Preparation of natural rubber based semi-IPNs superabsorbent and its adsorption behavior for ammonium.

In this study, a natural rubber (NR) based amphiphilic semi-interpenetrating polymer network (semi-IPN) superabsorbent hydrogel was designed and synthesized with natural rubber-graft-poly (acrylic acid-co-acrylamide) [NR-g-P(AA-co-AM)] network and linear poly (diallyldimethyl ammonium chloride) (PDADMAC). Through a series of characterization and test, the structure, morphology, thermal properties, biodegradation, and swelling properties of NR-g-P(AA-co-AM)/PDADMAC were determined. Subsequently, NR-g-P(AA-co-AM)/PDADMAC was used for ammonium adsorption to remove ammonium nitrogen in aqueous solution. The adsorption behavior of the absorbent was also studied. Results showed that the maximum water absorbency of NR-g-P(AA-co-AM)/PDADMAC was 112.04 ± 6.55 g/g and water retention capacity of soil with the superabsorbent was 115.62 ± 2.08%. The NH4+ adsorption quickly reached equilibrium and the maximum adsorption capacity was 13.02 mmol g-1 calculated from Langmuir isotherm model. The results suggest that the product is efficient for ammonium removal and can be used as water-retaining agents.

Poly-acrylic acid grafted natural rubber for multi-coated slow release compound fertilizer: Preparation, properties and slow-release characteristics.

In this study, a novel multi-coated slow release compound fertilizer based on natural rubber (NR) was prepared and characterized. Firstly, NR was grafted with poly-acrylic acid by in-situ radical solution polymerization to synthesize poly-acrylic acid grafted natural rubber (NR-g-PAA), the reaction conditions were optimized to increase the water absorption properties of NR-g-PAA. Through a series of characterization and test, the structure, morphology, thermal properties and biodegradability of NR-g-PAA were determined. Subsequently, a multi-nutrient fertilizer core was fabricated with urea, KH2PO4, and attapulgite by pan granulation. Then the fertilizer core was coated by NR as the inner layer and NR-g-PAA as the outer layer. Meanwhile, the slow release behavior of the compound fertilizer in soil was also studied. Results showed that the maximum water absorbency of NR-g-PAA is 744.00 ± 14.38%. The release rate of N, P and K in 30 days for NR/NR-g-PAA coated fertilizer was about 54.35 ± 1.49%, 51.18 ± 2.15% and 44.37 ± 1.38%, respectively, showing that the nutrient element release can last for >30 days. Overall, the novel method introduced in this study can inform the development of NR based controlled release fertilizers.

Stable expression of foot-and-mouth disease virus protein VP1 fused with cholera toxin B subunit in the potato (Solanum tuberosum).

The expression vector, pBI121CTBVP1, containing the fusion of the foot and mouth disease virus (FMDV) VP1 gene and the cholera toxin B subunit (CTB) gene was constructed by fused PCR and transferred into potato (Solanum tuberosum L.) by Agrobacterium-mediated transformation. Transformed plants were obtained by selecting on kanamycin-resistant medium strictly and regenerated. The transgenic plantlets were identified by PCR, Southern-blot and the production of fused protein was confirmed and quantified by Western-blot and ELISA assays. The results showed that the fused genes were expressed stablely under the control of specific-tuber patatin promoter. The expressed fused proteins have a certain degree of immunogenicity.

Immune response in cattle inoculated with the recombinant complete polyprotein of foot-and-mouth disease virus from Bombyx mori larvae.

The intact open reading frame (ORF) of foot-and-mouth disease virus (FMDV) Asia I/XJ strain was amplified by RT-PCR and inserted into the transfer vector pVL1393 to generate plasmid pVL-ORF. Bm-N cells were transfected with pVL-ORF and linearized Bm-BacPAK6 DNA, and the recombinant silkworm baculovirus Bm-ORF containing the full ORF of FMDV was obtained. The results of indirect immunofluorescence assay (IFA) showed that Bm-ORF could be expressed efficiently in Bm-N cell. After inoculating the early 5th instar larvae of silkworm, the polyprotein of FMDV could be detected by sandwich ELISA and empty capsid-like particles could be observed under the electron microscope. Expression products from silkworm were used as the antigen to immunize the cattle. The specific antibody was induced in all vaccinated animals. The immunized cattle were challenged with the virulent FMDV Asia I/XJ strain, two of the four cattle were completely protected and clinical symptoms were alleviated and delayed in the others. The results suggest that this strategy might be used to develop the new subunit FMDV vaccine.

