Plasma membrane of trout spermatozoa: I. Isolation and partial characterization

The plasma membrane from spermatozoa of rainbow trout was isolated by four techniques: sonication, hypotonic shock, mechanical homogenization after freeze-thawing, and nitrogen cavitation, in combination with continuous sucrose gradient centrifugation. Nitrogen cavitation (900 PSI, 20 min equilibration at 4°C) was the most effective technique.Following nitrogen cavitation, four bands were recovered in the sucrose gradient at densities 1.03, 1.05, 1.09 and 1.15 g/ml. Electron microscopy revealed membrane vesicles of various sizes in bands 1 to 3, while enzyme analysis revealed a 3.9 to 5.5-fold enrichment in 5'-nucleotidase and little contamination by lactate dehydrogenase (cytosol) and succinic dehydrogenase (mitochondria). Lipid analysis of bands 1 and 2 indicated a 6 to 7-fold enrichment in cholesterol and a cholesterol: phospholipid ratio of 0.59–0.70. Seven classes of phospholipids were present in bands 1–3 with no significant differences observed among bands. These data indicate that the vesicles (in bands 1 and 2) obtained after nitrogen cavitation are primarily plasma membranes. Membranes in band 3 appear to be slightly contaminated with nuclear membranes.Most of the plasma membrane proteins were acidic to neutral. The 2 main membrane proteins were 42 and 30 Kilodaltons.     

Sensitivity of Seeds to Chemical Mutagens,Detection of DNA Polymorphisms and Agro-Metrical Traits in M1 Generation of Coffee (Coffea arabica L.)

Journal of Crop Science and Biotechnology - Coffee (Coffea Arabica L.) is threatened by biotic and abiotic stresses. Nevertheless, the breeding of Arabica coffee is restricted due to its low...     

New method for reducing the binding power of sweet white wines

Sulfur dioxide is now considered to be a toxic chemical by most world health authorities. However, it remains an irreplaceable additive in enology for wine conservation, combining antioxidant and antibacterial properties. Sweet white wines from botrytized grapes retain particularly high SO 2 levels due to their high sulfur dioxide binding power. This paper presents a new method for reducing this binding power by removing some of the carbonyl compounds responsible, which are naturally present in these wines. The main carbonyl compounds responsible for the SO 2 binding power of sweet wines were removed, that is, acetaldehyde, pyruvic acid, 2-oxoglutaric acid, and 5-oxofructose. The method retained was selective liquid-solid removal, using phenylsulfonylhydrazine as a scavenging agent. The scavenging function was grafted on different classes of porous polymer supports, and its efficiency was evaluated on sweet white wines under conditions intended to conserve their organoleptic qualities. The results obtained showed that the method was efficient for removing carbonyl compounds and significantly reduced the binding power of the wines. Sensory analysis revealed that this process did not deteriorate their organoleptic qualities.     

Genetic variability of rhizobacteria from wild populations of four Lupinus species based on PCR-RAPDs§

The metabolic capacities of rhizosphere bacteria can depend on intraspecific genetic variability at strain level. We sampled bacteria from the rhizosphere of three populations of four different Lupinus species at two growth stages (flowering: GS1 and fruiting: GS2). Isolates were identified to the genus level by classical biochemical tests. The most abundant genera found were Bacillus, Aureobacterium, Cellulomonas, Pseudomonas, and Arthrobacter. Genetic divergence of rhizobacteria was tested by PCR-RAPDs. The genetic distances were low, with mean values of 37 % for Bacillus, 25 % for Aureobacterium, 46 % for Cellulomonas, 16 % for Pseudomonas, and 23 % for Arthrobacter. Aureobacterium, the most abundant genus, predominated in the rhizosphere of all populations and at both growth stages (GS1 and GS2) of L. angustifolius. The Aureobacterium strains consisted of 11 groups with 90 % similarity indexes. The cluster analysis of these groups shows that strains isolated from different lupin species and sampling times have extraordinary low diversity indexes, or are even identical. This fact, together with the low genetic distance detected in the rhizosphere, reveals a clear specificity in the plant-bacteria interaction. This specificity could be related with several aspects of plant physiology.     

