• Title/Summary/Keyword: Sperms

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Technique to Generate Sterile Males of Striped Fruit Flies, Zeugodacus scutellata, using Electron Beam Irradiation and their Application to Genetic Control (전자빔 조사를 이용한 호박꽃과실파리의 불임 수컷 제조 기술과 유전방제 응용)

  • Kim, Yonggyun;Al Baki, Md. Abdullah;Kwon, Gimyeon
    • Korean journal of applied entomology
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    • v.59 no.1
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    • pp.25-35
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    • 2020
  • The striped fruit fly, Zeugodacus scutellata, is endemic in Korea, but it has been regarded as one of the serious quarantine pests throughout the world. Sterile insect release technique (SIT) has been used to eradicate quarantine fruit flies. This study developed a technique to generate sterile males and applied SIT to control Z. scutellata. First of all, the reproductive systems of Z. scutellata adults were examined with fluorescent microscope. Polytrophic ovaries comprises of around 100 follicles with developing oocytes. Each follicle contains an oocyte with several nurse cells and are surrounded with follicular epithelium. Oocyte development began at 10 days after adult emergence (DAE) and formed chorionated oocytes after 20 DAE. On the other hand, male testes were well developed just after adult emergence. The vas deferens was filled with motile sperms. To generate sterile males, different doses (0~1,000 Gy) doses of electron beam were irradiated to 3~5 days old pupae of Z. scutellata. When male pupae were irradiated with electron beam at 200 Gy, they developed and mated with females without any significant difference compared to untreated males. Although the untreated females mated with the 200 Gy-irradiated males laid eggs, no eggs did not hatch. The 200 Gy-irradiated males were then applied to untreated male and female flies in a density ratio of 1:9 (untreated males : treated males). The laid eggs suffered significant infertility. These results suggest that electron beam-irradiated pupae at 200 Gy resulted in male sterility and the resulting males would be applied to SIT.

Histological and Histochemical Studies on the Epididymal Region and Deferent Ducts of the Drakes by the Age in Weeks (오리 부고환(副睾丸) 및 정관(精管)의 주령별(週齡別) 조직학적(組織學的) 및 조직화학적(組織化學的) 연구(硏究))

  • Lee, Jae-Hong;Ha, Chang-Su
    • Korean Journal of Veterinary Research
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    • v.23 no.2
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    • pp.137-148
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    • 1983
  • This study was made for the better information of the male reproductive system on the meat-type drake, Cherry Belly X White Golden. The epithelium of ductules of epididymal region and deferent duct were observed histologically and histochemically with the progress of their development. India-ink absorbability on the luminal epithelium was also investigated after the administration of India-ink. The results are as follows; 1. Rete testis and various round ductules in immature form appeared in epididymis within 6 weeks after hatching, and simple cuboidal and simple columnar epithelium were found in the epithelia of the ductules within 8 weeks after hatching. Larger ductules were found on epididymal surface which was in the developing stage near to the immature efferent ductule. From 10th to 20th week, various ductules appeared in epididymis, and developing form of efferent ductules were much more increased on epididymal surface. The luminal epithelium of the ductules were composed of ciliated simple columnar and pseudostratified ciliated columnar cells. At the same time, deferent duct appeared. From the 21th week, various ductules in epididymis became abruptly matured. Lumen of rete testis was lined by simple squamous or simple cuboidal epithelium, and that of efferent ductules, having many folds and being larger than any others were lined by pseudostratified ciliated columnar epithelium in which ciliated columnar cells, non-ciliated cells(clear cells) and basal cells were noted. Connecting tubules of star shaped lumen were composed of pseudostratified ciliated columnar epithelium in which ciliated columnar cells, nonciliated cells, and basal cells were observed. The luminal surface of epididymal ducts was smooth and has thick pseudostratified columnar epithelium which was composed of high columnar cells and basal cells. From 26th week after hatching, sperm pooling was started in various ductules. 2. From 4th to 10th week, simple cuboidal epithelium of deferent duct transformed to simple columnar epithelium with the progress of aging. At the basement of epithelium, clear round cells were noted. From 12th to 20th week, high columnar cells with enlongated nucleus were noted on the luminal border of deferent ducts, forming folds of pseuclostratified columnar epithelium. From 20th week, the deferent duct started to have septa in it's lumen and composed mainly of pseudostratified columnar epithelium, and round cells disappeared. From 20th week, the lumen diameter of deferent duct became wider with the progress of aging, but there was no difference among the values of lumen diameter in upper, middle, and lower part of deferent ducts. At 26th week, the pooling period of sperms in deferent ducts, the lumen diameter became rapidly widen, especially in the lower part of deferent ducts. Thickness of muscular layer of ductus deferens showed gradual growth within 24 weeks but did abrupt thickening from 26th week. 3. Saliva resistant PAS granules were dotted on the top of nucleus in efferent ductules epithelium but the amount of the granules were little in the connecting ductules's epithelium. The granules reactive to acid phosphatase were abundant in the some epithelial cells of efferent ductules and connecting ductules, especially above the nucleus of cells. The granules reactive to alkaline phosphatase were noted on the luminal border of efferent ductules. Parts of free border of efferent ductules and middle portion of deferent ducts were stained slightly by alcian blue technique. India ink granules were found mainly in the epithelium of efferent ductules but were few in that of connecting ductules.

