Use of VP60 RT-PCR to overcome the limitation of haemagglutination inhibition diagnosis of rabbit viral haemorrhagic disease

[EN] Rabbit viral haemorrhagic disease (RVHD) is a highly contagious, highly fatal, peracute and acute viral disease of both wild and domestic rabbits caused by rabbit haemorrhagic disease virus (RHDV). Testing for haemagglutination (HA) activity in processed liver samples is one of the cornerstones for rapid diagnosis of RHDV outbreaks in national rabbitries. However, RHDV isolates exhibiting no HA activity are increasingly being reported. The extent of deviation from classical HA activity patterns for RHDV strains in Egypt has not been investigated. This study compared the HA activity patterns of samples collected from 61 RHDV outbreaks that occurred between 1999 and 2005 to determine whether dependence on HA test (HAT) for diagnosis of RHDV outbreaks needs to be reviewed. All samples were confirmed RHDV positive using SDS-PAGE and western blotting. Using slide HAT, only 36.1% of samples were positive (22 samples). Plate HAT conducted at 4 0C detected an additional 16 positive samples bringing the total HA-positive samples to 38 (62.3%). Plate HAT conducted at 22 0C failed to detect additional positive samples. The majority of samples detected after plate HA testing (62.5%) had HA titres comparable to those obtained from slide-HA-positive samples, indicating that the difference in HA activity is dependent on the nature of the HA antigen rather than its presence. Direct detection of HA activity failed in 37.7% of samples despite the presence of classical signs, pathology, and being RT-PCR positive for three different VP60 regions. Experimental infection of seronegative rabbits with 9 HA negative RHDV samples showed that 5 isolates were in-fact HA positive, while only 4 isolates remained HA negative. The increased detection of viruses lacking HA activity and the low HAT sensitivity mandates the use of molecular techniques for rapid confirmation of RHDV diagnosis in the Egyptian environment.This work was partially funded by the Veterinary Serum and Vaccine Research Institute, Abbasia, Cairo, Egypt, and by Faculty of Veterinary Medicine, Cairo University, Giza, Egypt.Shakal, M.; Khelfa, D.; Salman, O.; Yousif, A.; Salwa, E. (2011). Use of VP60 RT-PCR to overcome the limitation of haemagglutination inhibition diagnosis of rabbit viral haemorrhagic disease. World Rabbit Science. 19(1):11-20. doi:10.4995/wrs.2011.813SWORD112019

Poly(D,L-lactide-coglycolide) particles containing gentamicin: pharmacokinetics and pharmacodynamics in Brucella melitensis-infected mice

Drug delivery systems containing gentamicin were studied as a treatment against experimental brucellosis in mice. Micro- and nanoparticles prepared by using poly(D,L-lactide-coglycolide) (PLGA) 502H and microparticles made of PLGA 75:25H were successfully delivered to the liver and the spleen, the target organs for Brucella melitensis. Both polymers have the same molecular weight but have different lactic acid/glycolic acid ratios. Microparticles of PLGA 502H and 75:25H released their contents in a sustained manner, in contrast to PLGA 502H nanoparticles, which were degraded almost completely during the first week postadministration. The values of the pharmacokinetic parameters after administration of a single intravenous dose of 1.5 mg/kg of body weight of loaded gentamicin revealed higher areas under the curve (AUCs) for the liver and the spleen and increased mean retention times (MRTs) compared to those for the free drug, indicating the successful uptake by phagocytic cells in both organs and the controlled release of the antibiotic. Both gentamicin-loaded PLGA 502H and 75:25H microparticles presented similar pharmacokinetic parameter values for the liver, but those made of PLGA 75:25 H were more effective in targeting the antibiotic to the spleen (higher AUCs and MRTs). The administration of three doses of 1.5 mg/kg significantly reduced the load associated with the splenic B. melitensis infection. Thus, the formulation made with the 75:25H polymer was more effective than that made with 502H microspheres (1.45-log and 0.45-log reductions, respectively, at 3 weeks posttreatment). Therefore, both, pharmacokinetic and pharmacodynamic parameters showed the suitability of 75:25H microspheres to reduce the infection of experimentally infected mice with B. melitensis

