DNA vaccine encoding avian influenza virus H5 and Esat-6 of Mycobacterium tuberculosis improved antibody responses against AIV in chickens

The H5 gene of avian influenza virus (AIV) strain A/chicken/Malaysia/5744/2004(H5N1) was cloned into pcDNA3.1 vector, and Esat-6 gene of Mycobacterium tuberculosis was fused into downstream of the H5 gene as a genetic adjuvant for DNA vaccine candidates. The antibody level against AIV was measured using enzyme-linked immunosorbent assay (ELISA) and haemagglutination inhibition (HI) test. Sera obtained from specific-pathogen-free chickens immunized with pcDNA3.1/H5 and pcDNA3.1/H5/Esat-6 demonstrated antibody responses as early as 2 weeks after the first immunization. Furthermore, the overall HI antibody titer in chickens immunized with pcDNA3.1/H5/Esat-6 was higher compared to the chickens immunized with pcDNA3.1/H5 (p < 0.05). The results suggested that Esat-6 gene of M. tuberculosis is a potential genetic adjuvant for the development of effective H5 DNA vaccine in chickens.     

Comparison of the pathogensis of two isolates of <Emphasis Type="Italic">Besnoitia caprae</Emphasis> in inbred BALB/c mice

This study was performed to evaluate the infectivity of bradyzoites of two Besnoitia caprae isolates, BC-1 and BC-2, to inbred BALB/c mice. Each group of inbred BALB/c mice was inoculated intraperitoneally with 1 × 10³, 1 × 10⁴, 1 × 10⁵, 5 × 10⁵ and 1 × 10⁶ of one of the two isolates of B. caprae bradyzoites. The mice were monitored daily for a period of 40 days for survival. After death of each mice, several passages from its peritoneal washing and tissues were analyzed using ribosomal DNA-specific PCR assay. Marked differences in pathogenicity between the isolates were seen. All the inbred BALB/c mice infected with BC-2 survived but all the mice that were administered with 1 × 0⁵, 5 × 10⁵ and 1 × 10⁶ BC-1 bradyzoites were died within 4–9 days post-infection (DPI). Histopathological examination of the tissues of the dead mice revealed hyperemia and necrosis with presence of mononuclear and polymorphonuclear cell infiltration in myocardium, spleen and intestines together with interstitial pneumonia and peritonitis. All inbred BALB/c mice in the 1 × 10³ and 1 × 10⁴ groups of BC-1 inoculated mice survived and they were euthanized after 40 DPI. Chronic inflammation with infiltration of mononuclear cells was evident in myocardium, spleen, alveolar septa of the lungs of most of the examined tissues with hemorrhagic enteritis in the mice infected with 1x10⁶ bradyzoites. The mice infected with different doses of BC-2 were euthanized after 40 DPI and no lesion was seen in histopathological sections of their organs. All peritoneal washings and examined tissues were PCR positive in BC-1 group. This experiment is the first report to show inbred BALB/c mice as a relevant model for B. caprae and demonstrates that this strain of inbred BALB/c mice is a suitable animal model for biological studies and examination of pathogenesis for this species of Besnoitia. The present findings also provide evidence for significant differences between the two isolates of B. caprae.     

Fasciation in <Emphasis Type="Italic">Crassula argentea</Emphasis>: molecular identification of phytoplasmas and associated antioxidative capacity

The present study reports on phytoplasma induced fasciation in Crassula argintea (Crassulaceae). DNA was extracted from symptomless and fasciated tissues and amplified by nested PCR using universal primers P1/P7 followed by R16F2n/R16R2 produced amplicons of 1.2 Kb. The nucleotide sequence analyses of the amplicons indicated that fasciated plants were infected by phytoplasma. Phylogenetic analysis placed the Crassula fasciation phytoplasmas in 16SrII-D group. Histochemical staining for reactive oxygen species indicated that phytoplasma infected (PI) tissues possess significantly higher levels of hydrogen peroxide (H₂O₂) rather than superoxide (O₂ ^·-) as compared with symptomless tissues. PI tissues were also associated with a significant increase in antioxidant enzyme activities (catalase, peroxidase, polyphenol oxidase, and glutathione reductase) and electrolyte leakage as compared with symptomless tissues.     

