Acute infection of encephalomyocarditis (EMC) virus in Syrian hamsters.

One-month-old Syrian hamsters of the APA and Std: golden strains were inoculated intraperitoneally with 10(5) PFU/head of the D variant of encephalomyocarditis (EMC) virus and examined virologically and pathologically up to 7 days after inoculation. APA hamsters developed apparent hyperglycemia due to pancreatic islet cell damage while Std:golden hamsters did not. Hamsters of both strains showed clear histopathologic changes in the testis with prominent viral replication as well as in the brain, heart and exocrine pancreas. The susceptibility to EMC virus-infection was higher in males than in females and in APA than in Std: golden hamsters.     

Homolog of FlhDC, a master regulator for flagellum synthesis: required for pathogenicity in Erwinia carotovora subsp. carotovora

 Erwinia carotovora subsp. carotovora (Ecc) is a causal agent of soft-rot diseases in a wide variety of plants. Here, we have isolated nonmotile mutants in Ecc by in vivo insertional mutagenesis using a transposon Tn5. The sequence disrupted by the Tn5 insertion in YMU1 and YMU5 mutants was highly homologous to that of flhC and flhD genes, respectively. They are involved in the initiation of the expression of flagellum-related genes in many gram-negative bacteria such as Escherichia coli and Salmonella. With electron microscopy, the flhC and the flhD homolog mutants were shown to be aflagellate. Furthermore, the virulence of these mutants was greatly reduced in Chinese cabbage and potato compared to that of the parental strain. These results suggest that flagellar formation is required for the pathogenicity of Ecc. Received: November 5, 2002 / Accepted: December 2, 2002 Acknowledgments This research was supported in part by Grant-in-Aid (12052210) and by a grant from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (13073).     

Including an additional systematic environmental effect within a generation in an evaluation model improves accuracy of prediction of breeding values in a closed herd of pigs

The present study evaluated the advantage of mixed‐model techniques over a selection index under different magnitudes of an additional systematic environmental effect (ASEE) in terms of accuracy of prediction and expected genetic gain. The data attempted to simulate a closed herd in a pig breeding program. The base population (G₀) consisted of 10 males and 50 females. Six generations (G₀ to G₅) were selected by using a selection index of three traits without overlapping. Additional systematic environmental constants with four levels in a generation were assigned from a uniform distribution at different ranges. Breeding values of animals in the last generation (G₅) were estimated on the basis of an index of individual phenotype (SI‐U), SI‐U adjusted for ASEE using a least‐squares mean (SI‐A), best linear unbiased prediction using an animal model excluding ASEE (AM‐E), and an animal model including ASEE (AM‐I). Accuracy of prediction and expected genetic gain were larger by the animal model than by the selection index, even if heritability of the traits selected was high and ASEE was set to zero. When ASEE was zero, the accuracy of prediction and expected genetic gain given by SI‐U and AM‐I were similar to those given by SI‐A and AM‐E, respectively. However, the differences in accuracy and expected gain between SI‐U and AI‐A and between AM‐I and AM‐E increased as the range of ASEE increased. It was concluded that selection based on an animal model was more effective than index selection, even if the herd environment was uniform and traits with high heritability were selected, and that it should be always included in an evaluation model, however slight any systematic environmental effect may be in a closed herd.     

Improving management for higher reproduction accelerates genetic improvement in closed herd of swine

The present study compared responses to selection at different conception rates and litter sizes at weaning in a simulated closed herd in a swine breeding program. The base population consisted of 10 males and 50 females, and 10 generations of selection was practiced by using individual phenotype or best linear unbiased prediction of breeding values for a trait with heritability (h²) of either 0.2 or 0.5. The probability of conception in a single mating was assumed to be 0.8, 0.9 or 1.0. Litter size at weaning was sampled randomly from a normal distribution with mean 8, 10 or 12 and variance 8.1225. Genetic response increased by approximately 6% for h² = 0.2 and approximately 5% for h² = 0.5 at generation 10 when conception rate was increased from 0.8 to 1.0. However, litter size at weaning did not affect response to selection. In conclusion, improving conception rate by environmental management increases genetic response indirectly in a breeding program of a closed swine herd.     

Laboratory diagnosis of canine GM2-gangliosidosis using blood and cerebrospinal fluid.

In the present study, laboratory techniques were used to diagnose canine GM2-gangliosidosis using blood and cerebrospinal fluid (CSF) that can be collected noninvasively from living individuals. Lysosomal acid beta-hexosaminidase (Hex) was measured spectrofluorometrically using 4-methylumbelliferyl N-acetyl-beta-D-glucosaminide and 4-methylumbelliferyl 7-(6-sulfo-2-acetamido-2-deoxy-beta-D-glucopyranoside) as substrates. Main isoenzymes A and B of Hex in leukocytes were also analyzed using cellulose acetate membrane electrophoresis. GM2-ganglioside in CSF was detected and determined quantitatively by using thin-layer chromatography/enzyme-immunostaining method with anti-GM2-ganglioside antibody. In normal dogs, Hex activities could be determined in leukocytes, serum, and CSF and the total activities were markedly reduced in all the enzyme sources in a dog with Sandhoff disease. Electrophoresis of a leukocyte lysate from a normal dog showed that the Hex A and Hex B were not separated distinctively with formation of a broad band, whereas there were no bands in electrophoresis of a lysate from a dog with Sandhoff disease, showing a deficiency in the total enzyme activity. GM2-ganglioside could be detected and determined quantitatively in as little as 100 microl of canine CSE GM2-ganglioside in CSF in a dog with Sandhoff disease increased to 46 times the normal level. In conclusion, the methods in the present study are useful for diagnosis of canine GM2-gangliosidosis. These techniques enable definitive and early diagnosis of canine GM2-gangliosidosis even if tissues and organs cannot be obtained.     

