Influence of pesticides on the activity and kinetics of invertase,urease and acid phosphatase enzymes

The effects of four pesticides (three herbicides, atrazine, paraquat, glyphosate, and an insecticide, carbaryl) on the activity and kinetics of an invertase (from yeast), a urease (from jack bean) and an acid phosphatase (from potato) were investigated. Glyphosate and paraquat showed a marked activation effect on invertase activity. From the dependence of V_max and K_m parameters on pesticide concentrations, a mixed-type activation mechanism was suggested. The kinetic behaviour of urease and acid phosphatase appeared to be uninfluenced by both pesticides. Methanol, used as solvent for both carbaryl and atrazine, inhibited invertase, urease and acid phosphatase with, respectively, competitive, non-competitive and uncompetitive mechanisms. The extent of the inhibition was different, as estimated by the values of the inhibition constant K₁. An additional inhibition effect of urease and invertase activities was achieved in the presence of carbaryl, whereas no influence was detected on the activity and kinetics of acid phosphatase. Finally, atrazine was shown to alter the kinetics of the enzymes only at higher concentrations.     

Comparative pathology and pathogenesis of spontaneous and experimentally induced fibropapillomas of green turtles (Chelonia mydas)

Tumor biopsy samples from 25 Floridian and 15 Hawaiian green turtles (Chelonia mydas) with spontaneous green turtle fibropapillomatosis (GTFP) and from 27 captive-reared green turtles with experimentally induced GTFP were examined microscopically to differentiate the histologic features that result from GTFP pathogenesis and those that result from incidental factors that may vary according to geographic region. Common histologic features for spontaneous and experimentally induced tumors included fibroblast proliferation in the superficial dermis, epidermal acanthosis and hyperkeratosis, epidermal basal cell degeneration with dermal-epidermal cleft formation, spinous layer degeneration with intraepidermal vesicle and pustule formation, and ulceration. Visceral tumors, found in eight of 10 (80%) free-ranging turtles with cutaneous disease that were examined after death, had extensive interstitial fibrous proliferation. The presence of spirorchid trematode eggs and associated foreign body granulomas, common secondary findings within spontaneous tumors, varied by geographic location, and these findings were not observed in experimentally induced tumors. Eosinophilic intranuclear inclusions and intranuclear herpesvirus-associated antigen immunoreactivity were found in 18 of 38 (47%) experimentally induced cutaneous tumors and nine of 119 (7.5%) spontaneous tumors from Floridian but not Hawaiian turtles. The possible involvement of GTFP-associated herpesvirus in the pathogenesis of epidermal degenerative changes and GTFP pathogenesis is discussed.     

Humoral response (IgG) of goats experimentally infected with Fasciola hepatica against cysteine proteinases of adult fluke

The use of cysteine proteinases from Fasciola hepatica adult flukes for the serodiagnosis of caprine fasciolosis by means of an indirect ELISA test was studied. Two proteolytic fractions from adult fluke homogenates, with apparent molecular weights of 28 and 34 kDa (P28 and P34 respectively), were characterised as cysteine proteinases using azocasein assays and gelatin gel analysis. Both P28 and P34 fractions were electroluted and used as antigens in two different indirect ELISA tests. Serum IgG levels against P28 and P34 in goats given an experimental primary infection with 200 metacercariae or in goats given two experimental infections with 200 metacercariae were determined and compared with those observed in an uninfected control group. ELISA tests using both cysteine proteases showed a rapid and consistent detection of specific IgG in all experimentally infected goats. The IgG response to P28 was the first to be detected as early as 2-3 weeks post-infection and remained elevated throughout the experiment. The response to P34 was detected later (4-6 wpi) and disappeared in some animals at 18 wpi, while flukes were still present in the bile ducts. No significant differences were observed between the anti-P28 and anti-P34 IgG responses between animals receiving a primary or a challenge infection. The results of our study, although preliminary, are promising since the P28 ELISA described here may be a reliable method for the immunodiagnosis of F. hepatica infection in goats.     

Evaluation of a specific immunochemotherapy for the treatment of canine visceral leishmaniasis

The efficacy of specific immunochemotherapy against Leishmania infantum infection in dog was studied. The effects on transmission of the disease, as well as the cellular and humoral immune response were examined. The treated animals showed a significant reduction in the infection rates that were detected in Phlebotomus perniciosus females fed on the dog. The humoral immune response, assayed with an indirect immunofluorescence antibody test (IFAT), did not show significant variations under the influence of the therapy. The characterisation of the peripheral blood mononuclear cells (PBMC) using flow cytometry indicated a significant increase in the proportion of T lymphocytes, especially of CD4/TcR(alpha)(beta)(+) and CD4/CD45RA(+) cells, without showing evidence for modifications in the other leukocyte subsets. Cellular lymphoproliferation studies indicated a lack of a specific response to soluble leishmanial antigen (SLA), but the non-specific lymphoproliferative capacity assayed with phytohemagglutinin (PHA) was maintained.     

