Polypeptide composition of fraction 1 protein from parasexual hybrid plants in the genus Nicotiana

Analysis of the subunit polypeptide composition of Fraction 1 proteins gives information on the expression of both nuclear and chloroplast genomes; the large subunits of the protein are coded by chloroplast DNA, whereas the small subunits are coded by nuclear DNA. Fraction 1 protein isolated from the leaves of parasexual hybrid plants derived from the fusion of protoplasts of Nicotiana glauca and N. langsdorffii contains the small subunit polypeptides of both parent species and the large subunit polypeptides of only N. glauca. Fraction 1 protein isolated from the leaves of a hybrid plant obtained after the uptake of chloroplasts of N. suaveolens by protoplasts of white tissue of a variegating mutant of N. tabacum contains the large subunit polypeptides of both N. suaveolens and N. tabacum, as well as the small subunit polypeptides of both these species.     

Effect of endocytic and metabolic inhibitors on the internalization and intracellular growth of Brucella abortus in Vero cells.

Uptake, transfer to rough endoplasmic reticulum, and intracellular growth of Brucella abortus were studied in Vero cells treated with endocytic and metabolic inhibitors. Infection of Vero cells was suppressed when inhibitors of energy metabolism (iodoacetate, dinitrophenol), receptor-mediated endocytosis (monodansylcadaverine, amantadine, methylamine), or endosomal acidification (chloroquine, ammonium chloride, monensin) were added to the inoculum. Inhibition was not observed when these drugs were added after the inoculation period. Infection of Vero cells by B abortus was inhibited by dibutyryl-cyclic adenosine monophosphate and Vibrio cholerae enterotoxin, but was stimulated by dibutyryl-cyclic guanosine monophosphate and escherichia coli heat-stable enterotoxin a. Uptake of B abortus by Vero cells was not prevented by colchicine, but was abolished by cytochalasin B. Uptake of heat-killed B abortus and noninvasive E coli was similar to that of viable brucellae. Intracellular growth of B abortus was not affected by cycloheximide. Results indicate that: B abortus may be internalized by a receptor-mediated phagocytic process; transfer of B abortus from phagosomes to rough endoplasmic reticulum may require endosomal acidification; and replication of B abortus within the rough endoplasmic reticulum may not depend on protein synthesis by the host cell.     

Tomato yellow leaf curl Sardinia virus and its vector Bemisia tabaci in Sicilia (Italy): present status and control possibilities*

Tomato yellow leaf curl Sardinia begomovirus (TYLCSV) appeared in Sicilia (IT) in 1988, creating great threats to agriculture and causing huge losses, especially in south‐eastern areas of the island, where protected tomato cultivation is widespread. Towards the mid‐1990s, a reduction occurred in the virus epidemics, probably due to new approaches which have been applied to rational control of its vector, the whitefly Bemisia tabaci. More recently, TYLCSV has increased again, creating great concern among local tomato producers and stimulating new research. Besides studies on natural enemies of the vector, aiming to investigate their role and distribution, the main current research lines in Sicilia concern the possibility of reducing both whitefly activity, using photoselective plastics as covers, and virus damage, by growing tolerant tomato genotypes.     

Biofilm bacteria: formation and comparative susceptibility to antibiotics

The Calgary Biofilm Device (CBD) was used to form bacterial biofilms of selected veterinary gram-negative and gram-positive pathogenic bacteria from cattle, sheep, pigs, chicken, and turkeys. The minimum inhibitory concentration (MIC) and minimum biofilm eradication concentration (MBEC) of ampicillin, ceftiofur, cloxacillin, oxytetracycline, penicillin G, streptomycin, tetracycline, enrofloxacin, erythromycin, gentamicin, tilmicosin, and trimethoprim-sulfadoxine for gram-positive and -negative bacteria were determined. Bacterial biofilms were readily formed on the CBD under selected conditions. The biofilms consisted of microcolonies encased in extracellular polysaccharide material. Biofilms composed of Arcanobacterium (Actinomyces) pyogenes, Staphylococcus aureus, Staphylococcus hyicus, Streptococcus agalactiae, Corynebacterium renale, or Corynebacterium pseudotuberculosis were not killed by the antibiotics tested but as planktonic bacteria they were sensitive at low concentrations. Biofilm and planktonic Streptococcus dysgalactiae and Streptococcus suis were sensitive to penicillin, ceftiofur, cloxacillin, ampicillin, and oxytetracycline. Planktonic Escherichia coli were sensitive to enrofloxacin, gentamicin, oxytetracycline and trimethoprim/ sulfadoxine. Enrofloxacin and gentamicin were the most effective antibiotics against E. coli growing as a biofilm. Salmonella spp. and Pseudomonas aeruginosa isolates growing as planktonic populations were sensitive to enrofloxacin, gentamicin, ampicillin, oxytetracycline, and trimethoprim/sulfadoxine, but as a biofilm, these bacteria were only sensitive to enrofloxacin. Planktonic and biofilm Pasteurella multocida and Mannheimia haemolytica had similar antibiotic sensitivity profiles and were sensitive to most of the antibiotics tested. The CBD provides a valuable new technology that can be used to select antibiotics that are able to kill bacteria growing as biofilms.     

