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 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.     

Effects of three esca-associated fungi on Vitis vinifera L.: IV. Diffusion through the xylem of metabolites produced by two tracheiphilous fungi in the woody tissue of grapevine leads to esca-like symptoms on leaves and berries

Two naphthalenone pentaketides (scytalone and isosclerone) and α-glucans (pullulan) are produced in vitro and in planta by Phaeomoniella chlamydospora (Pch) and Togninia minima (Tmi), two tracheiphilous fungi associated with the ‘esca’ disease of grapevines. The possible role of such fungal metabolites in inducing symptoms on leaves and berries was studied in a vineyard of Vitis vinifera cv. Italia located in southern Italy. During early spring, pruning off two to four branches per vine allowed samples of xylem sap to be collected. Vine bleeding, assessed as ml day⁻¹ vine⁻¹, reached its maximum at bud burst and stopped within 28 days. The total amount of sap collected from healthy vines was about a quarter of that from esca-affected vines. During the same period, the leaf water potential of diseased vines increased progressively (i.e. showed less negative values), indicating a dysfunction in water and nutrient supply to the new growth. Both fungi were isolated from the xylem sap and from the woody tissue of branches and the trunk of diseased vines. Conidia isolated from the sap showed a high germination rate (>90%). Bioactive concentrations of the two pentaketides were detected in xylem sap, leaves and berries at various stages of seasonal growth. Exopolysaccharides, including pullulan, were found in the xylem sap. Absorption of culture filtrates of Pch and Tmi, as well as weak solutions of purified preparations of scytalone, isosclerone or pullulan, on detached leaves and berries caused symptoms similar to those shown by the esca-affected vines in the field.     

<Emphasis Type="Italic">In vitro</Emphasis> screening for sunflower (<Emphasis Type="Italic">Helianthus annuus</Emphasis>L.) resistant calli to <Emphasis Type="Italic">Diaporthe helianthi</Emphasis> fungal culture filtrate

Diaporthe helianthi the causal agent of sunflower (Helianthus annuus) stem canker, causes significant reductions in yield and oil content in most sunflower-growing areas. With the aim of enhancing host resistance, we selected in vitro sunflower calli against culture filtrates of two pathogen isolates (7/96 and 101/96). This technique may be an effective and rapid tool to discriminate the most virulent D. helianthi isolate and to screen for host resistance in the early stage of a breeding programme. Further investigation on the mechanisms involved in defence pathways showed no induction of salicylic acid and pathogenesis-related proteins in calli, indicating that the host resistance is not associated with Systemic Acquired Resistance but probably other biochemical mechanisms.     

Ineffectiveness of pruning to control citrus huanglongbing caused by <Emphasis Type="Italic">Candidatus</Emphasis> Liberibacter americanus

The huanglongbing (HLB) disease of citrus trees, caused by Candidatus Liberibacter asiaticus and Ca. Liberibacter americanus, was first reported in Brazil in March, 2004. The presence of the disease has caused serious concerns among growers. Pruning experiments were conducted to determine if removal of symptomatic branches or the entire canopy (decapitation) would eliminate infected tissues and save HLB-affected trees. Pruning was done in five blocks on a total of 592 3- to 16 year-old ‘Valência’, ‘Hamlin’ or ‘Pêra’ sweet orange trees showing no symptoms or with two levels of symptom severity. Ten decapitated trees per block were caged and all trees were treated with insecticides to control the psyllid vector, Diaphorina citri. Mottled leaves reappeared on most symptomatic (69.2%) as well on some asymptomatic (7.6%) pruned trees, regardless of age, variety, and pruning procedure. Presence of the pathogen (Ca. Liberibacter americanus) in all symptomatic trees was confirmed by PCR. In general, the greater the symptom severity before pruning the lower the percentage of trees that remained asymptomatic after pruning.     

Evaluation of Feed Attractants in Juvenile Senegalese Sole,Solea senegalensis

The Senegalese sole, Solea senegalensis, is a flatfish with increasing interest in the European aquaculture. However, one of the main limitations found in this species is related to the scarce attraction to commercial aquafeeds, thus determining low feed intake in fish farms. The aim of this study was to evaluate four ingredients of marine origin (fish meal hydrolysate [FMH], polychaete meal [POM], mussel meal [MUM], and squid meal [SQM]) as potential attractants for juvenile Senegalese sole. Four experimental feeds containing 10% of the potential attractants, plus a control diet without these ingredients, were evaluated. A total of 150 fish were distributed into ten 40‐L tanks (duplicate tanks per feed). A Latin square design was carried out, and hence all the experimental diets were offered to each tank in successive periods of 7 d. Fish were fed twice a day (morning and afternoon), and the feeding behavior of the animals was assessed by direct observations. Behavioral pattern assessed was classified as distant orientation, near orientation, or continuation of intake. The acceptability index for each behavioral phase and experimental feeds used was established for each observation. No behavioral differences could be noticed between the morning and the afternoon feedings. FMH showed the greatest attractant potential than among the ingredients assessed, and consequently this ingredient yielded the higher feed intake rates. On the other hand, POM was not the best attractant for juvenile Senegalese sole.     

The effect of being housed with adult female flock-mates on the age at puberty of autumn-born ewe-lambs

Veterinary Research Communications - Twenty-five ewe-lambs born in November were used to measure the effect of the presence of cyclic or non-cyclic adult ewes on the age at first fertile estrus....