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Effect of $\beta$-Mercaptoethanol on In Vitro Maturation of Porcine Folliculay Oocytes and Development of Porcine IVM/IVF Embryos ($\beta$-Mercaptoethanol의 첨가배양이 돼지난포란의 체외성숙과 배발달에 미치는 영향)

  • 한만희;이경본;천행수;박병권;서길웅;이규승
    • Korean Journal of Animal Reproduction
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    • v.27 no.2
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    • pp.125-133
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    • 2003
  • The present study was carried out to examine the effect of $\beta$-Mercaptoethanol ($\beta$-ME) on in vitro maturation (IVM) of porcine follicular oocytes and oxygen concentration with $\beta$-ME on in vitro development (IVD) of porcine IVM/IVF embryos. The results were summarized as follows. 1. The rates of nuclear maturation, penetrated oocytes, polyspermic oocytes, pronucleus formation and mean numbers of the penetrated sperms were not significantly different using NCSU-23 maturation media for 0, 25, 50 and 100 $\mu$M $\beta$-ME (P>0.05). 2. The rates of blastocyst formation at day 7 after in vitro fertilization were higher in oocytes matured with 25 $\mu$M $\beta$-ME (25.4$\pm$0.9%) than in those matured with 0 (14.5$\pm$1.6%), 50 (17.3$\pm$1.7%) and 100 $\mu$M (12.4$\pm$1.3%) (P<0.05). However, no differences ware found in total cell numbers of blastocyst among the treatments. 3. The rates of blastocyst formation at day 7 after in vitro fertilization were higher in the NCSU-23 Culture medium With 25 $\mu$M $\beta$-ME (23.6$\pm$2.8%) than in those Cultured With 0 (15.4$\pm$4.4%), 12.5 (17.5$\pm$2.3%) and 50 $\mu$M $\beta$-ME (18.6$\pm$2.1%) Under the 5% and 20% $O_2$ Concentrations (P<0.05). However, no differences was found in total cell numbers of blastocyst among the treatments. These results suggested that the addition of 25 $\mu$M $\beta$-ME in the IVM/IVD media were effective on the porcine embryo production. However, the rates of blastocyst formation and total cell numbers of blastocyst at day 7 of porcine IVM/IVF embryos were not significantly different in the NCSU-23 culture medium under 5% and 20% 02 concentrations.

Production of In-Vitro Fertilized Embryos and Result of Transfer with Fresh or Frozen Embryos for Hanwoo and Holstein Cattle (한우 및 젖소에서 체외 수정란 생산과 신선 및 동결 수정란 이식 결과)

  • Kim Y. J.;Kim H. C.;Seo S. H.;Jeong K. N.;Kim Y. S.;Lee H. R.;Shin D. S.;Jo S. W.;Kim S. H.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.79-87
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    • 2005
  • The ovaries from Hanwoo and Holstein were collected from labattoir and transferred to laboratory. Oocytes were aspirated and incubated in $CO_2$ incubator for 24 hours for maturation. Oocytes were coincubated with the sperms for 5 hours. Cleaved oocytes were selected 48 hours after coincubation and half of the medium was changed newly every 48 hour until blastocyst formation. Cleavage rate and blastocyst rate were investigated according to different breeds and different status of cumulus cells surrounding the oocytes. Blastocysts were either transferred to the recipients or frozen until use. The result of embryo transfer with fresh or frozen embryos was investigated. The rate of male offspring following embryo transfer was also investigated. The rate of cleavage was $66.4\%$ for Hanwoo and $62.4\%$ for Holstein oocytes. The rate of cleavage according to status of oocyte was shown highest in the oocytes completely surrounded with cumulus cells and lowest in denuded oocytes for both Hanwoo and Holstein oocytes. The rate of blastocyst from cleaved oocytes was $40.6\%$ for Hanwoo and $36.9\%$ for Holstein. The rate of pregnancy/delivery following embryo transfer with fresh IVF embryos was $57.2\%$ for Hanwoo and $53.3\%$ for Holstein. The rate of pregnancy/delivery following embryo transfer with frozen IVF embryos was $40.9\%$ for Hanwoo and $36.4\%$ for Holstein. The rate of male calf produced by embryo transfer was $63.6\%$ for Hanwoo and $50.0\%$ for Holstein.

