An inactivated vaccine to control the current H9N2 low pathogenic avian influenza in Korea

The H9N2 subtype low pathogenic avian influenza is one of the most prevalent avian diseases worldwide, and was first documented in 1996 in Korea. This disease caused serious economic loss in Korea''s poultry industry.In order to develop an oil-based inactivated vaccine, a virus that had been isolated in 2001 (A/chicken/Korea/01310/2001) was selected based on its pathogenic, antigenic, and genetic properties. However, in animal experiments, the efficacy of the vaccine was found to be very low without concentration of the antigen (2⁷ to 2¹⁰ hemagglutinin unit). In order to overcome the low productivity, we passaged the vaccine candidate virus to chicken eggs. After the 20th passage, the virus was approximately ten times more productive compared with the parent virus. For the most part, the passaged virus maintained the hemagglutinin cleavage site amino acid motif (PATSGR/GLF) and had only three amino acid changes (T133N, V216G, E439D, H3 numbering) in the hemagglutinin molecule, as well as 18 amino acid deletions (55-72) and one amino acid change (E54D) in the NA stalk region. The amino acid changes did not significantly affect the antigenicity of the vaccine virus when tested by hemagglutination inhibition assay. Though not complete, the vaccine produced after the 20th passage of the virus (01310 CE20) showed good protection against a homologous and recent Korean isolate (A/chicken/Korea/Q30/2004) in specific pathogen- free chickens.The vaccine developed in this study would be helpful for controlling the H9N2 LPAI in Korea.     

Molecular characterization and genogrouping of VP1 of aquatic birnavirus GC1 isolated from rockfish Sebastes schlegeli in Korea

The cDNA nucleotide sequence of genome segment B encoding the VP1 protein was determined for the aquatic birnavirus GC1 isolated from the rockfish Sebastes schlegeli in Korea. The VP1 protein of GC1 contains a 2,538 bp open reading frame, which encodes a protein comprising 846 amino acid residues that has a predicted MW of 94 kDa. The sequence contains 6 potential Asn-X-Ser/Thr motifs. Eight potential Ser phosphorylation sites and 1 potential Tyr phophorylation site were also identified. GC1 contains the Leu-Lys-Asn (LKN) motif instead of the typical Gly-Asp-Asp (GDD) motif found in other aquatic birnaviruses. We also identified the GLPYIGKT motif, the putative GTP-binding site at amino acid position 248. In total, the VP1 regions of 22 birnavirus strains were compared for analyzing the genetic relationship among the family Birnaviridae. Based on the deduced amino acid sequences, GC1 was observed to be more closely related to the infectious pancreatic necrosis virus (IPNV) from the USA, Japan, and Korea than the IPNV from Europe. Further, aquatic birnaviruses containing GC1 and IPNV have genogroups that are distinct from those in the genus Avibirnaviruses and Entomo-birnaviruses. The birnavirusstrains were clustered into 5 genogroups based on their amino acid sequences. The marine aquatic birnaviruses (MABVs) containing GC1 were included in the MABV genogroup; the IPNV strains isolated from Korea, Japan, and the USA were included in genogroup 1 and the IPNV strains isolated primarily from Europe were included in genogroup 2. Avibirnaviruses and entomobirnaviruses were included in genogroup 3 and 4, respectively.     

Effect of placenta feeding to sows on plasma biochemical composition and growth of their piglets