Characteristics of Peri-urban Dairy Herds of Bobo-Dioulasso (Burkina Faso)

Peri-urban dairy cattle farms within 50 km of Bobo-Dioulasso were studied to assess herd type, disease incidence, management, feeding and breeding strategy. Out of 417 cattle farmers, 42% had dairy objectives and were studied. Among these peri-urban dairy farmers, 60% were settled, 36% semi-settled, and 4% transhumant. In total, they held 4558 dairy cows, of which 32% lactated during the study. The prevalence of mastitis (55%) increased (p<0.05) with herd size. Advanced strategies for supplementary feeding and breeding were most frequent in small herds (<30 cattle). None of the large herds (>60 cattle) had advanced breeding strategies. Bulls and cows were culled at younger ages in herds with better breeding strategy. Overall, this resulted in higher individual milk offtake in small herds (2.46 L/day) compared with large herds (1.25 L/day). Pure breeds were rarely used, and the presence of Bos taurus baoulé naturally selected for trypanotolerance was low. The prevalence of trypanosomosis (40%) in herds dominated by Bos indicus zebu and Méré (Zebu x Baoulé) is an argument for maintenance of biodiversity and selection of Baoulé for milk production.     

Comparison of diagnostics techniques in an outbreak of infectious laryngotracheitis from meat chickens

Various diagnostics techniques were compared for their ability to detect infectious laryngotracheitis (ILT) during an outbreak in chickens aged between 4 and 21 wk. Gross lesions ranged from excess mucus to accumulation of fibrinonecrotic exudate in the larynx and trachea. Syncytial cells with intranuclear inclusion bodies were found in sinus, conjunctiva, larynx, trachea, lung, and air sac. Virus isolation in chicken embryos was attempted in every case. Negative-stain electron microscopy detected herpesvirus in only 6% of the cases. Yet, isolation of ILT virus in the chorioallantoic membrane was presumed by histology in >20% of the samples and confirmed by fluorescent antibody (FA) in 35% of the embryos inoculated with conjunctivas or tracheas from affected birds. Overall, results from histology and FA tests were highly correlated. FA test has the advantage over histology of being diagnostically specific for ILT virus. Polymerase chain reaction was the most sensitive test and detected the viral DNA even in cases where histology and FA were negative. ILT virus DNA was quantified by real-time polymerase chain reaction (Re-Ti ILTV). Histologic and FA results from larynx and trachea were negative if the concentration of the viral DNA was < or =4 of log10. A viral DNA concentration higher than log10 4, as determined by Re-Ti ILTV, was required for clinical ILT to be manifested.     

Bioactive cinchona alkaloids from Remijia peruviana

Three known Cinchona alkaloids of the quinine type, quinine (1), cupreine (2), cinchonine (3), and the possible artifact cinchonine-HCl (3-HCl), along with two new ones, acetylcupreine (4) and N-ethylquinine (5), have been isolated from the bark of Remijia peruviana (Rubiaceae). Their stereochemical structures were established by high resolution NMR spectroscopy. Alkaloids 2-4 had antifeedant effects on Leptinotarsa decemlineata with varying potencies. Compound 4 was cytotoxic to both insect Sf9 and mammalian CHO cells after 48 h of incubation, while 3-HCl had stronger and selective cytotoxicity to Sf9. Quinine 1 had a moderate to low effect on Trypanosoma cruzi. Tumoral cells were also affected by these alkaloids, with 4 and 3-HCl being the most cytotoxic to all the cell lines tested. Overall, the 8R, 9S configurations, as in 3 and 3-HCl, as well as the C-6'acetylated alkaloid 4, with an 8S, 9R configuration, showed stronger biological effects.     

Embolism induced by winter drought may be critical for the survival of <Emphasis Type="Italic">Pinus sylvestris</Emphasis> L. near its southern distribution limit

Introduction   

Scots pine populations in the SE “Sistema Ibérico” range suffered a severe defoliation in the S face of the crown in isolated trees of thinned stands. This process was detected at the end of the winter 2001–2002.     

Protective Effects Against Erwinia amylovora Induced by hrp Mutants in the Whole Plant are Only Partially Mimicked in Cultivated Cells

A virulent strain of Erwinia amylovora, the causal agent of fire blight of Maloideae, and two of its non-virulent hrp mutants (a secretory and a regulatory mutant) were inoculated into apple cell suspensions either alone or in mixed inoculations. In single inoculations, death of 4- to 5-day-old apple cells occurred only when the concentration of the virulent strain of E. amylovora reached a threshold inoculum concentration of 10⁴CFUml^–1, while high concentrations of the hrp mutants were unable to kill apple cells. When mixed inoculated with the virulent parental strain, both hrp mutants protected apple cells from death caused by the virulent strain. The protective effect was associated with a decrease in the population level of the virulent strain and it was dependent on the non-virulent to virulent inoculum concentration suggesting a competition between the non-virulent mutant and the virulent strain. However, no differential protective ability between the two types of mutants could be noticed, contrary to previous results obtained with apple seedlings or apple flowers in which the regulatory mutant was significantly more effective than the secretory mutant. In conclusion, inoculation of apple cell cultures with E. amylovora does not seem to be a model suitable for investigating mechanisms leading to protection.