Development of a canine trypsin-like immunoreactivity assay system using monoclonal antibodies

The radioimmunoassay (RIA) for trypsin-like immunoreactivity (TLI) is one of the most sensitive and specific tests for detecting exocrine pancreatic insufficiency (EPI). An abnormally low serum TLI concentration (<2.5 ng/ml) indicates end-stage EPI. Although RIA methods can be used to detect canine serum TLI, these procedures are beyond the capabilities of most veterinary clinics and general laboratories. Using monoclonal antibodies (mAbs), we developed an enzyme-linked immunosorbent assay (ELISA) for canine TLI and incorporated it into an immunochromatographic test (ICT) for the diagnosis of EPI. The ELISA was linear over TLI concentrations of 1-100 ng/ml. Levels of intra-assay coefficients of variance (CVs) were 1.8-6.1%, inter-assay CVs were 5.1-9.8%, and the recovery of TLI added to two samples of canine serum ranged from 89 to 111 and 93 to 108%, respectively. Good correlation (correlation coefficient, 0.974) occurred between the TLI values obtained by the ELISA method and those by RIA from 56 clinical samples. Serum TLI values in clinically healthy dogs ranged from 7.8 to 29.2 ng/ml by ELISA, and those from dogs with EPI were 0.0-0.6 ng/ml. The values were 0.0-287.4 ng/ml for dogs with pancreatitis, and those from dogs with gastrointestinal disease were 5.5-58.9 ng/ml. The only statistically significant difference (P<0.01) occurred between the TLI level of healthy dogs and those with EPI. The ICT kit showed high reproducibility, and the TLI values yielding negative results differed significantly (P<0.01) from those returning positive results. The ICT kit yielded negative results (indicating EPI) from clinical serum samples with TLI concentrations of 0.0-4.1 ng/ml by ELISA. Both the ELISA and ICT kit are useful tools in the diagnosis of canine EPI.     

Susceptibility of liver proliferative lesions in heterozygous p53 deficient CBA mice to various carcinogens

To investigate the liver tumorigenic sensitivity to various carcinogens in heterozygous p53 deficient [p53 (+/-)] CBA mice and their wild-type littermates [p53 (+/+) mice], 71 p53 (+/-) and 74 p53 (+/+) CBA mice (male, 6-12 weeks of age) were given diet containing 4,000 or 0 ppm flumequine (FL) for 26 weeks or a single intraperitoneal injection of 5 mg/kg body weights dimethylnitrosamine (DMN) at start of the study in Exp. 1, diet containing 6,000 or 0 ppm di(2-ethylhexyl)-phthalate (DEHP) for 26 weeks in Exp. 2, or diet containing 12,000, 6,000 or 0 ppm phenolphthalein (PhP) for 26 weeks in Exp. 3. All surviving animals of these groups were killed after completion of treatment of the test substances for 26 weeks. In the FL groups, the incidences of hepatocellular altered foci in p53 (+/-) mice, the multiplicities of those in p53 (+/-) and p53 (+/+) mice were significantly increased as compared to the corresponding control groups. The incidences and multiplicities of altered foci in the DMN groups were higher than those in the corresponding control groups in p53 (+/-) and p53 (+/+) mice, but no significant differences were indicated between the groups. There were no significant differences in the incidences, multiplicities and proliferating cell nuclear antigen labeling indices of altered foci in the FL or DMN groups between p53 (+/-) and p53 (+/+) mice. There were no significant differences in the incidences and multiplicities of altered foci between the DEHP or PhP and control groups. The present results suggest that p53 gene knocked out heterozygously does not enhance the chemical hepatocarcinogenesis in CBA mice.     

Detection and molecular characterization of bovine leukemia virus in beef cattle presented for slaughter in Egypt

Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis, the most common neoplastic disease of cattle worldwide and a serious problem for the cattle industry. Previous studies have shown the molecular prevalence of BLV and the coexistence of BLV genotype-1 and -4 in Egyptian dairy cattle; however, the molecular characteristics of BLV in Egyptian beef cattle are unknown. Therefore, we collected blood samples of 168 beef cattle from slaughterhouses in three governorates in Egypt. Based on BLV-CoCoMo-qPCR-2 targeting long terminal repeats and nested PCR targeting the env-gp51 gene, the BLV provirus infection rates were found to be 47/168 (28.0%) and 42/168 (25.0%), respectively. Phylogenetic analysis based on 501 bp of the BLV env-gp51 gene from 42 BLV isolates revealed that at least six distinctive strains (b, e, f, g, x, and z) were prevalent in cattle across the examined regions. Furthermore, phylogenetic analysis of the 420 bp sequence of the BLV env-gp51 region of the six strains against 11 known genotypes showed that the strains b, e, f, and g were clustered into genotype-1, and strains x and z were clustered into genotype-4. Our results also indicated that strains b and x exist in both dairy and beef cattle in Egypt. The present study is the first to detect and genotype BLV among beef cattle in Egypt.     

