• Journal of Veterinary Clinics
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• v.39 no.6
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• pp.319-325
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• 2022

Two-port laparoscopic ovariectomy (Lap-OVE) has been performed in small dogs, using 3-mm and 5-mm portal sites, and is associated with reduced surgical stress and postoperative pain. However, extension of the incision is often needed to extract the ovaries. In this study, we aimed to minimize invasiveness by using smaller-sized cannulas as well as a novel technique for ovary extraction. Lap-OVE was performed on six, healthy female dogs (range, 3 to 7.2 kg) using two 3-mm midline portals. The middle finger of a size M nitrile glove was cut at its base and sterilized preoperatively. The ovary was suspended at the body wall using a 1-0 blue nylon needle, and the ovarian pedicle and ligaments were transected using a 3-mm bipolar forceps. To facilitate the glove passing through the 3.9-mm port, it was turned inside out to expose the smooth inner surface, before being inserted into the abdominal cavity with an applicator. Both ovaries were placed inside, and the mouth of the glove was exteriorized through the port with a laparoscopic grasping forceps. The ovaries were morcellated inside the glove, using Adison-Brown tissue forceps and iris scissors, which enabled safe extraction without incision enlargement. Median incision lengths were 4.3 mm (3.5-mm cranial cannula) and 4.8 mm (3.9-mm caudal cannula). An advantage of this procedure was that there was no need for skin sutures. In conclusion, using our novel technique, sutureless Lap-OVE was possible in small dogs using two 3-mm portal sites without additional incision.

• Korean Journal of Veterinary Research
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• v.37 no.1
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• pp.71-78
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• 1997

This study was designed to investigate the number of the growing and mature follicles following gonadotrophin treatments for superovulation in mature rats. Eighteen mature rats (Sprague-Duwely, initially 190~230gm) were randomly alloted into 3 groups. One group was control group, another FSH-treated group was injected intramuscularly with 0.5 units of follicular stimulating hormone (FSH) / rat, and third PMS and HCG-treated group was intramuscularly injected with 20~25IU of pregnant mare serum (PMS) / rat and then at the 48 hrs later, with 20~25IU of human chorionic gonadotrophin (HCG) / rat. The uteri and ovaries of rats were collected and then were observed grossly and serial sections of paraffin embedding ovaries were stained with H-E. Number of ovarian follicles by following 3 grades of large, middle and small follicles from secondary and tertiary follicles were investigated by LM photography of preparations. Small follicles were classified as secondary follicles of preantral follicles with more than 2 layers of granulosa cells surrounding the oocyte and middle follicles were classified as secondary follicles with early signs of antral cavity or with more than one small cavity on either side of the oocytes and large follicles were classified as tertiary follicles with a single medium sized antral cavity or large well-formed antral cavity. In gross findings, the uteri were slightly swelling in FSH-treated group and markedly swelling or filled with fluid in the uterine lumen in PMS and HCG-treated group. In histological findings, the shape and size of the follicles were diverse in middle and large follicles of FSH-treated group and PMS and HCG-treated group, and proportion of atretic follicles was increased in FSH-treated group and PMS and HCG-treated group than those in control group. The uteri of FSH-treated group and PMS and HCG-treated group were hypertropied or filled with fluid in the lumens and walls of uteri. The wall tissue layers were flattened and their blood and lymph vessels were dilated. The mean number of follicle per ovary in control group were appeared to be $17.1{\pm}5.6$($14.0%{\pm}4.6%$), $37.8{\pm}9.1$($30.9{\pm}7.4%$) and $67.6{\pm}30.1$($55.2{\pm}24.6%$) respectively at large, middle and small follicles and total number of these 3 grade follicles were appeared to be $122.5{\pm}40.0$. The mean number of follicle per ovary in FSH-treated group were appeared to be $22.8{\pm}7.0$($17.4%{\pm}5.3%$), $43.4{\pm}6.6$($33.2{\pm}5.1%$) and $64.5{\pm}13.0$($49.3{\pm}9.9%$) respectively at large, middle and small follicles and total number of these 3 grade follicles were appeared to be $130.7{\pm}16.6$. The mean number of follicle per ovary in PMS and HCG-treated group were appeared to be $29.7{\pm}11.0$($16.3%{\pm}6.0%$), $61.9{\pm}17.2$($33.9{\pm}9.4%$) and $91.1{\pm}28.2$($49.9{\pm}15.4%$) respectively at large, middle and small follicles and total number of these 3 grade follicles were appeared to be $182.6{\pm}32.7$. The above findings reveal that large follicles were increased 29.8% in FSH-treated group and 73.7% in PMS and HCG-treated group than those in control group and in histologic findings, proportion of atretic follicles were more increased in ovaries with more number of more developing follicles.

