A new fluorometric method for determination of chlortetracycline

A new fluorometric method is presented, based on data showing a linear relationship between CTC concentration and intensity of fluorescence of aluminum-CTC chelate, and little interference due to presence of decomposition products of CTC. A fairly good agreement was obtained between CTC values assayed by the microbiologic method and those by the proposed fluorometric method. Moreover, this method can be employed for field assay of CTC concentration in ice (Figs. 1,2, and Tables 1,3). A precise determination by using Beckman spectrophotometer : To a 10 cc aliquot of sample and each of standard solution series (1-5 mcg per cc) and 1 cc of a 1 % potassium alum (KA1(SO_4)_2?12H_2O) solution in M/2 acetate buffer of pH 4.6. Allow it to stand for 20 to 60 minutes at room temperature. Measure the fluorescence at a wave length of 500 mu of the sample when irradiated by an excitation light of 365 mu or 436 mu. Measure likewise the fluorescence of a series of standard solutions added with the alum reagent. Subtract the blank value from each measured intensity of fluorescence, and prepare a standard curve by plotting the intensity against the concentration of CTC. By employing this standard curve find the CTC concentration of the sample solution corresponding to its intensity of fluorescence which is corrected for the blank value. Approximate determination : The following methods are developed for the field work. (1) Test tube method : Place 10 cc of a supernatant of a thawed ice sample in a flat bottom test tube of equal diameter (about 27 mm), and add 1 cc of the alum reagent. After allowing it to stand for 5 to 10 minutes, observe the fluorescence from the top of the tubes irradiated with the black ray lamp from the side. Estimate CTC concentration of the ice sample by comparing the fluorescence of the sample with those of the standard series treated in the same way as the sample. (2) Portable nephelo-colorimeter method : Add a 1 cc aliquot of the alum reagent to both a 10 cc aliquot of supernatant of a thawed sample and a standard solution (4 or 2 mcg/cc depending on CTC content of sample). After allowing them to stand for 10 minutes, place the sample in one of the comparison tube (A) and the standard in the other tube (B), set the bottom of the inner tube for the tube (B) at a depth of 30 mm and by evenly irradiating with the long wave ultra-violet lamp and matching the intensity for tube (A) against that for tube (B), find the reading, h, for tube (A) where the intensity of the sample is identical with that of the standard. The content of CTC (mcg/cc) can be calculated by a formula, S×30/h, where S is the CTC concentration of the standard solution employed.サリチル酸およびサリチル酸アミドのような螢光性有機物はアルカリ,アルカリ土類および稀土類などの金属と酷塩をつくつて,発螢光性が強くなる.一方CTCはMgCl_2およびCaCl_2の溶液中で黄色の螢光を発し,またCa, Mgのような金属とキレート化合物をつくることが知られている.因つて著者はCTCのある種の金属キレート化合物は強い螢光を発するのではないかと想像し,分光光度計を用いてまず種々の金属イオンによる螢光度の差異をみたところ,アルミニウムイオンの存在によつてCTCの螢光が最も強くなることを見出したので,定量法の決定に必要な種々の条件を検討し一新定量法を決定することができた

The influence of repeated bleeding and anesthetization on change of blood sugar of fish

