Characterization of biotype T ofBemisia tabaci associated withEuphorbia characias in Sicily

The T biotype ofBemisia tabaci (Gennadius, 1889), a population found colonizingEuphorbia characias L. plants on the Nebrodi-Peloritani mountains in Sicily, was biologically characterized. The minimum development time was 29.7 days at 28°C. Based on the regression of 1/dayvs T, the rate of development was calculated as 0.00206, the theoretical lower temperature threshold for development as 9.3°C, and the sum of effective temperatures as 485.1. At 25° C, egg-to-adult development was significantly shorter onDatura stramonium (30.1 days) than on eitherEuphorbia pulcherrima orEuphorbia characias (35.6 and 35.4 days, respectively). The fourth instar nymphs grown onD. stramonium had the typical oval outline and seven pairs of dorsal setae located on cone-like processes, often barely visible. The fourth instar nymphs and their pupal cases grown onE. characias had the outline deformed by the presence of hairs on the lower surface of the leaf. The pupae onD. stramonium were significantly larger (both longer and wider) than those reared onE. characias; on both host plants, female pupae were significantly larger than male ones. Analysis of variance showed that width of females onD. stramonium was significantly larger than the width of those reared onE. characias. Attempts at courtship between T- and Q-biotypes were observed, but adults from different biotypes were never seen mating. Only males were obtained from the seven heterologous crossing attempts, either way, whereas homologous, control breeding produced males and females. The T biotype was able to transmitTomato yellow leaf curl Sardinia begomovirus (TYLCSV) fromD. stramonium toD. stramonium, from tomato to tomato and from tomato toD. stramonium. Attempts to transmit TYLCSV fromD. stramonium to tomato were unsuccessful. The transmission efficiency was significantly lower when tomato was the test plant. The diverse biology and ecology of the T biotype confirm that it is genetically different from most Mediterranean biotypes. This paper is dedicated to the dear memory of our colleague Dr Claudio Arnò, who died on March 12, 2004, while this paper was being written.     

Effects of certain herbicides on the oxidative decarboxylation of tryptophan by peroxidase

Non-enzymic and peroxidase-catalysed oxidative decarboxylations of tryptophan (TPP) were studied. The in-vitro rate of enzymic reaction was affected by various herbicides at low concentrations (10^?5?10^?4_M): dinoseb, 2,4-D, dichlobenil and benazolin acted as inhibitors; on the other hand, chlorpropham, bromacil, diphenamid and 4-ethylamino-6-isopropylamino-1,3,5-triazin-2-ol (hydroxyatrazine) were stimulatory. The results of in-vivo experiments showed that chlorpropham and 2,4-D changed the activity of peroxidase from Brassica germinated seeds in vivo, as they do in vttro. Determinations of consumed TPP were carried out either by spectro-photometry or by chromatography.     

Clinical, haematological and biochemical findings in foals with neonatal Equine herpesvirus-1 infection compared with septic and premature foals.

A retrospective multicentre study comparing historical, clinical, haematological, acid-base and biochemical findings of foals with Equine herpesvirus-1 (EHV-1) infection, septicaemia or prematurity was performed to determine if early diagnosis of EHV-1 foals was possible. Fifty-three foals were studied and were assigned to one of 2 groups: herpes positive (n = 14) or herpes negative (n = 39). The latter group included 20 septic, 11 premature, and 8 premature and septic foals. The presence of herpes antigen was confirmed by immunoperoxidase histochemical staining of tissues from necropsied foals. A nonparametric statistical analysis followed by a backwards elimination logistic regression was performed to establish a model at a P value of <0.05. All herpes positive foals died, while 47% (9/19) of the septic foals survived. Based upon our analysis, herpes positive foals were more likely to have total white blood cell counts less than 3 x 10(9)/l and to be icteric as compared to the septic and premature foals. Despite profound hepatic necrosis in the herpes positive foals, liver enzymes were not elevated and were not significantly different from the controls.     

