Construction and immunogenicity studies of recombinant fowl poxvirus containing the S1 gene of Massachusetts 41 strain of infectious bronchitis virus

The spike 1 (S1) surface glycoprotein of infectious bronchitis virus (IBV) is the major inducer of the generation of virus neutralizing antibodies, and the administration of purified S1 has been shown to elicit a protective immune response against virulent virus challenge. On the basis of these observations, recombinant fowl poxvirus (rFPV) containing a cDNA copy of the S1 gene of IBV Mass 41 (rFPV-S1) was constructed and its immunogenicity and vaccine potential were evaluated. Initially, rFPV-S1 was shown to express the S1 in vito by indirect immunofluorescence staining and western blot analyses. Later, in vivo expression was demonstrated by the detection of IBV-specific serum immunoglobulin G and neutralization antibodies in the sera of chickens immunized with rFPV-S1. That the recombinant virus elicited anti-IBV protective immunity was indicated by the manifested, relatively mild clinical signs of disease, decreased titers of recovered challenge virus, and less severe histologic changes of the tracheas in virulent IBV Mass 41-challenged chickens previously receiving rFPV-S1 as compared with parental fowl poxvirus (FPV)-vaccinated control birds. In contrast, chickens immunized with either recombinant or parental FPV were resistant to a subsequent virulent FPV challenge. As to a preferred method of immunization, wing web administration appeared to be superior to the subcutaneous route because a greater percentage of birds vaccinated by the former protocol exhibited an anti-IBV humoral immune response. Thus, rFPV-S1 has potential as a poultry vaccine against both fowl pox and infectious bronchitis.     

Characterization of Capsaicinoids and Antioxidants in Hot Peppers as Influenced by Hybrid and Harvesting Stage

Studies were conducted to investigate the accumulation pattern of capsaicinoids and antioxidants such as carotenoids, ascorbic acid and phenolic compounds in three hot pepper hybrids at five different harvesting stages: immature green, mature green, color break, red ripe and dried fruit. Capsaicin and dihydrocapsaicin contents were maximum at mature green stage in both Sky Red (61.30 and 43.76 mg/100 g) and Wonder King (43.93 and 26.16 mg/100 g) hybrids, whereas Maha hybrid reached top values at color break stage (39.13 and 24.20 mg/100 g). The accumulation of total carotenoids showed an 8-fold increase from red ripe (12 mg/100 g) to dried fruit stage (96 mg/100 g), while a noticeable decline by 76 % was observed for ascorbic acid at same harvesting stages (150 vs. 36 mg/100 g, respectively). The three hot pepper hybrids showed great variations in the evolution of total phenolic contents during harvesting stages. Overall, the mature green stage was ideal to acquire maximum pungency due to capsaicinoids, while peppers at red ripe stage were best sources of ascorbic acid and dried fruits contained higher levels of total carotenoids.     

Development and survival of Haemonchus contortus infective larvae derived from sheep faeces under sub-tropical conditions in the Potohar region of Pakistan

Assessment on the development and survival of Haemonchus contortus larvae was made to evaluate the influence of microclimatic variables viz., relative humidity (%), temperature (°C), rainfall (mm) and intensity of sunlight (hrs). Pasture plots in a sub-tropical area, Pakistan were contaminated with H. contortus eggs at the start of every month. The plots were sampled on fortnightly basis after contamination for three consecutive months. The overall pattern indicated the most favorable conditions for survival, development and translation to herbage was during the wet season from July to October and the least favorable was during the dry season from April to June. Peak infective larvae (L₃) recovery was during the 15–45 day interval for each plot and the lowest number was during the 75–90 day interval. Herbage was collected in the morning, afternoon and evening and greatest recovery of L₃ was in the morning time and least in the afternoon. The number of L₃ on pasture was directly related to the pattern of rainfall and relative humidity with a significant (P < 0.05) positive correlation and temperature and intensity of sunshine were negatively correlated (P < 0.05). The results suggest that pastures can remain infective for up to 2 months and become relatively clean by 90 days after contamination. Thus, long pasture rest periods, especially during the high risk wet season, may be a helpful strategy to reduce the infection levels.     

