Serological characterization of viruses isolated from experimental mucosal disease

Four cattle persistently infected with non-cytopathogenic bovine viral diarrhoea-mucosal disease (BVD-MD) virus and 3 normal controls were challenge exposed to cytopathogenic BVD-MD viruses that are antigenically different from the persistent viruses. Two of the persistently infected cattle developed mucosal disease and became moribund on postinoculation days (PID) 28 and 14, respectively; one developed severe and chronic diarrhoea and became moribund on PID 32; and the other remained healthy until the end of the experiment (PID 150). All control cattle showed transient fever, but no diarrhoea, and recovered from infection. Cytopathogenic viruses were isolated from blood of all cattle early in infection (PID 5-10) and from carcasses at necropsy. The former viruses were antigenically identical with the challenge viruses. On the other hand, the antigenicity of the cytopathogenic viruses isolated from carcasses at necropsy were different from that of the challenge viruses but similar to that of the non-cytopathogenic persistent viruses. Three of 4 persistently infected cattle, but not the calf which became moribund on PID 14, produced serum neutralizing (SN) antibodies to the challenge viruses, but not to the persistent viruses and the cytopathogenic viruses isolated from carcasses at necropsy. Control cattle produced SN antibodies to both the challenge and persistent viruses.     

Lineage classification of canine inheritable disorders using mitochondrial DNA haplotypes

To estimate the maternal effects of dog breeds using mitochondrial DNA(mtDNA) haplotypes in the dogs with several clinical disorders, 600 base pairs of mtDNA D-loop region were amplified from 365 dogs and were determined for mtDNA sequences. The diversity of the 600-bp sequences was classified into 64 haplotypes, including 46 newly discovered haplotypes, and the haplotypes were grouped into four clusters I to IV. Lineage analysis using the mtDNA haplotype indicated that each dog breed genetically comprises one or a few mtDNA haplotypes. When the relationship between genetic background and occurrences of clinical diseases was estimated, canine lineage analysis using mtDNA haplotype revealed that the disorders distributed in the dominant mtDNA haplotypes of each dog breed, but no disorder closely associated with mtDNA haplotypes was detected.     

Evaluation of Meat and Bone Meal as Replacement for Protein Concentrate in the Practical Diet for Sutchi Catfish,Pangasius hypophthalmus (Sauvage 1878), Reared under Pond Condition

An 80‐d feeding trial was conducted to evaluate meat and bone meal (MBM) as replacement of protein concentrate (PC) in practical diets for sutchi catfish, Pangasius hypophthalmus. Triplicate groups of juvenile sutchi catfish (initial weight 4.80 g) were stocked at the rate of 20,000 fish/ha in 12 earthen ponds (30 m² each). Four isocaloric diets (4.12 kcal/g) were formulated by replacing 0 (Diet 1, control), 33 (Diet 2), 67 (Diet 3), and 100% (Diet 4) PC protein with MBM. The growth parameters [final weight, % weight gain, and specific growth rate (SGR)] of fish fed Diets 1, 2, and 3 were not significantly different (P > 0.05) from each other but were significantly (P < 0.05) higher than those in Diet 4. The protein content in fish fed Diet 4 was significantly (P < 0.05) lower than the others. The total production of fish in different treatments ranged between 1917 and 2648 kg/ha/80 d. A simple economic analysis determined that the highest net profit (Tk. 52,965/ha) was obtained with Diet 2 and the lowest (Tk. 27,127/ha) with Diet 4 having 100% PC protein replacement. From the results of the study it is concluded that MBM can substitute up to 67% PC protein in catfish diet without hampering the growth and feed utilization.     

Development of an on-site plum pox virus detection kit based on immunochromatography

Sharka disease, caused by plum pox virus (PPV), is the most serious viral disease of stone fruit trees. Among the eight known strains of the virus, PPV-D is the most important due to its recent global spread. Although enzyme-linked immunosorbent assay (ELISA) is the most common approach for diagnosing sharka, it involves time-consuming steps and requires expensive equipment and trained technicians. In this study, an on-site PPV detection kit based on immunochromatography was developed using polyclonal antibodies against the coat protein (CP) of a PPV-D isolate. The immunochromatographic (IC) assay kit was as sensitive as a commercial ELISA system for detecting Japanese PPV-D isolates. Moreover, it was easy to use (a one-step procedure), and results could be obtained on-site within 15 min without special laboratory equipment. The IC assay kit detected the virus from every aerial part of symptomatic Japanese apricot trees. In a detailed study of viral localization in leaves, the most suitable plant parts for use in the IC assay were symptomatic mesophyll tissues and the region from the petiole to the main vein. A positive reaction was also observed using the CP of other major (PPV-M and PPV-Rec) and minor (PPV-EA, PPV-W, and PPV-T) strains.     

