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.     

Nutrient intake of horses in Thoroughbred and Standardbred stables

SUMMARY Twenty-five Thoroughbred (TB) and 25 Standardbred (SB) stables were visited to determine their feeding practices. The ingredients of the main feed of the day for a mature gelding of average size in full training were weighed at each stable. Nutrient content of diets was calculated using published data for the individual ingredients. Results are expressed as mean±sd. The estimated body weight of TB horses was 493±34 kg and 437±32 kg for SB horses. There was considerable variation in diet composition and nutrient intake between stables. The TB trainers fed 11.0±2.4 kg and SB trainers 11.8±2.5 kg per day. The concentrate component of the diet weighed 7.8±1.6 and 7.7±2.3 kg for TB and SB stables, respectively, and the roughage component for TB horses 3.3±1.4 and SB horses 4.1±1.4 kg per day. The digestible energy intake of horses at TB stables was 129±29 MJ per day and at SB stables 132±31 MJ per day. Crude protein intake of TB horses was 1452±363 g and SB horses 1442±338 g per day. There were differences in some feeding practices at TB and SB stables. Standardbred trainers fed more roughage than TB trainers. Standardbred trainers fed chaffed lucerne (alfalfa) and cereal hays as the major roughage, whereas TB trainers fed more hay. The major hay type fed by TB trainers was lucerne, whereas many SB trainers preferred clover hay. Both trainers fed oats as the major grain, but TB trainers fed slightly more maize (corn) than SB trainers. The SB trainers fed barley as part of the concentrate component of the diet, whereas TB trainers usually fed boiled barley and linseed oil in winter only. Although many trainers used vitamin and mineral supplements, this appeared unnecessary in many Instances, especially with respect to Iron. Calcium and NaCI supplementation was necessary for some diets. We concluded that while there was a wide range in feed intake and diet composition for both TB and SB horses, average nutrient intakes were similar to National Research Council (1989) recommendations for horses performing intense work.     

Fatty acid profiles and sensory and carcass traits of tissues from steers and swine fed an elevated monounsaturated fat diet

Twelve Angus X Hereford steers were assigned to either a control high-energy diet or a test diet consisting of 20% rapeseed at the expense of 20% corn. Twelve pigs were allotted to a control diet and two test diets containing either 10 or 20% canola oil (CO). Both CO and oil in the rapeseed contained 60 to 64% oleic acid. Cattle fed rapeseed exhibited little effect from the diet due to apparent indigestibility of the rapeseed. Total saturated fatty acids decreased from 40% in adipose tissue of the control pigs to 15% in the 20% CO-fed pigs. The ratio of monounsaturated to saturated fatty acids (M/S) increased from 1.19 in adipose tissue from control pigs to 3.63 with the addition of 20% CO to the diet. In muscle, the M/S ratio increased from 1.21 in control pigs to 2.46 in the 20% CO treatment group. The percentage of the saturated fatty acids in muscle decreased from 42% in the control to 23% in the 20% CO treatment. Significant increases in "oiliness" and decreases in fat firmness were observed when increasing levels of canola oil were fed. Sensory traits, cooking loss and shear-force values of pork chops were similar among treatment groups. In conclusion, monounsaturated fatty acid content can be elevated substantially in pork without adversely influencing the quality of the meat, thus producing a product perceived to be more healthful by the consumer.     

Tissue reaction to Cysticercus bovis in the lung of artificially infected cattle

The tissue reaction to Cysticercus bovis in the lung of cattle with an experimental infection was an inflammatory rim originating in the immediate vicinity of the cysts. The cysts recovered at days 83 and 102 p.i. contained living cysticerci. The rim was composed either of a layer of high histiocytes organized in palisades (at day 83 p.i.), or a lyer of flat histiocytes (at day 102 p.i.). The outer layer of the rim consisted of fibroblasts, reticular cells and a different number of eosinophil- and neutrophil luekocytes. On the periphery, the rim was formed by granulation tissue infiltrated with lymphoplasmocytes. At the border between the layers of the inflammatory rim there were conspicuous foci of a necrotic appearance typical of a tissue reaction to C. bovis.     

Evaluation of resistance to Phytophthora cinnamomi in seed-grown trees and clonal lines of Eucalyptus marginata inoculated in lateral branches and roots

Seed-grown trees and six clonal lines of 3·5–4·5-year-old Eucalyptus marginata (jarrah) growing in a rehabilitated bauxite mine site in the jarrah forest were underbark-inoculated on lateral branches (1995) or simultaneously on lateral branches and lateral roots (1996) with isolates of Phytophthora cinnamomi in late autumn. Individual seedlings from which the clonal lines were derived had previously been assessed as either resistant (RR) or susceptible (SS) to P. cinnamomi . At harvest, the acropetal lesion and colonization lengths were measured. Overall, the length of colonization in roots and branches was more consistent as a measure of resistance than lesion length, because colonization length recorded the recovery of P. cinnamomi from macroscopically symptomless tissue ahead of the lesion which, on some occasions, was up to 6 cm. In both trials, one RR clonal line was able to contain the P. cinnamomi isolates consistently, as determined by small lesion and colonization lengths in branches and roots. In contrast, the remaining two RR clonal lines used in both trials were no different from the SS line in their ability to contain lesions or colonization. These latter two RR lines may therefore not be suitable for use in rehabilitation of P. cinnamomi -infested areas. Differences in lesion and colonization lengths among P. cinnamomi isolates occurred only in the 1995 trial. Colonization and lesion lengths in branches were up to eight times greater in 1996 than in 1995, but the relative rankings of clonal lines were consistent between trials. Although colonization was always greater in branches than roots, the relative rankings of the lines were similar between branch and root inoculations. Branch inoculations are a valid option for testing the resistance and susceptibility of young jarrah trees to P. cinnamomi .     

