Detailed physicochemical characterization of the 2S storage protein from rape (Brassica napus L.)

Chromatographic, chemical, and spectroscopic techniques were used to characterize the physicochemical properties of napin purified by preparative chromatography. The molar extinction coefficient was determined (epsilon = 0.56), and static and dynamic light scattering measurements enabled the average molecular weight (M(w) = 13919), the second virial coefficient (A(2) = 23.95 x 10(-)(5) mol cm(3) g(-)(2)), and the hydrodynamic radius (R(H) = 1.98 nm) to be determined. No conformational changes were observed by fluorescence and circular dichroism measurements in different buffers at pH 3, 4.6, 7, and 12, confirming the high pH stability of this protein. From MALDI-TOF analysis and after enzymatic digestion, it was found that this purified sample, extracted from the rapeseed variety Express, contained mainly isoform 2SS3_BRANA.     

Biological activity of peanut (Arachis hypogaea) phytoalexins and selected natural and synthetic Stilbenoids

The peanut plant (Arachis hypogaea L.), when infected by a microbial pathogen, is capable of producing stilbene-derived compounds that are considered antifungal phytoalexins. In addition, the potential health benefits of other stilbenoids from peanuts, including resveratrol and pterostilbene, have been acknowledged by several investigators. Despite considerable progress in peanut research, relatively little is known about the biological activity of the stilbenoid phytoalexins. This study investigated the activities of some of these compounds in a broad spectrum of biological assays. Since peanut stilbenoids appear to play roles in plant defense mechanisms, they were evaluated for their effects on economically important plant pathogenic fungi of the genera Colletotrichum, Botrytis, Fusarium, and Phomopsis. We further investigated these peanut phytoalexins, together with some related natural and synthetic stilbenoids (a total of 24 compounds) in a panel of bioassays to determine their anti-inflammatory, cytotoxic, and antioxidant activities in mammalian cells. Several of these compounds were also evaluated as mammalian opioid receptor competitive antagonists. Assays for adult mosquito and larvae toxicity were also performed. The results of these studies reveal that peanut stilbenoids, as well as related natural and synthetic stilbene derivatives, display a diverse range of biological activities.     

Isolation of Babesia divergens from carrier cattle blood using in vitro culture

Babesia divergens, the main causative agent of bovine babesiosis in Western Europe, was isolated from naturally infected cattle. Ninety-six blood samples were examined by means of an in vitro culture technique in sheep erythrocytes: 19 of them were collected from animals in the acute phase of the disease with visible parasitemia on blood smears, while the 77 remaining animals showed no microscopically detectable parasites. B. divergens was cultured from the 19 first blood samples as well as from 31 samples collected from asymptomatic animals. The time period before parasites could be detected in the culture varied in the latter samples from 6 to 20 days. The effects of sampling condition (anticoagulant used) and storage length were tested. A good correlation was obtained between immunofluorescent antibody test and culture, with identical results (positive or negative) for 89.6% of the samples collected from asymptomatic animals. The sensitivity of the in vitro culture method was determined and was about 10 parasites/mL of whole blood from three independent experiments performed with three different isolates, confirming its suitability to detect and culture diverse B. divergens isolates from carrier cattle. The parasites could indeed be isolated 9 months after the acute babesiosis phase in the blood of naturally infected animals. The 50 isolates collected in this study were successfully subcultured, cryopreserved and resuscitated using the same culture medium. The in vitro isolation of B. divergens from asymptomatic carrier cattle was achieved and will allow the analysis of parasite diversity within cattle herds.     

Lippia alba (Verbenaceae) hydrolate as sedative of tambaqui (Colossoma macropomum) juveniles in simulated transport conditions

Lippia alba hydrolate (LAH) is produced as a secondary product in essential oil extraction. The objective of this study was to evaluate the use of LAH in a simulation of the transport of tambaqui (Colossoma macropomum) juveniles. A series of 17 hr simulations were performed using load densities of 30, 60 and 90 juveniles per L and four LAH concentrations of 0.4%, 1%, 2% and 5%, with a control of just water. Fish survival, temperature, dissolved oxygen, conductivity, pH, ammonia excretion, alkalinity water hardness and net Na⁺, K⁺, Cl^‐ fluxes, were all evaluated before and after the transport simulations. Mucous cell densities in the gills were also determined microscopically, post simulation. The results showed that addition of 5% LAH achieved better survival during the simulation than the water control. Dissolved oxygen was significantly lower with an increased load density. Also, conductivity, NH₄ excretion and net ionic fluxes (K⁺ and Cl^?) were lower with 5% LAH, although mainly where higher load densities were tested. Mucous cell density was lower in LAH concentrations of 2% and 5%, again with higher load densities. In view of these findings, it is suggested that 5% LAH is an effective sedative for use in the transport of tambaqui, mainly with high load densities. This work is the first study of LAH and highlights its potential applications in aquaculture management.     

Lethal concentration of Cymbopogon citratus (Poaceae) essential oil for Dolops discoidalis and Argulus sp. (Crustacea: Argulidae)

This study investigated the acute toxicity (LC_50-24 hr) effects of the essential oil of Cymbopogon citratus for adult Argulus sp. and Dolops discoidalis, before and during oviposition. In vitro acute toxicity (LC_50-24 hr) was tested using 20, 40, 60, 80, 100, 120, 140 and 160 μg/L of C. citratus essential oil, and two control groups (one with cultivation tank water and one with cultivation tank water + alcohol) were used. Specimens of Argulus sp. and D. discoidalis submitted to acute toxicity were evaluated using histological procedures. The major chemical compounds of C. citratus essential oil were geranial (47.5%), neral (35.6%) and myrcene (6.7%). The LC_50-24 hr for Argulus sp. adults was 67.97 μg/L, while for D. discoidalis it was 59.55 µg/L. In the oviposition of both species of argulids, maximum mortality began with treatments of 140 μg/L, while the LC_50-24 hr for Argulus sp. and D. discoidalis was 83.98 μg/L and 82.48 μg/L, respectively. In both argulid species exposed to C. citratus essential oil, morphological alterations were observed only in the eyes, and they occurred in the ommatidium and rhabdomeres and were dependent on the concentration of C. citratus essential oil and the parasite species.     

Pathogenic Botryosphaeriaceae associated with <Emphasis Type="Italic">Mangifera indica</Emphasis> in the Kimberley Region of Western Australia

Members of the Botryosphaeriaceae, in particular Lasiodiplodia theobromae, Neofusicoccum parvum, N. mangiferum and Botryosphaeria dothidea, commonly cause stem cankers, dieback and stem end rot of mangoes worldwide. In the current study, eight taxa of Botryosphaeriaceae were identified as canker-associated fungi, pathogens, potential pathogens or endophytes of mangoes in the Kimberley, Australia. These include Neoscytalidium novaehollandiae, Ne. dimidiatum, Pseudofusicoccum adansoniae, P. ardesiacum, P. kimberleyense, Lasiodiplodia sp. 1, L. iraniensis and L. pseudotheobromae. The pathogenicity of a selection of these species toward fruit and branches was tested. All were pathogenic to mango in comparison to the control, with Lasiodiplodia spp. being the most pathogenic. It appears that either geographic isolation or the unique growing conditions in the Kimberley may have provided an effective barrier to the acquisition or establishment of known botryosphaeriaceous pathogens. Wounds caused by mechanical pruning may provide an entry point for infection, whilst severe pruning may increase plant stress.