• Journal of Food Hygiene and Safety
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• v.27 no.1
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• pp.42-49
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• 2012

This study was carried out to investigate the toxicity of food Red No.2 in the Sprague-Dawley (SD) female rat for 4 weeks. SD rats were orally administered for 28 days, with dosage of 500, 1,000, 2,000 mg/kg/day. Animals treated with food Red No.2 did not cause any death and show any clinical signs. They did not show any significant changes of body weight, feed uptake and water consumption. There were not significantly different from the control group in urinalysis, hematological, serum biochemical value and histopathological examination. In conclusion, 4 weeks of the repetitive oral medication of food Red No.2 has resulted no alteration of toxicity according to the test materials in the group of female rats with injection of 2,000 mg/kg. Therefore, food Red No.2 was not indicated to have any toxic effect in the SD rats, when it was orally administered below the dosage 2,000 mg/kg/day for 4 weeks.

• Journal of Dairy Science and Biotechnology
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• v.34 no.2
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• pp.99-116
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• 2016

We herein performed animal safety assessment in accordance with Good Laboratory Practice (GLP) regulations with the aim of developing sialic acid from glycomacropeptide (hereafter referred to as "GMP") as an index ingredient and functional component in functional foods. GMP is a type of whey protein derived from milk and a safe food, with multiple functions, such as antiviral activity. A test substance was produced containing 7% (w/w) sialic acid and mostly-hydrolyzed whey protein (hereafter referred to as "7%-GNANA") by enzymatic treatment of substrate GMP. The maximum intake test dose level was selected based on 5,000 mg/kg/day dose set for male NOEL (no-observed-effect-level) and female NOAEL (no-observed-adverse-effect-level) determined by a dose-range finding (DRF) test (GLP Center of Catholic University of Daegu, Report No. 15-NREO-001) that was previously conducted with the same test substance. To evaluate the toxicity of a repeated oral dose of the test substance in connection with the previous DRF study, 1,250, 2,500, and 5,000 mg/kg of the substance were administered by a probe into the stomachs of 6-week-old SPF Sprague-Dawley male and female rats for 90 d. Each test group consisted of 10 male and 10 female rats. To determine the toxicity index, all parameters, such as observation of common signs; measurements of body weight and food consumption; ophthalmic examination; urinalysis, electrolyte, hematological, and serum biochemical examination; measurement of organ weights during autopsy; and visual and histopathological examinations were conducted according to GLP standards. After evaluating the results based on the test toxicity assessment criteria, it was determined that NOAEL of the test substance, 7%-GNANA, was 5,000 mg/kg/day, for both male and female rats. No animal death was noted in any of the test groups, including the control group, during the study period, and there was no significant difference associated with test substance, as compared with the control group, with respect to general symptoms, body weight changes, food consumption, ophthalmic examination, urinalysis, hematological and serum biochemical examination, and electrolyte and blood coagulation tests during the administration period (P<0.05). As assessed by the effects of the test substance on organ weights, food consumption, autopsy, and histopathological safety, change in kidney weight as an indicator of male NOAEL revealed up to 20% kidney weight increase in the high-dose group (5,000 mg/kg/day) compared with the change in the control group. However, it was concluded that this effect of the test substance was minor. In the case of female rats, reduction of food consumption, increase of kidney weight, and decrease of thymus weight were observed in the high-dose group. The kidney weight increased by 10.2% (left) and 8.9% (right) in the high-dose group, with a slight dose-dependency compared with that of the control group. It was observed that the thymus weight decreased by 25.3% in the high-dose group, but it was a minor test substance-associated effect. During the autopsy, botryoid tumor was detected on the ribs of one subject in the high-dose group, but we concluded that the tumor has been caused by a naturally occurring (non-test) substance. Histopathological examination revealed lesions on the kidney, liver, spleen, and other organs in the low-dose test group. Since these lesions were considered a separate phenomenon, or naturally occurring and associated with aging, it was checked whether any target organ showed clear symptoms caused by the test substance. In conclusion, different concentrations of the test substance were fed to rats and, consequently, it was verified that only a minor effect was associated with the test substance in the high-dose (5,000 mg/kg/day) group of both male and female rats, without any other significant effects associated with the test substance. Therefore, it was concluded that NOAEL of 7%-GNANA (product name: Helicobactrol) with male and female rats as test animals was 5,000 mg/kg/day, and it thus was determined that the substance is safe for the ultimate use as an ingredient of health functional foods.

