Characterization of an endopolygalacturonase Gene cppg1 from Phytopathogenic Fungus Chondrostereum purpureum

Chondrostereum purpureum, a phytopathogenic fungus, produces endopolygalacturonase (endoPG) which has been suggested to have a causal role in the silver-leaf symptom of apple trees. In this paper, we detected C. purpureurn-derived endoPG at the infection sites using ELISA with a polyclonal antibody against endoPG I. A gene encoding endoPG I and its homolog were also isolated from the C. purpureum genome. The endoPG I gene was designated as cppg1. The cppg1 gene is the first fungal endoPG gene reported in the Basidiomycetes. Received 31 May 2000/ Accepted in revised form 13 September 2000     

Effect of intravenous injection of acetate on the pancreas of sheep

The effect of an intravenous injection of acetate on plasma insulin and glucagon concentration was examined in conscious sheep. Sodium acetate (312 to 5000 mumol kg-1 bodyweight) injection increased plasma insulin concentrations in a dose-dependent manner. Plasma glucagon concentration was not affected by doses up to 1250 mumol kg-1. Doses of 2500 and 5000 mumol kg-1 produced significant increases (P less than 0.05), but not in a dose related manner. The results of this study indicate that the receptive mechanism of the A cell in the sheep pancreas to acetate might be different from that of the B cell.     

Evaluation of coral bleaching condition <Emphasis Type="Italic">in situ</Emphasis> using an underwater pulse amplitude modulated fluorometer

ABSTRACT:   Photosynthetic activities of hermatypic corals (28 genera, c . 68 species and c. 92 specimen) were measured in November 2000 at Sekisei lagoon, Okinawa, Japan, using an underwater pulse amplitude modulated fluorometer. Maximum quantum yield (Fv/Fm) ranged from 0.61 to 0.70 and had an average value of 0.664 (standard deviation [SD], 0.0289). Following the measurement of Fv/Fm, quantum yield (Fm'-F)/Fm' at eight-leveled actinic-light intensities were obtained from all specimens. Values of electronic transport rates (ETR) were nearly the same under low photosynthetic active radiation (PAR; µmol quanta/m² per s) condition and varied with increasing PAR. The value of ETR at 100 µmol quanta/m² per s ranged from 21.3 to 30.0 and had an average of 27.8 (SD, 2.09). In August 2001, a coral bleaching event was observed in Sekisei Lagoon. During the event, photosynthetic activities of 21 specimens of eight species (seven genera) were measured. By applying two indicators, Fv/Fm and ETR at 100 µmol quanta/m² per s obtained in August 2000, 18 of 21 specimens were suggested under the bleaching process. In case of Porites lutea , seven colonies were visually evaluated for their condition of bleaching and were found to have two 'normal', three 'slightly bleached' and two 'heavily bleached'. From the density of zooxanthellae, colonies evaluated 'normal' or 'slightly bleached' did not show an apparent difference. However, they were clearly identified by Fm/Fv and ETR at 100 µmol quanta/m² per s.     

RESUME:Studies on the Chemical Control of Aonidiella aurantii (Mask.)

Aonidiella aurantii (Mask.) which was accidentally introduced in Sicily in 1968 has since become a dangerous pest requesting special treatments in citrus groves. Experiments have been carried out on Citrus lemon to evaluate the efficacy of several organophosphorous compounds partly in mixture with white oil and this latter applied sole (cf. table 1). The results obtained against young females are set out in table 2; they show excellent results with almost all compounds whereas the same pesticides had a rather poor effect on mature females (cf. table 3).
It is concluded that the replacement of white oils by organophosphorous compounds is not justified providing that the control is directed against larvae and young females. This means that a proper timing is essential to obtain a satisfactory mortality, thus avoiding the use of detrimental broad acting pesticides.     