Development of a foot-and-mouth disease virus serotype A empty capsid subunit vaccine using silkworm (Bombyx mori) pupae.

Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals that inflicts severe economic losses in the livestock industry. In 2009, FMDV serotype A caused outbreaks of FMD in cattle in China. Although an inactivated virus vaccine has proven effective to control FMD, its use may lead to new disease outbreaks due to a possible incomplete inactivation of the virus during the manufacturing process. Here, we expressed the P1-2A and the 3C coding regions of a serotype A FMDV field isolate in silkworm pupae (Bombyx mori) and evaluated the immunogenicity of the expression products. Four of five cattle vaccinated with these proteins developed high titers of FMDV-specific antibody and were completely protected against virulent homologous virus challenge with 10,000 50% bovine infectious doses (BID(50)). Furthermore, the 50% bovine protective dose (PD(50)) test was performed to assess the bovine potency of the empty capsid subunit vaccine and was shown to achieve 4.33 PD(50) per dose. These data provide evidence that silkworm pupae can be used to express immunogenic FMDV proteins. This strategy might be used to develop a new generation of empty capsid subunit vaccines against a variety of diseases.

FMD subunit vaccine produced using a silkworm-baculovirus expression system: protective efficacy against two type Asia1 isolates in cattle.

Cattle vaccinated with a single dose of subunit vaccine containing the capsid and 3C proteinase coding regions of foot-and-mouth disease virus (FMDV) Asia I/HNK/CHA/05 strain were protected when challenged 28 days later with a homologous virus. Here, the 50% bovine protective dose (PD(50)) test was performed to assess the potency of the subunit vaccine. When challenged with two Chinese isolates, the subunit vaccine could achieve 6.5 PD(50) (challenged with Asia I/HNK/CHA/05 strain) and 5.2 PD(50) (challenged with Asia I/JSL/05 strain) per dose.

Expression of foot-and-mouth disease virus capsid proteins in silkworm-baculovirus expression system and its utilization as a subunit vaccine.

BACKGROUND: Foot-and-mouth disease (FMD) is a highly contagious disease of livestock that causes severe economic loss in susceptible cloven-hoofed animals. Although the traditional inactivated vaccine has been proved effective, it may lead to a new outbreak of FMD because of either incomplete inactivation of FMDV or the escape of live virus from vaccine production workshop. Thus, it is urgent to develop a novel FMDV vaccine that is safer, more effective and more economical than traditional vaccines. METHODOLOGY AND PRINCIPAL FINDINGS: A recombinant silkworm baculovirus Bm-P12A3C which contained the intact P1-2A and 3C protease coding regions of FMDV Asia 1/HNK/CHA/05 was developed. Indirect immunofluorescence test and sandwich-ELISA were used to verify that Bm-P12A3C could express the target cassette. Expression products from silkworm were diluted to 30 folds and used as antigen to immunize cattle. Specific antibody was induced in all vaccinated animals. After challenge with virulent homologous virus, four of the five animals were completely protected, and clinical symptoms were alleviated and delayed in the remaining one. Furthermore, a PD(50) (50% bovine protective dose) test was performed to assess the bovine potency of the subunit vaccine. The result showed the subunit vaccine could achieve 6.34 PD(50) per dose. CONCLUSION: The results suggest that this strategy might be used to develop the new subunit FMDV vaccine.

High expression of foot-and-mouth disease virus structural protein VP1 in tobacco chloroplasts.

A tobacco chloroplast expression vector, pTRVP1, containing the foot-and-mouth disease virus (FMDV) VP1 gene and the selective marker aadA gene, was constructed and transferred to tobacco by biolistic method. Three resistant lines were obtained through spectinomycin selection, and each transgenic line was subjected to a second round of spectinomycin selection. PCR and PCR southern blot analysis revealed that the VP1 gene had integrated into the chloroplast genome. Western blot and quantification ELISA assays indicated that the VP1 gene was expressed in tobacco chloroplasts and accounted for 2-3% of total soluble protein. This suggested that plant chloroplasts were an efficient expression system for the potential production of recombinant antigens in plants.