• Korean Journal of Veterinary Service
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• v.25 no.1
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• pp.53-73
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• 2002

It has been reported that degranulation of mast cells in rats, rabbits and dog was observed after dosing xylazine hydrochloride(Xh) which has been widely used as sedative, analgesic and muscular relaxant. Therefore, this experiment was conducted to examine the relations between Xh and histamine release and to identify the action of ${\alpha}$-adrenoceptors which exists on the suface of mast cells. 1. The content of histamine within serum was measured with HPLC by performing the O-phthalaldehyde(OPA) fluorescent derivation. The pretreatment method had a little modification from the conventional method. The pretreament was carried out in the following method. 0.2$m\ell$ of serum and 1$m\ell$ of butanol were added to mixed together and then the liquid was centrifugally separated at 4$^{\circ}C$ and 2,000 rpm for 3 minutes. 0.4$m\ell$ of 0.1N HCl and 1.6$m\ell$ of heptane were added to 0.8$m\ell$ of supernatant taken from the liquid, and they were mixed together. This mixture was also centrifugally separated at 4$^{\circ}C$ and 2,000 rpm for 5 minutes. The supernatant was thrown away and the OPA fluorescent derivation was carried out with 0.2$m\ell$ of the lower liquid then, 5 minutes after mixing 400${\mu}\ell$ of 0.1N HCl, 120${\mu}\ell$ of 1N NaOH and 40${\mu}\ell$ of 0.1% OPA in the 0.2$m\ell$ of the lower liquid,120${\mu}\ell$ of 3.57N H$_3$PO$_4$ was added to the mixed liquid, and the liquid, was mixed again and syringe-filtered. Then, the measurement was done with HPLC in the 30 : 70(ν/ν) ratio of 0.004M KH$_2$PO$_4$: CH$_3$CN, flow rate of 1.0$m\ell$/min., and a wavelength of λex= 350nm and λem=444nm at the column temperature of 27$^{\circ}C$, using the fluorescence detector. 2. The content of histamine in each laboratory animal appeared to be higher in such an order as rabbit, rat, guinea pig, dog, Korean indigenous goat, swine, Korean indigenous cattle, Holstein, and mouse, of which the individual mean values${\pm}$standard deviation were 2.0668 ${\pm}$ 0.6049. 0.4999 ${\pm}$ 0.2278, 0.4241 ${\pm}$ 0.1974, 0.1054 ${\pm}$ 0.0556, 0.1028 ${\pm}$ 0.0276, 0.0972 ${\pm}$ 0.0513, 0.0872 ${\pm}$ 0.0373, 0.0717 ${\pm}$ 0.0379, and 0.0706 ${\pm}$ 0.0366, respectively. 3. The content of histamine was measured at the moments of 15-, 30-, 60-, 120-minutes after inoamuscular injection of 20mg/100kg Xh into two to 4 years old Holstein weighing 600∼700kg. The result showed that there was a significant increase at the times of 30- and 90-minutes after injection(p<0.05). 4. Intramuscular injection of 3mg/10kg Xh was given to crossbred pug dogs weighing 2.5∼4.3kg. The content of histamine was measured at the times of 30-, 60-, 90- and 120-minutes after injection. The result revealed that there was a significant increase at the times of 60-and 90-minutes after injection(p<0.05). 5. Intramuscular injection of 10mg/$m\ell$∼25mg/$m\ell$ Xh in concentration of 0.1$m\ell$ was applied to Korean indigenous goat over 5 months old. Then, the content of histamine was measured at the times of 15-, 30-, 60- and 90-minutes after injection. A significant increase was shown at the times of 30- and 60-minutes after injection(p<0.05). 6. The content of histamine was measured at the moments of 30- and 60-minutes after intramuscular injection of 0.1-0.2$m\ell$ Xh (20mg/$m\ell$) into male rabbits weighting 2.5-4kg. A significant increase was found at the moment of 60 minutes after injection(p<0.001). 7. After administering Xh to the mast cell taken from the abdominal cavity of mouse, the content of histamine was measured. The result showed that the higher the concentration, the more significantly the content of histamine was increased(p<0.05). 8. Compound 48/80 was administered in concentration of 5$\mu\textrm{g}$/$m\ell$ and 10$\mu\textrm{g}$/$m\ell$ to the mast cell picked from the abdominal cavity of mouse. The result showed that there was a significant increase in the content of histamine in case of the concentration of 10$\mu\textrm{g}$/$m\ell$(p<0.05). It was found to be about 10,000 to 500,000 times stronger than the Xh. 9. After premedication of 1mg/kg of yohimbine hydrochloride as ${\alpha}$$_2$-adrenergic antagonist to rabbits, the Xh was administered to them. The result was that the value of histamine within serum was decreased significantly(p<0.001). 10. After premeditation of 1mg/kg of prazosin hydrochloride as ${\alpha}$$_1$-adrenergic antagonist to rabbits, the Xh was administered to them. It was found that the value of histamine within serum was decreased significantly(p<0.005). 11, Prazosin hydrochloride and yohimbine hydrochloride as ${\alpha}$$_1$-adrenergic antagonist, respectively, and ${\alpha}$$_2$-adrenergic antagonist were administerd. In this case, the value of histamine within serum was decreased significantly(p<0.0001). As the results, when the Xh is administered to various kinds of animals, the amount of histamine release within serum is increased. In view of the results so far achieved, it is concluded that Xh acted on both a$_1$-adrenoreceptor and ${\alpha}$$_2$-adrenoreceptor induces the degranulation of mast cell.

