Selection,characterization and genetic analysis of laboratory mutants of <Emphasis Type="Italic">Botryotinia fuckeliana</Emphasis> (<Emphasis Type="Italic">Botrytis cinerea</Emphasis>) resistant to the fungicide boscalid

Resistance to the fungicide boscalid in laboratory mutants of Botryotinia fuckeliana (Botrytis cinerea) was investigated. The baseline sensitivity to boscalid was evaluated in terms of colony growth (EC₅₀ = 0.3–3 μg ml⁻¹; MIC = 10–30 μg ml⁻¹) and conidial germination (EC₅₀ = 0.03–0.1 μg ml⁻¹; MIC = 1–3 μg ml⁻¹) tests. Mutants were selected in vitro from wild-type strains of the fungus on a fungicide-amended medium containing acetate as a carbon source. Mutants showed two different levels of resistance to boscalid, distinguishable through the conidial germination tests: low (EC₅₀ ∼ 0.3 μg ml⁻¹, ranging from 0.03 to 1 μg ml⁻¹; MIC > 100 μg ml⁻¹) and high (EC₅₀ > 100 μg ml⁻¹) resistance. Analysis of meiotic progeny from crosses between resistant mutants and sensitive reference strains showed that resistant phenotypes were due to mutations in single major gene(s) inherited in a Mendelian fashion, and linked with both the Daf1 and Mbc1 genes, responsible for resistance to dicarboximide and benzimidazole fungicides, respectively. Gene sequence analysis of the four sub-units of the boscalid-target protein, the succinate dehydrogenase enzyme, revealed that single or double point mutations in the highly conserved regions of the iron-sulphur protein (Ip) gene were associated with resistance. Mutations resulted in proline to leucine or phenylalanine replacements at position 225 (P225L or P225F) in high resistant mutants, and in a histidine to tyrosine replacement at position 272 (H272Y) in low resistant mutants. Sequences of the flavoprotein and the two transmembrane sub-units of succinate dehydrogenase were never affected.     

Evaluation of Compost Maturity,Hydrophysical and Physicochemical Properties: Indicators for Use as a Component of Growing Media

The cocomposting of agricultural waste is a new management priority in Tunisia. In this study, four composts were evaluated by comparing the changes in measured hydrophysical and physicochemical properties and phytotoxicity. The organic wastes used were almond shell (AS), sesame bark (SB), olive husk (OH), and green and wood wastes (GW and WW, respectively). Composts CI and CII were composed of AS/SB and OH/SB, respectively, at a ratio of 75/25 (wet weight basis). CIII consisted of OH, SB and CAS (coarsely-ground almond shell used as a bulking agent) at a ratio of 55/25/20. Finally, CIV was composed of 25%SB+9%CAS+18%GW+48%WW. The composts studied were characterized by basic pH and an electric conductivity (EC) value ranging from 1.6 to 2.4 mS/cm. The organic matter contents (OM) and C/N ratios of composts ranged from 20 to 46% and from 10 to 21%, respectively. Based on hydrophysical analyses, composts CI, CIII and CIV, containing AS, were shown to have a porosity and a water content of 10-26% and 10-20%, respectively. The phytotoxicity of composts was studied on the basis of cress seed germination. Results revealed that differences in properties are mainly related to the nature of composted waste and that some of these composts are compatible for use as constituents in growing media for horticultural soilless cultures.     

Seasonal Variations in Proximate and Fatty Acid Composition of Viscera of Sardinella aurita,Sarpa salpa,and Sepia officinalis from Tunisia

Seasonal variations of the proximate composition and fatty acid profiles in viscera from Sardinella aurita, Sarpa salpa, and Sepia officinalis were studied. Significant seasonal variations were observed in the amounts of moisture, lipid, protein, and ash between species. Viscera protein content undergoes large fluctuations. This tendency is different from results observed for the edible parts or for the whole body of many marine species. Ash content also showed significant differences. Lipid contents varied with seasons, in a proportionally inverse manner to water contents. Fatty acid composition showed significant differences from October to December. Interestingly, the highest total omega-3 contents were comparable to many commercial marine fish oils.     

Detection of Water and Sediments Pollution of An Arid Saltern (Sfax, Tunisia) by Coupling the Distribution of Microorganisms With Hydrocarbons

We investigated the coupling of abundance of bacteria, phytoplankton and ciliates with hydrocarbons in the surface water and sediments of five interconnected ponds in the arid Sfax solar salterns. This study aimed at determining the potential sources of hydrocarbons and the effects of salinity gradients on microorganism metabolism. Hydrocarbon analysis was performed by gas chromatography (GC-FID) and gas chromatography coupled with mass spectrometry (GC-MS). The GC-FID allowed the detection of aliphatic hydrocarbons and n-alkanes ranging from n-C₁₃ to n-C₃₀. Total aliphatic hydrocarbon concentrations varied from 92.5 mg. l^?1 in the first pond (having marine characteristics) to 661.1 mg. l^?1 in the last pond (crystallizer) (316.8?±?120.1 mg. l^?1) for water samples and from 26.7 to 127.8 μg. g^?1 dry weight for sediment samples. The GC-MS enabled us to detect halogenated hydrocarbons (bromoalkanes and chloroalkanes) and n-alkenes. The distribution of n-alkanes indices coupled to several environmental factors suggests that a major fraction of hydrocarbons resulted from both prokaryotic (bacteria) and eukaryotic (protists) developments. A low hydrocarbon fraction might be petrogenic.     