• Korean Journal of Ichthyology
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• v.6 no.2
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• pp.237-243
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• 1994

This work was conducted to study sex differentiation in the black sea bream, Acanthopagrus schlegeli (Bleeker), using a histological method for the appearance of primordial germ cell, formation of primitive gonads, differentiation of female and male from newly hatched larva to the ovotestis stage of fish. The 3~4 primordial germ cells of $6.8{\sim}7.2\;{\mu}m$ in size, which were buried under fibrous mesenchymal tissue between gut duct and notochord of pre-larva with a total length (T.L.) of 2.4 mm at 3 days after hatching. The proto-gonial cells were located in the epithelium of the coelom attached with pigment cells of juvenile with 6.4 mm in T.L. at 21 days after hatching. In juvenile of 20.8 mm in T.L. at 59 days after hatching, the proto-gonial cells were migrated to the retro-peritoneum through the lineshaped primitive gonad composed of fibrous mesenchymal tissue. In juvenile of 7.8 em in T.L. at 186 days after hatching, the mitotic division of proto-gonial cell appeared in the lineshaped primitive gonad having many eosinophilic granule cells and abundant fibrous connective tissue. In juvenile of 9.5 em in T.L. at 254 days after hatching, the gonad was occupied by abundant fibrous connective tissue, bundles of spermatocyte and spermatid. In juvenile of 10.5 cm in T.L. at 13 months after hatching, the gonad was divided into cortical layer and medullary layer. The former was composed of bundles of a few spermatocytes and proto-gonial cells, the latter was filled with the fibrous mesenchymal tissue and a few proto-gonial cells. In juvenile of 14.7 em in T.L. at 16 months after hatching, the gonad was separated into ovarian part and testicular part by the fibrous connective tissue. The ovarian part is consisted of ovarian cavity and oocytes of perinucleolus stage. The testicular part was occupied by spermatogonia in the cyst.

• Korean Journal of Animal Reproduction
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• v.21 no.3
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• pp.255-264
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• 1997