Soybean stem fly,Melanagromyza sojae (Diptera: Agromyzidae), on Sumatra: Seasonal incidence and the role of parasitism

Abstract

The density of M. sojae and percentage parasitism was studied in unsprayed soybean at 18 farmers’ field sites in Aceh, North Sumatra and West Sumatra provinces, Indonesia, during 1992 and 1993. M. sojae generally infested soybean throughout the season; infestation was initially low, reached its peak from the fifth till the eighth week after planting and declined towards the end of the season. A complex of seven hymenopterous parasiloids had a high impact on the pest. Parasitism levels built up alongside with host density and remained high until just before harvest. The eucoilid Gronotoma sp. was the most prevalent species during the early and mid‐season; a complex of pteromalids became dominant towards the end of the season. In soybean planted successively at 2‐week intervals, the parasitism level in the first‐planted crop built up slowly, whereas the parasitism level in the third‐planted crop was high from the early crop stage onwards, which suggests that the third‐planted crop benefited from parasitism build‐up in the earlier‐planted crops. The role of parasitoids in controlling beanflies in unsprayed soybean is discussed.     

Influence of variety on the within-plant distribution of cassava green spider mite (Acari: Tetranychidae), and leaf anatomical characteristics and chemical components in relation to varietal resistance

The within-plant distribution of the cassava green spider mite, Mononychellus tanajoa Bondar, and the anatomical characteristics and the chemical components relating to varietal resistance of cassava, were studied using 11 cassava genotypes with varying levels of resistance for two dry seasons and one wet season. The results show that M. tanajoa aggregates on the top leaves of cassava at low levels of resistance as compared with a more even within-plant distribution at higher levels of resistance. Thus, for accurate sampling of mite populations, it might be important to consider young and old leaves, not just the young leaves, in fields containing resistant and susceptible cassava genotypes. Correlations between the anatomical characteristics and mite population density and damage scores were not consistent across months, either within or over seasons. This suggests that leaf anatomical characteristics may not be important in the varietal resistance of cassava to M. tanajoa. During the dry season, mite population density was positively correlated with leaf nitrogen, potassium and phosphorus and negatively correlated with leaf calcium and fat. Only calcium was negatively associated with mite damage at the peak of the dry season (January 1993 and 1994). Breeding cassava genotypes with high levels of leaf calcium and fat, and low levels of leaf nitrogen, potassium and phosphorus, may improve the level of resistance to M. tanajoa.     

Perturbation of the EphA2-EphrinA1 System in Human Prostate Cancer Cells by Colonic (Poly)phenol Catabolites

The Eph tyrosine kinase receptors and their ephrin ligands play a central role in human cancer as their deregulated expression induces tumorigenesis with aggressive phenotypes. To evaluate their potential contribution to EphA2-ephrinA1 modulation, several colonic catabolites of dietary (poly)phenolics, known to be generated in vivo, were screened using an ELISA-based binding assay. Some of the catabolites inhibited the binding in a dose-dependent manner (IC(50) values from 0.26 to 43 μM). Functional studies on prostate adenocarcinoma cells revealed that pyrogallol and protocatechuic acid specifically antagonized ephrinA1-Fc-induced EphA2 phosphorylation at concentrations that were not cytotoxic. The active concentrations of pyrogallol appear to be close to what can be reached in vivo under physiological conditions. Finally, because of the roles played by the Eph-ephrin system not only in cancer development but also in neurodegeneration and diabetes, pyrogallol and protocatechuic acid are candidates for more detailed functional studies to elucidate their role in these pathophysiological processes.     

