This study was carried out to investigate the effects of Ca, BSA, heparin, semen storage and individual bull on motility and acrosome reaction of bovine fresh sperm and sperm stored in lactose-egg yolk solution(LES) at 5$^{\circ}C$ for 4hours, and the results obtained were as follows: 1. When sperm was incubated in SCS containing Ca, BSA, Ca + BSA, heparin, heparin + Ca, heparin + BSA, and heparin + Ca + BSA for 15 minutes, there was significant difference in sperm motility among the treatments, especially BSA showed significantly higher sperm motility than the others. Also there was significant difference in sperm acrosome reaction among the treatments, especially BSA and Ca + BSA showed significantly higher sperm acrosome reaction than the others. 2. Bull KNC 1 showed significantly higher sperm motility than KNC 1, HOL 1 and 2 in both fresh and stored semen, however KNC 1 showed significantly lower sperm acrosome reaction than KNC 1, HOL 1 and 2. Therefore, there was significant difference in sperm motility and acrosome reaction among individual bulls. 3. When KNC 1 and KNC 2 sperm were incubated in SCS and SCS + Ca, SCS + BSA, SCS + Ca + BSA, SCS + heparin, SCS + heparin + Ca, SCS + heparin + BSA, and SCS + heparin + Ca + BSA, there was significant difference in sperm motility among individual bulls, especially BSA in KNC 1 and BSA, Ca and Ca + BSA in KNC 2 showedsignificantly higher motility than the others. However, there was significant difference in sperm acrosome reaction among individual bulls, Ca in KNC 1 and Ca + BSA in KNC 2 showed higher acrosome reaction than the others.
During 2005~2009, current status on the occurrence and the management of the major disease, insect and mite pests were investigated in the non-chemical or organic cultured apple orchards in Korea. Numbers of certified organic or non-chemical apple orchards increased from 14 in 2005 to 78 in 2008. Severe damages on leaves and fruits were caused by the several diseases such as marssonina blotch, bitter rot, white rot, sooty blotch and flyspeck, and the several insect pests such as apple leaf-curling aphid, woolly apple aphid, oriental fruit moth and peach fruit moth on the almost certified organic or non-chemical pest control orchards. About 10 and 18 environmental-friendly materials were used to control diseases and insect or mite pests, respectively. But, lime sulfur and bordeaux mixture to diseases and machine oil, plant oil mixed with egg yolk, and pheromone mating disruptions to insect pests were effective under the adequate conditions.
Antioxidative activities of ethanol extracts of seed, branch, leaves, and aerial part of Crotalaria sessiflora L. were compared using in vitro experimental model. Solid contents of each extracts were 12.68, 16.28, 13.04, and 18.59 g/ 100 mL, and phenolic acid contents were $0.082{\pm}0.003,\;0.099{\pm}0.010,\;0.071{\pm}0.002,\;and\;0.094{\pm}0.011\;mg/mL$ in branch, aerial part, seed, and leaves, respectively. Extracts prepared from leaves showed highest electron-donating ability toward DPPH. SOD-liked activities were 78.95, 70.85, 74.65, and 87.49% in aerial part, branch, seed, an leaves respectively. Extracts prepared from leaves showed 59.39% inhibitory effect on peroxidation of egg yolk lecithin.
