Evaluation and characterization of a novel probiotic Lactobacillus pentosus PL11 isolated from Japanese eel (Anguilla japonica) for its use in aquaculture

In the present study, a potential Lactobacilli probiotics were isolated from Japanese eels (Anguilla japonica) and characterized and evaluated for their possible use in eel farming. Sixteen Lactobacilli were isolated from intestines of Japanese eels, using selective media. The lactobacilli strains (represented as PL1 to PL16) were screened by their ability to produce digestive enzyme. Among these, three strains (PL11, PL13 and PL16) producing four digestive enzymes (amylase, cellulase, protease and phytase) simultaneously were characterized further using API ZYM kit. From these, PL11 (Lactobacillu (L.) pentosus) was identified as potential probiotics candidate producing 15 enzymes among 20 tested. Further examination of biological activities of PL11 revealed tolerance against pH, artificial bile juice and antibacterial activity against several fish pathogenic bacteria. The in vitro competitive exclusion assay also revealed 88.4% reduction in adhesion of fish pathogen (Edwardsiella tarda) by PL11 to host intestinal mucus. In vitro incubation of Japanese eel foregut with Baclight‐labelled PL11 showed colonization of the enterocyte surface by confocal and scanning electron microscopy. In summary, PL11 isolated from eels could serve as a potential probiotics with acid and bile tolerance, production of digestive enzymes, antibacterial activity and inhibition of fish pathogen adhesion to intestinal mucus.     

Genetic Diversity and Population Structure in the Heavily Exploited Korean Rockfish,Sebastes schlegeli,in Korea

The Korean rockfish, Sebastes schlegeli, is a valuable and intensively exploited species in Korea. We discuss the genetic diversity and genetic structure of four Korean rockfish populations using eight microsatellite loci. In total, 161 different alleles from 138 individuals were observed. Average allele number per locus ranged from 2.5 to 23 and allelic richness varied from 13.38 to 14.63 within a population. Despite a long history of stocking practices, we found very high levels of polymorphism (mean heterozygosity = 0.810), which is comparable to other congeneric species. No significant difference in genetic diversity and molecular genetic variance (F_ST and R_ST) was observed among four local samples (P > 0.1). Little indication of contemporary inbreeding (F_IS= 0.051) or population structure (K = 1) was detected. This absence of differentiation may reflect high levels of gene flow along the coast of Korea. Our study demonstrates that rockfish in Korea should be managed as a single unit. Currently, the species does not appear to be genetically threatened, but the potential for a rapid loss of genetic diversity remains. This information on the genetic characteristics of Korean rockfish populations has important implications for fisheries management and conservation efforts, and will aid in the sustainable exploitation of the fishing resources and the preservation of biodiversity.     

Efficacy of live virus vaccines against infectious laryngotracheitis assessed by polymerase chain reaction-restriction fragment length polymorphism

The efficacy of four different commercial live vaccines (vaccines A, B, C, and D) against the infectious laryngotracheitis virus (ILTV) was assessed in specific-pathogen-free (SPF) chickens. SPF chickens were vaccinated intraocularly at 6 wk old with ILTV live vaccines and were challenged intratracheally with the N91B01 strain of virulent Korean ILTV 2 wk after vaccination. The immunity against ILTV live vaccines was assessed by the incidence of latent infection by the challenge virus in the chickens' tracheas and trigeminal ganglia, the reisolation rate of the challenge virus, and the clinical signs in the chickens challenged with the N91B01 strain of ILTV. The latent infection in chickens was assessed by nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Our data showed that the clinical signs and challenge virus isolation were negative in all chickens receiving four difference commercial ILTV live vaccines. The viral DNA of the vaccine strain, but not that of the challenge virus, was detected in chickens vaccinated with vaccine A by nested PCR-RFLP. The viral DNAs of both the vaccine and challenge strains were detected from chickens vaccinated with vaccines B, C, and D. This study showed that only vaccine A can protect chickens from latent infection with the field virulent ILTV. We speculate that the efficacy of infectious laryngotracheitis live vaccines to protect chickens from latent infection with virulent ILTVs can be assessed by nested PCR-RFLP analysis.     

Effects of dietary electrolyte balance on the chemistry of blood and urine in lactating sows and sow litter performance

One hundred fifty-three sows (average parity of 2.2) were used to determine the effects of dietary electrolyte balance (calculated as mEq/kg of diet for Na + K - Cl) on sows and their litters during lactation. The sows were fed corn-soybean meal-based diets (1.0% lysine, 1.0% valine, 0.95% Ca, and 0.80% P; as-fed basis) starting on d 109 of gestation and throughout the 21-d lactation experiment. Dietary electrolyte balance (dEB) was 0, 100, 200, 350, and 500 mEq/kg (as-fed basis), well above and below the dEB of 185 mEq/kg found in a simple corn-soybean meal-based lactation diet. To achieve the desired dEB, diets had the following: 1) 1.8% HCl (6 N) and 1.06% CaCl2, 2) 1.0% CaCl2, 3) 0.04% NaHCO3, 4) 1.29% NaHCO3, and 5) 2.54% NaHCO3 (as-fed basis). Increasing dEB increased blood pH (linear and quadratic effects, P < 0.001), partial pressure of carbon dioxide (linear effect, P < 0.001), HCO3- concentration (linear and quadratic effects, P < 0.001), and blood base excess (linear and quadratic effects, P < 0.001). However, increased dEB resulted in lower blood concentrations of K (linear and quadratic effects, P < 0.04), Cl (linear and quadratic effects, P < 0.001), and ionized Ca (linear and quadratic effects, P < 0.001). Changing dEB did not affect ADFI; water usage, litter weight gain; sow weight change; sow backfat change; percentages of CP, lactose, and fat in the milk; percentage of sows returning to estrus; days to estrus; and number of pigs born alive in the subsequent litter (P = 0.06). However, piglet survivability to d 10 and overall was greatest with the lower dEB treatments (linear effect, P < 0.05). The pH (linear and quadratic effects, P < 0.001) and colony forming units of total bacteria (linear effect, P < 0.03) in the urine increased as dEB of the diet was increased. In conclusion, dEB had pronounced effects on the physiological status of sows and decreasing dEB below that in a simple corn-soybean meal-based diet decreased bacterial counts in the urine and increased piglet survivability. However, milk composition, sow and litter weights at weaning, and subsequent rebreeding performance of the sows were not affected by dEB.