A long‐term study of the effects of SLC12A1 homozygous mutation (g.62382825G>A,p.Pro372Leu) in Japanese Black cattle

Recessive missense mutation in the solute carrier family 12, member 1 (SLC12A1) gene (g.62382825G>A) is associated with hydrallantois, which is the accumulation of fluid in the allantoic cavity of a pregnant animal, and usually causes fetal death in Japanese Black cattle. However, the symptoms of a homozygote with this mutation that do not result in fetal death have not previously been tracked and evaluated. In the present study, we observed a homozygote with the SLC12A1 risk allele over a long‐term period. The calf did not show any obvious clinical symptoms, although it did exhibit a slight growth retardation that accompanied mild calciuria. At 28 months of age, the homozygote showed renal dysfunction, which in turn resulted in hydronephrosis. The time course of the symptoms was consistent with the phenotype of Bartter syndrome in humans. Additionally, the risk heterozygous genotype did not any effects on carcass traits, which indicates that eliminating the risk allele would not have any unfavorable effects. Therefore, we emphasize that both the fetal‐ and late‐stage symptoms associated with the SLC12A1 risk allele compromise animal welfare, and consequently may result in severe economic losses for individual farmers if the SLC12A1 risk allele is not eliminated from the population.     

The effect of magmatic activity on hydrothermal venting along the superfast-spreading East pacific rise

A survey of hydrothermal activity along the superfast-spreading (approximately 150 millimeters per year) East Pacific Rise shows that hydrothermal plumes overlay approximately 60 percent of the ridge crest between 13 degrees 50' and 18 degrees 40'S, a plume abundance nearly twice that known from any other rige portion of comparable length. Plumes were most abundant where the axial cross section is inflated and an axial magma chamber is present. Plumes with high ratios of volatile ((3)He, CH(4), and H(2)S) to nonvolatile (Mn and Fe) species marked where hydrothermal circulation has been perturbed by recent magmatic activity. The high proportion of volatile-rich plumes observed implies that such episodes are more frequent here than on slower spreading ridges.     

Lipid class and fatty acid composition of phospholipids from the gonads of skipjack tuna

ABSTRACT:   Lipid class and fatty acid composition of phospholipids from the gonads of skipjack tuna were examined to evaluate effective utilization of the processing of by-products. The predominant phospholipids in the ovaries were phosphatidylcholine (PC; 47.9%), phosphatidylethanolamine (PE; 19.3%) and lyso-phosphatidylcholine (LPC; 19.1%). In contrast, those in the testes were PC (40.1%), PE (29.3%) and phosphatidylserine (PS; 9.6%). The percentage of docosahexaenoic acid (DHA) was markedly high at more than 50% in LPC of the ovaries, and PE and PS of the testes were also high. The percentages of DHA at sn-position 2 of the predominant phospholipids, except for PC in the testes, were more than 60%, in particular PE in the testes was remarkably high at 81.9%. After storage for 2 days at 5°C, the LPC content in the ovaries increased twofold and the DHA level of LPC was the same as before, though the contents of other phospholipids decreased.     

Photosynthetic traits and growth of Quercus mongolica var. crispula sprouts attacked by powdery mildew under free-air CO2 enrichment

To predict the performance of coppice forests with Japanese oak (Quercus mongolica var. crispula) in future changing environment, we studied the growth, photosynthesis, and powdery mildew (Erysiphe alphitoides) infection of sprouts of Japanese oak under free-air CO₂ enrichment. Elevated CO₂ reduced powdery mildew infection in both leaves of the shoot emerged in spring (1st flush) and the lammas and proleptic shoots (2nd flush) of sprouts. We observed significant increase in the net photosynthetic rate at growth CO₂ concentration (i.e., 370 and 500 μmol mol^?1 for ambient and elevated CO₂ treatments, respectively) in both 1st and 2nd flush leaves of sprouts grown under elevated CO₂. On the other hand, no significant increase in net photosynthetic rate under elevated CO₂ was found before cutting. The photosynthetic activity of 2nd flush leaves in the sprouts under ambient condition was greatly reduced by severe infection to powdery mildew. Growth of sprouts was enhanced in the elevated CO₂ condition. We conclude the growth enhancement in Japanese oak sprouts under elevated CO₂ in the present study was achieved not only by physiological response (i.e., photosynthetic stimulation) but also by disease interaction.     

Inoculation with N2-generating denitrifier strains mitigates N2O emission from agricultural soil fertilized with poultry manure

Pelleted poultry manure is recommended for use with agricultural soil as a replacement for chemical fertilizers; however, application of the manure stimulates nitrous oxide (N₂O) emission from the soil through denitrification. To mitigate the N₂O emission caused by application of pelleted poultry manure, soil microcosms were set up; each microcosm was inoculated with one of the following N₂-generating denitrifier strains previously been isolated from paddy soil: Azoarcus, Dyella, Dechloromonas, Niastella, and Burkholderia. When pelleted poultry manure was incubated on its own, N₂O was produced by denitrification. In contrast, N₂O emission was significantly lowered when the manure was inoculated with most of the N₂-generating strains. In soil microcosms, N₂O was emitted during incubation after application of the pelleted manure, while N₂O flux was significantly lowered when the soil was inoculated with Azoarcus sp. KS11B, Niastella sp. KS31B, or Burkholderia sp. TSO47-3 on the 12th day of incubation. In addition, when pelleted manure was inoculated with the strains prior to application in the soil microcosms, the level of N₂O emission was significantly lowered to ca. 40–60 % that from the non-inoculated control. Our study provides the prototype of a technique that uses microbial technology to mitigate N₂O emission from agricultural soil fertilized with pelleted poultry manure.