Analysis of antibody response by temperature-sensitive measles vaccine strain in the cotton rat model

Measles virus (MeV) vaccine strain, AIK-C, is temperature sensitive (ts), which is thought to be associated with attenuation of virus pathogenicity. In this study, replication and antibody response were examined in cotton rats using viruses carrying different forms of the P gene, which is responsible for the ts phenotype of strain AIK-C and its parental Edmonston strain. When cotton rats were inoculated intranasally, ts viruses neither replicated in lungs, nor reproducibly generated an antibody response. When inoculated intramusculary (i.m.), however, ts strains raised an antibody titer in all animals. This response was not observed when ultraviolet-inactivated virus was used. ts virus, inoculated i.m., was recovered from cotton rat drainage lymph nodes. These results suggest that ts virus, inoculated i.m., could replicate in the cotton rat, presumably at the superficial lymph node, and induce an antibody response. Therefore, cotton rats can serve as a small-animal model for investigating immune responses to safer ts vaccine, as well as recombinant vaccine using AIK-C as a vector for protection against other infectious agents.     

Pathogenicity of highly pathogenic avian influenza viruses of H5N1 subtype isolated in Thailand for different poultry species

Highly pathogenic avian influenza (HPAI) viruses of the H5N1 subtype have caused several rounds of outbreaks in Thailand. In this study, we used 3 HPAI viruses isolated in Thailand in January 2004 from chicken, quail, and duck for genetic and pathogenetic studies. Sequence analysis of the entire genomes of these isolates revealed that they were genetically similar to each other. Chickens, quails, domestic ducks, and cross-bred ducks were inoculated with these isolates to evaluate their pathogenicity to different host species. A/chicken/Yamaguchi/7/04 (H5N1), an HPAI virus isolated in Japan, was also used in the chicken and quail studies for comparison. All four isolates were shown to be highly pathogenic to chickens and quails, with 100% mortality by 10(6) EID50 inoculants of the viruses. They caused sudden death in chickens and quails within 2-4 days after inoculation. The mean death times (MDT) of quails infected with the Thai isolates were shorter than those of chickens infected with the same isolates. Mortality against domestic and cross-bred ducks ranged from 50 to 75% by intranasal inoculation with the 10(6) EID50 viruses. Neurological symptoms were observed in most of the inoculated domestic ducks and appeared less severe in the cross-bred ducks. The MDTs of the ducks infected with the Thai isolates were 4.8-6 days post-inoculation. Most of the surviving ducks infected with the Thai isolates had sero-converted until 14 dpi. Our study illustrated the pathobiology of the Thai isolates against different poultry species and would provide useful information for improving control strategies against HPAI.     

Highly invasive intracranial malignant schwannoma in a rat

A highly invasive intracranial malignant schwannoma containing several masses was detected in a 28-week-old male Crl:CD(SD) rat. Macroscopically, 3 masses were noted in the cranial cavity; one was present at the bottom of the cranial cavity and involved the trigeminal nerve, and the other two were in the parietal bone. Histologically, each mass consisted of fusiform cells with interlacing fascicular, wavy and nuclear pseudopalisading arrangements and round cells with cystic lesions. The tumor cells invaded not only the brain but also the parietal bone. In the brain, the tumor cells infiltrated diffusely into the leptomeningeal and perivascular spaces and parenchyma, in which the tumor cell morphology and invasive pattern closely resembled those of malignant astrocytoma and malignant reticulosis. Immunohistochemically, the tumor cells in the masses showed positive reactions for both S-100 protein and GFAP, while those in the cerebral invasion sites were negative for GFAP and less positive for S-100 protein. Electron microscopically, a single basal lamina layer and short intricate cell processes were confirmed in the tumor cells. From these results, the present tumor was diagnosed as a malignant schwannoma arising in the cranial cavity, probably originating from the trigeminal nerve. The present tumor is considered to be a relatively unique malignant schwannoma based on its growth and invasion patterns.     