Evidence that a Leaf-Disk Test Allows Assessment of Isolate-Specific Resistance in Brassica oleracea Crops Against Downy Mildew (Peronospora parasitica)

A rapid resistance/susceptibility test for Peronospora parasitica (downy mildew) was established by inoculating leaf-disks of four Brassica oleracea accessions. Several conditions were tested: disk disinfection or not, agar medium with or without nutrients and with 50 or 100 ppm of benzimidazole. Using disinfected disks placed on agar (no nutrient and benzimidazole at 50 or 100 ppm), the responses of leaf-disks to four isolates were similar to those obtained using the classical cotyledon test, whereas undesired contaminations occurred in all other conditions. The possible effect of the particular leaf used for obtaining the disks was also studied. In each incompatible interaction tested, disks were resistant whatever the leaf used. In compatible interactions, susceptible phenotypes were observed on disks derived from the six lowest leaves, but disks from upper leaves were resistant. The genetic basis of resistance in a F1 hybrid broccoli was assessed, by testing six isolates on an F2 population derived from this hybrid. The cotyledon test only allows inoculation of two isolates per seedling, whereas many isolates can be tested on each plant by using leaf-disks. The segregation of the resistance to each of the six isolates was analysed: two dominant genes (tightly linked) control resistance to all isolates (one to five isolates; the other to only one isolate).     

Evaluation of β‐carotene assimilation in leopard geckos (Eublepharis macularius)

Although leopard geckos (Eublepharis macularius) are commonly kept under human care, their vitamin requirements are largely unknown. Many invertebrate preys display a low vitamin A concentration; thus, gut‐loading insects with vitamin A or carotenoids is a common practice. The objective of this prospective experimental study was to investigate whether dietary supplementation with β‐carotene, including prey gut‐loading, leads to sufficient vitamin A hepatic storage and prevents epithelial squamous metaplasia development in leopard geckos. Ten clinically healthy female leopard geckos were randomly divided in two groups with various supplementations: a group receiving vitamin A supplementation and a group receiving β‐carotene. Insects were gut‐loaded continuously with a supplement containing vitamin A or β‐carotene, depending on the group. Oral supplementation with cod liver oil or carrot juice was administered weekly to each lizard from “vitamin A group” and “carotenoid group” respectively. After 10 weeks of supplementation, surgical hepatic biopsies were obtained in three geckos of each group while the two remaining geckos were euthanized to undergo complete necropsy. Hepatic vitamin A concentration was determined for each lizard (n = 10) by ultra‐performance liquid chromatography. Histopathology revealed hepatocellular vacuolization and vitellogenic follicles in five females. Epithelial squamous metaplasia was not observed in any of the geckos. Hepatic vitamin A concentration was significantly higher in the carotenoid‐supplemented group than in the vitamin A‐supplemented group (p = 0.03). Our results suggest that in leopard geckos, dietary supplementation with β‐carotene allows sufficient vitamin A hepatic storage.     

Detection and genetic diversity of porcine rotavirus A,B and C in eastern Australian piggeries

Rotaviruses (RV) have a high prevalence in piggeries worldwide and are one of the major pathogens causing severe diarrhoea in young pigs. RV species A, B, and C have been linked to piglet diarrhoea in Australian pig herds, but their genetic diversity has not been studied in detail. Based on sequencing of the structural viral protein 7 (VP7) RVA G genotypes G3, G4 and G5, and RVC types G1, G3, G5, and G6 have been identified in Australian piggeries in previous studies. Although occurrence of RVB was reported in Australia in 1988, no further genetic analysis has been conducted. To improve health management decisions in Australian pig herds, more information on RV prevalence and genetic diversity is needed. Here, 243 enteric samples collected from 20 pig farms within Eastern Australia were analysed for the presence of RV in different age groups using a novel PCR-based multiplex assay (Pork MultiPath™ enteric panel). RVA, RVB, and RVC were detected in 10, 14, and 14 farms, respectively. Further sequencing of VP7 in selected RV-positive samples revealed G genotypes G2, G5, G9 (RVA), G6, G8, G14, G16, G20 (RVB), and G1, G3, G5, G6 (RVC) present. RVA was only detected in young (<10 weeks old) pigs whereas RVB and RVC were also detected in older animals (>11 weeks old). Interestingly, RVB and RVC G-type occurrence differed between age groups. In conclusion, this study provides new insights on the prevalence and diversity of different RV species in pig herds of Eastern Australia whilst demonstrating the ability of the Pork MultiPath™ technology to accurately differentiate between these RV species.