Mitochondrial DNA analysis of Nepalese domestic dwarf cattle Lulu*

Dwarf Lulu cattle, the only Bos Taurus type of cattle in Nepal, are raised under severe environments in the mountainous zone of that country. In the present study, the body measurement traits, cytogenetic and molecular genetic characteristics of the Lulu cattle are investigated. Blood samples were collected from 31 animals in four villages (altitudes 2590–3550 m) in the southern part of Mustang. The Lulu cattle had a normal karyotype with 2n = 60, XY or XX. Only one male examined had a large submetacentric X‐chromosome and a small submetacentric taurine type Y‐chromosome. The mitochodrial DNA (mtDNA) genotypes were analyzed by PCR mediated restriction fragment length polymorphisms, displacement (D)‐loop region PCR mediated single strand conformation polymorphisms, and D‐loop region sequences. Many base substitutions were found in the D‐loop region, suggesting that the Lulu cattle originated from at least 10 maternal lines. Three types of mtDNA from these cattle were found, the Bos taurus type (n = 23), the Bos indicus type (n = 6), and the Bos grunniens type (n = 2). In the village at the lowest altitude, four of the five cows were of the Bos indicus type. These results indicated that mtDNA types of the Lulu cattle mostly belong to Bos taurus, but have been hybridized with Bos indicus cattle in lower‐elevation regions in their maternal lineage.     

Nitrogen fertilization affects yields and storage compound contents in seeds of field-grown soybeans cv Enrei (Glycine max. L) and its super-nodulating mutant En-b0-1 through changing N2 fixation activity of the plants

ABSTRACT

We compared the protein and oil contents, seed yields, and natural abundance of ¹⁵N (σ¹⁵N) of seeds from the plants of the cultivar Enrei, which has moderate nodulating ability (Enrei) with those of its two near-isogenic lines (NILs), a super-nodulating genotype of Enrei, i.e., En-b0-1, and a non-nodulating genotype of Enrei, i.e., En1282. Plants of these three genotypes were grown on four types of field plots with different types of urea coated slow-release nitrogen (N) fertilizers (CUSLNFs) which delivered N in different manners during plant growth . The seed yield of the En1282 plants was similar to that of the Enrei plants but much greater than that of the En-b0-1 plants when the plants were grown on the field to which a CUSLNF with a long lifespan was applied. The amounts of protein per seed were positively correlated with those of oil per seed in the case of En1282 plants irrespective of the field N conditions. The seed protein contents were proportional to the seed weight in both En1282 and Enrei plants. Such a relationship was not observed in the En-b0-1 plants, and the protein and oil contents in the seeds of En-b0-1 plants were negatively correlated with each other. These observations suggested that the N supply to maturing seeds was a key to the protein contents in the En1282 and Enrei plants and that the C supply to maturing seeds was a key to the protein contents in the En-b0-1 plants. The comparison of the σ¹⁵N values, protein contents, and seed yields of seeds from En1282 plants suggested that the Enrei plants assimilated considerable amounts of N from the soil during the late plant growth. We concluded that high N₂ fixation activity depressed the amounts of oil per seed and changed the protein and oil contents in soybean seeds.     

A survey of antimicrobial resistance in Escherichia coli isolated from wild sika deer (Cervus nippon) in Japan

We examined the antimicrobial susceptibility of 848 Escherichia coli isolates from 237 feces samples of wild sika deer (Cervus nippon) captured between 2016 and 2019 in 39 of the 47 prefectures of Japan. Five of the 237 wild sika deer (2.1%) carried E. coli with resistance to at least one antimicrobial, and all the resistant isolates showed resistance to tetracycline. The resistant isolates contained antimicrobial resistance genes that were similar to those in E. coli derived from humans and farm animals. Although wild sika deer are not currently likely to be a source for the transmission of antimicrobial resistance in Japan, they can potentially mediate antimicrobial resistance spread by coming into contact with humans, animals, and their surroundings.     

Characterization of a novel nicotinamide adenine dinucleotide-cytochrome b5 reductase mutation associated with canine hereditary methemoglobinemia

Hereditary methemoglobinemia associated with nicotinamide adenine dinucleotide-cytochrome b5 reductase (b5R) deficiency is a rare autosomal recessive disorder in animals. Recently, nonsynonymous b5R gene (CYB5R3) variants have been reported to be associated with canine and feline hereditary methemoglobinemia. However, the underlying molecular mechanisms of canine and feline methemoglobinemia caused by these nonsynonymous variants have not yet been reported. Previously, we reported a Pomeranian dog family with hereditary methemoglobinemia, carrying CYB5R3 mutation of an A>C transition at codon 194 in exon 7, replacing an isoleucine residue with leucine (p.Ile194Leu). In this study, we investigated the enzymatic and structural properties of the soluble form of wild-type and Ile194Leu canine b5Rs to characterize the effects of this missense mutation. Our results showed that the kinetic properties of the mutant enzyme were not affected by this amino acid substitution. The secondary structure of the wild-type and Ile194Leu b5Rs detected by circular dichroism showed a similar pattern. However, the mutant enzyme exhibited decreased heat stability and increased susceptibility to trypsin hydrolysis. Moreover, the thermostability and unfolding measurements indicated that the mutant enzyme was more sensitive to temperature-dependent denaturation than the wild-type b5R. We concluded from these results that unstable mutant enzyme properties with normal enzymatic activity would be associated with hereditary methemoglobinemia in the Pomeranian dog family.