This study was carried out to investigate the effects of liquid boar sperm stored at 4$^{\circ}C$ on sperm motility, normal acrosome, and in-vitro fertilization and culture of pig oocytes matured in-vitro. The sperm-rich fraction (30~60 ml) of ejaculate was collected into an insulated vacuum bottle. Semen was slowly cooled to room temperature (20~23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min at 800$\times$g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of lactose, egg yolk and N-acetyl-D-glucosamine (LEN) diluent to provide 1.0$\times$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$ and preserved for 5 days to examine sperm motility and normal acrosome. The medium used for oocyte maturation was modified tissue culture medium (TCM) 199. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 6 h in 500 ${mu}ell$ mTBM fertilization media with 0.2, 1, 5 and 10$\times$10$^{6}$ /ml sperm concentration, respectively. At 6 h after IVF, oocytes were transferred into 500 ${mu}ell$ Hepes-buffered NCSU-23 culture medium for further culture of 6, 48 and 144 h. There were significant differences in sperm motility and normal acrosome among preservation days and incubation times, respectively. The rates of sperm penetration and polyspermy were higher in 5 and 10$\times$10$^{6}$ sperm/ml than in 0.2 and 1$\times$10$^{6}$ sperm/ml. Male pronuclear formation was lower in 0.2$\times$10$^{6}$ sperm/ml than in 1, 5 and 10$\times$10$^{6}$ sperm/ml. Mean numbers of sperm in penetrated oocyte were highest in 10$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. The rate of blastocysts from the cleaved oocytes (2~4 cell stage) was highest in 1$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend 1$\times$10$^{6}$ ml sperm concentration for in-vitro fertilization of pig oocytes.
Tran Thi Huyen;Ha Phuong Trang;Nguyen Thi-Ngan;Bui Dinh-Thanh;Le Pham Tan Quoc;Trinh Ngoc Nam
Fisheries and Aquatic Sciences
/
v.26
no.3
/
pp.204-215
/
2023
The thermolabile haemolysin (tlh) of Vibrio parahaemolyticus (Vptlh) from V. parahaemolyticus is a multiple-function enzyme, initially describes as a haemolytic factor activated by lecithin and phospholipase A2 enzymatic activity (Shinoda, 1991; Vazquez-Morado, 2021; Yanagase et al., 1970). Until now, the tlh structure has hypothesized including N-terminal and C-terminal domain, but what domain of the Vptlh structure does the haemolytic activity has not been refined yet. In this study, a 450-bp VpTLH nucleotide sequence of the entire Vptlh gene encoded the C-terminal domain cloned firstly to examine its responsibility in the activity of the Vptlh. The C-terminal domain fused with a 6-His-tag named the His-tag-VpC-terminal domain was expressed successfully in soluble form in the BL21 (DE3) PlysS cell. Remarkably, both expression and purification results confirmed a high agreement in the molecular weight of the His-tag-VpC-terminal domain was 47 kDa. This work showed the His-tag-VpC-terminal domain lysed the erythrocyte membranes in the blood agar and the phosphate buffered saline (0.9%) media without adding the lecithin substrate of the phospholipase enzyme. Haemolysis occurred at all tested diluted concentrations of His-tag-VpC-terminal domain (p < 0.05), providing evidence for the independent haemolytic activity of the His-tag-VpC-terminal domain. The content of 100 ㎍ of the His-tag-VpC-terminal domain brought the highest haemolytic activity of 80% compared to that in the three remaining contents. Significantly, the His-tag-VpC-terminal domain demonstrated not to involve the phospholipase activity in Luria-Bertani agar supplemented with 1% (vol/vol) egg yolk emulsion. All results proved the vital responsibility of the His-tag-VpC-terminal domain in causing the haemolytic activity without the required activation by the phospholipase enzyme. Raw extracts of Phellinus igniarus and Phellinus pipi at 10-1 mg/mL inhibited the haemolytic activity of the His-tag-VpC-terminal domain from 67.7% to 87.42%, respectively. Hence applying the His-tag-VpC-terminal domain as a simple biological material to evaluate quickly potential derivatives against the Vptlh in vivo conditions will accessible and more advantageous than using the whole of the Vptlh.