Studies on the Biological Effects of Chemosterilant, Hempa, on the Rice Weevil (Sitophilus oryzae 1.) and Transmission of Sterility (화학적 불임유기물질 Hempa가 쌀바구미(Sitophilus oryzae L.)에 미치는 생물학적 영향 및 불임성의 전달에 관한 연구)

  • Shim Jai Wook
    • Korean journal of applied entomology
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    • v.12 no.1
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    • pp.1-21
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    • 1973
  • Some experiments were conducted to investigate the effects of the chemosterilant, hempa, on the biology of the rice weevil, Sitophilus oryzae L., and the transmission of the lethal factors in the progeny. One to three days old adult males were fed on the wheat grains treated with concentrations of 0.0625, 0.125, 0.25, and $0.5\%$ of hempa water solution. The effects of the treatment on the mortality, longevity, and the performance of oviposition were examined for the Pl generation, and the hatchability and mortality in the postembryonic development were also tested in the $F_1,\;F_2,\;BC_1,\;F_3,\;and\;BC_2$ generations to analyze the inheritance of the lethal factors. The results obtained were summarized as follows. (1) The average longevity of the treated males were ranged from 26.6 to 30.4 days, and indicated no statistical differences. (2) The mortality of the treated males were ranged between $3.3\%\;and\;13.3\%$ and showed no statistical significance. (3) The overall mean number of eggs laid by a female mated to a treated male with concentrations of 0.0625, 0.125, 0.26 and $0.5\%$ were 3.78, 4.05, 3.75 and 3.61 for the respective treatments, and they were not differ significantly from those of control which were 3.60 per female per 3 day period. The unmated female laid 1.91 in the same period, and significantly differ from those in other experimental groups. (4) The overall mean hatchability of the eggs laid by the females mated with males that had been treated with various concentrations of hempa were 86.82, 64.77, 53.47, 40.33 and $24.78\%$ for the respective concentrations of 0, 0.0625, 0.125, 0.25 and $0.5\%$. The hatchability decreased with the increasing concentrations. (5) The minimum hatchabilities were obtained from the eggs laid in the period of 10-12 days after treatment, then the hatchability increased showing some recovery. The recovery seemed to be very much delayed for the males which had been treated with the greater concentrations. Such a difference in hatchability might be related with the sensitivity of the developmental stages of the sperms, and broader spectrum in the stages and severer effects seemed to be associated with the increased concentrations. (6) The overall mean of larval mortality in the $F_l$ generation were 6.55, 17.89, 27.40, 35.42 and $52.17\%$ for the respective concentrations of 0,0.0625, 0.125,0.25 and $0.5\%$. And there was a tendency to increase in the mortality with the increase of concentrations. (7) The correlation coefficients between per cent sterile eggs and larval mortality for the experimental plots of 0.125, 0.25 and $0.5\%$ treatments showed r=+0.83 and +0.85, respectively, and it seemed to be close correlation between the lethal effects on the embryonic and post-embryonic developments. (8) Since the $SC_{50}$ of the sterile eggs was $0.133\%$ and $SC_{50}$ of the larval mortality was $0.565\%$, it was considered that tile lethal factors expressed more in the egg stages than the larval stages. (9) The ratio of female to male in the $F_l$ adults showed 100 : 125, 100 : 108 and 100 : 124 for the plots of 0.125, 0.25 and $0.5\%$ treatments, respectively. And it n·as considered that the sex ratio distortions might occur with the higher concentrations. (10) When the F, males originated 1.on the eggs had been laid by p, in the period of 16-18 days after treatment, were crossed to normal females $(BC_1)$ and made sib matings $(F_2)$, the per cent sterile eggs of the $BC_1$ generation were 13.88 and $33.04\%$ , and were 31.01 and $38.73\%$ for the $F_2$generation with the plots of 0.0625 and $0.125\%$ treatment, respectively. And these seemed to be a results of the $F_1$ individuals are carrying some chromosomal aberrations (11) The larval mortality was the highest in the $F_2$ plot and followed the female backcross plot, and the least in the male backcrosses. (12) The proportions of 1st and 2nd instar larvae among the larval development at tile 17th day after oviposition were 10.98, 27.26, 32.98 and $15.73\%$ in the normal female $\times$ normal male, $F_1$ female$\times$normal male, normal $female \;\times F_1$ male and $female \;\times F_1$ male plots, respectively. It was considered that the larval development might be delayed by the treatment in the 2nd generation. (13) Per cent larval mortality and sterile eggs were greater in the $F_2$ sib mating plots $(F_3)$ than both of $F_2$ backcrosses. Therefore, it seemed that some of the recessive lethal mutations might affect in the further generations. (14) The sterility, induced by the treatment of chemosterilant, hempa, was considered as the result of the dominant lethal mutations due to chromosomal aberrations such as translocation and/or deletion. The effects of these lethal factors seemed to be inherited tip to 3rd generation after treatment.