In the glucose tolerance test (GTT) and insulin glucose tolerance test (IGTT) with fish, the change of blood sugar content is determined by a repeated bleeding method or a one-time bleeding method. It is desirable that the change of blood sugar is found through samples of blood taken repeatedly at regular intervals from a given fish. It is to be expected that the blood sugar will increase in proportion to the rapid shock caused by handling and repeated bleeding. The present study, therefore, w as carried out to determine the influence of repeated bleeding and anesthetization on the change of blood sugar content of the red sea bream (Chrysophrys major), to determine the difference of influences among anesthetics, and to compare the change of blood sugar content in the GTT and IGTT which was obtained by a repeated bleeding method and that obtained by a one-time bleeding method. The results obtained can be summarized as follows: 1) The blood sugar content of the red sea b ream determined by repeated bleeding without anesthetization increased remarkably each time a bleeding was taken, and there was a large difference between the change of blood suger content in individual fish. 2) The increase of blood sugar obtained by repeated bleeding on a given fish anesthetized with urethane (5,000 ppm), or quinaldine (20 ppm), or MS-222 (100 ppm) was lower than that obtained by repeated bleeding without anesthetization. The increase of blood sugar obtained by repeated bleeding with MS-222 anesthetization was the least. 3) The average blood sugar content obtained from three different fish at each t ime by the one-time bleeding method with urethane anesthetization was always in the range from 50-75 mg/dl. 4) The changes of blood sugar content in the GTT and IGTT obtained from a given fish by repeated bleeding with MS-222 anesthetization showed the same curve as those obtained from numerous fish by a one-time bleeding method with urethane anesthetization, but the quantities of increase and decrease of blood sugar content obtained by both methods were different.同一個体より反復採血した場合のマダイの血糖変動,その血糖変動に対する麻酔の影響,麻酔剤の種類による影響の違い,同一魚体より反復採血して求めた血糖変動と多数魚体からそれぞれ1回採血して求めた値を集積してえた血糖変動との異同について検討し,次の結果を得た. 1. 麻酔しない同一魚体から反復採血すると,採血回数が増すにしたがつて血糖が著しく増加した.また血糖上昇に大きな個体差が認められた.　2.　Urethane(5000ppm),Quinaldine(20ppm)およびMS-222(100ppm)で麻酔した同一魚体より反復採血した場合の血糖増加は,麻酔しないで反復採血した場合のそれよりかなり小さかつた.特にMS-222で麻酔し反復採血した場合の血糖上昇はゆるやかであつた. 3. 各測定時にUrethaneで麻酔した3個体からそれぞれ1回採血して求めた平均血糖値は,常に50～75mg/dlの範囲にあり,ほとんど一定の値を示した. 4. MS-222で麻酔し,同一魚体から反復採血して求めたグルコースあるいはインシュリン・グルコース負荷後の血糖変動は,多数個体のそれぞれから1回採血して求めた負荷後の血糖変動と同じ傾向を示したが,その血糖の上昇程度あるいは降下程度にかなりの差が認められた

魚類血糖におよぼす反復採血および麻酔の影響

In the glucose tolerance test (GTT) and insulin glucose tolerance test (IGTT) with fish, the change of blood sugar content is determined by a repeated bleeding method or a one-time bleeding method. It is desirable that the change of blood sugar is found through samples of blood taken repeatedly at regular intervals from a given fish. It is to be expected that the blood sugar will increase in proportion to the rapid shock caused by handling and repeated bleeding. The present study, therefore, w as carried out to determine the influence of repeated bleeding and anesthetization on the change of blood sugar content of the red sea bream (Chrysophrys major), to determine the difference of influences among anesthetics, and to compare the change of blood sugar content in the GTT and IGTT which was obtained by a repeated bleeding method and that obtained by a one-time bleeding method. The results obtained can be summarized as follows: 1) The blood sugar content of the red sea b ream determined by repeated bleeding without anesthetization increased remarkably each time a bleeding was taken, and there was a large difference between the change of blood suger content in individual fish. 2) The increase of blood sugar obtained by repeated bleeding on a given fish anesthetized with urethane (5,000 ppm), or quinaldine (20 ppm), or MS-222 (100 ppm) was lower than that obtained by repeated bleeding without anesthetization. The increase of blood sugar obtained by repeated bleeding with MS-222 anesthetization was the least. 3) The average blood sugar content obtained from three different fish at each t ime by the one-time bleeding method with urethane anesthetization was always in the range from 50-75 mg/dl. 4) The changes of blood sugar content in the GTT and IGTT obtained from a given fish by repeated bleeding with MS-222 anesthetization showed the same curve as those obtained from numerous fish by a one-time bleeding method with urethane anesthetization, but the quantities of increase and decrease of blood sugar content obtained by both methods were different.同一個体より反復採血した場合のマダイの血糖変動,その血糖変動に対する麻酔の影響,麻酔剤の種類による影響の違い,同一魚体より反復採血して求めた血糖変動と多数魚体からそれぞれ1回採血して求めた値を集積してえた血糖変動との異同について検討し,次の結果を得た. 1. 麻酔しない同一魚体から反復採血すると,採血回数が増すにしたがつて血糖が著しく増加した.また血糖上昇に大きな個体差が認められた.　2.　Urethane(5000ppm),Quinaldine(20ppm)およびMS-222(100ppm)で麻酔した同一魚体より反復採血した場合の血糖増加は,麻酔しないで反復採血した場合のそれよりかなり小さかつた.特にMS-222で麻酔し反復採血した場合の血糖上昇はゆるやかであつた. 3. 各測定時にUrethaneで麻酔した3個体からそれぞれ1回採血して求めた平均血糖値は,常に50～75mg/dlの範囲にあり,ほとんど一定の値を示した. 4. MS-222で麻酔し,同一魚体から反復採血して求めたグルコースあるいはインシュリン・グルコース負荷後の血糖変動は,多数個体のそれぞれから1回採血して求めた負荷後の血糖変動と同じ傾向を示したが,その血糖の上昇程度あるいは降下程度にかなりの差が認められた