Fermentability of grape must after inhibition with dimethyl dicarbonate (DMDC)

Dimethyl dicarbonate (DMDC) was added to grape must and to synthetic media and results showed that, at 20 degrees C, 150 mg.L(-)(1) DMDC completely inhibited the fermentation of a grape must that was previously inoculated with 10(6) cells.mL(-)(1) Saccharomyces bayanus and Saccharomyces uvarum. Brettanomyces intermedius, Candida guilliermondii, Hansenula jadinii, Hansenula petersonii, Kloeckera apiculata, Pichia membranaefaciens, and Saccharomyces cerevisiae were inhibited by 250 mg.L(-)(1). Candida valida was inhibited in the presence of 350 mg.L(-)(1), whereas Hanseniaspora osmophila, Saccharomycodes ludwigii, Schizosaccharomyces pombe, and Zygosaccharomyces bailii required 400 mg.L(-)(1). Delay of fermentation (but not inhibition) was noted in the presence of 400 mg.L(-)(1) for the following cultures: Brettanomyces anomalus, Hanseniaspora uvarum, Metschnikowia pulcherrima, Schizosaccharomyces japonicus, Torulaspora delbrueckii, and Zygosaccharomyces florentinus. Acetobacter aceti and Lactobacillus sp. were completely inhibited using 1000 and 500 mg.L(-)(1) DMDC, respectively. The fermentation of a grape must inoculated with 10(6) cells.mL(-)(1) of different wine yeasts was delayed for 4 days after the prior addition of 200 mg.L(-)(1) of DMDC; 200 mg.L(-)(1) DMDC did not show any residual inhibitory effect after 12 h, nor did 300 mg.L(-)(1) 24 h after the addition. In cellar experiments, indigenously contaminated grape musts (with and without skins) showed a delay in fermentation of 48 h after the addition of only 50 mg.L(-)(1) DMDC. The possibility of using DMDC (as pure grade as commercially available) in grape must as a disinfectant for the decontamination of musts indigenously contaminated with wild yeast should be considered seriously, despite its apparent low solubility in water.     

Characterization of "lettucine", a serine-like protease from Lactuca sativa leaves, as a novel enzyme for milk clotting

In this work we focused on the characterization of a novel plant rennet purified from lettuce leaves (Lactuca sativa L. cv Romana). The lettuce protease, lettucine, showed trypsin-like, SV8-like, and caseinolytic activities. Although the enzyme did not recognize peptides having hydrophobic amino acid residues in the P(1) position of the target bond, it did show milk-clotting activity, suggesting that different bonds rather than the Phe(105)-Met(106) of the kappa-casein might be cleaved, still inducing milk-clotting. The enzyme exhibited proteolytic activity toward alpha-casein, beta-casein, kappa-casein, and milks with different fat contents, with the highest activity observed with partially skimmed milk, total casein, and alpha- and kappa-casein. SDS-PAGE studies showed that lettucine cleaved alpha-casein, beta-casein, and kappa-casein. In particular, we showed that alpha-casein breakdown occurred even though total casein or milks were supplied, suggesting that the lettuce enzyme is able to operate a significant disorganization of the casein's micellar structure. Moreover, the proteolytic activity of the enzyme analyzed under various technological parameters, such as temperature and pH, indicated that the lettuce enzyme is highly consistent with the milk-clotting process.     

Arylsulphatase activity in soils amended with crop residues: Kinetic and thermodynamic parameters

Laboratory experiments were performed to determine the Michaelis-constants (K_m values) and maximum reaction velocities (V_max values) of arylsulphatase in a clay-loam and a sandy clay-loam soil. When crop residues were buried in either soil, the K_m and V_max values were altered markedly; the changes depended not only on the soil properties but also on the type of crop residue incorporated in the soils. This was confirmed at various temperatures, and data obtained were used to calculate the activation energies (E_a), and changes in enthalpy (ΔH_a) and entropy (ΔS_a) of activation. Whereas these properties were also affected by incorporating crop residues, the free energies of activation (ΔG_a) values, were similar in both soils and derived soil samples.     

Genetic variability and divergence among Italian populations of common ash (Fraxinus excelsior L.)

The level of genetic variation throughout the Italian range of common ash (Fraxinus excelsior L.) was estimated using six microsatellite markers. High levels of allelic diversity was detected. The levels of expected heterozygosity for each of the populations ranged from 0.726 to 0.871, with an average of 0.798, and indicated that populations have a high level of genetic variation. A general and significant homozygote excess was found at most loci in all populations, with an overall mean F _IS of 0.284. Possible explanations for such situations are discussed. Only 4.9% of the total diversity was attributable to differentiation among populations. Although divergence among pedo-climatic regions explained only a small part of the variance it was possible to observe some partial clustering of populations belonging to the same regions. The contribution of the results in relation to the definition of the most appropriate strategies to collect forest reproductive material is discussed.