Serogrouping of United States and some African serotypes of bluetongue virus using RT-PCR

The diagnostic potential of RT-PCR for detection of bluetongue virus (BTV) ribonucleic acid (RNA) sequence in cell culture and tissue samples from infected ruminants from United States, Sudan, South Africa and Senegal, was evaluated. The non structural protein 1 (NS1) gene of North American BTV serotype 11 was targeted for PCR amplification. The United States BTV serotypes 2, 10, 11, 13 and 17 and the Sudanese BTV serotypes 1, 2, 4 and 16 and BTV serotype 4 from South Africa and BTV serotype 2 from Senegal were studied. RNAs from all BTV field isolates used in this study, propagated in cell cultures, were detected by the described RT-PCR-based assay. The first specific 790bp BTV PCR products were amplified using a pair of outer primers (BTV1 and BTV2). Specificity of the PCR products was confirmed by a nested amplification of a 520bp PCR product using a pair of internal (nested) primers (BTV3 and BTV4). The BTV PCR products were visualized on ethidium bromide-stained agarose gels. Amplification products were not detected when the RT-PCR-based assay was applied to RNAs from closely related orbiviruses including, epizootic hemorrhagic disease virus (EHDV) prototypes serotypes 1, 2, 4; RNA from Sudanese isolate of palyam orbiviruses serogroup and total nucleic acid extracts from uninfected Vero cells. Application of the nested BTV RT-PCR to clinical samples resulted in amplification of BTV RNA from blood and serum samples from goats experimentally infected with BTV4 and from naturally infected sheep, goats, cattle and deer. The results of this study indicated that this RT-PCR assay could be applied for rapid detection of BTV, in cell culture and clinical samples from susceptible ruminants during an outbreak of the disease, in the United States and African.     

Effects of hypertonic dextrose and paraffin solution as non‐antibiotic treatments of clinical endometritis on reproductive performance of high producing dairy cows

The objective of this study was to compare the reproductive performance of cows affected by clinical endometritis (CE) following treatment with an intrauterine infusion of 50% dextrose solutions (DEX) and liquid paraffin (LP) as alternative therapies with routine treatments including PGF_2α injection and intrauterine infusion of oxytetracycline (OTC). Moreover, the reproductive indices of cows treated with endometritis were compared with those of healthy ones. At postpartum reproductive tract examination (28–35 DIM) in one Iranian dairy farm, cows with CE without any selection were assigned to four groups: (a) OTC, a common treatment in Iran, was administered (5 g) to 396 cows, (b) PGF₂α (PG) was injected to 496 cows, (c) dextrose solution (DEX): intrauterine infusion of 50% dextrose solution (200 ml) was done in 427 cows, and (d) liquid paraffin (LP) was administered (100 ml) to 423 cows via intrauterine route. We further assigned 2,233 clinically healthy cows to the control group. The incidence of endometritis was 41.6% in this study. Based on the results of reproductive indices including median days to first AI, days open (DO), first service conception rate, conception rate in 2nd and 3rd services, conception rate in all three services, pregnancy rate < 100 days and pregnancy rate < 200 days, except for median days to first AI in other reproductive indexes, reproductive performance was significantly lower in LP group compared with the healthy cows and other treatment groups (p < 0.05). Except for the first service conception rate and proportion of cows pregnant before 100 days in milk (DIM), there existed no significant difference between the DEX group and the control as far as reproductive performance is concerned (p ≥ 0.05). The first service conception rate was recognizably lower in DEX group compared with OTC and PG (p < 0.05). In conclusion, the use of a non‐antibiotic special solution of dextrose 50% is a good alternative to antibiotic agents concerning the treatment of CE in dairy cows.     

Inheritance of resistance to sunflower downy mildew races 1, 2 and 3 in cultivated sunflower

Sunflower downy mildew (SDM) caused by Plasmopara halstedii, is a major disease of sunflower. Eleven resistance genes have been identified, but allelic relationships among these genes are not clear. This study examined the inheritance and allelic relationships of genes conferring resistance to SDM races 1, 2 and 3 (virulence phenotypes 100, 300 and 700, respectively) and confirmed a twelfth resistance gene. Three USDA Plant Introductions, AMES 3235, PI 497250, and PI 497938, and three released lines, RHA 266, RHA 274 and DM‐2 were studied. RHA 266 has only the Pl₁ gene for race 1 resistance. Digenic inheritance of resistance was found in AMES 3235, PI 497250, and RHA 274. These lines have the Pl₁ and Pl₁₂ genes, conferring resistance to race 1, and the Pl₂ and Pl₁₁ genes, conferring resistance to race 2. DM‐2 and PI 497938 have Pl₁₂ (but not Pl₁ for resistance to race 1, the Pl₁₂ gene (but not the Pl₂) for resistance to race 2, and Pl₅ for resistance to race 3. These resistance genes will serve as a foundation for future gene designations and genetic diversity studies of resistance to SDM.     