Induction of Immunogenic Response in BALB/c Mice by Live and Killed Form of Recombinant Lactococcus lactis Displaying EG95 of Echinococcus granulosus

Background:CE is a zoonotic parasitic infection caused by Echinococcus granulosus worldwide and is associated with economic losses among livestock animals. EG95 is an immunogenic antigen from the E. granulosus. Lactococcus lactis has been prested as a safe vehicle for antigen delivery. The goal of this study was to design a novel L. lactis strain displaying EG95 as a vaccine delivery system. Methods:The eg95 encoding gene fragment fused to the M6 anchoring protein was cloned into the pNZ7021 vector, and L. lactis NZ9000 displaying recombinant EG95 was constructed. The expression of an approximately 32-kDa EG95 protein was confirmed by Western blotting and immunofluorescence analysis. The immune responses were evaluated in BALB/c mice immunized orally and subcutaneously with the live and killed recombinant L. lactis, respectively. Results:Total IgG level in mice immunized with heat-killed recombinant L. lactis (pNZ7021-eg95) significantly increased compared to the control group. sIgA was significantly higher in mice received live recombinant L. lactis (pNZ7021-eg95) compared to the control mice. Splenic lymphocytes from immunized mice represented the high levels of IFN-γ and the low-levels of IL-4 and IL-10. Conclusion:Our results indicate that immunization with EG95-expressing L. lactis can induce both specific humoral and cellular immune responses in mice. Key Words: Echinococcus granulosus, Lactococcus lactis, Immunization, Vaccines     

Stimulating and toxicity effects of nickel on growth,yield, and fruit quality of cucumber supplied with different nitrogen sources

Limited information is available on biological effects of various levels of nickel (Ni) (deficiency to toxicity levels) on growth and yield of certain crops, particularly vegetables. In this sand‐culture study, we investigated the effects of four levels of Ni (0, 50, 100, and 200 μM) on growth, yield, and fruit‐quality attributes of two cucumber cultivars (Cucumis sativus L. cvs. Super Dominus and Negin) supplied with urea or NH₄NO₃ as nitrogen source. Addition of 50 μM Ni to the nutrient solution resulted in a significant increase of shoot and root dry‐matter yield of cv. Negin although this increase was greater in the urea‐fed plants than those fed with NH₄NO₃. In both cultivars, addition of 50 μM Ni increased urease activity and thereby decreased the urea concentration in the urea treatment. Addition of 100 and 200 μM Ni caused a significant decrease in root and shoot growth of cucumber although this decrease was insignificant for cv. Super Dominus in the 100 μM treatment. The highest fruit yield, total soluble solids (TSS), and fruit firmness were achieved at the 50 μM Ni treatment. Regardless of nitrogen source, Ni addition proportional to the concentration used increased leaf Ni concentration and fruit acid ascorbic concentration. The concentration of Ni required for optimum growth and yield of cucumber varied with cultivars. The level of 50 μM was sufficient for optimum growth of cv. Negin in nutrient‐solution culture while lower concentration of Ni was required for cv. Super Dominus. While the beneficial effects of sufficient levels of Ni on growth and yield of urea‐fed plants was greater than with NH₄NO₃‐fed plants, the toxic effects of Ni in these plants were also greater.     

Barseem (Trifolium alexandrnum) protein concentrate in broiler rations

Animal protein in broiler rations was replaced with Barseem Protein Concentrate (BPC). The standard group having blood, meat and fish meal as sources of animal protein showed an average weight gain of 1655 g. Replacement of blood meal with BPC resulted in a slight decrease in the average weight gain (1541 g) of broilers. Average weight gain of broilers increased to 1775 g and 1671 g when fish or meat meal was replaced with BPC in rations. Complete replacement of animal protein with BPC adversely affected growth and average weight gain decreased to 1443 g. Supplementation with lysine or methionine when animal protein was replaced with BPC did not improve growth of the broilers.