The purpose of this study was attempted to investigate the number changes of the growing and mature follicles in ovary following repeats of pregnant mare serum gonadotropin(PMSG) treatments for superovulation in nulliparous rats. Thirty two rats(Sprague-Dawely, about 200~250 gm) were randomized into 4 groups. Control group rats were sacrified at estrus phase confirmed by vaginal smear. PMSG-treated group 1 rats, PMSG-treated group 2 rats and PMSG-treated group 3 rats were sacrified at 48 hrs after injection once with PMSG 25 IU, after 2 repeated injection by a week interval, and 3 repeated injection, respectively. The uteri and ovaries of rats were removed and weighed and then were observed grossly and serial sections of all ovaries and some sections of uteri by paraffin embedding were stained with H-E. Number of ovarian follicles about 3 grades of small, middle and large follicles from seondary and follicles were investigated by LM photographies of ovary preparations. The criteria of the small, middle, and large follicles were based as small follicle with preantral follicles with 2~4 layers of granulosa cells surrounding the oocyte, as secondary follicles with more than 5 layers of granulosa cells and early signs of antral cavity or with small clefts on either side of the oocytes, and as tirtiary follicles with a single medium sized antral cavity or large well-formed antral cavity, respectively. In gross findings, the wall of the uteri in control group were thin, and those in 3 PMS-treated group were markedly thickened and some uterine lumen of those filled with fluid. In histological findings, the walls of the uteri from 3 PMSG-treated groups were hypertrophied and their blood and lymph vessels were dilated than those of control group. The ovaries fo 3 PMSG-treated groups were more increased in size and the cortexes were more developed and increased in width but there are no difference of development and changes in 3 PMSG-treated groups. The weight of the uteri and ovaries per rat in PMSG -treated group 1, 2 and 3 were a, pp.ared to be significantly increased 171.4$\pm$47.6%, 162.3$\pm$43.9%, 206.9$\pm$30.4%, respectively than those of control groups. The mean number of follicle per ovary in control group were a, pp.ared to be 17.1$\pm$3.5, 46.2$\pm$14.5, and 74.3$\pm$22.7 at large, middle and small follicles, respectively and total number of these 3 grade follicles per ovary were a, pp.ared to be 137.7$\pm$31.7. The mean number of follicle per ovary in PMSG-treated group 1 were a, pp.ared to be 25.6$\pm$7.3, 78.1$\pm$29.9, and 83.2$\pm$34.0, at large, middle and small follicles, respectively and total number of these 3 grade follicles were a, pp.ared to be 187.5$\pm$58.8. The mean number of follicle per ovary in PMS-treated group 2 were a, pp.ared to be 21.9$\pm$5.2, 67.8$\pm$16.8, and 68.0$\pm$14.9 at large, middle and small follicles, respectively and total number of these 3 grade follicles were a, pp.ared to be 157.7$\pm$26.2. The mean number of follicle per ovary in PMS-treated group 3 were a, pp.ared to be 21.7$\pm$4.8, 61.5$\pm$17.0, and 59.7$\pm$16.2 at large, middle and small follicles, respectively and total number of these 3 grade follicles were a, pp.ared to be 143.5$\pm$29.6. The number of follicles in PMSG-treated group 1 a, pp.ared to be more number than other 2 PMSG-treated gruops and tended to be decreased by frequency of PMSG-treatment.

• The Korean Journal of Malacology
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• v.17 no.1
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• pp.45-55
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• 2001

Gametogenes, reproductive cycle, first sexual maturity(biological minimum size), sex ratio and larval development of the marsh clam Corbicula japonica were investigated monthly by histological observations. Samples were collected in brackish water of Gokgang stream, Kyungsangbuk-Do, Korea, from August 1997 to July 1998. Sexuality of Corbicula japonica is dioecious and the species are an oviparous clam. The gonads are irregularly arranged from the sub-region of mid-intestinal gland in visceral cavity to reticular connective tissue of foot. The ovary is composed of a number of ovarian sac which are branched arborescent. Oogonia actively proliferate along the germinal epithelium of ovarian sac, in which young oocytes are growing. The testis is composed of a number of testicular tubules, and the epithelium of the tubule has function of germinal epithelium, along which spermatogonia actively proliferate. A great number of undifferentiated mesenchymal tissue and eosinophilic granular cells are abundantly distributed between developing oocytes and spermatocytes in the early developmental stages. With the further development of the ovary and testis these tissue and cells gradually disappear. Then the undifferentiated mesenchymal tissue and eosinophilic granular cells are considered to be related to the growing of the oocytes and spermatocytes. The spawning period is from July to September, and the main spawning occur between July and August when seawater temperatures reach above 22$^{\circ}C$. The reproductive cycle of this species can be divided into five successive stages; early active (February to April), late active (May to July), ripe (June to September), partially spawned (July to September), degenerative (September to October) and resting stage (October to February). Percentages of first sexual maturity of female and male clams ranging in length from 10 mm to 12 mm are over 50% and 100% for clams over 16.0 mm in shell length. Fertilized eggs or Corbicula japonica were 80-90 ${\mu}{\textrm}{m}$ in diameter. In the early embryonic development of C. japonica, the appearance of polar body, trochophore and D-shaped veliger were observed around 40 min., 27 hours and 4 days after spawning, respectively, at a water temperature of 26.5-28.$0^{\circ}C$. The size of larvae of early umbo stage was about 185-210 ${\mu}{\textrm}{m}$ in shell length, 160-180 ${\mu}{\textrm}{m}$ in shell height around 7 days after fertilization. The correlation of relative growth between the culture day (D) and shell length (SL) was expressed by the following simple formula from D-shaped veliger to metamorphosing stage; SL = 13.300D + 209.36($r^2$= 0.9078).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.1
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• pp.35-43
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• 1996

