Shifts in disease resistance induced by growth regulators

Experiments on the effect of various compounds with plant growth regulating activity led to the hypothesis that conditions inhibitory to indoleacetic acid (IAA) action or leading to a decrease in the IAA level in cucumber seedlings would be unfavourable for the development of cucumber scab, caused byCladosporium cucumerinum. Susceptibility decreased as the result of treatment with growth retardants, which cause an increase in IAA-oxidase activity in the plants, whereas application of indoleacetic acid or compounds expected to decrease the rate of oxidative breakdown of IAA in the seedlings resulted in an increase of susceptibility. Growing the plants under various periods of illumination also influenced both the susceptibility of the plants and the IAA-oxidase activity in the hypocotyl tissue.To obtain further information about the relation between the increase in resistance and the increase in the rate of IAA oxidation, the effect of one compound, viz. L-threo-β-phenylserine (abbr. phenylserine) was studied in more detail. The rate of oxidative breakdown of IAA was increased in extracts of hypocotyls and cotyledons of plants treated with phenylserine as compared with extracts of tissue of untreated plants. Extracts of phenylserine-treated plants were found to contain a higher cofactor and/or a lower inhibitor content than those of control plants. The substances involved were not identified, but this result may indicate a shift in the concentrations of various phenols in the tissue.The question must remain open whether a shift in the concentration of phenols some-how effected by phenylserine treatment determines both degree of susceptibility and rate of IAA oxidation independently, or whether the rate of IAA breakdown and the resulting change in IAA content in the plants is directly related to the degree of susceptibility.     

New potyvirus isolated from <Emphasis Type="Italic">Cryptotaenia japonica</Emphasis>

 A potyvirus, for which the name Japanese hornwort mosaic virus (JHMV) is proposed, was isolated from Japanese hornwort plants (Cryptotaenia japonica) with mosaic disease symptoms. The virus was used to inoculate mechanically 34 plants belonging to 33 species of 10 families. Of these species seven from two families were infected. Faint chlorotic spots appeared on the inoculated leaves of Chenopodium quinoa and C. amaranticolor, but no systemic infection occurred in these plants. JHMV systemically infected only Umbelliferae plants; they did not infect 26 other species in eight families. JHMV was transmitted in a nonpersistent manner by aphids (Myzus persicae). The virus was a flexuous rod-shaped particle about 750 nm in length. Sequencing the nucleotides in the 3′ terminal region of JHMV revealed that the coat protein contains 280 amino acids with a molecular mass of 32.2 kDa. The nucleotide sequence of the coat protein of JHMV had the highest similarity with that of Zantedeschia mosaic virus (83.3%) compared to those of other potyviruses (57.0%–64.9%). An antiserum against JHMV reacted strongly with JHMV and weakly with Potato virus Y. These results indicate that JHMV is a new potyvirus. Received: September 9, 2002 / Accepted: November 7, 2002 RID="*" ID="*" The nucleotide sequence determined in this work appears in the DDBJ/EMBL/GenBank nucleotide sequence databases with the accession number AB081518     

Characterization of a feline homologue of the alphaE integrin subunit (CD103) reveals high specificity for intra-epithelial lymphocytes

The characteristics of a feline homologue of the alphaE integrin (CD103), defined by two murine monoclonal antibodies, Fe7.1B8 (IgG1) and Fe7.2D8 (IgG1), are described. These antibodies recognized 75% of intra-epithelial (range 59-88%) and 40% of lamina proprial (range 28-46%) T cells of the intestinal mucosal tissue of the small intestine in contrast with approximately 2% of peripheral blood lymphocytes. Both antibodies immunoprecipitated a 180 kDa protein from biotinylated feline intra-epithelial mucosal leukocytes consistent with the alphaE integrin subunit in conjunction with a 120 kDa protein consistent with the beta7 subunit. The nucleotide sequence of feline alphaE integrin, generated from molecular cloning of the feline alphaE encoding cDNA, is also reported. This feline molecule shares 72% sequence homology with human and 69% homology with murine and rat counterparts. Homology includes the presence of an X (extra) domain, that appears unique to alphaE molecules as described for human, rat and mouse, as well as areas of homology common to other alpha integrins. Of note is a typical I (inserted) domain, the presence of seven repeat regions, and highly conserved sequences in the cytoplasmic tail. Transfection studies demonstrated that both antibodies recognized an extracellular component which encompassed the X and I domains of the cloned alphaE integrin subunit. These studies demonstrate that the pattern of tissue distribution, biochemical characteristics, and cDNA sequence of the feline alphaE integrin subunit are largely similar to that described for other species.     

A rapid and simple method for the separation of pure lymphocytes from horse blood

A method for the separation of pure and viable lymphocytes and granulocytes from the same blood sample in horses was reported. By centrifuging equine heparinized blood at 100 xg for 10 min at room temperature (r.t.), the resulting supernatant plasma was an almost pure (97.71 +/- 0.30%; n = 15) suspension of highly viable (98.72 +/- 0.28%) lymphocytes. When sodium citrate was used as an anticoagulant, lymphocyte suspensions collected in the same manner showed lower purity (87.89 +/- 1.59%; n = 9) and higher yields (56.56 +/- 3.89%, n = 9 versus 36.11 +/- 2.23%, n = 15). Where needed, a further centrifugation at 250 xg for 3 min (r.t.) of heparinized lymphocyte preparations removed an average of 87.39% (n = 15) contaminating platelets. A suspension of 85.96 +/- 2.20% pure granulocytes (93.23 +/- 1.74% neutrophils; n = 14) with minimal contamination by erythrocytes and high viability (93.11 +/- 1.26%) was obtained by performing a flash red blood cell lysis on the white-greyish layer resulting from the centrifugation of the heparinized blood samples. Among the several methods available, the procedure described herein is easy, rapid, cheap and reproducible.     

