Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

Effects of Dietary Supplementation of Barodon on Growth Performance,Innate Immunity and Disease Resistance of Juvenile Olive Flounder,Paralichthys olivaceus,Against Streptococcus iniae

This study was conducted to evaluate the effects of dietary supplementation of Barodon, an anionic alkali mineral complex, on growth, feed utilization, humoral innate immunity and disease resistance of olive flounder. A basal experimental diet was used as a control and supplemented with 0.1, 0.2, 0.3, 0.4, or 0.5% Barodon. Triplicate groups of fish (26.4 ± 0.2 g) were fed one of the diets to apparent satiation twice daily for 10 wk. The growth performance was enhanced (P < 0.05) linearly and quadratically in fish fed diets containing Barodon compared with that in fish fed the control. Feed utilization was significantly improved by Barodon supplementation. Serum lysozyme and antiprotease activities were increased quadratically in Barodon fed groups. Also, significantly higher superoxide dismutase activity was found in Barodon‐fed fish. Dietary supplementation of 0.1–0.3% Barodon resulted in significant enhancement of fish disease resistance against Streptococcus iniae. The findings in this study indicate that dietary supplementation of Barodon can enhance growth, feed utilization, innate immunity, and disease resistance of olive flounder and that the optimum level seems to be 0.1% in diets.     

Distribution and migration of Japanese common squid, Todarodes pacificus, in the southwestern part of the East (Japan) Sea

Data on squid catches of Korean and Japan, water temperatures at depth of 100 m, and night-visible images of fishing boats collected in the East (Japan) Sea from 1970 to 1999 were analyzed to examine the distribution and migration of the Japanese common squid, Todarodes pacificus, in the southwestern part of the East (Japan) Sea. The main fishing grounds detected from squid catch in each grid (0.5° latitude × 0.5° longitude) by Korean squid fishery and night-visible images provided by the Defense Meteorological Satellite Program (DMSP) Operational Linescan System (OLS) were situated in the southwestern part of the East (Japan) Sea. The distribution and migration route was illustrated from squid catch in each grid and DMSP OLS image. In years of high catches, the fishing grounds were situated mainly between Ulleung Island and the eastern coastal waters of Korea, while in years of low catches they were situated between Ulleung Island and the Yamato Bank in the central East (Japan) Sea. The center of fishing activity began moving northward from around the Korea/Tsushima strait to the northern boundary of the Tsushima Warm Current in March, reaching into the Yamato Bank in September, and then returning to the strait by February. The northward and southward migration routes differed; the northward migration route occurred closer to the mainland coast of Korea than the southward migration route did. This work suggests T. pacificus begin their northward migration almost 2 months earlier than previously suggested.     

The role of flatfishes in the organization and structure of the eastern Bering Sea ecosystem

We evaluated the role of flatfishes in the organization and structure of the eastern Bering Sea ecosystem using the Ecopath/Ecosim approach. As basic input data for the Ecopath/Ecosim model, we used estimates of biomass from bottom trawl surveys and age-structured population models, production/biomass (P/B) ratio, consumption/biomass (Q/B) ratio, diet composition (DC), and fisheries harvests for each component of species or species groups. We estimated the trophic level of each component, niche overlaps among flatfishes, and the impacts of competition and predation on flatfish species in the eastern Bering Sea ecosystem. Based on those estimates, we developed the tropho-dynamic structure of the ecosystem, and the model was used to simulate ecological effects of fishery exploitation patterns. No single flatfish species appeared to have a profound and uniquely important role in the organization and structure of the ecosystem. Instead, the most important component among the guild of flatfish species appeared to be yellowfin sole Pleuronectes asper, which had greater biomass than other flatfish and a relatively diverse diet among the small flatfish species. Pacific halibut Hippoglossus stenolepis, Greenland turbot Reinhardtius hippoglossoides, and arrowtooth flounder Atheresthes stomias were important keystone predators in the eastern Bering Sea ecosystem together with some groups of marine mammals and sea birds. Intra flatfish complex cannibalism was not observed, however, substantial diet overlaps were common in the flatfish guild system.     

THE USE OF IMIDOCARB DIPROPRIONATE FOR THE TREATMENT OF THEILERIA SERGENTI INFECTIONS OF CATTLE

SUMMARY Imidocarb diproprionate (IMDP) was used therapeutically at a dose of 1.2 mg IMDP/kg of body weight against Theileria sergenti infections in Hereford heifers on the island of Jeju-Do, Republic of Korea. In the first experiment IMDP was seen to be at least as effective in reducing T. sergenti parasitaemias as primaquine phosphate (primaquine), the drug normally used on the island. In the second and third experiments IMDP was shown to reduce T. sergenti parasitaemias by about 80%. No drug is known to be completely effective against T. sergenti infections but the suggestion is made that IMDP may be used in preference to primaquine for treatment of acutely ill animals since it also has the advantage of eliminating Babesia ovata, another element of the piroplasmosis syndrome on the island.     

Risk factors for L. monocytogenes contamination of dairy products in Switzerland, 1990-1999

Our purpose was to identify the main hazards associated with the spread of Listeria monocytogenes in dairy products in Switzerland and to determine the changes in predominant serotypes of the isolates, using databases on dairy-processing and environments from the Swiss Dairy Research Station during the years 1990-1999. Overall, of 76,271 samples collected, 3722 (4.9%) were positive for the presence of L. monocytogenes. Cheese-ripening facilities had the highest proportion of positive samples (7.6%), followed by small-scale local dairies (4.4%). By sample type, the highest proportion of positive samples (9.5%) was observed in water samples used for cheese-washing, followed by cheese-surface swabs (5.0%). During the 10-year period, no positive samples were obtained from cream, ice cream, milk powder, yogurt, or fresh cheese. Of 3722 L. monocytogenes isolates, 1328 (35.7%) were serologically typeable. Serotypes 1/2a, 1/2b, and 4b accounted for 92.7% of the 1328 isolates. Until 1995, the most-prevalent serotype was 1/2b (annual proportional prevalence 39.3-72.2%)--whereas since 1996, 1/2a was the most prevalent (34.7-54.7%). During 1996-1999, serotype 1/2a increased by 88%, compared to the average of 1990-1995. In the final random-effect multivariable logistic model, the strongest predictor of a positive culture was samples from cheese-ripening plant (OR=1.54; 95% CI: 1.14, 2.08) and the second-strongest predictor was samples collected by someone who was employed by the plant (OR=1.48; 1.29, 1.71). Hard and semi-hard cheeses were more likely to be associated with serotype 1/2b and soft cheeses with serotype 1/2a.     

Temporal,environmental and management factors influencing the epidemiological patterns of sea lice (Lepeophtheirus salmonis) infestations on farmed Atlantic salmon (Salmo salar) in Scotland

In association with industry, an extensive database has been established on measurements of sea louse counts on farmed Atlantic salmon for the years 1996 to 2000 from 33 Scottish fish farms. These data include extensive counts on the sea louse species, Lepeophtheirus salmonis, at various stages of the life cycle and in particular the chalimus and mobile stages. There has been considerable speculation as to what factors might affect the abundance of sea lice, much of which is based on limited evidence. Our analyses show that there is tremendous variation in sea louse infestation patterns from year to year, whereas stock type, geographical region and coastal exposure do not appear to affect mean levels of abundance. In contrast, treatments lead to pronounced cycles of sea louse infestation with peaks and troughs at 3-week intervals, and these interventions are important if the sea louse levels on fish are to be controlled. There was no evidence of water temperature affecting the mean annual abundance of sea louse infestation.