Isolation and characterization of immortalized porcine aortic endothelial cell lines

Primary porcine endothelial cells have a limited life span in culture. After four to five passages, they tend to de-differentiate and eventually reach senescence. The aim of this work was to establish immortalized porcine aortic endothelial cell lines (AOCs) to facilitate in vitro studies of different pathological process involving the endothelium. Primary porcine aortic endothelial cells (PAECs) were transfected with a plasmid containing the SV40 genome and selected on the basis of morphological and phenotypical features. Flow cytometry analysis demonstrated uptake of acetylated low density lipoproteins (Ac-LDL) and constitutive expression of SLA class I, CD29, CD31, CD41/61, CD80/86, CD46, SWC3, and LAMP-1 antigens by all analyzed lines and showed little differences to primary cells. The functional similarity between primary and immortalized endothelial cells was demonstrated in a cytotoxicity assay using a human natural killer cell line (NKL) as effector. The AOCs cell lines should be valuable tools for in vitro study of the human immune response against pig endothelial cells. In addition, they would be very useful to gain insight in the pathogenesis of some viral haemorrhagic diseases of pig such as African swine fever (ASF) or classical swine fever (CSF).     

Evaluation of 65% permethrin spot-on for prevention of canine visceral leishmaniasis: effect on disease prevalence and the vectors (Diptera: Psychodidae) in a hyperendemic area

A study was designed to examine the effect of 65% permethrin spot-on on the prevalence of canine visceral leishmaniasis and the abundance of sand flies in two neighborhoods in Corumbá, Mato Grosso do Sul, Brazil known to have a high prevalence of visceral leishmaniasis. An enrollment survey was conducted to determine the prevalence of visceral leishmaniasis. Area residents were provided with information about the project; the study area was defined, and all dogs (160 in Cristo Redentor and 230 in Popular Velha) identified in the study area were enrolled. Three 65% permethrin spot-on treatments (June 15-25, July 13-15, and August 10-12) were administered to 150, 110, and 99 dogs, respectively, in Popular Velha, according to label recommendations. Dogs in Cristo Redentor were untreated controls. Visceral leishmaniasis was diagnosed periodically by indirect immunofluorescence assay. A reduction in canine visceral leishmaniasis prevalence was observed at the Popular Velha site. The infection rate for treated dogs 1 month following the final treatment was approximately 50% reduced from that observed before treatment(19.3% vs 9.6%). Conversely, the infection rate at the control site was more than 80% higher at the September sampling than that observed pretreatment (4.1% vs 7.4%). Similar numbers of sand flies were captured and identified from both sites throughout the study. The results suggest that regular use of 65% permethrin during months of high risk for canine visceral leishmaniasis can be a useful strategy for reducing the prevalence of this disease in hyperendemic areas. It should be stressed, however, that the success of this strategy depends not only on the efficacy of the product itself but also on the adoption of other control measures and on economic variables, considering the low purchasing power of the populations living in higher-risk neighborhoods.     

Volume Contents

Volume Contents

Volume Contents     

Interactions between Plasmopara helianthi, Glomus mosseae and Two Plant Activators in Sunflower Plants

Interactions between Plasmopara helianthi, Glomus mosseae and two plant activators DL--amino-n-butyric acid (BABA) and CGA 245704 (acibenzolar-S-methyl (BTH)) in sunflower plants susceptible to downy mildew were studied in four experiments using different methods of treatment and pathogen inoculation. Both chemicals were applied as soil drenches and foliar sprays, whereas P. helianthi infection was obtained by root and cotyledon inoculations of the seedlings. Soil drenches at the rates of 50 and 100mgkg^–1 soil of BABA and BTH given 1 and 3 days before P. helianthi inoculation, respectively to mycorrhizal plants, provided moderate protection against the pathogen (about 50–55%). Morphological changes and decrease in mycorrhizal colonization in roots of BTH-treated plants and in BTH-treated mycorrhizal plants were also observed. Delay in the emergence and reduction of the root systems were more evident at the highest concentration but decreased with time. These effects were absent with the BABA treatment.Foliar spray treatment of BABA and BTH, applied at 4000 and 200µgml^–1, respectively (1 day post-inoculation) to mycorrhizal plants provided good protection (about 80%) against P. helianthi foliar infections. No effects on mycorrhizal colonization or on root systems were observed. In vitro tests on the effect of the compounds on the mycorrhizal fungus showed that the germination of G. mosseae sporocarps increased with BABA treatment whereas it was greatly inhibited by BTH treatment.     

Improvements in somatic embryogenesis protocol in Feijoa (Acca sellowiana (Berg) Burret): Induction,conversion and synthetic seeds

Pineapple guava (Acca sellowiana) syn. Feijoa sellowiana, a Brazilian indigenous Myrtaceae is under domestication in South Brazil. Previous works showed that this species is responsive to somatic embryogenesis and recalcitrant to conventional methods of clonal propagation. In the present work it was evaluated the role of components of culture medium in the induction and development of somatic embryos. The technology of synthetic seeds was also evaluated. Zygotic embryos were inoculated in LPm medium supplemented with 8 mM glutamic acid and 8 mM l-glutamine, 2,4-dichlophenoxiacetic acid (20 μM) and myo-inositol. For conversion of somatic embryos and synthetic seeds it was tested the effect of 6-benzylaminopurine and gibberellic acid combined or not with activated charcoal. The highest values for embryogenetic induction (100%) and number of somatic embryos/explant (113) were observed in the LPm medium supplemented with Glu (8 mM), and 2,4-D. The culture medium supplemented with BA (0.5 μM) and GA₃ (1 μM) and activated charcoal (1.5 g L⁻¹) enhanced the conversion of somatic embryos to plantlets. Pre-germinated somatic embryos encapsulated in sodium alginate with BA (0.5 μM) and GA₃ (1 μM) developed radicles. The use of synthetic seed was a requisite for the survival of plantlets.