Atlantic salmon fed a nutrient package of surplus methionine,vitamin B12, folic acid and vitamin B6 improved growth and reduced the relative liver size,but when in excess growth reduced

Feeding plant‐based diet through smoltification of Atlantic salmon requires verification of the optimal level of 1C nutrients. Here, we fed Atlantic salmon plant‐based diets containing three different surplus amounts of the 1C nutrients; methionine, cobalamin (vitamin B12), pyridoxine (vitamin B6) and folic acid during 6 weeks in fresh water, through smoltification, followed by 3 months on‐growing period in salt water. The three diets were fed to fish dispersed in triplicate tanks throughout the experiment. Mean start body weight was 32 g. Dietary methionine levels in the diets were 6.7, 9.2 and 11.7 g/kg. Dietary B6 was 6.75, 8.45 and 11 mg/kg. Cobalamin was 0.16, 0.18 and 0.20 mg/kg. While dietary folic acid was 2.9, 4.8 and 6.3 mg/kg, diets are referred to as low, medium and high 1C diet. All other amino acids were similar between diets. The results showed no differences in growth or feed utilization in the fresh water period, but following the on‐growing salt water period, differences between diets occurred. The fish fed the medium 1C diet showed better growth, as compared to fish fed the low or high 1C diet (p = .009). The medium 1C fed fish showed a relative lower liver weight compared with fish fed low or high 1C diet (p = .025). Condition factor was better in fish fed the medium and high 1C diet as compared to those fed the low 1C diet (p = .0006). As expected, free methionine in liver, plasma and muscle increased by dietary methionine inclusion. Surplus vitamins only had minor effect on tissue concentrations. Based on these findings, we conclude that the micronutrient and methionine level presented in the medium 1C diet improved the growth, liver size and condition factor; however, more research is needed to evaluate the optimal requirement level for each of the 1C nutrients.     

Identification and primary structures of eel type I collagen proα1, proα2 and proα3

Many bony fish type I collagens have a characteristic third chain designated as α3(I). However, much less is known about the primary structure and distinction of the proα(I) chains. Their cDNAs were cloned by RT-PCR from the muscle tissue of Japanese eel, Anguilla japonica. Three cDNAs coding for the triple-helical domain of fibrillar collagen were identified as proα1(I), proα2(I) and proα3(I) chains by sequencing selected tryptic peptides isolated from eel type I collagen subunit chains, α1(I), α2(I) and α3(I). Eel proα3(I) had high amino acid sequence identity (81 %) to its proα1(I). The distribution of seven Cys residues in the C-propeptide of proα3(I) was identical to that of proα1(I). There was a third Cys residue at the 1,268th position from the N-terminus in proα1(I), though a supposed Cys residue at the 1,264th position in proα3(I) was replaced by a Ser residue. Similar replacement has been observed in the proα3(I) chains of trout and zebrafish. These combined results suggest that replacement of the Cys residue allows for the identification of fish collagen proα(I) previously not identified as proα3(I).     

Effects of thyroid hormone on gonadotropin-induced steroid production in medaka,Oryzias latipes,ovarian follicles

Blood and ovarian samples were collected at intervals of 4h prior to spawning time from medaka (Oryzias latipes) that were maturationally synchronized with artificial photoperiod (14h light: 10h dark). Plasma estradiol-17β (E₂) levels increased rapidly from 16h before spawning and peaked at 8h before spawning. Follicle-enclosed oocytes (ovarian follicles) at different stages of development were isolated from the ovaries and used to study the in vitro effects of thyroid hormone (triiodothyronine; T₃) on pregnant mare serum gonadotropin (GTH)-induced E₂ production. GTH at a concentration of 100 IU/ml stimulated E₂ production by ovarian follicles collected between 32 and 16h before spawning. At 32h before spawning, T₃ (5 ng/ml) administered along with GTH (100 IU/ml) resulted in a 3.5 fold increase in E₂ production, compared with GTH administered alone. These results suggest that T₃ can act on ovarian follicles directly to modulate GTH-stimulated E₂ production in the medaka.