The objective of the present study was to investigate the effects on the plasma biochemical composition and growth of piglets of placenta‐fed sows. The experimental animals were a total of 21 sows (L × LW) of third parity, and each sow was suckled by 11 ± 1 piglets. Sows fed without placenta were considered as the control (C), treatment 1 (T1) were placenta‐fed from days 1–7 after farrowing and treatment 2 (T2) were placenta‐fed from day 7 before farrowing to day 7 after farrowing. Glucose of piglets was lower (P < 0.05) in T2 than C and T1 on day 10. Total cholesterol of piglets was higher (P < 0.05) in T2 than C on day 4. Fe of piglets was higher (P < 0.05) in T2 than C on days 4, 20 and 30. Unsaturated iron‐binding capacity was lower (P < 0.05) in T2 than C on days 4 and 30. Immunoglobulin G of piglets was higher (P < 0.05) in T1 and T2 than C on days 4 and 10. Daily gain of piglets was higher (P < 0.05) in T1 than C and T2 on day 4 when daily gain was calculated. Total daily gain increased (P < 0.05) by prolongation of placenta feeding period. The results indicate that feeding placenta to sows may improve the plasma biochemical composition and growth of their piglets.     

The radioprotective effects of the hexane and ethyl acetate extracts of Callophyllis japonica in mice that undergo whole body irradiation

The radioprotective activity of extracts from the red seaweed Callophyllis (C.) japonica was investigated in mice that underwent whole-body exposure to gamma radiation. A methanol extract of C. japonica and its fractions [hexane, ethyl acetate (EtOAc), butanol and the remaining H₂O] were used. Each fraction (100 mg/kg body weight) was administered intraperitoneally (i.p.) 2 times into the BALB/c mice, once at 1 and once at 24 h before exposure to 9 Gray (Gy) of gamma radiation. Pre-irradiation administration of the hexane and EtOAc fractions saved the mice, with their survival rates being greater than 80% at 30 days post-irradiation; the mice that were pretreated with the other fractions showed survival rates lower than 20% over the same time period. To examine the effect of each C. japonica fraction on the survival of intestinal and bone marrow stem cells, the number of intestinal crypts and bone marrow cells in the gamma-irradiated mice were examined. Pre-treatment of mice (i.p., 100 mg/kg body weight at 1 and 24 h before irradiation) with the hexane or EtOAc fraction prior to 6-Gy irradiation significantly protected the number of jejunal crypts and bone marrow cells at 9 days after irradiation. These findings suggest that certain extracts from C. japonica, when they are administered prior to irradiation, play an important role in the survival of irradiated mice, and this is possibly due to the extracts protecting the hematopoietic cells and intestinal stem cells against gamma irradiation.     

Effects of aluminum on the reduction of neural stem cells,proliferating cells,and differentiating neuroblasts in the dentate gyrus of D-galactose-treated mice via increasing oxidative stress

Aluminum (Al) accumulation increases with aging, and long-term exposure to Al is regarded as a risk factor for Alzheimer''s disease. In this study, we investigated the effects of Al and/or D-galactose on neural stem cells, proliferating cells, differentiating neuroblasts, and mature neurons in the hippocampal dentate gyrus. AlCl₃ (40 mg/kg/day) was intraperitoneally administered to C57BL/6J mice for 4 weeks. In addition, vehicle (physiological saline) or D-galactose (100 mg/kg) was subcutaneously injected to these mice immediately after AlCl₃ treatment. Neural stem cells, proliferating cells, differentiating neuroblasts, and mature neurons were detected using the relevant marker for each cell type, including nestin, Ki67, doublecortin, and NeuN, respectively, via immunohistochemistry. Subchronic (4 weeks) exposure to Al in mice reduced neural stem cells, proliferating cells, and differentiating neuroblasts without causing any changes to mature neurons. This Al-induced reduction effect was exacerbated in D-galactose-treated mice compared to vehicle-treated adult mice. Moreover, exposure to Al enhanced lipid peroxidation in the hippocampus and expression of antioxidants such as Cu, Zn- and Mn-superoxide dismutase in D-galactose-treated mice. These results suggest that Al accelerates the reduction of neural stem cells, proliferating cells, and differentiating neuroblasts in D-galactose-treated mice via oxidative stress, without inducing loss in mature neurons.     