The appearance of the primordial germ cells (PGC's), early gonadogenesis, sex differentiation and sex ratio of the embryo in the viviparous teleost, Ditrema temmincki were investigated by using photomicroscopy. The PGC's were first observed in the fibrous mesenchymal tissue located between the early alimentary tract and the dorsal body wall in the late embryonic development stage. During the period from the hatching to the individual total length (TL) of 4.0 mm the PGC's were evenly distributed in the fibrous mesenchymal tissue between alimentary tract and body wall. But the period of TL 5.0 mm mesenchymal tissue separated from the dorsal body wall, the PGC's moved to the posterior mesenchymal tissue and formed the primitive gonad. During the early gonadogenesis, differentiation of the testis and ovary were distinguished from the arrangement of the germ cells and somatic cells. Gonad of embryo in TL 10.0 mm can be separated into the ovary and testis by external morphology. The testis had a separated form which was consisted with two lobes, and the ovary had a fused form in half-posterior part. In the testicular differentiation of the embryo, histological pattern of the seminiferous tubule appeared when TL of the embryo was to be 25.0 mm, for the seminal vesicle was formed In the individual TL of 30.0mm. The testis of the embryo with TL of 45.0 mm was similar to that of the adult fish in the external and internal structures. In the ovarian differentiation, formation of the ovigerous folds and the ovarian cavity were clearly observed when the TL reached to 30.0 mm. The ovary from the individual with TL of 60.0 mm was differentiated into a similar ovary as seen in the adult fish in the external and internal structure. Right before parturition the total length of the individual was approximately 63.0 mm of which the individual embryo has an ovary containing the oocytes in the chromatin nucleolus stage, or a testis containing the spermatogonia, respectively. And the embryonic sex ratio of female to male was 1.65 : 1. Ditrema temmincki is dioecism and the pattern of sex differentiation is belonged to a differentiation type.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.5
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• pp.413-431
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• 1992

Gonadal development, fertilization and egg development in the maternal body and reproductive cycle of ovoviviparous rockfish, Sebastes inermis, were investigated histologically. Gonadosomatic index(GSI) of male and female were increased from September and reached maximum values in December. In the male, GSI decreased from January, but in the female maintained high values till February and decreased from March. Hepatosomatic index(HSI) was related to GSI conversely. In both sex, HSI increased from February and reached maximum in August as the gonad were degenerating and resting, and began to decrease from September as gonad were glowing. This ovoviviparous rockfish copulates in December. Fertilization with sperms maintained between ovulated oocytes in the ovary occurs in January mainly. Egg development in the ovarian cavity and discharging of hatched preiarva occurs from January to February. The reproductive cycle includes the successive stages: Growing(September), Mature (October-November), Ripe and Fertilization(Decembr-Janua), Egg development and Discharging of hatched larva(January-February), Degeneration and Resting(February-August). According to the frequency distribution of egg diameter and histological observation, the ovoviviparous rockfish discharged the prelarva at a time in a spawning season. The sexual maturation is first attained at 2 ages. All females and males reaches first maturity at body length of 17.1cm and 15.1cm respectively. The mean number of the embryos increased with the increase of the total length of female.