Evaluation of serological tests for the diagnosis of Neospora caninum infection in dogs: optimization of cut off titers and inhibition studies of cross-reactivity with Toxoplasma gondii

Diagnosis of Neospora caninum infection in dogs is based on serological assays such as the indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assays (ELISA). This study evaluated two serological tests (IFAT and ELISA) for the detection of IgG antibodies to N. caninum in 300 serum samples of dogs through the optimization of cut off titers by using the two-graph receiver-operating characteristic (TG-ROC) curve. In addition, the identification of major cross-reactive antigens with Toxoplasma gondii was investigated by inhibition ELISA and immunoblotting (IB) assays. IFAT and ELISA results showed 74% agreement, with a good negative concordance (P(neg)=0.83), but a poor positive concordance (P(pos)=0.42). The great majority (86%) of sera with positive concordant results (IFAT+/ELISA+) recognized at least two out of three N. caninum immunodominant antigens, particularly the 29-32 and 35-37 kDa bands. Optimization of cut off titers in IFAT and ELISA was performed considering the reactivity to at least two out of three N. caninum immunodominant antigens as infection markers, obtaining a titer of 50 for IFAT and 200 for ELISA. Seropositivity to N. caninum was significantly associated with T. gondii-seropositive samples, particularly in ELISA (55.4%). Inhibition ELISA curves for N. caninum showed a partial heterologous inhibition, indicating some degree of cross-reactivity between N. caninum and T. gondii antigens. Inhibition IB assays showed a moderate heterologous inhibition for N. caninum antigens above 45-50 kDa. These results indicate that ELISA should be used critically when crude tachyzoite antigen preparations are employed, due to possible cross-reactivity with other related parasites as T. gondii. Also, the cut off dilution of 1:50 in IFAT showed to be the most appropriated for N. caninum serology in dogs. Therefore, we suggest that N. caninum immunodominant antigens, specially the 17 and 29-32 kDa proteins, should be selected markers in serological assays for canine neosporosis.     

Development of new assessment methods for Acropora coral recruitment using coral settlement devices and holes of marine block

We developed new assessment methods for Acropora coral recruitment using coral settlement devices (CSDs) and holes of a marine block. Both devices were deployed in situ before mass spawning of Acropora corals. The CSDs were sampled after 3–4 months of spawning to measure early recruitment, whereas holes of the marine block were observed underwater or MB was sampled to observe holes after 1 year of spawning to measure the recruitment. By combining the results, we would know the recruitment (visually identified underwater) in reefs, including the amount of early recruitment (identified by stereomicroscopic methods), and the environmental condition of the coral reef. These results would help to predict the future of a reef. Nagura Bay had a high early Acropora recruitment; however, due to competition with algae and sedimentation, the recruitment of juvenile corals was extremely low. The reef would possibly be healthy and sustainable at sites where both the early recruitment and recruitment are high. However, if both are low, the reef will be in a critical condition where urgent restoration will be required.     

Essais de lutte chimique contre Aonidiella aurantii (Mask.)

Les essais de lutte chimique contre Aonidiella aurantii (Mask.) sur citronniers en Sicile ont été réalisés à l'aide d'une série de produits organo-phosphoriques en partie mélangés avec des huiles blanches, et compareés à l'efficacité des huiles blanches appliquées seules. II est apparu que les meilleurs résultats sont obtenus contre les larves et les jeunes femelles, alors que pour les femelles matures il n'a pu être enregistré une mortalité suffisante. L'analyse statistique n'a pas révélé de différences significatives entre l'efficacité de l'huile blanche et celle des autres produits testés.     

Enrofloxacin is able to control Toxoplasma gondii infection in both in vitro and in vivo experimental models