Fibroblast growth factor-10 maintains the survival and promotes the growth of cultured goat preantral follicles

The aim of the present study was to investigate the effects of fibroblast growth factor-10 (FGF-10) on the survival, activation (transition from primordial to primary follicles), and growth of goat preantral follicles cultured in vitro. Pieces of ovarian cortex were cultured for 1 and 7 d in the absence or presence of FGF-10 (0, 1, 10, 50, 100, and 200 ng/mL). Noncultured and cultured tissues were processed and analyzed by histology, transmission electron microscopy, and viability testing. Results showed that after 7 d, a greater percentage (79.9%) of morphologically normal follicles (containing an oocyte with regular shape and uniform cytoplasm, and organized layers of granulosa cells without a pyknotic nucleus) was observed when cultured with 50 ng/mL of FGF-10 when compared with other concentrations of FGF-10 (0 ng/mL, 67.3%; 1 ng/mL, 68.2%; 10 ng/mL, 63.3%; 100 ng/mL, 64.4%; 200 ng/mL, 52.7%). Ultrastructural analyses and viability testing using fluorescent markers confirmed the follicular integrity of FGF-10 (50 ng/mL)-treated fragments after 7 d of culture. After 7 d, all FGF-10 concentrations reduced the percentage of primordial follicles and increased the percentage of developing follicles. In the presence of 50 ng/mL of FGF-10, follicles increased in diameter after 7 d of culture when compared with other concentrations tested. In conclusion, this study demonstrates that FGF-10 maintains the morphological integrity of goat preantral follicles and stimulates the growth of activated follicles in culture. The culture conditions identified here contribute to the understanding of the factors involved in goat early follicular development.     

SONOGRAPHY OF THE EQUINE PALMAR METACARPAL SOFT TISSUES

It was hypothesized that ultrasonography may be a sensitive method for identifying pathologic changes in the tendons and ligaments of the palmar metacarpus of the horse. The palmar meta-carpi of equine cadavers and live horses were examined sonographically. The advantages and disadvantages of various ultrasound scanning techniques and the normal appearance of longitudinal and transverse palmar metacarpal sonograms are described.     

An evaluation of the nutritional value of alternative lipid sources to juvenile southern rock lobster, Jasus edwardsii

Experimental lobster feeds are currently based on fish meal and fish oil formulations, and although survival and growth similar to that of lobsters fed fresh blue mussels has been achieved, varying the protein level in previous experimental feeds has not increased growth beyond that of lobsters fed natural food. This experiment assessed the growth performance of lobsters fed pelleted feeds containing constant amounts of protein, lipid and energy where the lipid was provided by a range of oil-rich ingredients (fish oil, FO; fish oil with added soybean lecithin, FOL; canola oil, CO; tuna oil, TO; mussel meal, MM; and squid meal, SQM). Feed performance was assessed by lobster growth rate, survival, final biochemical composition, nutrient retention and nutrient efficiency. Twenty tanks containing 15 post-larval lobsters each (1.5 ± 0.04 g) were randomly allocated one of six test feeds in triplicate, and the two remaining tanks were fed freshly opened blue mussels (FRM) as a reference feed. Lobsters were fed daily to excess for 10 weeks. Final individual weights of whole body and digestive gland were measured, and tissue chemical composition analysed. There were no significant differences in survival (88.4 ± 3.3%), or specific growth rate (1.3 ± 0.1%.day^− 1) among the formulated feed fed lobsters, which were significantly lower than the survival (100 ± 0.0%) and SGR (2.2 ± 0.1%.day^− 1) of FRM fed lobsters. The SQM fed lobsters had a significantly lower lipid efficiency ratio and lipid productivity value than lobsters fed TO, FOL and MM feeds. The digestive gland lipid content (g.100 g wet tissue^− 1) of lobsters fed the feeds TO (3.7 ± 0.4), FO (3.5 ± 0.3) and SQM (2.2 ± 0.2) were significantly lower than lobsters fed feeds MM (9.9 ± 1.1), FOL (9.0 ± 2.3) and FRM fed lobsters contained most digestive gland lipid (12.3 ± 1.5).     

Nucleotide sequence and mRNA expression analysis of <Emphasis Type="Italic">β</Emphasis>-actin gene in the orange-spotted grouper <Emphasis Type="Italic">Epinephelus coioides</Emphasis>

The full-length cDNA, encoding the orange-spotted grouper β-actin and spanning 1920 bp including a poly (A) tail, was cloned from its brain cDNA library. The open reading frame encodes a protein of 375 amino acids. Sequence analysis indicated that it contained the typical structural features of cytoplasmic actins, and showed higher homology with other vertebrate β-actin than any other members of the actin family. The partial genomic sequence indicated that the organization of the β-actin gene in the orange-spotted grouper might also be conserved. Northern blot analysis indicated that it was expressed at high levels in the brain, spleen, adipose tissue, ovary, and liver, but at low levels in the gill filament and heart, and at a very low level in the kidney. The expression of β-actin gene in the skeletal muscle was barely detectable. These results indicated that the expression of the orange-spotted grouper β-actin gene showed significant variation in different tissues. Therefore, caution should be taken when using β-actin gene as an internal control in the normalization of gene expression among tissues. Whereas, semi-quantitative RT-PCR analysis indicated that treatment with 17α–methyltestosterone (MT) had little effect on the mRNA expression of β-actin gene in the in vitro incubated hypothalamus, pituitary, and ovary fragments of the orange-spotted grouper, suggesting β-actin can be used as an internal control for RT-PCR analysis of MT effects on gene expression in these tissues.