Effects of different forages on the chemical compositions and antiosteoporotic activities of velvet antlers

For this study, we aimed to assess the dose–response antiosteoporotic effects of the middle section of velvet antlers (VAs) from sika deers (Cervus nippon) fed with different types of fodders. VAs prepared from farmed sika deers fed with feed mixtures containing sorghum distillery residue (VA‐SDR) or without SDR (SDR replaced with hay, VA‐Hay) were divided into upper (VAU), middle (VAM) and basal (VAB) sections. The chemical constituents of the middle sections obtained from each VA type were compared, and their antiosteoporotic activities were evaluated using rats with ovaries removed surgically (ovariectomy, OVX). The VA‐Hay exhibited markedly increased iron and cysteine levels, whereas the VA‐SDR exhibited markedly increased level of alcoholic extract and testosterone. Both VA‐Hay‐ and VA‐SDR‐treated rats exhibited increased femur strength compared with the control group. However, VA‐SDR exhibited greater bone‐strengthening effects than did VA‐Hay. The serum osteocalcin and estradiol levels were significantly moderated in the VA‐Hay group alone. These results suggest that VA‐SDR and VA‐Hay prevent the loss of bone strength, and preserve trabecular architecture connectivity in an estrogen‐deficient state. However, differences in the chemical compositions of different forages may be responsible for the varying antiosteoporotic mechanisms observed. Thus, the addition of SDR in deer forage may enhance antiosteoporosis activity in VAs, and confer considerable economic and ecological benefits.     

Selection and characterization of broad‐spectrum antibacterial substance‐producing Lactobacillus curvatus PA40 as a potential probiotic for feed additives

Lactic acid bacteria were screened for potential probiotics for use as feed additives. We obtained 3,000 isolates from feces of: cattle, dogs, goats, and infants; milk; yogurt; cheese; fermented sausages; Kimchi; and Cheonggukjang and tested their antibacterial activity toward indicator pathogens, including Bacillus cereus, Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella enterica Enteritidis. We further tested their tolerance to artificial gastric juice (1% [w/v] pepsin, pH 2.5) and bile acid (0.1% [w/v] oxgall, pH 6.8). Six isolates exhibited strong antibacterial activity against indicator pathogens. The PA40 isolate from Kimchi exhibited marked resistance to artificial gastric juice and bile acid. The antibacterial substances produced by PA40 were stable to heat, pH, and enzymes. Strain PA40 was identified as a Lactobacillus curvatus strain using chemical tests and 16S rDNA sequencing and produced 248.4 mmol/L lactic acid after 48 hr of fermentative growth. The L. curvatus PA40 strain was also highly tolerant of the artificial gastrointestinal model system. Our results indicate that L. curvatus PA40 could be used as a potential probiotic feed additive.     

Effects of Lonicera japonica extract supplementation on in vitro ruminal fermentation,methane emission,and microbial population

Lonicera japonica (LJ; honeysuckle) is used in traditional folk medicine in Korea and is a rich source of ascorbic acid and phenolic components that are reported to have antioxidant and antibiotic properties. We performed an in vitro experiment to assess the effects of LJ extracts (LJE) on ruminal fermentation. Timothy hay (0.3 g dry matter [DM]) was incubated with buffer, ruminal fluid, and 0%, 3%, 5%, 7%, and 9% LJE. Batch culture fermentation was conducted separately for 12, 24, and 48 hr to determine gas production (GP), ruminal fermentation characteristics, and microbial population characteristics. The effects on GP were generally similar to those on DM degradability, with a linear decrease observed at 9% extract at 24 hr. NH₃‐N showed a linear increase with increasing extract concentrations at 12 hr, whereas a decrease was observed at 24 hr. Extract supplementation decreased methane (CH₄) production at 12, 24, and 48 hr. In addition, the abundance of fibrolytic bacteria and ciliate‐associated methanogen was reduced at all concentrations of extracts. These results indicate that LJE have the potential to serve as a ruminal fermentation modifier to suppress CH₄ production with minimal effects on nutrient digestion in the rumen.