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Effects of a Herb Drug Extracts on Mitigation of Reproductive Toxicity after a Continuous Dose of Dioxin in Mice (생약재제가 Dioxin의 연속투여 후 생식독성의 완화에 미치는 영향 관한 연구)

  • 김상근;김민수
    • Korean Journal of Animal Reproduction
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    • v.24 no.3
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    • pp.241-248
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    • 2000
  • In this study, we examined the number and motility of sperms, and also observed the changes in testes weight, and histological changes of several organs after 5 days of a continuous administration of dioxin (2,3,7,8-tetrachlorodibenzo-p-dioxin) of per oral administration of a herb drug extracts, which were administerated altermate days, to elucidate the effects of the a herb drug extracts on reproductive toxicity of dioxin. 1. The sperm numbers of dioxin-administered groups were 90.7$\pm$3.6~l18.5$\pm$3.6$\times$10/suup 6/$m\ell$ and 67.3$\pm$4.1~88.2$\pm$3.3$\times$10$^{6}$ $m\ell$ for 10~20 $\mu\textrm{g}$/kg and 30~40 $\mu\textrm{g}$/kg dosages of dioxin-administerated groups, respectively. Each dioxin-administered group showed prominently lower value than that of control group's which was 119.3$\pm$3.4~120.2$\pm$4.7 $\times$ 10$^{6.}$$m\ell$. 2. The sperm motility of each dioxin-administered group's also showed lower value than that of control group's which was 93.6$\pm$3.8~94.9$\pm$3.4%. The sperm motility of each dioxin-administerated group were 77.0$\pm$4.7~89.5$\pm$3.6% and 66.5$\pm$3.3 ~79.9$\pm$3.8% for 10~20 $\mu\textrm{g}$/kg and 30~40 $\mu\textrm{g}$/kg dosages of dioxin-administerated groups, respectively. 3. The sperm numbers of each group, which was administered a herb drug extracts, were 77.4$\pm$3.2~90.9$\pm$3.4$\times$10$^{6}$ $m\ell$, 78.0$\pm$3.3~105.0$\pm$4.2$\times$10$^{6}$ $m\ell$, 76.2$\pm$2.8~84.4$\pm$3.5$\times$10$^{6}$ $m\ell$ and 75.4$\pm$3.3~80.2$\pm$3.3$\times$10$^{6}$ $m\ell$ for extracts of green leaf, red ginseng, Kugija and Oume-administered groups respectively. And the sperm motility of each group were 63.4$\pm$3.8~77.0$\pm$4.0%, 65.5$\pm$4.1~87.4$\pm$3.8%, 64.3$\pm$4.2~69.8$\pm$4.2%, 66.3$\pm$3.9~66.0$\pm$4.0% for extracts of green leaf, red ginseng, Kugija and Oume extracts-administered groups, respectively. 4. The number and motility of sperm of control group were 119.3$\pm$3.4~120.2$\pm$4.7$\times$10$^{6}$ $m\ell$ and 93.0$\pm$3.5~96.1$\pm$3.5%, respectively. Red ginseng extracts- administered group seemed to be recovered than my other groups, and the green leaf extracts-administered group was shown to be the next. The Kugija and Oume extracts-administered groups didn't show to be recovered much. 5. Most a herb drug extracts-administered group except the red ginseng-administered group displayed prominently lower values of testes weights than that of control group's which was 0.15$\pm$0.01~0.16$\pm$0.01g. The red ginseng extracts-administered group seemed to be recovered conspicuously. 6. After 5 days of a continuous administration of 30 $\mu\textrm{g}$/kg dosage of dioxin followed by 3 weeks of per oral administration of green leaf, red ginseng, Kugija, or Oume extracts, histological findings showed that the liver, spleen, and testis of most a herb drug extracts-administered groups were damaged severely. By the way, the testes of red ginseng extracts-administered group seemed to recover compared to the other group's.ompared to the other group's.

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