Currently, toxoplasmosis is treated with sulfadiazine and pyrimethamine. However, this treatment presents several adverse side effects; thus, there is a critical need for the development and evaluation of new drugs, which do not present the same problems of the standard therapy. Enrofloxacin is a fluoroquinolone antibiotic known to control infection against several bacteria in veterinary medicine. Recently, this drug has demonstrated protective effects against protozoan parasites such as Neospora caninum. The present study aimed to determine the effect of enrofloxacin in the control of Toxoplasma gondii infection. For this purpose, human foreskin fibroblast (HFF) cells were infected with T. gondii RH strain and treated with sulfadiazine, penicillin/streptomycin, pyrimethamine, or enrofloxacin. Following treatment, we analyzed the infection index, parasite intracellular proliferation and the number of plaques. Additionally, tissue parasitism and histological changes were investigated in the brain of Calomys callosus that were infected with T. gondii (ME49 strain) and treated with either sulfadiazine or enrofloxacin. Enrofloxacin was able to reduce the infection index, intracellular proliferation and the number of plaques in HFF cells infected by T. gondii in comparison with untreated or penicillin/streptomycin-treated ones. Enrofloxacin was more protective against T. gondii in HFF infected cells than sulfadiazine treatment (P<0.001). In addition, pyrimethamine, enrofloxacin or the associations of sulfadiazine plus pyrimethamine, enrofloxacin plus sulfadiazine or enrofloxacin plus pyrimethamine-treatments were able to reduce the plaque numbers in HFF cells infected by T. gondii when compared to medium, penicillin/streptomycin or sulfadiazine alone. In vivo experiments demonstrated that enrofloxacin diminished significantly the tissue parasitism as well as the inflammatory alterations in the brain of C. callosus infected with T. gondii when compared with untreated animals. Based on our findings, it can be concluded that enrofloxacin is a potential alternative drug for the treatment of toxoplasmosis.     

Myeloid differentiation factor 88 is required for resistance to Neospora caninum infection

Neospora caninum is an intracellular parasite that causes major economic impact on cattle raising farms, and infects a wide range of warm-blooded hosts worldwide. Innate immune mechanisms that lead to protection against this parasite are still unknown. In order to investigate whether myeloid differentiation factor 88 (MyD88) is required for resistance against N. caninum, genetically deficient mice (MyD88^−/−) and wild type littermates were infected with live tachyzoites and the resistance to infection was evaluated. We found that sub-lethal tachyzoite doses induced acute mortality of MyD88^−/− mice, which succumbed to infection due to uncontrolled parasite replication. Higher parasitism in MyD88^−/− mice was associated with the lack of IL-12 production by dendritic cells, delayed IFN-γ responses by NKT, CD4⁺ and CD8⁺ T lymphocytes, and production of high levels of IL-10. MyD88^−/− mice replenished with IL-12 and IFN-γ abolished susceptibility as the animals survived throughout the experimental period. We conclude that protective IFN-γ-mediated immunity to N. caninum is dependent on initial MyD88 signaling, in a mechanism triggered by production of IL-12 by dendritic cells. Further knowledge on Toll-like receptor recognition of N. caninum antigens is encouraged, since it could generate new prophylactic and therapeutic tools to control parasite burden.     

Intraocular pressure,specular microscopy,and prostaglandin E2 concentration in dogs with mature and hypermature cataract

This study aimed to evaluate and correlate intraocular pressure (IOP), endothelial cell density (CD), and hexagonality (HEX), and the aqueous humor prostaglandin E₂ (PGE₂) concentration in dogs with mature (MG, n = 8) and hypermature (HG, n = 8) cataracts. Eight laboratory beagles with no ocular abnormalities were included as a control group (CG). The IOP was measured using a digital applanation tonometer. Noncontact specular microscopy was used to evaluate CD and HEX. Samples of aqueous humor were used to determine prostaglandin E₂ concentration using enzyme‐linked immunoassay. Data were compared by anova and Bonferroni's multiple comparison test, and possible correlations among the PGE₂ aqueous concentration and corneal endothelium cell parameters were assessed by Person′s test (P < 0.05). Average values of IOP (P = 0.45) and CD (P = 0.39) were not significantly different between MG, HM, and CG. Average values of HEX were lower, and PGE₂ concentration was increased in the MG and HG in comparison with CG (P < 0.05); however, such parameters did not change significantly between MG and HG (P > 0.05). PGE₂ values did not correlate with IOP, CD, and HEX in any group (P > 0.05). Although there were a small number of dogs studied, our results demonstrated that cataract progression from mature to hypermature did not have a significant change in PGE2 aqueous concentration, IOP, corneal endothelial cell count, or morphology. In addition, PGE₂ concentration was not correlated with parameters of the corneal endothelium or IOP in dogs with mature or hypermature cataracts.