-199 solution containing 10% FGS for 72~120 hours at 39$^{\circ}C$ in a 5% $CO_2$ incubator. The cultured nuclear transplant embryos were stained with Hoechst 33342 solution and the number of cells were counted by fluorescence microscopy. The successful injection rate of 8-, 16- and 32-cell-stageblastomeres into enucleated oocytes was 86.7, 91.0 and 93.9%, respectively. The electrofusion rate of 8-, 16- and 32-cell-stage blastomeres with enucleated oocytes was 93.3,89.3 and 79.0%, respectively. Development of blastomeres to blastocyst was similar with 8-,16- and 32-cell-stage donor nuclei(26.2, 25.8 and 26.6%, respectively, P<0.05). The mean number of cell cycle per day during in vitro culture in nuclear transplant embryos which received 8-, 16- and 32-cell- stage nuclei was 1.87, 1.81 and 1.43, respectively.
For a large sclase production of genetically identical or cloned animals, the effect of cryopreservation by vitrification on the post-thaw viability of nuclear transplant rabbit embryos were investigated. The embryos of 16-cell stage were collected from the mated does at 48 hours post-hCG injection, and they were synchronized to G1 phase of 32-cell stage were injected into enucleated recipient cytoplasms by micromanipulation. After culture until 20h post-hCG injection, the nuclear transplant oocytes were electrofused and activated by electrical stimulation. After in vitro culture for 48h, the nuclear transplant embryos developed to morula stage were cryoperserved with EFS solution by vitrification method. The forzen nuclear transplant embryos were thawed and cultured for 72h and the nuclear transplant of blastomeres under a fluorescence microscopy. The in vitro development to blastocyst of intact-fresh and intact-frozen 16-cell embryos was found to be 96.9 and 63.9%, respectively. The in vitro development to blastocyst of nuclear transplant and frozen-thawed nuclear transplant embryos was found to be 74.5 and 42.9%, respectively. Also, their mean blastomere numbers and mean cell cycles/day was 153 and 105, 145 and 1.34, respectively. From the above results it was concluded that the present cryopreservation by vitrification of nuclear transplant rabbit embryos might be useful though was decreased significantly.
Intravenous injection of Mori Radicis Cortex Water Extract (MWE) regularly caused the does-related, lowering of blood pressure in the rabbits anesthetized with urethane, and then did not show the cumulative effect and the tachyphylaxis. The hypotensive effects of MWE were inhibited by atropine, chlorisodamine, phentolamine and bethanidine, while not altered by diphenhydramine, propranolol and cyproheptadine. Atropine after chlorisondamine did not alter the effect of MWE. MWE potentiated the pressor effect of nore-pinephrine, but did not carotid occlusion was inhibited by previous administration of MWE. It is conclude that MWE elicits hypotensive action in the rabbit by the centrally induced cholinergic effect and the inhibitation of responses to sypathetic adrenergic nerve activation.
The effect of exogenous gonadotrophins on superovulation in rabbits was examined. One hundred and sixteen sexually mature California, Chinchilla and New Zealand White rabbits were randomly allocated to control (100 IU hCG), PMSG-treated (100 IU HCG following 150 IU PMSG) and FSH-treated groups (0.3 mg/head /12 h for 3 days followed by 100 IU hCG). All does were mated after hCG injection and were sacrificed or laparotomized within 1 to 4 days postcoitus for counting the number of ovulation points. The number of ovulations was higher in FSH-treated animals than in the control and PMSG-treated groups (37.2 vs. 10.4 and 14.5, p<0.05). Follicle haemorrhagicum was observed in many cases in the PMSG-treated group. No significant difference in ovulation number was observed between left and right ovaries regardless of gonadotropin treatment. In another experiment, 2-cell stage embryos were collected at 26 h postmating and blastomeres were separated by mechanical pipetting or gentle pressure with a fine glass needle. Aggregated or chimeric embryos were produced from two single blastomeres from two breeds, New Zealand White and Chinchlla, with different coat colors. All the embryos were cultured in Ham's F-10 medium supplemented with 1.5% BSA (bovine serum albumin fraction V) and 10% PRS (pregnant rabbit serum), and incubated in a humidified atmosphere with 5% $CO_2$ at $38^{\circ}C$. After development to morula or early blastocyst, the embryos were transferred into the oviducts of recipient does. Results showed that 7 out of 10 does (70%) receiving intact embryos (control) became pregnant and 41 kits were delivered. However, no pregnancy was obtained from the recipient of either denuded demi- or aggregated embryos. It is suggested that embryos without zona pellucida could not develop to term in rabbits.
Zea mays Linne belonging to Graminae family has been ordinarily made use of foods and fodder. Maydis Stigma, the stigma of maize, has been discarded without any special application with the exception of being applied as diuretic in the sphere of Chinese medicine, moreover, the pharmacological research toward Maydis Stigma as a medical herb has not been regretfully undertaken at all. The authors hereby paid attention to this point of view and made experiment to find out whether the water extract of Maydis Stigma might possess some pharmacological effect on the motility of the isolated rabbit uterus. The experiment was performed making the use of several agents related to the uterine motility such as acetylcholine, pilocarpine, epinephrine, atropine, barium chloride, quinine sulfate and oxytocin. The motility of the isolated rabbit uterus in Tyrode' solution was recorded with the electric kymograph according to Magnus method. The results of the experiment are as follows. 1. The motility of the isolated pregnant and nonpregnant rabbit uterus represents the tendency of gradual contractility in proportion to the concentration of ZW $10^{-4}$, $5{\times}10^{-4}$ and $10^{-3}$. 2. The uterine contractile effect of ZW $10^{-3}$ probably seems to antagonize the effect of epinephrine $10^{-6}$. 3. ZW $10^{-3}$ does not seem to have the significance with barium chloride, quinine sulfate and oxytocin respectively.
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