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.     

Multiple cytopenias associated with monocytic proliferation in a dog

An unusual combination of blood cytopenias and monocytic proliferation was observed in a dog. Initial hematologic findings included severe thrombocytopenia, neutropenia, mild nonregenerative anemia and apparently normal bone marrow. Subsequently, a severe persistent monocytosis developed and the bone marrow became populated with monocytes and cytophagic macrophages. Splenomegaly was due to reticuloendothelial hyperplasia and extramedultary hematopoiesis. Treatment consisted of splenectomy and azathioprine but the response was poor and the dog was euthanized. Postmortem examination revealed a hypocellular bone marrow which contained moderate numbers of monocytes and plasma cells. Neoplastic proliferation was absent in visceral organs. No definite diagnosis was established; chronic blood cell consumption, perhaps immune-mediated, may have been responsible for the extensive reticuloendothelial hyperplasia and cytophagia.     

Molecular cloning and tissue distribution of a short form chicken leptin receptor mRNA

In mammals, alternative splicing of the leptin receptor (LEPR) produces several C-terminal truncated isoforms that are believed to play a role in the transport, cellular internalisation and degradation of the hormone leptin. The chicken leptin receptor (chLEPR) is similar to its mammalian counterparts in terms of its intron/exon structure and conserved motifs. However, it is unknown whether the chLEPR also undergoes alternative splicing. To test this, structural analysis of intron 19 of the chLEPR, equivalent to the intron in which alternative splicing occurs in mammals, was combined with 3'-rapid amplification of cDNA ends (3'-RACE) to search for chLEPR splice variants. A 44-amino acid alternative exon 20 was identified that is spliced to generate a short isoform of the chLEPR (chLEPR-SF). Comparative sequence analysis of intron 19 identified two regions that are highly conserved between the chicken and mammals, indicating their possible importance as intronic elements in the regulation of alternative splicing of the LEPR in vertebrates. Tissue expression of the chLEPR-SF was lower and more restricted than that of the chLEPR long isoform. Collectively these data demonstrate that the chLEPR is alternatively spliced to produce at least one short isoform, as is the case in mammals.     

Hypertrophic osteopathy associated with infective endocarditis in an adult boxer dog

A 20-month-old, entire male boxer dog was presented with lethargy and intermittent shifting limb lameness. Diagnostic tests revealed aortic valve vegetations suggestive of infective endocarditis causing severe aortic outflow obstruction, and hypertrophic osteopathy of all four limbs. The dog was treated symptomatically and euthanased four days later. The association of infective endocarditis and hypertrophic osteopathy has been poorly documented in the veterinary literature. The pathogenesis of hypertrophic osteopathy is unknown; however, four theories have been put forth to explain this disease: pulmonary shunting, vagal nerve stimulation, humoral substances produced by neoplastic cells and megakaryocyte/platelet clump hypothesis.     

An analysis of sap flow in mountain ash (Eucalyptus regnans) forests of different age

Diurnal measurements of sap velocity were made in 50-, 90-, 150- and 230-year-old mountain ash (Eucalyptus regnans F. Muell.) forests in the North Maroondah catchment (southeast Australia) over the periods January 8, 1990 to April 4, 1990 and October 29, 1990 to April 16, 1991. Over the two periods, daily mean sap velocities for the four forests, in order of increasing age, were 11.5, 11.4, 9.9 and 11.8 cm h(-1) respectively. Daily mean sap velocity did not differ significantly among the 50-, 90- and 230-year-old plots. However, in the 150-year-old trees it was significantly smaller by an average of 14%. Sap velocity varied diurnally and also between positions within individual trees and among trees both within and between stands. Despite this variability, the sampling intensity and duration were sufficient to establish that behavior was highly correlated among individuals within plots. There was a significant decline with age in the overstory sapwood conducting area of these forests. In order of increasing age, the values were 6.7, 6.1, 4.2 and 4.0 m(-2) ha(-1), respectively. When combined with daily mean sap velocity, these data allowed the calculation of overstory water use. Over the experimental period, water use of the overstory decreased with age ranging, on average, from 1.86 mm day(-1) for the 50-year-old plot to 0.81 mm day(-1) for the 230-year-old plot. Mean daily water use for the two intermediate-aged forests was 1.67 and 1.00 mm day(-1), respectively. Annual water use decreased with forest age from 679 mm for the 50-year-old stand to 296 mm for the 230-year-old stand. This difference corresponds to 3.8 x 10(3) m(3) ha(-1). The annual water use of the intermediate-aged stands was 610 and 365 mm for the 90- and 150-year-old stands, respectively.     

Evaluation of Bacterial Effects on Folic Acid Loss in Fortified,Nixtamalized Corn Masa Flour

Many Mexican women are deficient in folic acid. Fortification of the corn tortilla could be an effective way to help increase the folic acid levels among the Mexican population. Previous studies have shown significant folic acid losses in the masa dough as it is held before baking. This loss in folic acid could be owing to utilization by lactic acid bacteria naturally present in the masa. The objective of this study was to determine the effect of bacteria native to corn masa on the folic acid content in masa. Bacteria in dough samples from six mills in Guadalajara, Mexico, were isolated and identified. Bacterial isolates were inoculated into sterile fortified corn masa flour, which was converted to masa and held at 56°C for 0, 3, and 6 h, replicating the conditions of freshly milled masa as held before baking. All samples, including the control, showed losses of folic acid between 66 and 79% w/w in the first 3 h of incubation. Because folic acid degradation in the sterile control sample was not different than the inoculated sample results, the decline in folic acid was not owing to bacteria (mainly Streptococcus spp.) present in the masa flour but appeared to be a chemical degradation related to time and temperature.