This experiment was designed to evaluate effects of freezing and frozen storage on survival of sanitary indicative bacteria in seafoods. Culture of bacteria such as Escherichia coli type I, Citrobacter freundii type I, Klebsiella aerogenes type I and Streptococcus faecalis was inoculated into homogenates of pollack, shrimp, and sardine frozen in a contact plate freezer at $-40^{\circ}C$ and chest freezer at $-20^{\circ}C$, stored at $-20^{\circ}C$, and then survival of the inoculated bacteria was determined over a period of 95 days. Coliform group was highly sensitive to freezing and frozen storage showing survival of about $2\%$ after 95 days of frozen storage at $-20^{\circ}C$, whereas Streptococcus faecalis was relatively resistant with $20\%$ survival rate. The sanitary indicative bacteria count was rapidly decreased in the early stage of frozen storage revealing 90 to $95\%$ loss of coliform group and 40 to $70\%$ loss in case of Streptococcus faecalis after 10 days storage. In determining recovery rate, most probable number (MPN) method gave more reproducible recovery of the tested strain than did the selected agar plate method.
Two vibrio sp. strains were isolated from disease catfish(silurus asotus). The present isolates were identified as Vibrio ordalii based on their biological and biochemical characteristics ; they were positive for acid production from glucose, Iactose, maltose, sucrose and salicine, while negative for arabinose, galactose, inocitol and xylose. They are named KL-1 and KL-2, KL-1 and KL-2 strains were similar to physiological characteristics ; growth was observed at pH 5 to 10 and in 0% to 6.0% NaCl. Two strains did not growth at a concentration above 7.0% NaCl and pH10. This bacterium was infected into health catfish hypodermically. Such injection was found to induce haemorrhagic ulcers very similar to those observed in naturally infected fish. At 24h post-infection, the red spot developed around the injection site and grew bigger to from a red sport area. At 120h post-infection, the muscle ncerosis was extended near the ventral fin. The seventy percent lethal dosage was appeared to water temperature at $25^{\circ}C$. Two strains were tested for drug senistiveity by plate method. KL-1 and KL-2 strains were sensitive to GM. K, N, S and SxT, but resistant to CF, L and VA.
Biogenic amines are generally formed through the decarboxylation of specific free amino acids by exogenous decarboxylases released by microbial species associated with the fish products and fermented feeds. This study was conducted to investigate the properties of e tuna waste regarding the control of degradation of biogenic amines (histamine, tyramine, tryptamine, putrescine, and cadaverine) that might be related with the anti-nutritional factor of the tuna waste that is used for manufacturing domestic fish meal. The values of pH and the salt content were 6.51, 3.35% in tuna waste and 5.58 and 5.83% in tuna fish meal, respectively. The strains and dominant bacteria tested in the tuna waste sample were 9.20, 9.29, 5.67, 7.82 and 7.58 log CFU/g of total bacteria, aerobic plate count (APC), total coliform (TC), Lactobacillus spp. and Bacillus spp., respectively. The main histamine forming-bacteria (HFB) in tuna waste were detected by silica gel thin-layer chromatography (TLC) and 7 histamine-forming bacterial species were isolated among microbes grown in selective medium. The histamine concentration was determined by detection of fluorescence of ο-phthaldialdehyde (OPA) derivatives using HPLC and the date were used to reconfirm the identities of the amine-producing bacteria. The 15 histamine- forming bacteria strains grown in trypicase soy broth (TSB) supplemented with 1% L-histidine (TSBH) were identified as Lactococcus(L.) lactis subsp. lactis, Klebsiella pneummonlae, L. garvieae 36, Vibrio olivaceus, Hafnia alvei and L. garvieae which were main dominant amine - producing strains, and Morganella morganii identified by 16S ribosomal RNA (rRNA) sequencing with PCR amplification. A Phylogenetic tree generated from the 16S rRNA sequencing data showed different phyletic lines that could be readily classified as biogenic amine forming gram-positive and negative bacteria.
Journal of the Korean Society of Fisheries and Ocean Technology
/
v.33
no.4
/
pp.352-359
/
1997
The visual acuity of filefish Stephanolepis cirrhifer was studied through a series of experiments by observing their responses to target plates. The fish were trained to respond to the target plates made of white acrylic resin with a vertical black line 5cm long in the center. The width of the black line ranged from 0.2mm to 8.0 mm. The line width was diminished unto the fish could no longer distinguish the line at a distance of 100 cm from the target plate. This was repeated under light intensities of 400, 20, 5, 3 and 1 lx at the water surface. Fish were rewarded with bait in front of the target plate if the fish went to the target date (i.e., success).The results show that the training effect of 1lensh had a success rate of over 80% and that the reach times to the target plates were 4~5 seconds over 210 experimental tunes. The success rate was high using the thick line with strong apparent contrast, but was low at the 1 lx. The visible critical width of line became thick with decreasing light intensity, 0.24mm at 400 lx, followed by 0.30mm at 20 lx, 0.40mm at 5 lx, 0.46 mm at 3 lx and 2.87mm at 1 lx. The apparent contrast for visible critical width of line increased with decreasing light intensity, 0.01 at 400 and 20 lx, 0.02 at 5 lx, 0.03 at 3 lx and 0.09 at 1 lx. The line acuity of filefish was best 1.21 at the 400 lx, followed by 0.97 at 20 lx, 0.73 at 5 lx, 0.63 at 3 lx and sharply decreased to 0.10 at 1 lx. The visible ranges for 1mm and 6mm in width of line were about 4.2 m and 25.0 m at the 400 lx light intensity and decreased m 1/14 times and 1/12 times of the 400 lx at 1 lx, respectively.