• Journal of Aquaculture
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• v.5 no.1
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• pp.29-67
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• 1992

Life cycle and seed production of the freshwater prawn, Macrobrachium nipponense, were studied and the results are as follows : 1. Larval development : Embryos hatched out as zoea larvae of 2.06 mm in mean body length. The larvae passed through 9 zoea stages in $15{\~}20$ days and then metamorphosed into postlarvae measuring 5.68 mm in mean body length. Each zoea stage can be identified based on the shapes of the first and second antennae, exo- and endopodites of the first and second pereiopods, telson and maxillae. 2. Environmental requirements of zoea larvae : Zoea larvae grew healthy when fed with Artemia nauplii. Metamorphosing rate was $65{\~}72{\%}$ at $26{\~}28\%$ and $7.85{\~}8.28\%_{\circ}Cl.$. The relationship between the zoeal period (Y in days) and water temperature (X in $^{\circ}C$) is expressed as Y=46.0900-0.9673X. Zoeas showed best survival in a water temperature range of $26{\~}32^{\circ}C$ (optimum temperature $28^{\circ}C$), at which the metamorphosing rate into postlarvae was $54{\~}72\%$ The zoeas survived more successfully in chlorinity range of $4.12{\~}14.08{\%_{\circ}}Cl.$, (optimum chlorinity $7.6{\~}11.6\;{\%_{\circ}}Cl.$.), at which the metamorphosing rate was $42{\~}76{\%}$. The whole zoeal stages tended to be longer in proportion as the chlorinity deviated from the optimum range and particularly toward high chlorinity. Zoeas at all stages could not tolerate in the freshwater. 3. Environmental requirements of postlarvae and juveniles : Postlarvae showed normal growth at water temperatures between $24{\~}32^{\circ}C$ (optimun temperature $26{\~}28^{\circ}$. The survival rate up to the juvenile stage was $41{\~}63{\%}$. Water temperatures below $24^{\circ}C$ and above $32^{\circ}$ resulted in lower growth, and postlarvae scarcely grew at below $17^{\circ}C$. Cannibalism tended to occur more frequently under optimum range of temperatures. The range of chlorinity for normal growth of postlarvae and juveniles was from 0.00 (freshwater) to $11.24{\%_{\circ}}Cl.$, at which the survival rate was $32{\~}35\%$. The postlarvae grew more successfully in low chlorinities, and the best growth was found at $0.00\~2.21{\%_{\circ}}Cl.$. The postlarvae and juveniles showed better growth in freshwater but did not survive in normal sea water. 4. Feeding effect of diet on zoea Ilarvae : Zoea larvae were successfully survived and metamorposed into postlarvae when fed commercial artificial plankton, rotifers, and Artemia nauplii in the aquaria. However, the zoea larvae that were fed Artemia nauplii and reared in Chlorella mixed green water showed better results. The rate of metamorphosis was $68\~{\%}75$. The larvae fed cow live powder, egg powder, and Chlorella alone did not survive. 5. Diets of postlarvae, juveniles and adults : Artemia nauplii and/or copepods were good food for postlarvae. Juveniles and adults were successfully fed fish or shellfish flesh, annelids, corn grain, pelleted feed along with viscera of domestic animals or fruits. 6. Growth of postlarvae, juveniles and adults : Under favorable conditions, postlarvae molted every five or six days and attained to the juvenile stage within two months and they reached 1.78 cm in body length and 0.17 g in body weight. The juveniles grew to 3.52 cm in body length and 1.07 g in body weight in about four months. Their sexes became determinable based on the appearance of male's rudimental processes (a secondary sex character) on the endopodites of second pereiopods of males. The males commonly reached sexual maturity in seven months after attaining the postlarvae stage and they grew to 5.65 cm in body length and 3.41 g in body weight. Whereas the females attained sexual maturity within six to seven months, when they measured 4.93 cm in body length and 2.43 g in body weight. Nine or ten months after hatching, the males grew $6.62{\~}7.14$ cm in body length and $6.68{\~}8.36$ g in body weight, while females became $5.58{\~}6.08$ cm and $4.04{\~}5.54$ g. 7. Stocking density : The maximum stocking density in aquaria for successful survival and growth was $60{\~}100$ individuals/$\ell$ for zoeas in 30-days rearing (survival rate to postlarvae, $73{\~}80{\%}$) ; $100{\~}300$ individuals/$m^2$ for postlarvae of 0.57 cm in body length (survival rate for 120 days, $78{\~}85{\%}$) ; $40{\~}60$ individuals/$m^2$ for juveniles of 2.72 cm in body length (survival rate for 120 days, $63{\~}90{\%}$) : $20{\~}40$ individuals/$m^2$ for young prawns of 5.2 cm in body length (survival rate for 120 days, $62\~90{\%}$) ; and $10\~30$ individuals/$m^2$ for adults of 6.1 cm in body length (survival rate for 60 days, $73\~100{\%}$). The stocking density of juveniles, youngs and adults could be increased up to twice by providing shelters.