Effects of Feeding Rate and Feeding Frequency on Survival, Growth, and Body Composition of Ayu Post-Larvae Plecoglossus altivelis

The effects of feeding rate and feeding frequency on survival, growth and body composition of ayu post‐larvae (0.15 g in body weight and 3.5 cm in total length) were investigated in this study. A factorial experimental design of two feeding rates (3 and 6% of body weight of fish per meal) five feeding frequencies (one meal in 2 d, one meal a day, two meals a day, four meals a day, and six meals a day) with three replicates was used. Survival of ayu post‐larvae was significantly (P 0.05) affected by feeding frequency but not by feeding rate. Survival of ayu improved linearly with feeding frequency at both feeding rates. Weight and length gains and specific growth rate (SGR) of ayu was significantly (P 0.05) affected by feeding frequency but not by feeding rate, with weight and length gains and SGR linearly elevated with increasing feeding frequency at both feeding rates. The greatest weight and length gains were observed in fish receiving six meals daily at both feeding rates; however, no significant difference in weight gain was observed among two, four, and six meals a day, or in length gain between four and six meals a day. Feed efficiency ratio (FER) was significantly (P 0.05) affected by both feeding rate and feeding frequency. FER linearly decreased with feeding frequency at both feeding rates or feeding rate in the same feeding frequency. When the total daily amount of feed supply was constant with various feeding frequencies at different feeding rates (one meal a day at 3% feeding rate and one meal in 2 d at 6% feeding rate, two meals a day at 3% feeding rate and one meal a day at 6% feeding rate, or four meals a day at 3% feeding rate and two meals a day at 6% feeding rate), improvement in survival, weight and length gains, and SGR was observed in fish with higher feeding frequency at lower feeding rate. Moisture, protein, and lipid content of fish were not significantly (P > 0.05) affected by either feeding rate or feeding frequency. However, lipid content of ayu linearly increased with feeding frequency at 6% feeding rate. The highest body lipid content was observed in fish receiving six meals daily at both feeding rates. Ash content of fish was significantly (P 0.05) affected by feeding frequency but not by feeding rate. Based on performance of ayu, it can be concluded that optimum feeding rate and feeding frequency for ayu post‐larvae (an initial weight of 0.15 g) were 3% per meal and four meals a day, respectively, under these experimental conditions.     

Protection of chickens from Newcastle disease with a recombinant baculovirus subunit vaccine expressing the fusion and hemagglutinin-neuraminidase proteins

Recombinant baculoviruses containing the fusion (F) and hemagglutinin-neuraminidase (HN) glycoprotein gene of the viscerotropic velogenic (vv) Newcastle disease virus (NDV) isolate, Kr-005/00, and a lentogenic La Sota strain of the NDV were constructed in an attempt to develop an effective subunit vaccine to the recent epizootic vvNDV. The level of protection was determined by evaluating the clinical signs, mortality, and virus shedding from the oropharynx and cloaca of chickens after a challenge with vvNDV Kr-005/00. The recombinant ND F (rND F) and recombinant HN (rND HN) glycoproteins derived from the velogenic strain provided good protection against the clinical signs and mortality, showing a 0.00 PI value and 100% protection after a booster immunization. On the other hand, the combined rND F + HN glycoprotein derived from the velogenic strain induced complete protection (0.00 PI value and 100% protection) and significantly reduced the amount of virus shedding even after a single immunization. The rND F and rND HN glycoproteins derived from the velogenic strain had a slightly, but not significantly, greater protective effect than the lentogenic strain. These results suggest that the combined rND F + HN glycoprotein derived from vvNDV can be an ideal subunit marker vaccine candidate in chickens in a future ND eradication program.     

Analysis of radiosensitivity of cancer stem‐like cells derived from canine cancer cell lines

Cancer stem‐like cells (CSCs) are self‐renewing cells comprising a small subpopulation in tumours, and generate differentiated progeny through asymmetric division. It has been shown that CSCs are resistant to ionizing radiation, and this feature could be one of the mechanisms of tumour recurrence after radiation therapy. Much attention has been focused on to target CSCs; however, difficult of isolating CSCs and lack of knowledge on their radiosensitivity have limited this kind of research in veterinary medicine. In the present study, sphere‐forming cells (SC), cultured using sphere formation method, were isolated from four type of canine tumour cell lines and evaluated if they have CSCs‐like properties by expression of CSCs markers (real‐time polymerase chain reaction) and capacity of tumorigenesis (xenograft transplantation in nude mice), and were assessed radiosensitivity (clonogenic survival assay) and DNA repair kinetics (immunofluorescence staining for p53‐binding protein 1) after X‐ray irradiation in comparison with the corresponding normal adherent culture cells (AC). All SCs were isolated using sphere formation and showed high gene expression of CD133 and tumorigenic ability as compared with AC. All SCs were significantly resistant against X‐ray irradiation as compared with AC. In addition, the amount of DNA double‐strand breaks after X‐ray irradiation were significantly lower in SC compared with the corresponding AC. These results indicate that SC isolated through sphere formation possess CSCs‐like characteristics and CSCs are important factor that affect radiosensitivity in canine tumours. In addition, radioresistance of CSCs may depend on reaction of DNA double‐strand break after X‐ray exposure.     

