The side-effects of pesticides used in integrated production of peaches in Brazil on the egg parasitoid Trichogramma cacoeciae Marchal (Hym., Trichogrammatidae)

Abstract: The side‐effects of six pesticides used on peaches in Brazil were tested on the hymenopteran egg parasitoid Trichogramma cacoeciae using four laboratory tests: (a) adult parasitoid exposure to fresh pesticide residue on glass plates (worse case); (b) direct spray of host eggs enclosing the parasitoid egg, larvae or pupae (less‐exposed life stages); (c) exposure of adults to pesticide residues on plant leaves at different intervals after application (persistence); (d) Dose–response pesticide exposures of adults on glass plates. Two dose rates were used: (1) The highest recommended field dosage (FD) and (2) the predicted initial environmental concentrations (PIEC). The results showed that the preparations greatly differed in their initial toxicity and persistence. The insecticide Valient^® (methoxyphenozide) and the fungicide Venturol^® (dodine) were considered harmless to T. cacoeciae adults as they fell into the class 1 category according to the guidelines of the International Organization for Biological Control (IOBC) when parasitoids were directly exposed to chemical residues. The insecticide/acaricide Assist^® (mineral oil) was slightly harmful at the rate of PIEC 0.4 (40% of FD) and moderately harmful at FD. Pesticides in the categories harmless and moderately harmful can be considered for use in integrated pest management (IPM). The fungicide/acaricide Kumulus DF^® (sulphur) and the insecticides Dipterex^® 500 (triclorfon) and Lebaycid^® 500 (fenthion) were harmful at both concentrations. In the persistence test, Assist^® was short lived and therefore may in special cases (i.e. reduced direct contact) be considered for use in IPM, but Kumulus^® DF, Dipterex^® 500 and Lebaycid^® 500 constantly reduced parasitism between 77 and 100% and were rated as persistent (more than 30 days). The direct spray of parasitized host eggs at intervals after parasitism showed that Assist^® and Kumulus^® DF were harmless to the parasitoid egg, larvae and pupae within the host eggs. Dipterex^® 500 was slightly harmful when sprayed one day after parasitism (parasitoid egg) and moderately harmful to the other two stages (larvae and pupae). Lebaycid^® 500 was harmful to the parasitoid egg and larvae and moderately harmful to the pupae. The dose–response test showed that Kumulus^® DF and Dipterex^® 500 were toxic to T. cacoeciae. Kumulus^® DF was harmful from 1 PIEC 0.4 to 0.125 PIEC 0.4 dosages and was slightly harmful with 0.0625 PIEC 0.4 dosage. Dipterex^® 500 was harmful to T. cacoeciae in all the dosages tested.     

TB trifusion antigen adsorbed on calcium phosphate nanoparticles stimulates strong cellular immunity in mice

Among various vaccine candidates, subunit vaccines play an important role in immune protection against tuberculosis (TB). Calcium phosphate (CP) is considered as a strong inorganic adjuvant due to its great potential in increasing immune responses. The purpose of this study was to evaluate specific immune responses following the administration of trifusion-CP nanoparticles. The physiochemical properties of these nanoparticles, including morphology, particle size, zeta potential and adsorption rate, were measured in vitro. Subcutaneous immunization was performed three times on days 0, 14, and 28. Two weeks after the last administration, IFN-gamma, IL-4, and TGF-beta levels were measured by indirect enzyme linked immunosorbent assay (ELISA). The trifusion protein was successfully adsorbed onto calcium phosphate nanoparticles. The mean sizes of the resultant trifusion- CPN and CPN were 97.84 ± 12.08 and 67 ± 11.85 nm, respectively. CPN containing trifusion had stronger ability to induce IFN-gamma than the control groups. IL-4 and TGF-beta secretions in trifusion and trifusion-CPN groups were higher than those in the PBS group. However, there was no significant (p > 0.05) difference in IL-4 and TGFbeta concentrations between trifusion group and trifusion- CPN group. Therefore, calcium phosphate nanoparticles are good candidates for immunization against TB because antigen can be easily adsorbed onto CPN and strong cellular immune responses against CPN-antigen can be stimulated.     

