Acute phase proteins and pesticide exposure

Gamma mobility C-reactive protein (CRP) level was determined in the sera of persons occupationally exposed to pesticides and controls in conjunction with serum protein analysis and other biochemical and enzymologic tests. Workers chronically exposed to dieldrin and pentachlorophenol showed significantly higher prevalence of CRP than the unexposed persons. In addition, the pentachlorophenol-exposed subjects revealed significantly elevated levels of total bilirubin and creatine phosphokinase, although the levels were within normal limits. The results suggest that chronic exposure to pentachlorophenol may have been responsible for the difference in the prevalence of CRP between the pentachlorophenol and control groups.     

Comparison of DT-diaphorases purified from the liver cytosol of untreated, and 3,4,5,3',4'-pentachlorobiphenyl- and 3-methylcholanthrene-treated rats

DT-Diaphorase purified from the liver cytosol of rats treated with a highly toxic PCB congener, 3,4,5,3',4'-pentachlorobiphenyl (PenCB), was compared to those from 3-methylcholanthrene (MC)-treated and untreated rats. Treatments with PenCB and MC resulted in about 8- and 7-fold increases of cytosolic DT-diaphorase activity, respectively. Purification of the enzyme preparations from untreated, and PenCB- and MC-treated rats were conducted by using DE-52, DEAE-Sephadex A-50, hydroxylapatite, and Bio-Gel P-150 column chromatographies. Both Sephadex G-100 gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that all of the final preparations from the three origins were homogeneous and had the same molecular weight of 59,000, consisting of two subunits with molecular weights of 30,000. Further studies on amino acid composition, Km value, optimum pH, and catalytic activities for various substrates also indicated that both PenCB- and MC-inducible DT-diaphorases were identical with that from the untreated rats. All three DT-diaphorases contained about 2 mol of FAD per mol of enzyme. Partial digestion of the enzymes by trypsin and subsequent analysis by HPLC revealed that the three preparations were indistinguishable. The identity among the three purified DT-diaphorases was finally confirmed by Ouchterlony immunodiffusion employing anti-serum raised against each enzyme preparation.     

Extension of Bacillus endospore gas dynamic heating studies to multiple species and test conditions

Aims: Shock wave–induced damage to a variety of Bacillus endospore species is studied for a wide range of postshock temperatures and test times in oxidative and non‐oxidative gas environments. Methods and Results: Bacillus atrophaeus and Bacillus subtilis endospores are nebulized into an aqueous aerosol, loaded into the Stanford aerosol shock tube (SAST) and subjected to shock waves of controlled strength. Endospores experience uniform test temperatures between 500 and 1000 K and pressures ranging from 2 to 7 atm, for either a short test time (～2·5 ms) or a relatively long test time (～45 ms). During this process, the bioaerosol is observed using in situ laser absorption and scattering diagnostics. Additionally, shock‐treated samples are extracted for ex situ analysis including viability plating and flow cytometry. For short test times, results are consistent with previous studies; all endospore species begin to lose the ability to form colonies when shock‐heated to temperatures above 500 K, while significant breakdown in morphology is observed for postshock temperatures above 700 K. Oxidative bath gases did not affect viability losses or morphological breakdown rates. Experiments with extended postshock test time showed increased viability loss with minimal morphological damage for shocks between 600 and 700 K. Conclusions: Genetic differences between B. subtilis and B. atrophaeus endospores do not confer noticeable gains in resistance to shock heating. Oxidative environments do not exacerbate shock‐induced damage to endospores. Extended test time experiments reinforce our hypothesis that a temperature/time‐dependent inactivation mechanism that does not involve morphological breakdown exists at low‐to‐moderate postshock temperatures. Significance and Impact of the Study: The methodology and experiments described in this paper extend the study of the interactions of endospores with shock/blast waves to new species and environmental conditions.     

