Formulation of a flowable liquid concentrate of Bacillus thuringiensis serotype H-14 spores and crystals as mosquito larvicide

Bacillus thuringiensis serotype H-14 spores and crystals, produced in 51 fermenters, were centrifuged and resuspended in emulsified palm olein to give 3.2 x 10(11) colony forming units (cfu)/ml. The suspension was mixed with a cassava-molasses-palm olein-charcoal (CMPC-2) mixture which served as the carrier, adhesive, dispersant and protectant. The final concentration of the formulation was 3.2 x 10(9) cfu/ml. The lethal concentrations capable of killing 50% of the test population (LC50) of CMPC-2 during 0, 1 and 2 years of storage at 32 +/- 4 degrees C were 0.056, 0.058 and 0.058 mg/ml respectively as against 0.054, 0.051 and 0.054 mg/ml for the Institut Pasteur Standard-1978 (IPS-78) during the corresponding period. The chi 2 tests showed that the results were homogeneous at P = 0.05. The relative potencies of the preparations were 964.3, 879.3 and 931 International toxic units (ITU) Aedes aegypti as compared with the 1000 ITU assigned to IPS-78. At 95% confidence limits there was no significant difference between the potencies of CMPC-2 and IPS-78. Field tests showed that CMPC-2 provided between 87.5 and 100% control of natural populations of Aedes spp. and Cutex spp. Sedimentation tests showed that CMPC-2 settled markedly during storage. This, therefore, required that the product be thoroughly shaken before use.     

Developmental changes in rat liver alcohol dehydrogenase

Hepatic alcohol dehydrogenase activity and mass content change coordinately during development in male rats. Enzyme activity and mass content increase continuously after birth to 100 and 80% of maximal values within 6 weeks (2.6 ± 0.4 μmole/min/g liver and 92 ± 20 μg/g liver), respectively. When expressed per milligram of soluble proteins, both parameters peak at 3 weeks (0.052 ± 0.002 μmole/min/mg protein and 2.0 ± 0.4 μg/mg protein) and then decrease gradually to plateau levels. These decreases probably arise from a “surge” in soluble liver protein levels that occurs after weaning. Similar developmental patterns also occur in female rats. These findings are the first quantitative measurements of this enzyme in developing animals.     

Structural-functional aspects of the postischemic recovery of the hepatocyte endoplasmic network in rats

The experiments on rats have shown that total hepatic ischemia reduces the content of microsomal cytochromes P-450 and b5 and causes amidopyrine and aniline disturbances over a 2-3-week post-ischemic period. The analysis of hepatocyte ultrastructure has revealed the interdependence of structural and functional changes in endoplasmic reticulum during recovery period. The damage of monooxygenase inducibility correlated with stable decline in the number of fixed ribosomes in post-ischemic period.     

DNA-Launched Alphavirus Replicons Encoding a Fusion of Mycobacterial Antigens Acr and Ag85B Are Immunogenic and Protective in a Murine Model of TB Infection

There is an urgent need for effective prophylactic measures against Mycobacterium tuberculosis (Mtb) infection, particularly given the highly variable efficacy of Bacille Calmette-Guerin (BCG), the only licensed vaccine against tuberculosis (TB). Most studies indicate that cell-mediated immune responses involving both CD4+ and CD8+ T cells are necessary for effective immunity against Mtb. Genetic vaccination induces humoral and cellular immune responses, including CD4+ and CD8+ T-cell responses, against a variety of bacterial, viral, parasitic and tumor antigens, and this strategy may therefore hold promise for the development of more effective TB vaccines. Novel formulations and delivery strategies to improve the immunogenicity of DNA-based vaccines have recently been evaluated, and have shown varying degrees of success. In the present study, we evaluated DNA-launched Venezuelan equine encephalitis replicons (Vrep) encoding a novel fusion of the mycobacterial antigens α-crystallin (Acr) and antigen 85B (Ag85B), termed Vrep-Acr/Ag85B, for their immunogenicity and protective efficacy in a murine model of pulmonary TB. Vrep-Acr/Ag85B generated antigen-specific CD4+ and CD8+ T cell responses that persisted for at least 10 wk post-immunization. Interestingly, parenterally administered Vrep-Acr/Ag85B also induced T cell responses in the lung tissues, the primary site of infection, and inhibited bacterial growth in both the lungs and spleens following aerosol challenge with Mtb. DNA-launched Vrep may, therefore, represent an effective approach to the development of gene-based vaccines against TB, particularly as components of heterologous prime-boost strategies or as BCG boosters.     

Polymorphism in Two Parasitoids Detected Using Random Amplified Polymorphic DNA Polymerase Chain Reaction

We used Chelex 100 chelating resin to prepare DNA for the polymerase chain reaction (PCR) from two species of Hymenopteran parasitoids, Trioxys pallidus and Diglyphus begini. Chelex 100 produces consistent DNA yields for both species, as measured with Hoescht dye fluorometry. Approximately twice as much DNA was obtained from individual D. begini wasps than from T. pallidus wasps, but there were no differences in yield between sexes. We used this DNA to perform random amplified polymorphic DNA (RAPD) analysis, a PCR technique that amplifies various regions of the genome using arbitrarily chosen 10-base primers. Of the 120 primers tested using T. pallidus, 92 produced a total of 342 scorable bands, 118 of which exhibited presence/absence polymorphism. Of the 25 primers tested using D. begini, 18 produced a total of 53 scorable bands, 30 of which exhibited presence/absence polymorphism. The level of genetic variation detected using this technique was greater than any found in Hymenoptera using allozymes. Scorable bands segregated as dominant Mendelian traits. Potential uses of RAPD-PCR in genetic analyses on parasitic Hymenoptera are discussed.