Dietary supplementation of β‐glucan improves growth performance,the innate immune response and stress resistance of red sea bream,Pagrus major

A 56‐day feeding trial was conducted to evaluate the effects of supplemented diets with β‐glucan (BG) at four levels [0 (D1), 250 (D2), 500 (D3) and 1000 (D4) mg BG kg^?1] on red sea bream, Pagrus major. The obtained results revealed a significant increase (P < 0.05) in final body weight, weight gain, specific growth rate, feed intake, body protein content, lysozyme activity and tolerance against low‐salinity stress test in all BG‐supplemented groups when compared with BG‐free group. Furthermore, D4 group resulted in a significant increase in feed efficiency ratio, protein gain, protein and lipid digestibilities, serum bactericidal activity and peroxidase content when compared with D1 group (P < 0.05). Haematocrit and plasma protein content in D3 group were significantly higher than those in D1 group (P < 0.05). Interestingly, BG supplementation decreased glutamic oxaloacetic transaminase (GOT) in D2 group and reactive oxygen metabolites in D2, D3 and D4 groups when compared with D1 group. Following low‐salinity stress test, significantly higher amounts of secreted mucus were observed in fish fed D2 and D4 diets than those from fish fed D1 diet (P < 0.05). In conclusion, the supplementation of BG improves growth, stress resistance and immune response of P. major.     

Lactic acid bacteria effective for regulating the growth of contaminant bacteria during the fermentation of <Emphasis Type="Italic">Undaria pinnatifida</Emphasis> (Phaeophyta)

ABSTRACT:   Lactic acid fermentation of seaweed is a recent topic and quite limited information is available on culture conditions. To know the suitable strains for use as a starter culture for seaweed fermentation, 14 lactic acid bacteria (LAB) strains, including 11 species, were tested in culture conditions prepared with or without salt. A commercial product of Undaria pinnatifida powder was used as a substrate for fermentation without sterilizing. Starter-suitability of the LAB strains was assessed from their predominance after culture. Among the tested strains, Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus casei and Lactobacillus rhamnosus showed high (>90%) predominance in their cultures, while control cultures prepared without inoculation of LAB did not show any detectable growth of acid producing bacteria and spoiled. A total of 102 strains not showing acid producing activity were isolated and characterized from spoiled cultures, and all the isolates were observed as Bacillus strains, including 64 strains (62.7%) and 16 strains (15.7%) of a Bacillus cereu s-related and B. fusiformis -related species, respectively. The Undaria substrate before fermentation contained culturable microorganisms at 1.4–3.1 × 10² CFU/g, but the Bacillus cereu s-related strain was not a major composition, suggesting a concern of selective growth of the Bacillus cereu s-related strain during the spoiled fermentation.     

Estimation of the outcrossing rate for annual Asian wild rice under field conditions

The Asian wild rice, Oryza rufipogon, has partial outcrossing behavior and shows high levels of genetic variation. To estimate an accurate outcrossing rate of annual form of O. rufipogon, two backcross lines (Lines R1 and R2) between Oryza sativa Nipponbare and O. rufipogon W630 were examined under field conditions. A chromosome survey confirmed that these lines had wild chromosomal segments at more than 92% of marker loci. As for the traits of glume, stamen and pistil, Line R1 showed similar floral morphology as that of O. rufipogon W630, whereas Line R2 had larger glumes. In 2005, 2006 and 2008, a total of 22 backcross plants were planted in the middle of wild rice plots. The successive progenies of each plant were examined using microsatellite markers that could clearly detect self-pollination and outcrossing. The outcrossing rates of Line R1 plants ranged from 4.04% to 25.50% with an average of 10.20%. This indicates that cross-pollination of wild rice is a chance event affected by many environmental factors. The outcrossing rates of Line R2 plants also varied, however, no significant difference was observed between the averages of Lines R1 and R2, suggesting that the glume sizes are not critical for outcrossing ability.     

Cloning of cDNAs encoding multiple subtypes of feline interferon-alpha from the feline epitherial cell line

Mammalian interferon (IFN)-alpha consists of a 23-amino acid signal peptide and a 166-amino acid mature protein. Feline (Fe) IFN-alpha has an extra unique molecule consisting of a 171-amino acid mature protein with a 5-amino acid insertion. We cloned eight new subtypes of cDNA encoding FeIFN- alpha from a feline epithelial cell line. Among all the FeIFN-alpha subtypes, including six that have previously been reported, the variations were found to be far less than those of IFN-alphas of other animals.     

Wind‐induced stock variation of the neon flying squid (Ommastrephes bartramii) winter–spring cohort in the subtropical North Pacific Ocean

Our examination of the neon flying squid (Ommastrephes bartramii) winter–spring cohort catch per unit effort (CPUE, an index of stock) revealed significant positive correlations with the interannual variations of observed chlorophyll‐a (Chl‐a) concentration and autumn–winter mixed layer depth (MLD) in the winter–spring feeding grounds of paralarvae and juveniles (130–170°E, 20–27°N). These correlations suggest the importance of integrated bottom‐up effects by the autumn–winter MLD for the neon flying squid stocks. However, the influence of autumn–winter MLD interannual variation in the forage availability for paralarvae and juveniles, i.e., particulate organic matter and zooplankton, has still been unclear. In this study, we use the lower trophic ecosystem model NEMURO, which uses the physical environmental data from the ocean reanalysis dataset obtained by the four‐dimensional variational (4DVAR) data assimilation method. The model‐based investigation enables us to clarify how the autumn–winter MLD controls the particulate organic matter and zooplankton abundance in the feeding grounds. Further, our investigation of the autumn–winter MLD interannual variation demonstrates that the stronger autumn wind in the feeding grounds develops a deeper mixed layer. Therefore, the deep mixed layer entrains nutrient‐rich water and enhances photosynthesis, which results in good feeding conditions for paralarvae and juveniles. Our results underline that the wind system interannual variation has critical roles on the winter–spring cohort of the neon flying squid stock.