Infestation of the white grub Dasylepida ishigakiensis (Coleoptera: Scarabaeidae) in hayfields on Miyako and Ishigaki Islands, Japan

To examine the local distribution of Dasylepida ishigakiensis Niijima et Kinoshita larvae (Coleoptera: Scarabaeidae) on Miyako Island and Ishigaki Island, a survey was conducted in hayfields on the two Islands in February and March 2011. Symptoms of white grub infestation, such as patches of wilted grass and thin, yellow grass stands, were observed in some of the 20 hayfields surveyed on Miyako Island. Three species of white grub were observed there: D. ishigakiensis, Anomala cpustulata cpustulata Matsumura in Hirayama, and Anomala albopilosa (Hope), in the ratio 92:7:1, respectively. The mean population density of D. ishigakiensis in the hayfields was 54.3 m^?2 and the maximum was 225 m^?2. No significant relationship was found between the damage levels of hay grass and the D. ishigakiensis larval densities. In contrast, no damage was observed in the 14 hayfields surveyed on Ishigaki Island, although some D. ishigakiensis larvae were found at a low density in 4 hayfields. In 2006, no D. ishigakiensis larvae were found in sugarcane fields on this island in a separately conducted intensive survey. These observations may suggest that D. ishigakiensis larvae are present in hayfields at a low density but that few individuals occur in sugarcane fields on Ishigaki Island.     

DNA from pre-erythrocytic stage malaria parasites is detectable by PCR in the faeces and blood of hosts

Following the bite of an infective mosquito, malaria parasites first invade the liver where they develop and replicate for a number of days before being released into the bloodstream where they invade red blood cells and cause disease. The biology of the liver stages of malaria parasites is relatively poorly understood due to the inaccessibility of the parasites to sampling during this phase of their life cycle. Here we report the detection in blood and faecal samples of malaria parasite DNA throughout their development in the livers of mice and before the parasites begin their growth in the blood circulation. It is shown that parasite DNA derived from pre-erythrocytic stage parasites reaches the faeces via the bile. We then show that different primate malaria species can be detected by PCR in blood and faecal samples from naturally infected captive macaque monkeys. These results demonstrate that pre-erythrocytic parasites can be detected and quantified in experimentally infected animals. Furthermore, these results have important implications for both molecular epidemiology and phylogenetics of malaria parasites. In the former case, individuals who are malaria parasite negative by microscopy, but PCR positive for parasite DNA in their blood, are considered to be “sub-microscopic” blood stage parasite carriers. We now propose that PCR positivity is not necessarily an indicator of the presence of blood stage parasites, as the DNA could derive from pre-erythrocytic parasites. Similarly, in the case of molecular phylogenetics based on DNA sequences alone, we argue that DNA amplified from blood or faeces does not necessarily come from a parasite species that infects the red blood cells of that particular host.     

Mating disruption by a synthetic sex pheromone in the white grub beetle Dasylepida ishigakiensis (Coleoptera: Scarabaeidae) in the laboratory and sugarcane fields

A serious sugarcane pest, Dasylepida ishigakiensis, remains in the soil during most of its life cycle except for a short period for mating. Mating disruption by an artificial release of the sex pheromone (R)-2-butanol (R2B), therefore, may be a feasible method to control this pest. We examined the effects of artificial release of R2B and its related compounds, (S)-2-butanol (S2B) and the racemic 2-butanol (rac-2B), on the mating success of this beetle both in the laboratory and in the field. In flight tunnel experiments, almost all males orientated towards a R2B-releasing source and 40% of them landed on the source. When the atmosphere was permeated with R2B, the frequency of males landing on the model was significantly reduced. Both rac-2B and S2B were less effective, but substantial reduction in landing success by males was achieved at higher rac-2B concentrations. R2B released from polyethylene dispensers in sugarcane plots greatly reduced not only the proportion of females mated with males but also the number of males caught by R2B-baited traps, indicating that male mate-searching behaviour was strongly affected by the released R2B. Similar inhibitory effects on male behaviour were also observed when tube- or rope-type dispensers released high rac-2B concentrations in the field. These results indicate that it would be highly possible to control D. ishigakiensis through the disruption of the sexual communication by releasing either synthetic R2B or rac-2B.