INTERACTIVE EFFECTS OF ZINC AND DIFFERENT NITROGEN SOURCES ON YIELD AND QUALITY OF ONION

and 2003 to investigate the interactive effect of Zn and different nitrogen sources, i.e., mineral nitrogen (ammonium sulfate) and organic nitrogen (poultry manure) on the growth and productivity of onion (Allium cepa) cv. Giza 20. Four levels of Zn application (0, 1, 2 and 3 ppm) in a sulfate form were foliar sprayed at 30 and 60 days after planting. Within each Zn application, three treatments of nitrogen dose were applied in different combination forms (mineral and organic sources). Nitrogen treatments were 100% mineral N; 75% mineral + 25% organic and 50% mineral + 50% organic. Organic-N source was poultry manure. All poultry manure treatments were applied during soil preparation. Data showed that all growth parameters of onion plants increased as the fraction of poultry manure increased. Zn had also a positive effect on all plant parameters except of plant length, and leaf dry weight. The interactive effect of Zn and poultry manure increased yield and quality of onion bulbs. The most effective treatments on bulb fresh weight, total yield and quality were the spraying with 1 or 2 ppm zinc and the application of N as 75% mineral and 25% organic, in both season

Elaboration and characterisation of novel low-cost adsorbents from grass-derived sulphonated lignin

AbstractThis study investigated the use of water-soluble sulphonated lignin (SL) extracted from grass, which has not been used before as a precursor of activated carbon (AC). Chemical activation of SL with three dehydrating salts (ZnCl2, KCl, Fe2(SO4)3·xH2O) at various salt concentrations (10%, 20%, 30%w/w), charring temperatures (600,700°C) and charring times (1,2h) has been carried out. The surface characteristics and removal efficiencies of cadmium, copper and zinc ions from aqueous solutions were affected by the activation conditions. The sulphonated lignin-based activated carbons (SLACs) with the highest specific surface area, total pore and micropore volume were produced at the lowest dehydrating salt concentration (10%w/w) and at 700°C and 2-h charring. These optimal sulphonated lignin-based ACs were named SLAC-ZC (optimal grass-derived SLAC activated by zinc chloride); SLAC-PC (optimal grass-derived SLAC activated by potassium chloride) and SLAC-FS (optimal grass-derived SLAC activated by ferric sulphate). The central composite design and surface response methodology of different SLACs characteristics showed that the optimal responses were achieved at the same operating conditions. These SLACs also achieved the highest removal efficiencies of Cd2+, Cu2+ and Zn2+ ions from aqueous solutions. The chemical activation had significantly increased the total porosity, microporosity and surface area of water-soluble SL. The activation mechanism depended on the used dehydrating salt where the porosity developed by the dehydration effect of ZnCl2, and by a series of hydrolysis and redox reactions for the other two salts. The results of this research demonstrated that water-soluble SL has a great potential as a novel precursor for the production of activated carbons

Dissection-independent production of Plasmodium sporozoites from whole mosquitoes

Progress towards a protective vaccine against malaria remains slow. To date, only limited protection has been routinely achieved following immunisation with either whole-parasite (sporozoite) or subunit-based vaccines. One major roadblock to vaccine progress, and to pre-erythrocytic parasite biology in general, is the continued reliance on manual salivary gland dissection for sporozoite isolation from infected mosquitoes. Here, we report development of a multi-step method, based on batch processing of homogenised whole mosquitoes, slurry, and density-gradient filtration, which combined with free -flow electrophoresis rapidly produces a pure, infective sporozoite inoculum. Human-infective Plasmodium falciparum and rodent-infective Plasmodium berghei sporozoites produced in this way are two-to threefold more infective than salivary gland dissection sporozoites in in vitro hepatocyte infection assays. In an in vivo rodent malaria model, the same P. berghei sporozoites confer sterile protection from mosquito-bite challenge when immunisation is delivered intravenously or 60-70% protection when delivered intramuscularly. By improving purity, infectivity, and immunogenicity, this method represents a key advancement in capacity to produce research grade sporozoites, which should impact delivery of a whole parasite based malaria vaccine at scale in the future.Host-parasite interactio