Development of a male sterile eggplant by utilizing the cytoplasm of Solanum virginianum and a biparental transmission of chloroplast DNA in backcrossing

To develop a male sterile line of eggplant (Solanum melongena L.) utilizing the cytoplasm of S. virginianum, an interspecific F₁ hybrid between S. virginianum and S. melongena ‘Senryo Nigou’ was continuously backcrossed to S. melongena ‘Uttara’ using ‘Uttara’ as a recurrent pollen parent and four backcross generations, BC₁, BC₂, BC₃ and BC₄ were produced. All the plants in backcross generations were anther indehiscent although the F₁ hybrid, S. virginianum and S. melongena were dehiscent. Pollen stainability with acetic carmine and in vitro germination ability of pollen in all the backcross progenies were quite lower than those of the parental species. Fruit set percentage, number of seeds per fruit and seed germination rate were high in all the backcross progenies. The present results indicate that anther indehiscent type of functional male sterile line of eggplant could be developed by utilizing the cytoplasm of S. virginianum. PCR-RFLP analysis of chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) were performed to identify the organelle inheritance. The F₁ hybrid and all the backcross progenies displayed maternal inheritance of mtDNA. The cpDNA of the examined single BC₁ plant exhibited the recombinant cpDNA pattern of the parental F₁ hybrid and ‘Uttara’, indicating occurrence of the biparental inheritance of cpDNA. As all the BC₂, BC₃ and BC₄ progenies showed the same recombinant cpDNA patterns of the BC₁ plant, the recombinant cpDNA might be stable and harmonize with the nuclear genome of S. melongena. The present male sterile line can contribute to expand the male sterility source of eggplant.     

Small urban woodlands as biodiversity conservation hot-spot: a multi-taxon approach

To evaluate the importance of urban woodlands to serve as potential sites for biodiversity conservation, we analysed bird, carabid beetle and small mammal community responses to urbanisation at different spatial scales. We analysed the relationships between the variations of the structure (species richness S, diversity H′ and dominance D) of animal communities of woodlands distributed along a rural–urban gradient, and the variations along this same gradient of (1) the vegetation within woodlands, (2) the landscape at 100 m and (3) 600 m around the woodlands. We identified the spatial scales whose variations along the gradient most affected each animal community structure, and characterised community responses to these variations. Our results showed that urbanisation affected taxa differently according to their dispersal ability. Carabid beetles, less mobile, seem to be sensitive to increasing fragmentation and built surfaces from periurban to town centre which could make their movement within the urban landscape difficult. Birds, mobile species, seem to be more sensitive to variations of the vegetation structure within woodlands from periurban to town centre that could affect their capacity to maintain in habitat patches. Although our study did not allow relating the small mammal community structure to urbanisation, it suggests that this taxa is sensitive to urban local disturbances. A relevant management scale of woodlands can be specified for each taxa conservation. Urban woodlands accommodate over 50% of the species present in periurban woodlands, and effective management could enhance this number. Woodlands seem to be a good choice for promoting biodiversity conservation in towns.     

Development of a DNA vaccine against chicken anemia virus by using a bicistronic vector expressing VP1 and VP2 proteins of CAV

In the present study, we describe the development of a DNA vaccine against chicken anemia virus. The VP1 and VP2 genes of CAV were amplified and cloned into pBudCE4.1 to construct two DNA vaccines, namely, pBudVP1 and pBudVP2-VP1. In vitro and in vivo studies showed that co-expression of VP1 with VP2 are required to induce significant levels of antibody against CAV. Subsequently, the vaccines were tested in 2-week-old SPF chickens. Chickens immunized with the DNA-plasmid pBudVP2-VP1 showed positive neutralizing antibody titer against CAV. Furthermore, VP1-specific proliferation induction of splenocytes and also high serum levels of Th1 cytokines, IL-2 and IFN-γ were detected in the pBudVP2-VP1-vaccinated chickens. These results suggest that the recombinant DNA plasmid co-expressing VP1 and VP2 can be used as a potential DNA vaccine against CAV.     

ISSR variations in eggplant (Solanum melongena L.) and related Solanum species

Inter-simple sequence repeat (ISSR) analysis was performed in eight cultivars of eggplant (Solanum melongena) and 12 accessions in eight related Solanum species to evaluate the applicability of this analysis for assessing the phylogenetic relationships and identifying cultivars. A total of 552 polymorphic amplified bands were obtained from 34 of the 100 primers tested, and the percentage of polymorphisms was 99.1%. Cluster analysis based on the ISSR markers classified the Solanum species into seven groups: (i) S. melongena; (ii) S. aethiopicum and S. anguivi; (iii) S. incanum; (iv) S. violaceum and S. kurzii; (v) S. macrocarpon; (vi) S. virginianum and (vii) S. torvum. Combining the ISSR markers obtained by a few of the 34 primers was enough for distinguishing of the eight cultivars of eggplant. This ISSR analysis was demonstrated to be available for the phylogenetic study and the cultivar identification.