Availabilities of Phosphorus Compounds as Dietary Phosphorus Sources for Red Sea Bream

Three experiments were conducted to compare the availabilities of eight phosphorus compounds as a phosphorus sources in the diet for red sea bream, Chrysophrys major. Phosphorus contents of the test diets were adjusted to 680 mg per 100g of diet in each experiment. The fish were reared on the test diets at 25℃ over a 77 day period (Exp. I), 62 day period (Exp. II), and an 88 day period (Exp. III). At the end of the feeding trial, ten fish from each group were selected for the chemical analyses of the blood serum, liver, and vertebrae. Sodium phosphates (mono-, di-, and tribasic), potassium phosphate monobasic, and calcium phosphate monobasic were more effective than calcium phosphates (di- and tribasic) to prevent the development of phosphorus deficiency symptoms. Calcium phytate was scarcely utilized as the dietary phosphorus source by red sea bream. Therefore, the water soluble phosphorus compounds, from which inorganic phosphorus is easily released, should be employed as the dietary phosphorus source for red sea bream

Requirement of Red Sea Bream for Dietary Na and K

Two experiments were conducted to examine the dietary requirement of red sea bream, Chrysopkvys major, for sodium and potassium supplementation. The fish were fed on two test diets with and without sodium supplementation (Exp. I), and potassium supplementation (Exp. II), over a 63 day period at 25℃. At the end of the feeding trial, histopathological examination of the tissues, hematological examinations, and chemical analyses of the liver and vertebrae were performed. No significant differences were recognized between the two groups in each experiment in the following determinations : the growth rate, feed efficiency, condition factor, and hepatosomatic index; the hemoglobin content, hematocrit value, red blood cell count, mean corpuscular hemoglobin, mean corpuscular volume, mean corpuscular hemoglobin concentration, mean corpuscular diameter, and percentage of immature erythrocytes ; the moisture, lipid, and glycogen content of the liver; and the lipid, ash, calcium, phosphorus, magnesium, sodium, and potassium content of the vertebrae. Furthermore, no pathological change was recognized in the organs of fish fed the diets without the sodium or potassium supplement. From these findings, it appears that sodium and potassium supplementation in the diet for red sea bream is not essential, even if these elements are scarcely contained in the diet

Effect of Starvation on Hematological Characteristics, and the Contents of Chemical Components and Activities of Enzymes in Blood Serum of Red Sea Bream

This study was conducted to determine the effect of starvation on the hematological characteristics, and the contents of chemical components and activities of enzymes in blood serum of red sea bream, Ckrysopkrys major. Wild fish were starved after the net fishing for 90 days at 13318℃. At each fixed day (0, 1. 2, 4. 9, 15, 30, 70 and 90th day), ten fish were presented for the hematological examinations and the biochemical analyses of blood serum. Hemoglobin content, hematocrit value, red blood cell count, and specific gravity of the whole blood increased early in the starving period and then decreased, and the fish exhibited a serious anemia. From the mean corpuscular constants (MCH. MCV and MCHC), the anemia was categorized as the hyperchromic microcytic type. Furthermore, it should be noted that the anemia was accompanied by the disappearance of immature erythrocytes. The contents of total protein and total cholesterol, and the activity of LAP in serum, also increased early in the starving period and greatly decreased later. BUN in serum was stable at a high level for some time after the abrupt increase, and decreased thereafter. The activities of GOT, GPT, and LDH in serum did not change till the 30th day, before the remarkable increase set in. Though the changes of the contents of glucose and total bilirubin, and the ALP activity in serum were recognized during the starvation, those levels were within the normal range

Studies on methods for keeping quality of \u22Kamaboko\u22, a cooked fish paste III : Effectiveness of chemical and physical treatments on the shelf-life of \u22Kamaboko\u22.