Journal of the Korean Society of Fisheries and Ocean Technology
/
v.23
no.1
/
pp.1-5
/
1987
The authors carried out an experiment to determine the vertical opening of the midwater trawl, which is the same used in the former experiment in this series of studies. To determine the vertical opening of otter board and front weight, three fish finders were used. A 200 KHz fish finder set on board the research vessel was used to sound the depth of water. A transmitter of 50 KHz fish finder was set through the shoe plate of otter board to determine the height of otter board from the sea bed, and a transmitter of another 50 KHz fish finder was set downwardly on the net pendant right before the front weight to determine the height of weight from the sea bed. The depth of otter board and weight were calculated by subtract the height of those from the depth of water, respectively. To determine the vertical opening of mouth, a transmitter of net recorder was set on the head rope and the vertical opening of that to ground rope was directly read on the recording paper. The results obtained can be summarized as follows: 1. The rate of the depth of otter board to the length of warp was in the range of 0.44 to 0.25, and the depth was linearly shoaled about 5m per 0.1m/sec of the towing speed or per 20rpm of the main engine. The rate of the observed depth to the calculated depth of otter board was in the range of 0.92 to 0.080 with a decreasing tendancy in accordance with the increase of towing speed. 2. The depth of head rope was 2 to 3m deeper than that of otter board, and the vertical opening of net mouth was in the range of 22 to 19m, with a decreasing tendancy in accordance with the increase of towing speed, 3. The difference of depth between front weight and otter board was about 20m and 22m respectively in the length of warp 100m and 150m without distinct change in accordance with the towing speed. The depth of front weight was 2 to 3m shallower than that of ground rope. 4. The changing range of depth of head rope according to the revolution of main engine was about 4m per 20rpm.
This study was performed to measure the mutagenicity of fish by cooking and storage. Mutagenicity of the fish extract was measured by Ames test(Salmonella typhimurium reversion assay with TA 100) in vitro and by micro-nucleus test in vivo. The fish samples screened in this study were white fish(Trichiurus, Croaker, Salted Croaker) and red fish(Saury pike, Mackerel, Yellowtail, Salmon). The number of revertants of red fish were significantly higher than that of white fish. And the mutagenicity of mackerel was higher than other red fish, so followed experiment was made by using the extract of mackerel. Mutagenicity of the samples cooked on microwave oven was the lowest, whereas there was no significant difference between the samples cooked on gas grill and the ones on electric grill. In the presence of S9 mixture, the methanol extract of mackerel showed 2∼4 times high values of mutagenicity in comparison with the extract without S9. The extract of mackerel cooked with various vegetable juices showed inhibitory effects on the mutagenicity in the order of green tea, ginger, and radish. Also, the number of revertants was increased in the stored samples. Mutagenicity of the samples stored in the refrigerator was higher than that of the freezer. In micronucleus test, the methanol extract treated with vegetable juice inhibited micro-nucleus formation in bone marrow by cyclophosphamide in the order of ginger, green tea, and radish. In TBA test, there was a tendency that TBA values were increased as the storage time increased. Also, the rancidity of sample were stored in the refrigerator was higher value than sample stored in the freezer. Samples cooked on microwave oven showed the highest value in rancidity. When the antioxidant effect of vegetable juice was measured by electron donating ability(EDA) of mackerel cooked with vegetable juice to DPPH, the samples treated with onion showed the highest value of EDA(%), and the samples treated with green tea, ginger and cabbage also showed the antioxidant effect.