Antioxidant systems and lymphocyte proliferation in the horse,sheep and dog

To better define the species-specific antioxidant systems and to ascertain the influence of the intracellular redox status on the immune system of different animal species, we determined lymphocyte glutathione peroxidase (GSHPx) activity, plasmatic glutathione levels (GSH) and the effect of H2O2 on the responsiveness of lymphocytes to proliferative stimuli. Among the three species considered, sheep presented the lowest plasmatic GSH and the highest lymphocyte GSHPx activity. On the contrary, dogs showed an inverted pattern (high GSH - low GSHPx). Horses displayed intermediate values for both parameters analysed. The effect of H2O2 on the proliferative capacity of lymphocytes was the same for all three species; the 200 microM dose in particular was strongly inhibiting. Each species, however, showed different rates of inhibition: sheep exhibited the highest sensitivity to the antiproliferative effect of H2O2. Our results confirmed that high H2O2 concentrations (200 microM) are noxious for the cellular functions of all animals; however this effect is mediated by a rigorously species-specific relationship between the intracellular reactive oxygen species (ROS) and the molecular systems involved in cell proliferation.     

Effect of 1-24ACTH administration on sheep blood granulocyte functions

The objective of the present study was to determine the efficiency of blood neutrophils (PMN) taken from sheep during acute stress. Ten healthy Charolle sheep were sampled before treatment (T0) and 1 (T1), 2 (T2), 24 (T24) and 48 (T48) hours after 1-24ACTH administration. Ten sheep serving as the controls were sampled at the same time intervals, using saline solution instead of 1-24ACTH. At each time sampling, rectal temperature, heart rate, cortisol, glucose, non-esterified fatty acids (NEFA), total and differential leukocyte counts were evaluated. PMN were isolated after centrifugation of whole blood and hypotonic lysis of RBC. Chemotaxis was evaluated on a modified Boyden chamber using a nitrate cellulose filter and both Zymosan activated serum (ZAS) and interleukin-8 (IL-8) as chemoattractants. Phagocytosis was measured using both non-opsonized latex beads and fluoresceinated yeasts opsonized with homologous serum. Superoxide (O(-)2) production was evaluated by measuring superoxide dismutase-inhibitable reduction of ferricytochrome C, and adherence by a colorimetric assay of acid phosphatase activity of adherent cells. The administration of 1-24ACTH induced an acute stress reaction, indicated by the presence of clinical, biochemical and hematological changes. Adherence significantly increased from T0 to T2 in treated sheep. This might be responsible for the depression of non-specific immunity in stressed animals. Studies using stressors other than 1-24 ACTH are needed to verify the influence of other components of the stress reaction on PMN functions.     

Feline leishmaniosis due to Leishmania infantum in Italy

A case of leishmaniosis in domestic cats (Felis catus domesticus) is described. The subject showed a nodular lesion on the eyelid. The diagnosis was achieved by serological, parasitological, and light and electron microscopic investigations. By molecular techniques the aetiological agent was identified as belonging to Leishmania infantum, the species implicated in human and canine leishmaniosis in southern Europe. A preliminary study on the prevalence of asymptomatic feline leishmaniosis, performed in the areas where the infected cat was identified, revealed a low seroprevalence of infection: only 1 (0.9%) of the 110 cat sera examined by indirect fluorescent antibody test was positive for anti-Leishmania antibodies. Because clinical signs in feline leishmaniosis are unspecific and similar to those observed in other diseases commonly found in this species, leishmaniosis must be added to the differential diagnosis by feline veterinary practitioners and adequate serologic and histopathologic investigations must be performed in endemic areas.     

Note: Pyrethroid resistance and possible involvement of glutathione<Emphasis Type="Italic">S</Emphasis>-transferases in<Emphasis Type="Italic">Helicoverpa armigera</Emphasis> from Turkey

An introductory study was conducted to investigate the pyrethroid resistance ofHelicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) strains in Turkey, collected from cotton fields in the Adana and Antalya provinces, through two different synthetic pyrethroid insecticides: lambda-cyhalothrin and esfenvalerate. In addition, the roles of glutathioneS-transferases (GSTs) in this resistance mechanism were analyzed. It was found that whereas resistance ratios for lambda-cyhalothrin (LD₅₀ levels) were 3- and 98-fold increased in the Adana and Antalya strains, respectively, esfenvalerate ratios were 3.3- and 92.3-fold increased in the Adana and Antalya strains, respectively, with respect to the susceptible strain. Furthermore, Adana and Antalya strains showed 2.4- and 2.9-fold higher GST activities than the susceptible strain, respectively. In the Antalya field strain, the minor increase in GST activity compared with the resistance levels implies that GSTs may be not greatly involved in this resistance. It also provides evidence that they could not be the only metabolic mechanism responsible for resistance to lambda-cyhalothrin and esfenvalerate inH. armigera from Turkey. http://www.phytoparasitica.org posting Nov. 16, 2006.     

Critical Use Exemption for methyl bromide for soil disinfestation: Italy’s experience with the European union process

Methyl bromide (MB) was phased out on January 1, 2005, and a Critical Use Exemption (CUE) process has been established in order to define the rules for allocating the needed amounts of the fumigant. This paper deals with the effects of the CUE process on Italian horticulture. The actual usage of the amounts of MB allocated to Italy for the year 2005 in the different areas and on the economically most important crops was monitored and critically evaluated. The usage pattern monitored shows that already in the first year after MB phaseout, Italy was able to replace it completely on crops such as zucchini, salad greens, basil and watermelon. However, on crops such as tomato, pepper, melon, eggplant, strawberry, and cut flowers, further work is required in order fully to replace MB. The emerging problems and research needs are discussed briefly.