In the previous papers, it was reported that the casing with cellophane, the irradiation by infrared and/or ultraviolet ray showed a marked prolongation of shelf-life of \u22 Kamaboko \u22. It was further noticed that when the preservatives were smeared on the surface, the combination of the preservatives and casing was rather inferior to the control group without casing. However, when the preservatives were mixed with the paste, the combination did not show any inferior effect to the control. The present paper deals with a comparison of the effectiveness between preservatives which were mixed with the paste and those which were applied on the surface, and further with the effectiveness of casing with cellophane, polyvinyl alcohol and polyvinyl acetate film and of infrared and/or ultraviolet ray irradiation all over the surface of \u22Kamaboko\u22. 1. When the preservatives were mixed with the paste, dehydroacetic acid and its sodium salt were effective in prolonging the storage life to six and four days, respectively, in contrast to two days in the control at 26℃. However, the preservatives which were smeared on the surface were not effective (See Tables 1 and 2). 2. Among the preservatives which were mixed with the \u22Kamaboko\u22, butyl p-hydroxybenzoate and o-hydroxybenzamide showed effectiveness to some extent on keeping quality (See Table 3). 3. The casing with polyvinyl alchol film gave rather an inferior result. The combination of the casing with polyvinyl alcohol film and the mixing with preservatives shortened the shelf-life compared to the control (See Table 4). 4. In the case of \u22 Kamaboko \u22 without the casing, irradiation by the infrared ray for short time prevented the growth of bacteria. However, irradiation by ultraviolet ray showed remarkable effect on inhibition of bacterial growth. Irradiation by ultraviolet ray twice daily was slightly more effective than that once daily. The combination of irradiation by ultraviolet ray twice daily and that by infrared ray every other day was effective in prolonging the storage life to five days in contrast to two days in the control without the irradiation (See Table 5, Figs. 1a, 1b and 1c). 5. Irradiation by ultraviolet ray over \u22 Kamaboko \u22 which was covered with cellophane or polyvinyl cetate was also effective. No difference was observed between the irradiation once daily and twice daily (See Table 6, Figs. 1a, 1b and 1c). 6. Daily irradiation by ultraviolet ray all over the surface of \u22Kamaboko\u22 which contained DHA at a 0.1 % level and covered with cellophane or polyvinyl acetate was able to prolong the storage life to 10 days in contrast to a storage life of two days in the control at 26℃ (See Tables 7 and 8).1. 無包装の板付無糖蒲鉾を用い,数種の防腐剤を塗布した場合と練り込んだ場合との効果を比較したところ, DHA及びDHA-NaはO.2%の濃度でいずれも蒲鉾の表面に塗布した場合は無効であつたが, 練り込んだ場合は鮮度保持に有効であった. 2. 数種の防腐剤を練り込んだ場合の効果を検討したところ, パラオキシ安息香酸ブチルェステル及びサリチル酸アミドが0.1或は0.2%の濃度で鮮度保持効果が認められた. またDHA或はDHA-Naとパラオキシ安息香酸ブチルエステル或は第四級アミンQA(BB)の相乗効果について検討したところ,殆んど認められなかつた. 3. ポリビニールアルコールで被覆すると無包装よりも悪く,また防腐剤の効果も一般に低下した. 4. 無包装の場合には, 短時間の赤外線照射はカビの発育を抑えるが, 細菌に対しては効果弱く, 逆に紫外線はカビに対して効果が弱いが, 細菌に対しては極めて有効であつた. また紫外線照射は1日1回よりも2回の方が多少有効であつた. 従つて30cm距離からの15W殺菌灯による15分間全面照射を2回毎日繰り返し, 之に隔日に赤外灯(500W,30cm距離)を5分間照射すると, 5日間鮮度が保持された. 5. セロファン或は醋酸ビニールで全面を覆うと, カビの二次的汚染が少いためか, 赤外線照射の効果は認められなかつたが, 紫外線照射は有効であつた. この場合には1日に1回照射した時と2回超射した時との間に殆んど差がなかつた. 6. DHAを1/1,000練り込み, セロファン或は醋酸ビニールで被覆し, 紫外線を毎日1回15分間全面照射すると, 少くとも26℃で10日間鮮度が保持された