Preparation of monodisperse poly(vinyl alcohol) (PVA) nanoparticles by dispersion polymerization and heterogeneous surface saponification

Monodisperse poly(vinyl alcohol) (PVA)/poly(vinyl acetate) (PVAc) nanoparticles with a skin-core structure were prepared through heterogeneous surface saponification of PVAc nanoparticles. For the preparation of PVAc nanoparticles with a uniform particle size distribution, vinyl acetate (VAc) was dispersion polymerized in a mixed solvent of ethanol and water using PVA with a low degree of saponification as a stabilizer. Increase of the amount of ethanol in media, the resulting PVAc nanoparticle size increases due to increasing solubility of VAc and oligomer PVAc. To preserve the sphericity and size uniformity of PVAc nanoparticles, we restricted saponification to the surface of the nanoparticles by using a small amount of aqueous sodium hydroxide solution. To determine the proper concentration of alkali solution for heterogeneous saponification, monodisperse PVAc nanoparticles were saponified with different concentrations of alkali solution at 25 °C for 0.5–3.0 h. The PVA/PVAc nanoparticles obtained by the heterogeneous saponification with 4 % (relative to the amount of the VAc) alkali solution for 2.0 h were uniformly shaped and monodispersed with diameter ranging from 428 to 615 nm. Transmission electron microscopy (TEM) confirmed the spherical nature and regular skin-core structure of the PVA/PVAc nanoparticles.     

Reduction of adult hippocampal neurogenesis is amplified by aluminum exposure in a model of type 2 diabetes

In this study, we investigated the effects of chronic aluminum (Al) exposure for 10 weeks on cell proliferation and neuroblast differentiation in the hippocampus of type 2 diabetic rats. Six-week-old Zucker diabetic fatty (ZDF) and Zucker lean control (ZLC) rats were selected and randomly divided into Al- and non-Al-groups. Al was administered via drinking water for 10 weeks, after which the animals were sacrificed at 16 weeks of age. ZDF rats in both Al- and non-Al-groups showed increases in body weight and blood glucose levels compared to ZLC rats. Al exposure did not significantly affect body weight, blood glucose levels or pancreatic β-cells and morphology of the pancreas in either ZLC or ZDF rats. However, exposure to Al reduced cell proliferation and neuroblast differentiation in both ZLC and ZDF rats. Exposure to Al resulted in poor development of the dendritic processes of neuroblasts in both ZLC and ZDF rats. Furthermore, onset and continuation of diabetes reduced cell proliferation and neuroblast differentiation, and Al exposure amplified reduction of these parameters. These results suggest that Al exposure via drinking water aggravates the impairment in hippocampal neurogenesis that is typically observed in type 2 diabetic animals.     

Vacuolar invertases in sweet potato: molecular cloning, characterization, and analysis of gene expression

Two cDNAs (Ib beta fruct2 and Ib beta fruct3) encoding vacuolar invertases were cloned from sweet potato leaves, expressed in Pichia pastoris, and the recombinant proteins were purified by ammonium sulfate fractionation and chromatography on Ni-NTA agarose. The deduced amino acid sequences encoded by the cDNAs contained characteristic conserved elements of vacuolar invertases, including the sequence R[G/A/P]xxxGVS[E/D/M]K[S/T/A/R], located in the prepeptide region, Wxxx[M/I/V]LxWQ, located around the starting site of the mature protein, and an intact beta-fructosidase motif. The pH optimum, the substrate specificity, and the apparent K(m) values for sucrose exhibited by the recombinant proteins were similar to those of vacuolar invertases purified from sweet potato leaves and cell suspensions, thus confirming that the proteins encoded by Ib beta fruct2 and Ib beta fruct3 are vacuolar invertases. Moreover, northern analysis revealed that the expression of the two genes was differentially regulated. With the exception of mature leaves and sprouting storage roots, Ib beta fruct2 mRNA is widely expressed among the tissues of the sweet potato and is more abundant in young sink tissues. By contrast, Ib beta fruct3 mRNA was only detected in shoots and in young and mature leaves. It appears, therefore, that these two vacuolar invertases play different physiological roles during the development of the sweet potato plant.     

Cloning and characterization of a plant defensin VaD1 from azuki bean

A recombinant mungbean defensin VrD1 was previously shown to exhibit antifungal and bruchid-resistant activity. To study the function and regulation of VrD1, genomic DNAs of plant defensins were isolated from Vigna radiata VC6089A and azuki bean Vigna angularis Kao Hsiung No. 6. The azuki bean defensin genomic DNA VaD1 was sequenced and converted to VaD1 cDNA. VaD1 defensin was purified from Vigna angularis Kao Hsiung No. 6 to apparent homogeneity. The complete amino acid sequence of the purified VaD1 was determined and was found to be exactly the same as the sequence deduced from VaD1 cDNA. VaD1 is a basic protein containing 46 amino acids with four conserved disulfide bonds and shares high sequence homology (78.3%) with VrD1. VaD1 inhibited the growth of Fusarium oxysporum, Fusarium oxysporum f. sp. pisi, Staphylococcus epidermidis, and Salmonella typhimurium. VaD1 also inhibited in vitro protein synthesis and bruchid larval development, but was less active than the recombinant VrD1.     