Resistance profiles of Botrytis cinerea populations to several fungicide classes on greenhouse tomato and strawberry in Lebanon

Tomato and strawberry are the most important protected crops in Lebanon and are seriously affected by grey mould disease, caused by Botrytis cinerea. In the present study, the fungicide sensitivity assays revealed medium to high frequencies of B. cinerea isolates resistant to benzimidazoles, dicarboximides, and anilinopyrimidines on tomato and strawberry. Fludioxonil- and boscalid-resistant mutants were uncommonly found at generally low frequency on both crops. Resistance to fenhexamid was detected in only one site on tomato but in most sites on strawberry with high frequencies, and the occurrence of resistance to QoI fungicides was ascertained on both crops. The majority of the tested isolates (>90%) exhibited multiple fungicide resistance, and isolates resistant to the seven antibotrydial fungicide classes were detected on strawberry in three locations. A high level of resistance was shown by B. cinerea mutants resistant to boscalid, fenhexamid, and QoI fungicides, while two levels of moderate and high resistance to anilinopyrimidines were identified. Genetic analysis revealed point mutations in the target genes commonly associated with resistance in B. cinerea isolates, with all mutants resistant to dicarboximides, fenhexamid, boscalid, and QoI fungicides carrying single-nucleotide polymorphims in BcOS1 (I365S/N, Q369P, and N373S), Erg27 (F412V/I), SdhB (H272R/Y), and cytb (G143A) genes, respectively. The general incorrect use of fungicides has caused the development and spread of fungicide resistance as a widespread phenomenon on protected tomato and strawberry in Lebanon. The implementation of appropriate antiresistance strategies is highly recommended.     

Golden Grey Mullet (Liza aurata) Alkaline Proteases: Biochemical Characterization,Application in the Shrimp Wastes Deproteinization,Laundry Commercial Detergents,and Preparation of Antioxidant Protein Hydrolysate

Digestive alkaline proteinases from golden grey mullet (Liza aurata) were extracted and characterized. The crude alkaline protease showed optimum activity at pH 8.0 and 60°C, and it was highly stable over a wide range of pH from 4.0 to 10.0, retaining more than 80% activity after incubation for 1 h at 4°C. The alkaline proteases showed extreme stability toward nonionic and anionic surfactants after preincubation for 1 h at 25°C and relative stability toward oxidizing agents. Additionally, the crude enzyme showed excellent stability and compatibility with various solid and liquid detergents. Further, proteases from golden grey mullet viscera were found to be effective in the deproteinization of shrimp wastes. The protein removal after 3 h at 45°C with an enzyme/substrate (E/S) ratio of 10 U/mg protein was about 76%. The golden grey mullet proteases were also shown to be efficient in the production of antioxidant protein hydrolysate.     

Growth performance, feed utilization, and body composition of Nile tilapia (Oreochromis niloticus L.) fed with differently heated soybean-meal-based diets

This study was undertaken in order to determine the effect of feeding heat-treated, defatted soybean meal (SBM) on growth, feed utilization, and body composition of Nile tilapia (Oreochromis niloticus). A control diet (SBM₀) of 378 g kg⁻¹ crude protein with 18.4 kJ g⁻¹ gross energy was formulated, and three diets identical to the basal diet were autoclaved for 10, 20, or 30 min. The autoclaved diets were named SBM₁₀, SBM₂₀, and SBM₃₀, respectively. Each diet treatment was applied to triplicate groups of 30 fish (2.45 ± 0.03 g) per tank (120 l). The fish were hand fed to satiation four times daily for 45 days. At the end of the feeding trial the fish fed with the SBM₃₀ diet had significantly (P < 0.05) higher weight gain and protein efficiency ratio than those fed with the other diets. No feed-related mortality was observed during the whole experimental period. Heating SBM for 30 min reduced trypsin inhibitor activity (TIA) and increased apparent protein digestibility (APD), and indicated significant differences (P < 0.05) among various treatments. No significant differences were found in carcass moisture, lipid, and ash of fish fed with different experimental diets. An increase in the body protein content of fish fed with diet SBM₃₀ was significantly (P < 0.05) higher than in all other experimental groups. The results of this study seem to indicate that autoclaving the SBM for 30 min improved its nutritional value in practical feeds for Nile tilapia fingerlings.