Effect of Starvation on Hematological Characteristics, and the Contents of Chemical Components and Activities of Enzymes in Blood Serum of Red Sea Bream

This study was conducted to determine the effect of starvation on the hematological characteristics, and the contents of chemical components and activities of enzymes in blood serum of red sea bream, Ckrysopkrys major. Wild fish were starved after the net fishing for 90 days at 13318℃. At each fixed day (0, 1. 2, 4. 9, 15, 30, 70 and 90th day), ten fish were presented for the hematological examinations and the biochemical analyses of blood serum. Hemoglobin content, hematocrit value, red blood cell count, and specific gravity of the whole blood increased early in the starving period and then decreased, and the fish exhibited a serious anemia. From the mean corpuscular constants (MCH. MCV and MCHC), the anemia was categorized as the hyperchromic microcytic type. Furthermore, it should be noted that the anemia was accompanied by the disappearance of immature erythrocytes. The contents of total protein and total cholesterol, and the activity of LAP in serum, also increased early in the starving period and greatly decreased later. BUN in serum was stable at a high level for some time after the abrupt increase, and decreased thereafter. The activities of GOT, GPT, and LDH in serum did not change till the 30th day, before the remarkable increase set in. Though the changes of the contents of glucose and total bilirubin, and the ALP activity in serum were recognized during the starvation, those levels were within the normal range

Requirement of Red Sea Bream for Dietary Na and K

Two experiments were conducted to examine the dietary requirement of red sea bream, Chrysopkvys major, for sodium and potassium supplementation. The fish were fed on two test diets with and without sodium supplementation (Exp. I), and potassium supplementation (Exp. II), over a 63 day period at 25℃. At the end of the feeding trial, histopathological examination of the tissues, hematological examinations, and chemical analyses of the liver and vertebrae were performed. No significant differences were recognized between the two groups in each experiment in the following determinations : the growth rate, feed efficiency, condition factor, and hepatosomatic index; the hemoglobin content, hematocrit value, red blood cell count, mean corpuscular hemoglobin, mean corpuscular volume, mean corpuscular hemoglobin concentration, mean corpuscular diameter, and percentage of immature erythrocytes ; the moisture, lipid, and glycogen content of the liver; and the lipid, ash, calcium, phosphorus, magnesium, sodium, and potassium content of the vertebrae. Furthermore, no pathological change was recognized in the organs of fish fed the diets without the sodium or potassium supplement. From these findings, it appears that sodium and potassium supplementation in the diet for red sea bream is not essential, even if these elements are scarcely contained in the diet

Effect of Starvation on Organ Weight and Chemical Component of Red Sea Bream

To determine the effect of starvation on the organ weight and chemical components of red sea bream, Chrysophrys major, wild fish were starved for 90-days at 13-18℃. At fixed days (0, 1, 2, 4, 9, 15, 30, 70 and 90th day). ten fish were selected for the determination of the fork length, weight, and chemical components of the whole body and organs. The condition factor decreased over the starving period. The weight of liver and intestine with adipose tissue declined drastically from the 9th day till the 30th day, and the weight of the heart decreased after the 30th day. However, the weight of spleen increased till the 15th day before the onset of the decline. A decreasing lipid content was first recognized in the liver, and then in the vertebrae and dorsal muscle. Glycogen in the liver increased till the 4th day, and decreased rapidly thereafter. Protein in the dorsal muscle was catabolized severely from the 9th day till the 30th day. The moisture contents of liver and dorsal muscle increased with the decline in lipid, glycogen, and protein content. Mineral composition of the vertebrae was not affected by the starvation