Laura Cogoi;Carla Marrassini;Elina Malen Saint Martin;Maria Rosario Alonso;Rosana Filip;Claudia Anesini
Journal of Pharmacopuncture
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v.26
no.4
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pp.338-347
/
2023
Objectives: Ilex paraguariensis (Aquifoleaceae) is cultivated to produce "yerba mate". Due to its nutritional, energizing, hypoglycemic and antioxidant effects, it is used in the elaboration of food, pharmaceuticals, and cosmetics. The oxidative stress related to protein glycation and production of advanced glycation end products (AGEs) leads to the development of several diseases. The objective of this work was to compare the antioxidant and anti-AGEs activity of a decoction of fruits (F) and leaves (L). Methods: The antioxidant activity was assayed by the DPPH assay and the inhibition of egg yolk lipid peroxidation (ILP), and anti-AGEs activity, through the inhibition of the formation of fructosamine (IF), β-amyloid (Iβ), protein carbonylation (IC) and AGEs (IA). Polyphenols were quantified by HPLC. Results: Maximum response ± SEM: For F 0.01 ㎍/mL: IF = 42 ± 4%, IC = 17 ± 2% and for 10 ㎍/mL: IA = 38 ± 4%, Iβ = 67 ± 7%. For L 0.1 ㎍/mL: IF = 35 ± 2%, IC = 19 ± 2% and for 100 ㎍/mL: IA = 26 ± 3%, Iβ = 63.04 ± 2%. The DPPH IC50 = 134.8 ± 14 ㎍/mL for F and 34.67 ± 3 ㎍/mL for L. The ILP IC50 = 512.86 ± 50 ㎍/mL for F and 154.8 ± 15 ㎍/mL for L. By HPLC L presented the highest amounts of flavonoids and caffeoylquinic acids. F and L showed strong anti-AGEs activity, affecting the early stages of glycation at low concentrations and the late stages of glycation at high concentrations. The highest activity for both F and L was seen in the IF and Iβ. F presented the highest anti-AGEs potency. L presented the highest antioxidant potency, which was related to the highest content of polyphenols. Conclusion: The fruits of I. paraguariensis could be a source of antioxidant and anti-AGEs compounds to be used with medicinal purposes or as functional food.
Chung-Kug Park;Seon-Yeong Hwang;Dae-Hong Kim;Seung-Jun Heo;Jae-Min Park
Korean Journal of Ichthyology
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v.36
no.1
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pp.1-9
/
2024
This study investigated the early life history of the Boleophthalmus pectinirostris living in the southern coastal Yeoja-man and compared the results with the same Gobiidae fishes. The brood stork used in the study were captured with bare hands in the tidal flats of Beolgyo-eup, Jeollanam-do, in June 2015. The amount of spawning was 411~11,688, and the eggs were short oval and the size was 1.40×0.72 mm. The time of hatched took 91 hours and 35 minutes at a water temperature of 25~27℃. Newly hatching larvae, the yolk sac had a total length of 3.02~3.31 (average 3.17±0.08, n=30) mm and did not eat rotifer. 4 days after hatching, the total length was 3.31~3.52 (3.43±0.07, n=30) mm, and as the mouth and anus opened, the fish transitioned to the preflexion larvae and fed. 14 days after hatching, the total length was 5.06~5.25 (5.16±0.06, n=30) mm, and the distal end of the vertebra was completely bent at 45° and the transitioned to the postflexion larvae. 41 days after hatching, the total length was 14.3~16.8 (15.4±0.85 mm, n=30), and the number of fins reached an integer of 5 first dorsal fins, 26~27 second dorsal fins, 24~27 anal fins, and 6 ventral fins, and the transitioned to the juveniles. As a result of the study, star-shaped melanophore were deposited from the front of the pectoral fin to the base of the caudal fin, which distinguished them in form from other postflexion larvae of Gobiidae fishes.