BORON DEFICIENCY IN RAINFED WHEAT IN PAKISTAN: INCIDENCE,SPATIAL VARIABILITY AND MANAGEMENT STRATEGIES

Boron (B) deficiency is potentially an important nutrient constraint in calcareous soils. We determined B deficiency incidence and spatial distribution in rainfed wheat (Triticum aestivum L.) in 1.82 Mha Pothohar plateau in Pakistan, its relationship with soil types, crop responses to B, and internal B requirement and B fertilizer use efficiency of wheat. Plant and soil analyses indicated deficiency in 64% of the 61 sampled fields; geostatistics-aided contour maps delineated B deficient areas. In rainfed field experiments, B use increased wheat yields up to 11%. Fertilizer requirement was 1.2 kg B ha^?1; critical B concentration (mg kg^?1) ranges were: young whole shoots, 4–6; flag leaves, 5–7. Boron uptake by wheat was 0.14–0.58% of applied dosage, leaving substantial residual impact. Highly cost-effective B use or B-efficient genotype adoption can enhance wheat productivity and grower-income. Such effective nutrient assessment and management approaches can be beneficially adopted elsewhere as well.     

BORON NUTRITION OF PEANUT GROWN IN BORON-DEFICIENT CALCAREOUS SOILS: GENOTYPIC VARIATION AND PROPOSED DIAGNOSTIC CRITERIA

Field studies were conducted to assess boron (B) requirement, critical concentrations in diagnostic parts based on yield response curves and genotypic variation by growing three peanut (Arachis hypogaea L.) cultivars (‘Golden’, ‘BARD-479’, ‘BARI-2000’) on two B-deficient calcareous soils. Boron application significantly increased pod yield of all the cultivars over control. Maximum pod yield increases were: ‘Golden’, 16?23%; ‘BARD-479’, 21?27%; and ‘BARI-2000’, 25?31%. The cultivars varied in B efficiency and cv. ‘Golden’ was the most B efficient (81?86%) while cv. ‘BARI-2000’ was the least efficient (76?80%). Boron requirements for near-maximum (95%) dry pod yield were 0.65 kg ha^?1 for ‘Golden’, 0.75 kg ha^?1 for BARD-479 and 0.80 kg ha^?1 for BARI-2000. Critical B concentrations in shoots and seeds were: ‘Golden’, 33 mg kg^?1 and 26 mg kg^?1; ‘BARD-479’, 38 mg kg^?1 and 31 mg kg^?1; and ‘BARI-2000’, 42 mg kg^?1 and 33 mg kg^?1.     

Fruit Characteristics and Detachment Parameters of Sour Cherry (Prunus cerasus L. cv. ‘Kütahya’) as Affected by Various Maturity Stages

The variability in fruit maturity and quality has difficulties in determining the optimal harvesting maturity. Therefore, maturity stage in harvest is crucial for both fruit quality and minimizing the fruit detachment force. This study investigates the fruit characteristics and detachment parameters of sour cherry (cv. ‘Kütahya’) in terms of different maturity stages. Sour cherries were grouped into five different maturity stages by observation according to their color and dimensions. The fruit mass and dimension increased as a function of maturity stage; however, the fruit mass remained almost same after the 4th stage. The highest values of total soluble solids and titrable acid contents of sour cherry were gained in the last stage of maturity. The value of chroma of fruit outer color was higher 50?% than that of fruit inner color in the 1st maturity stage. However, it was almost same in the 5th stage of maturity. Increasing fruit maturity caused a decrease in the values of fruit detachment force ranging from 4.25 to 1.78 N, giving the coefficient of determination of 0.947. Similarly, the ratio of fruit detachment force to fruit mass, detachment stress, and specific energy decreased as the fruit maturity increased. The harvest of sour cherry is done only once in the optimum time to decrease the harvest cost. Minimizing the variability in fruit maturity throughout the tree is essential for gaining the maximum profit and lower fruit detachment force in this convenient time. Results of this study indicated that 3rd?5th maturity stages were critical for harvest. In order to attain the maximum poundage per tree and color for the market the most of the fruits should be in these stages. Therefore, appropriate management strategies that could homogenize the maturity throughout the tree should be applied.     

