Immunocytochemical localization of the Bacillus sphaericus binary toxin components in Culex quinquefasciatus (Diptera: Culicidae) larvae midgut

This study describes the immunocytochemical localization of the Bacillus sphaericus 2362 binary toxin components, BinA and BinB, in Culex quinquefasciatus larvae that had been intoxicated with this entomopathogen. Ultrathin sections of C. quinquefasciatus larvae midgut embedded in the hydrophilic resin L.R. White were incubated with the antibodies anti-BinA or anti-BinB and then revealed with goat anti-rabbit IgG coupled to gold particles. Immunocytochemical detection demonstrated the presence of specific labeling in ultra-thin sections that had been incubated with the BinA antiserum. Gold particles were detected on the apical areas of cell membranes and inside the epithelial cell cytoplasm, particularly the mitochondria, of cells from the gastric caeca and posterior stomach in larvae exposed during 2 or 24 h to the entomopathogen. A similar labeling pattern was observed in ultrathin sections from both midgut regions when incubated with BinB antiserum.     

Effects of salinity on the growth and proximate composition of selected tropical marine periphytic diatoms and cyanobacteria

Marine periphytic cyanobacteria and diatoms have been examined as a potential source of feed supplement for rearing aquatic larvae in the aquaculture industry. Culture of the periphytic diatom Amphora sp., Navicula sp., Cymbella sp. and the cyanobacteria Oscillatoria sp. at different salinities showed significant changes in biomass and specific growth rates. Diatoms growth was significantly higher at 35 g L⁻¹, while for cyanobacteria growth was better at 25 g L⁻¹. Significantly higher levels of protein and lipid were found in diatoms at low salinities (15–25 g L⁻¹) and an increase in carbohydrate at high salinities (30–35 g L⁻¹). Conversely, cyanobacteria showed a significantly higher lipid content at 30–35 g L⁻¹ compared with other salinity levels but no significant changes were observed in the protein and carbohydrate contents at different salinity levels. The present findings can be taken into consideration when culturing marine periphytic Amphora sp., Navicula sp., Cymbella sp. and Oscillatoria sp. to provide appropriate levels of protein, lipid and carbohydrate as feed supplement as well as for bioremediation in aquaculture.     

CONTRAST-ENHANCED ULTRASONOGRAPHY IN NORMAL CANINE LIVER. EVALUATION OF IMAGING AND SAFETY PARAMETERS

Contrast-enhanced ultrasonography, a new imaging modality in veterinary medicine, can provide data on tissue perfusion. The objective of this study was to use the ultrasonographic contrast agent SonoVue to evaluate various transit time indices in the normal canine liver, to examine the effect of anesthesia on these parameters, and to evaluate the safety of this agent in dogs. The liver of 11 healthy dogs was studied by ultrasound during an intravenous bolus injection of SonoVue. Each dog underwent the examination twice, first with and later without the use of anesthesia. A time-intensity curve was generated from a selected region of interest within the liver from each scanning session. Ratios derived from peak enhancement, time to peak enhancement, up-slope and full-width half-maximum (FWHM) of the curve were calculated from the time-intensity curves, and are reported. There were no statistically significant differences (P > 0.05) in peak enhancement, up-slope and FWHM between dogs that were anesthetized and dogs that were not. Time to peak enhancement, however, was significantly shorter when the dogs were anesthetized than when they were nonanesthetized (P < 0.05). There were no biologically significant changes in clinical laboratory findings. This study indicates that contrast-enhanced ultrasound using SonoVue gives reproducible liver perfusion data, and appears to be a safe and well-tolerated agent for use in dogs. When considering normal values, the use of anesthetic drugs has to be considered.     

