Long-Term Observation of Fogwater Composition at two Mountainous Sites in Gunma Prefecture, Japan

We made serial observations on acid fog at Mt. Akagi and Mt. Haruna, Japan for 5 years. The altitudes of the sampling sites were 1500 m (Mt. Akagi) and 1200 m (Mt. Haruna) above sea level, and the sites were approximately 30 km apart. The average liquid water content (LWC) at Mt. Akagi and Mt. Haruna was 74 mg m^?3 and 63 mg m^?3, respectively. The pH of fogwater was 2.72–7.14 (mean 3.71) at Mt. Akagi and 2.94–6.58 (mean 3.73) at Mt. Haruna. Our long-term observations indicate that there was no significant difference in the chemical components in fogwater at both sites except for ammonium ion. However, there were some cases where the chemical components of fogwater at each site were differed remarkably even in the concurrent fog event. Nitrate and sulfate ions contributed to acidification of fogwater at Mts. Akagi and Haruna and 95% of sulfate ion in the fogwater originated from air pollution. Ammonia gas in the air was the main neutralizer of acidity in fogwater. When absorption of excessive nitric acid gas over ammonia gas in the air occurred, the pH of fogwater was lowered. Our back trajectory analysis indicated that the fogwater at Mt. Akagi was mainly affected by an air mass from the Kanto Plain, including Tokyo, while the fogwater at Mt. Haruna was influenced by an air mass from large, western cities, such as Nagoya and Osaka, as well as Tokyo.     

Comparative sequence analysis of four myosin heavy chain isoforms expressed in porcine skeletal muscles: Sequencing and characterization of the porcine myosin heavy chain slow isoform

A nucleotide sequence including the full coding region for the myosin heavy chain (MyHC) slow isoform was determined from the longissimus skeletal muscle. The deduced amino acid sequence was 1935 residues, which was the same length as the human and rat MyHC-slow isoforms. The porcine MyHC-slow isoform showed 97.6% and 97.4% amino acid identities to the human and rat isoforms on their entire regions, respectively. The functional regions were also highly conserved among mammalian MyHC-slow isoforms. Amino acid substitutions between the porcine MyHC-slow and MyHC-fast isoforms were concentrated on the functional regions. Loop 1, the controlling region of nucleotide binding and release, was conserved among the fast isoforms, but not between the slow and fast isoforms. Loop 2, a part of the actin binding region, was not conserved among any of the isoform types, and the most substitutions in this region were found in the slow isoform. The myosin essential light chain binding region was conserved among the fast isoforms, except for some substitutions in the 2b isoform, but was clearly different in the slow isoform. The myosin regulatory light chain binding region was conserved among the fast isoforms, but not between the slow and fast isoforms. These results indicate that the functional difference between the porcine MyHC-slow and -fast isoforms are controlled by the sequence diversity at the four functional regions compared.     

Detection of the esp gene in high-level gentamicin resistant Enterococcus faecalis strains from pet animals in Japan

We investigated the prevalence of the esp gene and the susceptibility to gentamicin in Enterococcus faecalis and E. faecium strains obtained from pet animals. Nine of 30 E. faecalis and 2 of 38 E. faecium strains from the pet animals had the esp gene. Three esp-positive E. faecalis strains, which were isolated from two dogs and a cat, showed gentamicin MICs of > or =256 microg/ml and harbored the high-level gentamicin resistance (HLGR) gene, aac(6')-Ie-aph(2')-Ia. Of the nine esp-positive E. faecalis strains, five, including the three strains with the HLGR gene, were closely related by numerical analysis of PFGE patterns. Longitudinal investigation needs to elucidate whether the HLGR gene was incorporated into a subpopulation of the esp-positive E. faecalis.     

Lymphocyte blastogenic responses to inciting food allergens in dogs with food hypersensitivity

Lymphocyte blastogenic responses against food allergens in dogs with food hypersensitivity were evaluated in this study. Eleven dogs with food hypersensitivity, based on food elimination and oral food provocation tests and allergic responses to food allergens, were examined by various tests such as intradermal testing, antigen-specific IgE testing, and lymphocyte blastogenic responses. The number and kinds of food allergens identified as positive by these tests were compared with the offending food allergens that were found in an oral food provocation test. In 9 (82%) of the 11 dogs with food hypersensitivity, there was close agreement for positive allergens between the results of lymphocyte blastogenic responses and oral food provocation test; however, there was little agreement for intradermal and IgE testing of the positive allergens with those of the oral food provocation test (11% and 31%, respectively). In the 9 dogs, the stimulation indices of lymphocyte blastogenic responses increased to 2.0-10.1 upon food provocation but decreased significantly to 0.7-1.4 upon feeding the elimination diet until clinical signs disappeared. These results indicate that lymphocyte blastogenic responses may fluctuate because of exposure to offending food allergens in dogs with food hypersensitivity. Lymphocytes reactive to food allergens may play an important role in the pathogenesis of food hypersensitivity in dogs.     