The 26S proteasome in garlic (Allium sativum): purification and partial characterization

The 26S proteasome (multicatalytic protease complex, MPC) was purified from fresh garlic cloves (Allium sativum) to near homogeneity by ion exchange chromatography on DEAE-sephacel, gel filtration on Sepharose-4B, and glycerol density gradient centrifugation. Two alpha-type (20S proteasome "catalytic core") subunits were identified by the direct sequencing of peptide fragments (mass fingerprint analysis, Mass Spectrometry Lab, Stanford University) or the sequencing of a cloned cDNA generated using a garlic cDNA library as the template; these subunits were found to have a high homology to those from other plants. Polyacrylamide gel electrophoresis under denaturing conditions separated the garlic MPC into multiple polypeptides having molecular masses in the range of 21-35 (components of the 20S catalytic core) and 55-100 kDa (components of the 19S regulatory units). The banding pattern of the garlic MCP is similar to that of spinach and rat liver with minor differences in some components; however, polyclonal antibodies against mammalian proteasomes failed to significantly stain the enzyme from garlic. This is the first work to identify the garlic proteasome.     

Zinc requirement of corn grown on two calcareous soils of Pakistan

In greenhouse studies, corn (Zea mays L.) growth increased with Zn fertilization of two alkaline calcareous soils. Zinc concentration and total uptake increased with Zn application. Very high correlations were recorded between plant tissue Zn concentration, total Zn uptake and soil Zn levels determined by DTPA and AB-DTPA soil tests. Correlation between Zn concentration in plants and relative yield was poor. However, close relationships were revealed between extractable soil Zn and relative yield. Near maximum dry matter yield of corn was associated with a fertilizer rate of 2 mg Zn/kg soil. Plant tissue Zn-requirement was 27 mg/kg in 15 days old plants and 32 mg/kg in corn shoots of 40 day age. Critical soil test Zn level was 1.2 mg/kg by DTPA and 1.7 mg/kg by AB-DTPA method. Use of AB-DTPA soil test is suggested for evaluating Zn status of calcareous soils.     

Rapid detection of group I avian adenoviruses by a loop-mediated isothermal amplification

A loop-mediated isothermal amplification (LAMP) assay was optimized for the rapid detection of Group I avian adenoviruses. A set of six primers was designed from the DNA sequences of hexon genes from Group I avian adenovirus. The assay was performed in a water bath for 60 min at 63 C, and the amplification result was visualized by adding a fluorescence dye reagent or by inspecting the white sediment. The results showed that the LAMP assay could detect all 12 serotypes of Group I avian adenovirus and nine Guangxi Group I avian adenovirus isolates. This avian adenovirus Group I-specific LAMP assay could detect 238 copies of avian adenovirus. No cross-reactions were detected using the LAMP assay with avian adenoviruses type II and III or with other avian viruses. The ability of LAMP to detect Group I avian adenovirus isolates was further evaluated with 184 cloacal swab samples from poultry. In total, 72 out of 184 cloacal swab samples from poultry were identified as positive by LAMP, whereas 45 out of 184 were identified as positive by conventional PCR test. The Group I avian adenovirus specific LAMP results were further confirmed by real-time PCR. This specific LAMP method holds promise as a rapid and specific diagnostic assay for detection of samples from birds suspected of adenovirus infection.