• Journal of Aquaculture
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• v.10 no.2
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• pp.179-187
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• 1997

Early development growth and sexual phenomena of the hybrid between male spotted flounder (Verasper variegatus) and female olive flounder (Paralichthys olivaceus) were investigated. The hybrid produced was newly called the "Bumnupchi" in Korean name, "Mottled flounder" in English name. Fertilization rate, hatching rate and early survival rate of the mottled flounder were 56.9~72.3%, 86.7~92.5% and 89.1~96.0%, respectively. At 50th day and 240th day after hatching, they were 2.1$\pm0.2cm\;and\;22.9\pm2.1cm\;in\;total\;length, \;0.10\pm0.002g\;and\;179.0\pm38.5g$ in body weight, respectively. Eye position of the mottled flounder was mixed with both dextral (77.7%) and sinistral type (22.3%). Gonads were differentiated into ovarian type and testis type at 80th day after hatching, and its ratio was 1:1 (P>0.05). Most Gonads seemed to be sterile without having germ cell, whereas only 3.6% of investigated indiciduals turned out to have oocytes in the gonad. When the induction of sex reversal were attempted by daily oral administration of $17\beta$-methyltestosterone (0.5mg/kg fish) andestradiol-17$17\beta$(0.5mg/kg fish)from 40th to 100th day after hatching, both testis and ovary type were induced 81.9% and 87.7% of each hormone-treated fish, respectively.fish, respectively.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.5
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• pp.373-377
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• 2000

This study Investigated the effects of $estradiol-l7{\beta} (E_2)$, $17{\alpha}-methyltestosterone$ (MT) and high temperature (WT) on gonadal sex differentiation in black rockfish, Sebastes srhlegeli. fish were reared to oral adminstration of E, at nominal concentrations of $20, 40 and 60 {\mu}g/g diet$, and MT at nominal concentration of 20 and $50 {\mu}g/g$ diet from 56 days to 77 days after parturition. In the treatment of WT, water temperature of the breeding tanks ranged $27.0{\pm}0.5^{\circ}C$ more than $10^{\circ}C$ approximately, in comparison to the control and other experimental group, In the process of sex differentiation until 56 days after parturition, gonads were composed of mostly gonia cells, sexually undifferentiated. In contrast, 128 dal's after parturition, the ovaries were composed of ovarian cavity and lamellae, and oogonia and perinucleolus oocytes were distributed in the lamellae of ovary, and the testes were composed of a number of seminiferous tubule, and spermatogonia were distributed in the seminiferous tubule, and also melanophore scattered in the matrix layer of testis. In the sex ratio, more females than male were observed from $E_2$. treatment groups when compared to the control, but more males than females were observed from MT and WT treatment groups when compared to the control. In the results of the present study, the concentration and kinds of the sex steroid hormones, and also the rearing high temperature caused to the factor of sex determination in the process of sex differentiation of black rockfish.

• Korean Journal of Ichthyology
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• v.19 no.2
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• pp.107-111
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• 2007

Sex differentiation of the brown croaker Miichthys miiuy (Basilewsky) is described from hatching to the 120th day post-hatching (dph) (water temperature $24^{\circ}C$). Primordial germ cells (PGCs) were observed on the 20th dph (10.4 mm total length (TL), 0.14 g body weight (BW), and began to protrude into the peritoneal cavity from the 40th dph (19.4 mm TL, 0.39 g BW). On the 65th dph (31.3 mm TL, 0.93 g BW, $1,560D^{\circ}$ (degree-days)), initial ovarian differentiation was identified by the PGCs with condensed chromatin, and their transformation into meiotic oocytes. By the 120th dph (4.60 mm TL, 1.38 g BW, $2,880D^{\circ}$), the oocytes were in the perinucleolus stage and had increased from 20 to $40{\mu}m$ in diameter. While ovaries gradually grew after sex was differentiated, testes continued to multiply from the 65th dph. On the 80th dph (37.9 mm TL, 1.39 g BW, $1,920D^{\circ}$), the beginning of testis lobule formation was indicated by the occurrence of spermatogonial cysts enveloped by somatic cells in some of the testes. On the 120th dph, the testis lobules of some of the fish contained all germ cell stages through to the spermatocytes. Therefore, the sex differentiation type of the brown croaker is identified as gonochoristic.