A regional evaluation of chromium tripicolinate supplementation of diets fed to reproducing sows

A cooperative research study involving 353 litters was conducted at three stations to determine the effects of graded levels of supplemental Cr from chromium tripicolinate (CrPic) on reproductive performance of sows and preweaning performance of their pigs. Primiparous and multiparous sows were fed fortified corn-soybean meal diets with supplemental levels of 0, 200, 600, or 1,000 ppb Cr (as-fed basis). Each station used at least three of the supplemental Cr levels, with two of those levels being 0 and 200 ppb. Station effects were observed for sow gestation weight gain, lactation weight change, lactation feed intake, litter size at birth and weaning, and pig weight at birth and weaning (P = 0.001 to 0.087). Supplemental Cr increased the number of pigs born live per litter (9.49, 9.82, 10.94, and 10.07; quadratic, P = 0.05) and sow lactation weight change (-0.2, 0.8, -4.1, and -3.9 kg; linear, P = 0.01) but decreased individual birth weight of total pigs born (1.61, 1.57, 1.47, and 1.56 kg; quadratic, P = 0.10). Tissues were obtained from a subset of sows from one station after they had completed three parities on the study. The content of Cr in the adrenal gland (16.4, 20.0, 34.0, and 48.4 ppb), kidney (35.8, 56.4, 132.6, and 176.0 ppb), and liver (22.8, 37.4, 87.6, and 92.2 ppb) was increased linearly (P = 0.001 to 0.005) by increasing CrPic supplementation. The results suggest that the supplementation level that maximizes the biological response is above that currently allowed. Additionally, supplementation of Cr at 1,000 ppb (five times currently permitted supplementation levels) was not detrimental to sow performance, even when fed continuously for three parities. There may be merit to continued research to evaluate higher supplementation rates.     

Comparison of virulence of various hantaviruses related to hemorrhagic fever with renal syndrome in newborn mouse model

The virulence of hantaviruses that are antigenically related but have different genetic characteristics from the prototype of hantavirus, Hantaan (HTN) virus, was examined in newborn mice. The H5 and B78 strains of the Amur (AMR) genotype, the Bao14 strain of the Far East (FE) genotype, and the 76-118 strain of HTN virus were inoculated subcutaneously (1focus-forming unit; FFU) into newborn mice. All of the AMR and FE genotype viruses inoculated mice were died by 16 days post-infection (dpi) and 21 dpi, respectively, while 50% of the HTN virus inoculated mice survived until 30 dpi. The AMR and FE genotype viruses inoculated mice had high viral titers in the lung (1.3x10(6) to 1.3x10(8) FFU/gram [g] tissue) , brain (2.1x10(7) to 1.2x10(9) FFU/g tissue), and kidney(2.5x10(5) to 1.6x10(7) FFU/g tissue), and showed a detectable level of antibodies (titers 1:16-1:32) at 14 dpi. In contrast, the HTN virus infected mice had viruses only in the lungs at low titers (1.1-5.3x10(5) FFU/g tissue). Observations of body-weight changes revealed that the AMR and FE genotype viruses inoculated mice had lower growth rates than the HTN virus inoculated mice. These data suggest that the AMR and FE genotype viruses are more virulent than the HTN virus in newborn mice.     

Identification of CpG oligodeoxynucleotide sequences that induce IFN-gamma production in canine peripheral blood mononuclear cells

Oligodeoxynucleotides containing the cytosine-phosphate-guanine (CpG) motif (CpG-ODNs) have been shown to induce T(H)1 immune responses in animals. Since the sequences of CpG-ODNs that induce T(H)1 responses are considered to vary among animal species, it is necessary to identify effective CpG-ODNs in each animal. In order to identify the sequences of CpG-ODNs that induce T(H)1 responses in dogs, mRNA expression and protein production of IFN-gamma were examined in peripheral blood mononuclear cells (PBMCs) from healthy dogs treated with 11 kinds of synthetic CpG-ODNs. One of the 11 CpG-ODNs (No. 2 CpG-ODN, 5'-GGTGCATCGATGCAGGGGGG-3') was shown to significantly increase mRNA expression and protein production of IFN-gamma in canine PBMCs in a manner dependent on the sequence of the CpG motif. This CpG-ODN also enhanced the expression of IL-12 p40 mRNA in canine PBMCs, whereas expression of IL-12 p35, IL-18, and IL-4 mRNAs was not induced by this CpG-ODN. These results indicate that this CpG-ODN was able to produce IFN-gamma by induction of T(H)1-skewed immune response in dogs. CpG-ODNs may be useful for inducing prophylactic and therapeutic immunity against allergic diseases, viral infection, and tumors in dogs.     

A novel repeated sequence located on the bovine Y chromosome: its application to rapid and precise embryo sexing by PCR

A novel repeated sequence specific to male cattle was identified and named S4. S4 is a highly repetitive sequence and is a 1.5 kb repeating unit that contains various internal repeated sequences. FISH analysis showed that S4 is localized on the whole long arm and the proximal region of the short arm of the Y chromosome. We found that a PCR primer set for S4 amplified a male-specific 178 bp product in addition to a 145 bp product common to both male and female cells. Although the origin of the 145 bp product is unknown, it acts as a positive internal control in practical embryo sexing. Due to the high copy number of S4, PCR required only 0.5 pg purified DNA for accurate amplification. This made it possible to reduce the amount of biopsy sample required for embryo sexing and thus result in less damage to embryos manipulated. These studies indicate that embryo sexing based on the S4 sequence is accurate and sensitive.