Seroconversion to avian influenza virus in free-range chickens in the Riverland region of Victoria

Background Since 2005, H5N1 avian influenza (AI) has spread from South-East Asia to over 60 different countries, resulting in the direct death or slaughter of over 250,000,000 poultry. Migratory waterfowl have been implicated in this spread and in Australia there have been numerous isolations of low-pathogenicity AI virus from wild waterfowl and shorebirds. The Department of Human Services, Victoria maintains 10 sentinel free-range chicken flocks in the Riverland at locations that are populated by large numbers of waterfowl known to carry a range of strains of AI. Objective This study analysed historical samples collected in 1991–94 and 2003–06 from the library of serum samples for antibodies against AI to assess the potential for transfer of AI virus from wild waterfowl to free-range poultry. Results Of the 2000 serum samples analysed, 17 were positive for antibodies against AI and 87 were suspect, with a clustering of positive and suspect results in the years 1994, 2003 and 2004. There was also a clustering of positive samples at the site of the Barmah flock. Nine sequential sets of sera from individual chickens with at least one positive result were identified. Analysis of these sequential sets showed that infection was acquired on site but that the antibody response to AI infection was short-lived and was no longer detectable at 8 weeks after the positive finding. Conclusion The surveillance of sentinel chickens is a potential avenue for monitoring the circulation of AI viruses and could provide an early warning system for the commercial poultry industries.     

Carcass characteristics of Hanwoo (Korean cattle) from different sex conditions, raising altitudes and slaughter seasons

This study was carried out to investigate the carcass characteristics of Hanwoo (Korean cattle) from different sex conditions, raising altitudes and slaughter seasons. The total number of cattle used in this study was 3608 heads which comprised of 1336 bulls and 1660 steers. The data was analyzed according to sex condition (bull and steer), raising altitude (low: 0–100 m and high: 700–800 m) and slaughter season (spring, summer, autumn and winter). At 24 h post-slaughter, the carcasses were weighed and evaluated for carcass traits according to the Korean carcass grading standard by an official grader. Carcass weight, ribeye area, yield index and grade, and meat color, firmness and maturity scores of carcass from bulls were significantly higher (P < 0.001) than those from steers. Inversely, the backfat thickness, marbling score and quality grade of carcass from steers were significantly higher (P < 0.001) than those from bulls. The maturity score of carcass from a high area was significantly higher (P < 0.01) than that from a low area. The ribeye area of carcass from summer was significantly smaller (P < 0.05) than that from other seasons. The maturity score of carcass from the summer season was highest among that from other seasons and the lowest was that from the winter season. Marbling score and carcass quality grade from the winter season were highest among those from other seasons and the lowest were those from the autumn season. There were no significant interactions between sex condition and raising altitude on carcass traits except the ribeye area. There were significant interactions between sex condition and slaughter season on marbling score and carcass quality grade. There were no significant interactions between raising altitude and slaughter season on all of the carcass traits. There were significant interactions among sex condition, raising altitude and slaughter season on meat color score. It was concluded that sex condition affected muscle and fat depositions on the carcass, raising altitude affected maturity and slaughter season affected ribeye area, maturity and marbling.     

Experimental reactivation of latent canine herpesvirus-1 and induction of recurrent ocular disease in adult dogs

Latent canine herpesvirus-1 (CHV-1) infection is common in domestic dogs, but recrudescent CHV-1 diseases are poorly characterized. To determine if administration of an immunosuppressive dosage of prednisolone to adult dogs latently infected with CHV-1 results in recurrent ocular disease, adult beagles with and without experimentally induced CHV-1 latent infection were divided into groups: group 1 latently infected and administered prednisolone, group 2 latently infected and administered placebo, and group 3 not latently infected and administered prednisolone. Prednisolone (3.0 mg/kg/day) was administered to dogs in groups 1 and 3 for seven consecutive days beginning on study day 1. Samples for CHV-1 polymerase chain reaction and serum neutralization (SN) assays were collected, and physical, ophthalmologic, and in vivo ocular confocal microscopic examinations were performed at intervals for 42 days. Bilateral ocular disease (i.e., conjunctivitis or keratitis) was detected in 83% of group 1 dogs between study days 3 and 18. In vivo confocal microscopic abnormalities included conjunctival leukocyte infiltration and corneal leukocyte infiltration, abnormal epithelial cell morphology, and Langerhans cell infiltration. Ocular viral shedding was detected in 50% of group 1 dogs on study days 10 and 13. Fourfold elevations in CHV-1 SN titers were detected in 100% of group 1 dogs by study day 14. Dogs in control groups did not develop clinical ocular disease (P < 0.05), CHV-1 titer elevations (P < 0.005), or viral shedding. Administration of an immunosuppressive dosage of systemic prednisolone to adult dogs latently infected with CHV-1 may result in viral reactivation and ocular disease recrudescence.