Jeong, Yon Na;Park, Su-Jin;Lee, Sang Hoon;Choi, Youngmin;Choi, Kap Seong;Chun, Jiyeon
Korean Journal of Food Science and Technology
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v.52
no.4
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pp.317-324
/
2020
In order to create the national food nutrient database, a total of 41 animal foods (ham, seafood, edible insects and eggs) were analyzed for their vitamin B12 content and the applied immunoaffinity-HPLC was verified. Ham vitamin B12 contents were 0.30-0.65 ㎍/100 g. Seafood showed relatively high vitamin B12 level, where the values of fermented clam were the highest (26.80 ㎍/100 g) followed by fermented pollack roe. Vitamin B12 was not detected in silkworm pupae and beetles, while relatively high levels were found in the two-spotted cricket imago (6.70 ㎍/100 g). Chicken and quail egg yolk had roughly 100- and 30-times higher vitamin B12 levels as compared to their egg white. Vitamin B12 contents in quail and chicken eggs were significantly enhanced by boiling (p<0.05). Results based on accuracy (97-102% recovery) and precision (<5% RSD) indicate that this study provides reliable vitamin B12 information on animal foods consumed in Korea.
Annual reproductive cycle of the Damselfish, Chromis notatus collected monthly at the four coastal areas around Chejudo, Korea are studied on the bases of histological observations of gonadal tissue and various quantitative variables including gonadosomatic index (GSI), fatness, egg diameter composition and the first maturity. The ovary consisted of a pair of saccular structure with many ovarian sacs. Oogonia proliferated along the germinal epithelium of the ovarian sac. Young oocytes with basophilic cytoplasm showed several nucleoli along the nuclear membrane. When the oocytes reached about $450{\mu}m$ in diameter, nucleus migrate toward the animal pole, nuclear membrane disappeared and most of cytoplasm were filled with yolk materials and oil drops. After ovulation, residual follicle and growing oocytes remaining in the ovarian sacs degenerated. But early young oocytes without follicle layer were not degenerated, and growing continuously till the next year. The testis consisted of a pair of lobular structures in the right and left were united in the posterior seminal vesicle. Cortex of testis was composed of many sperm ducts connected with lobuli. GSI began to increase from March, starting season of longer day length and higher water temperature, and reached the maximum value between June and August. It began to decrease from September with the lowest value appearing between October and February without any evident variation. The annual reproductive cycle could be devided into five successive stage : growing(April to Many), mature(May to August), ripe and spent (June to August) and recovery and resting stage(September to March). The spawning peak occurred from June to August. According to the frequency distribution of egg diameter, Chromis notatus was a polycyclic species to spawn twice or more in a spawning season. Fatness, correlated with gonadal phases, was remarkably decreased by spawning. Percentage of the first maturity . in femate and male fish ranging from 7.0 to 7.9 cm were $50\%$ and from 9.0 to 9.9 cm in total length $100\%$.
To determine the actual hygienic status of domestic chicken meats sold in public markets (conventional markets and department stores), microbial contamination levels (Total cells, Coliforms and Staphylococcal cells) and zoonotic pathogens (Salmonella species, Campylobacter species, Listeria species, and Staphylococcus aureus) isolation tests were conducted. Chicken meats and eggs tested were collected from the conventional markets (Si-Jang) and department-stores located in Seoul and Kyung-gi regions in 1996. In total cells and coliforms contamination tests, chicken meats sold in department stores were much lesser contamination status than those of Si-Jang, but staphylococcal cells level was much more higher than that of conventional markets. Salmonella isolation frequency was investigated as $68.8\%$, but Campylobacter jejuni and Listeria monocytogenes isolation frequency were appeared both $64.0\%\;and\;63.3\%$. In case of eggs sold in public markets, one of S. gallinarum strain $(0.7\%)$ was isolated only on the egg-shell part among the four-hundred and fourty-six. In comparison with foreign imported chicken meats, there were no big differences in microbial contamination status. On the other hand, both Salmonella and L. monocytogenes were isolated only in the chicken wings from Korea and China, but not from U.S.A. This data suggest that more hygienic control system in order to produce the safe and hygienic chicken meats and eggs is need in our country as soon as possible.
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