Phenotypic characterisation of Phaeoacremonium and Phaeomoniella strains isolated from grapevines: enzyme production and virulence of extra-cellular filtrate on grapevine calluses

Extra-cellular enzyme production of different Phaeoacremonium spp. and Phaemoniella chlamydospora isolates were used to assay the possibility of inter-specific characterisation. Isolates of Phaeoacremonium aleophilum, Phaeoacremonium angustius, P. viticola and Ph. chlamydospora were grown on solid media and the activities of extra-cellular amylases, lipases, proteases, cellulases, xylanases, laccase, polygalacturonase, pectate lyase, lignin peroxidase, manganese peroxidase, urease and chitinase were assayed. Phaeoacremonium species showed activities of a larger number of enzymes and also enzyme activity was frequently higher suggesting that Phaeoacremonium can be more virulent. To assay if the produced extra-cellular enzymes could reflect the virulence capacity of the two genera, calluses of Vitis vinifera L. (cvs. Baga and Maria Gomes) and of a rootstock (R3309) were inoculated with filtrated culture liquid medium of three isolates of Ph. chlamydospora and one of P. angustius. Filtrates from all strains decreased callus growth and membrane integrity, while soluble protein content of calluses decreased with the strains CAP 054 and 1AS. P. angustius (CAP 054) induced the more severe symptoms in all genotypes. Water content decreased together with an increase of osmolality in both cultivars but not in rootstock suggesting that osmorregulatory capacity is more affected in cultivars. Data show that: (1) Phaeoacremonium and Phaeomoniella genera have different patterns of extra- cellular enzymatic production; (2) these fungi produce extra-cellular compound(s) that induce(s) senescence symptoms in plant cells inhibiting callus proliferation; (3) among the strains tested in plant calluses the most virulent isolate (CAP 054) also produced higher amounts of some extra-cellular enzymes; (5) rootstock calluses were less sensitive to inoculation than grapevine calluses.     

<Emphasis Type="Italic">Physalis ixocarpa</Emphasis> and <Emphasis Type="Italic">P. peruviana</Emphasis>, new natural hosts of <Emphasis Type="Italic">Tomato chlorosis virus</Emphasis>

Tomato chlorosis virus causes yellow leaf disorder epidemics in many countries worldwide. Plants of Physalis ixocarpa showing abnormal interveinal yellowing and plants of Physalis peruviana showing mild yellowing collected in the vicinity of tomato crops in Portugal were found naturally infected with ToCV. Physalis ixocarpa and P. peruviana were tested for susceptibility to ToCV by inoculation with Bemisia tabaci, Q biotype. Results confirmed that ToCV is readily transmissible to both species. The infection was expressed in P. ixocarpa by conspicuous interveinal yellow areas on leaves that developed into red or brown necrotic flecks, while P. peruviana test plants remained asymptomatic. Infected plants of both P. ixocarpa and P. peruviana served as ToCV sources for tomato infection via B. tabaci transmission. This is the first report of P. ixocarpa and P. peruviana as natural hosts of ToCV.     

Growth,Feed Utilization,and Glycemic Response in European Sea Bass,Dicentrarchus labrax,Juveniles Fed Carbohydrate of Different Complexities

The effect of dietary carbohydrate complexity on growth, feed utilization, and glycemia was studied in European sea bass juveniles. Four isonitrogenous (50% crude protein) and isolipidic (15% crude lipids) diets were formulated to contain 20% pregelatinized maize starch (PGS diet), dextrin (DEX diet), maltose (MAL diet), or glucose (GLU diet). No effect of dietary carbohydrate complexity on growth was noticed. Feed efficiency and protein efficiency ratio were lower in fish fed the GLU diet than in the other groups, whereas the opposite was observed for feed intake. Plasma glucose peaked 3–4 h after feeding in fish fed the MAL and GLU diets, whereas in fish fed the PGS and DEX diets the peak was reached 5–6 h after feeding. Peak plasma glucose concentration (13 mmol/L) was higher in fish fed the GLU diet than the other diets (9 mmol/L). Shorter hyperglycemia duration was observed in fish fed the MALT and GLU diets (6 h) than the PGS and DEX diets (10 h). Complex carbohydrates delayed plasma glucose concentration peak compared with simple sugars, whereas the opposite was observed for hyperglycemia duration. Overall, dietary maltose, dextrin, and starch were apparently better utilized as energy source than glucose by European sea bass juveniles.