Humoral immune responses of black-footed penguins (Spheniscus demersus) after DNA-mediated immunization with a beta-galactosidase reporter gene.

Humoral immune responses of black-footed penguins (Spheniscus demersus) to DNA-mediated immunization with a beta-galactosidase reporter gene expression plasmid were evaluated. Six male and 6 female adult penguins received either test plasmid, pCMV-beta, containing the beta-galactosidase gene or control plasmid, pCI, lacking a gene for expression. Three birds from each group were used previously in a diluent control group and given one injection of sterile saline. All samples were screened for anti-beta-galactosidase antibodies by indirect enzyme-linked immunosorbent assay with anti-chicken immunoglobulin G as secondary antibody. Antibodies to beta-galactosidase were detected in the sera of pCMV-beta-inoculated penguins, with a peak response on day 21. Antibody titers of the test plasmid group versus both control groups on days 21, 28, and 42 differed significantly. These results demonstrate that black-footed penguins can be safely transfected with the gene encoding beta-galactosidase and will mount a humoral response against the in vivo-expressed protein. Knowledge from this initial study can be applied to the development of DNA-mediated vaccines against specific infectious diseases of penguins.     

Utilization of free and protein-bound lysine in the Japanese quail

The utilization of dietary lysine for protein synthesis is affected by the digestibility of protein-bound lysine, by its intestinal resorption and by its oxidative catabolism. The approach chosen in this paper enables a comparison of the cumulative effect of these processes on the utilization of free and protein-bound lysine, respectively. The principle of the approach is based on a quantification of the expiration of 14C-labelled carbon dioxide after an oral administration of a diet, which contains L-(U-14C)-lysine either as a free amino acid or bound to yeast proteins. During an adaptation phase cockerels of the Japanese quail received a diet based mainly on ground wheat and wheat gluten. This diet was supplemented either with yeast proteins or with a mixture of L-amino acids which simulates the composition of the yeast proteins. In the main experiment the expiration of labelled carbon dioxide was measured during 240 minutes after the administration of the corresponding labelled diets. Just before treatment the animals were either in the postprandial phase or in a state of slight hunger. The maximum of expiration of labelled carbon dioxide occurred around the 60th minute after administration of the corresponding labelled diets. The cumulative expiration of labelled carbon dioxide, expressed in per cent of the radioactive dose used, amounts to 15.5% and 14.3% for free and protein-bound lysine, respectively. The utilization of both forms of lysine in the Japanese quail is lower than in broilers.     

Contribution of propylene glycol-induced Heinz body formation to anemia in cats

Propylene glycol (PG) is a common preservative and source of synthetic carbohydrates in soft-moist pet foods. Propylene glycol was fed to cats for 5 weeks at concentrations found in commercial diets (1.6 g/kg of body weight; 12% of diet on a dry-weight basis) and for 3 weeks at concentrations exceeding usual intake (8 g/kg; 41% of diet). There was a dose-dependent increase in Heinz body percentage to 28% in cats fed the low dose of PG and to 92% in cats fed the high dose. Erythrocyte half-life, measured using [14C]-cyanate hemoglobin (Hb), decreased significantly (P less than 0.05) by 18.8% and 60% in cats fed the low and high PG doses, respectively. The PCV in cats fed the low dose was unaffected, whereas cats fed the high dose had a mean (+/- SEM) decrease in PCV from 33.5 +/- 1.05% to 26.3 +/- 1.45%, accompanied by punctate reticulocytosis and bone marrow erythroid hyperplasia. A dose-dependent increase in iron pigment was found in the liver and spleen of all cats. In cats fed the low dose of PG, erythrocyte reduced glutathione concentration actually increased from 7.02 +/- 0.56 to 9.74 +/- 0.69 mumol/g of Hb, but decreased to 2.96 +/- 0.27 mumol/g of Hb in cats fed the high dose. There was no significant increase in methemoglobin concentration. These results indicated that PG cannot be considered innocuous even at concentrations consumed by cats eating commercial diets. Heinz body-induced acceleration of RBC destruction develops in a dose-dependent manner, so that cats with greater food intake, ie, lactating queens and nursing kittens, are at greater risk for development of PG-induced Heinz body hemolytic anemia.     

Oesophagogastric parakeratosis in the growing pig: effects of the physical form of barley-based diets and added fibre

In four experiments a total of 288 individually fed pigs were given barley-based diets for about 100 days from about 20 kg liveweight. Fine grinding of barley increased the number and severity of oesophagogastric lesions. Pelleting a diet based on coarsely ground barley had a similar effect. Coarser grinding of the barley and substituting small proportions of oat husk, but not of bran, gave lower incidences and severities of lesions. The performance responses of the pigs differed and give a framework for deciding on the balance to be struck between optimal performance and the risk of lesion development.     

A specific DNA probe which identifies Babesia bovis in whole blood.

A genomic library of Babesia bovis DNA from the Mexican strain M was constructed in plasmid pUN121 and cloned in Escherichia coli. Several recombinants which hybridized strongly to radioactively labeled B. bovis genomic DNA in an in situ screening were selected and further analyzed for those which specifically hybridized to B. bovis DNA. It was found that pMU-B1 had the highest sensitivity, detecting 25 pg of purified B. bovis DNA, and 300 parasites in 10 microliters of whole infected blood, or 0.00025% parasitemia. pMU-B1 contained a 6.0 kb B. bovis DNA insert which did not cross-hybridize to Babesia bigemina, Trypanosoma evansi, Plasmodium falciparum, Anaplasma marginale, Boophilus microplus and cow DNA. In the Southern blot analysis of genomic DNA, pMU-B1 could differentiate between two B. bovis geographic isolates, Mexican strain M and Thai isolate TS4. Thus, the pMU-B1 probe will be useful in the diagnosis of Babesia infection in cattle and ticks, and in the differentiation of B. bovis strains.