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.     

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.     

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.     

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.     

BORON NUTRIPRIMING IMPROVES THE GERMINATION AND EARLY SEEDLING GROWTH OF RICE (ORYZA SATIVA L.)

Boron (B) is one of the important micronutrient required for rice in particular during reproductive growth. A laboratory study was conducted to explore the potential of boron nutripriming in improving the germination and early seedling growth of rice. Seeds of fine grain aromatic rice cultivar Super Basmati were primed in aerated B solution (0.001, 0.01, 0.1 and 0.5% w/v) while untreated dry seeds were taken as control. Seed priming in 0.001 and 0.1% B solutions improved the time to 50% germination, germination energy, final germination percentage, mean germination time, and germination index. Beyond this concentration either there was no effect or an adverse effect on rice seeds. In the cases of radicle length, plumule length and secondary roots priming more diluted B solution, i.e., 0.001% proved better than rest of the treatments as suppression in these three traits was observed by other B treatments than control. Seed priming in relatively concentrated B solution, i.e., 0.5% completely suppressed the germination and growth.     

Aflatoxins and ochratoxin A contamination in rice,corn and corn products from Punjab,Pakistan

In present study, a total of 275 retail cereal samples including rice, corn and corn products was analyzed for the presence of aflatoxins and ochratoxin A (OTA) using HPLC equipped with a fluorescence detector. The data has shown that 38 out of 68 samples of rice, 37/105 of corn and 43/102 samples of corn products have been found contaminated with AFs, average level of AFB₁ and total AFs, 8.23 and 19.54, 7.90 and 12.08, and 5.47 and 7.85 μg/kg, respectively. Furthermore, 34, 28 and 26 numbers of rice, corn and corn product samples were found to be contaminated with OTA with an average level of 12.94, 5.29 and 3.69 μg/kg, respectively. The samples of rice, corn and corn products found above the permissible European Union (EU) limit for AFB₁ and total AFs were 18, 13 and 14 and 28, 14 and 20%, respectively; however, the samples of rice, corn and corn products above the EU limit were 40, 14 and 15%, respectively.     

Contribution of Nitrogen Fixed by Mung Bean to the Following Wheat Crop

Crop rotation is a supportive management practice in which legumes greatly improve the growth and yield of subsequent cereal crops, particularly wheat. In this study, effects of mung bean (Vigna radiata)?? on the yield and quality of grains of following wheat crop were determined. An experiment was carried out with two sets of wheat (Triticum aestivum) crops: one grown following mung bean (Vigna radiata) crop and the other grown after a fallow period. Concentrations of macronutrients, sugar, protein, amino acids, and phytohormones of wheat were determined. The grain protein concentration of wheat was improved if cultivated after mung bean, and nitrogen and other macronutrients of wheat were increased significantly. In case of phytohormone contents in the wheat crop, abscisic acid concentration showed no change, but the concentrations of gibberellin increased significantly by 41% and indole acetic acid by 30% as a result of crop rotation. Sugar content in wheat cultivated after mung bean BRM-318 showed 10% increase and wheat protein content increased by 17% and 20%, respectively. The present study demonstrated that crop rotation simulated yield and improved the nutritional value of wheat grain compared to wheat which followed the fallow period.