Tang Lijie

High-level prokaryotic expression of envelope exterior of membrane protein of porcine epidemic diarrhea virus

Abstract The truncated fragment M′ gene, encoding the exterior of the viral envelope protein of PEDV, was subcloned into prokaryotic expression vector pGEX-6p-1. The recombinant plasmid pGEX-6p-M′ was constructed and transformed into E. coli BL21(DE3)pLysS for expression. SDS-PAGE analysis showed recombinant truncated M′ protein was highly expressed by pGEX-6p-M′ and the product fusion protein GST-M′ reached 45% in the total bacteria proteins with the analysis of software AlphaImager2200. The preliminary purified recombinant protein was evaluated for its antigenicity and reactivity through Western blotting and indirect enzyme-linked immunosorbent assay (ELISA) with monoclonal antibody against M protein of PEDV and porcine polyclonal anti-PEDV antiserum as the primary antibody. The results indicated the recombinant truncated M′ protein should be candidate as a feasible recombinant diagnostic reagent.

Laboratory Observations Regarding Different Instars of Cyclosainsulana （Costa, 1834） （Araneidae） During Developmental Stages

Abstract The current experiment was conducted to find out the optimal conditions for mass rearing and developmental changes of Cyclosainsulana. The lab. conditions were maintained at (27±2)°C and (65±5)% RH. The clear perplex cages and natural diet consisting of the aphids, larvae of the house fly and larvae of drosophila were used for rearing. C. insulana took (123.12±7.26) days to develop from eggs to adults passing through eight instars under prevailing vivo conditions. The eggs were greenish white in color with average size of 0.57 mm ±0.034. The eggs spent (7.52±1.64) days in emergence. Maximum number of spiderlings survived at the 5th instar (84%) and minimum at the 1st instar (34%). The measurements of different body parts including the total body length, cephalothorax and pedipalps of the both male and female C. insulana were done with the help of micrometer and presented as mean± SD. The measurements varied in the each developing instar. It was concluded that spiders were difficult to rear in the lab. conditions and each developing stage which was regarded as instars showed variations in size colors and body characteristics.

鱼源植物乳杆菌表达IPNV VP2-VP3重组蛋白及其口服免疫程序

为比较鱼源乳酸菌表达系统口服疫苗在不同免疫程序下诱导鱼免疫应答水平的差异，确定鱼源乳酸菌表达系统口服免疫虹鳟幼鱼的免疫程序。本研究构建了重组表达IPNV VP2-VP3蛋白的鱼源植物乳杆菌L1212，将重组菌pPG612-VP2-VP3/L1212包裹颗粒饲料，口服免疫虹鳟幼鱼。免疫程序分为连续免疫组、免疫1 d后间隔1 d再免疫1 d组、连续免疫4 d后32 d加强免疫1次组和间隔免疫后32 d加强免疫1次组。免疫后进行尾动脉采血，间接ELISA方法检测各组血清抗体效价，免疫接种66 d后，腹腔注射IPNV，计算各组相对免疫保护率。确定颗粒饲料按照1 mL/g 比例与108 CFU/mL 重组菌pPG612-VP2-VP3/L1212和2.5%海藻酸钠混合，于20 °C烘干制备口服疫苗。间隔免疫组血清抗体效价和攻毒保护率均显著高于其他组，并且免疫2次的间隔免疫组的血清抗体效价和攻毒保护率均显著高于免疫1次间隔免疫组。采用重组菌包被颗粒饲料饲喂虹鳟幼鱼，间隔免疫的免疫程序和免疫保护率优于连续免疫的免疫程序，对IPNV的入侵起到保护作用。

Abundance and Fluctuation in Spider Diversity in Citrus Fruits from Located in Vicinity of Faisalabad Pakistan

Abstract Spiders for the present study were collected from different fruit gardens (i.e. citrus) located at various localities (i.e., Tehsil Samundri, Jaranwala, Tandlianwala and Faisalabad) of District Faisalabad, Pakistan. Spiders belonging to six families and 33 species were captured from the two fruit gardens during the one year of this study. The citrus fruits garden was found to be best populated habitat as compared to other fruit garden. These sites were sampled by using pitfall traps; each month for five consecutive days from September 2010 to March 2011. As a result, 1 054 specimens were captured representing six families viz: lycosidae, thomosidae, gnaphosidae, saltisidae, araneidae and clubionidae. Lycosidae was more abundant, while clubionidae was less diverse during the study. Maximum population fluctuation among the spider specimens showed during the months from September and October, while the least abundance of spider specimens was reordered during June, November and December. Maximum taxonomic diversity was recorded from September to November, with the peak in September. It was concluded during these three months, when the citrus and guava gardens were attacked by the most of the pest insects. During the months of July and November diversity was moderate and mutually comparable, while in June and December, it was the least. This study contributed to the identification of spider diversity in the agro-ecosystem which could be used in the biological pest control.

Assessment of Phosphorus Levels in Small Ruminants as Affected by Summer and Winter

An experiment was carried out to assess the phosphorus status of free grazing goats at Faisalabad, Pakistan. Samples were collected fortnightly during summer and winter seasons of 2010 from soil and plants. The highest (118±0.54 mg • kg−1) levels of feces phosphorus were recorded in lactating goats during winter and (9.87±0.99 mg • kg−1) in urine of male during winter. Similarly maximum (71.0±0.88 mg • kg−1) phosphorus concentration was observed in the plasma of lactating animals. Milk contained (31.0±0.36) mg • L−1 in winter while during the months of summer the highest values recorded in forages, soils, canal and tube well waters were (755±1.98) mg • kg−1, (785±4.98) mg • kg−1, (0.97±8.78) mg • L−1 and (4.12±0.55) mg • L−1, respectively. It was revealed from the current results that fecal matter, forage, milk, tube well and canal water contained lower amounts of P, while P levels in blood plasma was found within the critical limits. Therefore, phosphorus supplementations were required in the area under experimentation to meet the requirements of the animals for their normal growth.