This study was conducted to evaluate microorganism contamination of food utensils and service facilities in school and to prevent hazards by food poisoning occurrence. As a result, the highest number of microorganism growth plate ($12.3{\pm}2.6$) was detected in total bacteria test plate, and also observed $10.3{\pm}3.9$ growth plates in Staphylococcus aureus test plate and $9.5{\pm}3.9$ growth plates in E. coli and coliform bacteria test plate. But we could detect to the lowest number of growth plates ($1.5{\pm}1.0$) in Vibrio test plate. We also assessed that floors were appeared to the highest microorganism contamination rate in food utensils and service facilities. Therefore, $4.5{\pm}0.6$ growth plates was detected in pre-operation floor and $4.3{\pm}1.0$ growth plates in floor. And high level of microorganism contamination also observed in tables as $3.3{\pm}1.0$ growth plates in cooking table and $3.0{\pm}0.0$ growth plates in dining table. The level of microorganism contamination of food utensils such as kitchen knife, cutting board, and food tray were lower than that in food service facilities. We analysed microorganism contamination according to purpose of use in kitchen knifes and cutting boards. The microorganism contamination rate in fish kitchen knife ($2.0{\pm}0.8$) and fish cutting board ($1.3{\pm}1.5$) were slightly higher than that of others purpose of use. As a result of microorganism identification, various strains of microorganism were contaminated in food service facilities and some strains could detected more than two times. Especially, Staphylococcus aureus was repeatedly identified in cooking table, trench, and kitchen knife. Bacillus cereus was identified in kitchen knife, and then Pseudomonas fluorescens and Pseudomonas aeruginosa were also detected in food utensils and service facilities as known to food spoilage microorganisms. Klebsiella pneumoniae was detected four times repeat, which widely distribute natural environment as normal bacterial flora but sometimes cause acute pneumonia. These results suggest that food utensils and service facilities are contaminated with not only major food poisoning microorganisms such as Staphylococcus aureus, but also food spoilage microorganisms. Taken together, strict personal hygiene control and efficient food service facilities management will be needed to enhance food safety in school feeding and to improve student health.
A scanning electron microscopic study on the glochidium and glchidial encystment of Anodonta grandis on the guppy was conducted. The shape of the glochidium is apparently triangular and its averge size is 0.45mm X0.4mm when closed, The two glochidial shell valves are of the same size, kept together by a ligament of 120${\mu}{\textrm}{m}$ in length and 7 ${\mu}{\textrm}{m}$ in width. Each of the glochidial shell valves has a 16 ${\mu}{\textrm}{m}$ long hook sitdded with many spines on the superior face. A large area to the apex of the valve surrounding the base of the hook is provided with numerous small spines which become progressively smaller towards the periphery of the area, The external surface of the glochidial shell valve is covered with numerous small processes showing successive change in the shape and the pattern of destribution by part. Besides the processes, there are a number of niches scattered all over the exterior surface. The glochidial shell valve has two layers. One is the outer thin membrane bearing the processes and the niches and the others is the inner layer bearing numerous holes which any accessory structure and 2.65 ${\mu}{\textrm}{m}$ in diameter, emerges from a canal located at center of ventral plate of the mamtle, A total of three types of the hair cells are observed. In present artificial infection of the glochidium to the guppy, it took about three to four hours to complete an early cysts, During the period of encystment, The epithelial cells of the host fish actively migrated toward the attached glochidium and covered it.
Background: Although an understanding of the proliferation and differentiation of fish female germline stem cells (GSCs) is very important, an appropriate threedimensional (3D) research model to study them is not well established. As a part of the development of stable 3D culture system for fish female GSCs, we conducted this study to establish a 3D aggregate culture system of ovarian cells in marine medaka, Oryzias dancena. Methods: Ovarian cells were separated by Percoll density gradient centrifugation and two different cell populations were cultured in suspension to form ovarian cell aggregates to find suitable cell populations for its formation. Ovarian cell aggregates formed from different cell populations were evaluated by histology and gene expression analyses. To evaluate the media supplements, ovarian cell aggregate culture was performed under different media conditions, and the morphology, viability, size, gene expression, histology, and E2 secretion of ovarian cell aggregates were analyzed. Results: Ovarian cell aggregates were able to be formed well under specific culture conditions that used ultra-low attachment 96 well plate, complete mESM2, and the cell populations from top to 50% layers after separation of ovarian cells. Moreover, they were able to maintain minimal ovarian function such as germ cell maintenance and E2 synthesis for a short period. Conclusions: We established basic conditions for the culture of O. dancena ovarian cell aggregates. Additional efforts will be required to further optimize the culture conditions so that the ovarian cell aggregates can retain the improved ovarian functions for a longer period of time.
This study aimed to compare the automated most-probable-number (MPN) TEMPO BC and the quantitative mannitol-egg yolk-polymyxin (MYP) plate methods for enumeration of Bacillus cereus in food samples known to be frequently contaminated. Food products that were naturally or artificially contaminated with B. cereus were analyzed by both methods. A difference of less than 1 log (CFU/g) between the two methods was noted in 95.3% samples. There were no significant differences in artificially contaminated products between the two methods in terms of $R^2$ values for sauce products, jorim products, fish products, etc. However, a significant difference was noted for sunsik, fermented soybean products, and products. The linear equation of naturally versus artificially contaminated food was $log_{(TEMPO\;BC)}=0.8453{\times}log_{(MYP\;plate\;agar)}+0.1642$. Statistical analysis of the results showed good agreement between the two methods. Due to growing interest in food safety, the use of the TEMPO BC method may increase. In response to this trend, the results from this study will offer valuable comparative data on the feasibility of existing methods and help develop new approaches for food safety testing.
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