Studies on the origin of biliary phospholipid. Effect of dehydrocholic acid and cholic acid infusions on hepatic and biliary phospholipids.

The correlation between the secretion of biliary phospholipid (PL) and bile acid suggests a regulatory effect of bile acid on PL secretion. Bile acids may influence PL synthesis and/or the mobilization of a preformed PL pool. The objective of this study was to determine the contribution of these two sources to biliary PL, by using an experimental protocol in which dehydrocholic acid (DHCA) and cholic acid (CA) were infused to manipulate biliary PL secretion. In control rats, there was a steady state in bile flow. PL secretion and the biliary secretion of newly synthesized phosphatidylcholine (PC). The specific radioactivity of PC in bile was significantly higher than in plasma, microsomes and canalicular membranes. DHCA infusion decreased biliary PC secretion rate by 80%, and secretion returned to normal values at the transport maximum of CA. The specific radioactivity of biliary PC was decreased by 30% by DHCA infusion and reached normal values during CA infusion. There were no significant changes in the specific radioactivity of PC in plasma or cellular organelles during infusion of bile acids. These data indicate that: (1) newly synthesized PC contributes a small percentage to biliary PC; thus a preformed pool (microsomal and extrahepatic) is a major source of biliary PL; (2) the contribution of the extrahepatic pool to the biliary PL may be more important than the microsomal pool.     

Liver cell plasma membrane lipids and the origin of biliary phospholipid.

The liver cell plasma membranes of fed male Wistar rats were separated into a fraction rich in bile canaliculi and the remainder of the plasma membrane. Electron-microscopically, the bile canalicular fraction consisted almost exclusively of intact bile canaliculi with thier contiguous membranes. The remaining plasma membrane fraction consisted primarily of vesicles and sheets of membranes essentially free from the bile canaliculi. The bile canalicular membrane fraction contained relatively more total lipid, cholesterol, and phospholipid, and relatively less protein. Although the phospholipid composition of the two fractions was the same, the specific activity of the bile canalicular membrane phosholipids, up to 12 h following in vivo administration of [2-3H]glycerol, was always significantly greater than that of the remaining plasma membranes, and showed a biphasic response not found in the latter. The specific activity of the phosphatidylcholine, phosphatidylethanolamine and lysophosphatidylcholine of the bile canalicular membranes rose to a peak within 40 min after administration of the label, fell sharply and then rose to a second peak after 120 min. The specific activity of the sphingomyelin and phosphatidylserine plus phosphatidylinositol of the bile canalicular membranes and of all the phospholipids of the remaining plasma membranes diphasic pattern but increased steadily to reach a maximum at 120 min. The specific activity of biliary phosphatidylcholine followed a pattern identical to that of the phosphatidylcholine, phosphatidylethanolamine and lysophosphatidylcholine of the bile canalicular membrane fraction. These results show that the average rate of turnover of phospholipid in the bile canalicular membranes is considerably greater than that in the remaining plasma membrane and other cell membrane fractions; they indicate that the phospholipid of the bile canalicular membranes exists in two or more pools, turning over a different rates; and they support the concept that biliary phospholipid is derived from the bile canalicular membrane. The results also suggest that bile canalicular phospholipid may be derived from two different sources, in contrast to the remainong plasma membrane.     

Central metabolism of beta-endorphin in different species of temperature acclimated rodents

The purpose of this study was to characterize the central metabolism/processing of beta-endorphin in wild, desert rodents and to study the effect of heat and cold acclimation on central metabolism/processing and beta-endorphin half-life. This report suggests that a shift occurs in the processing of beta-endorphin in desert rodents acclimated to heat and cold leading to an accumulation of gamma-type endorphins. The data support a significant difference in beta-endorphin half-life between the rodents and the laboratory white rat.     

Prevention of lithocholate--induced cholestasis by cycloheximide, an inhibitor of protein synthesis

Our previous investigations have shown that lithocholic acid (LCA)-induced cholestasis is associated with an increased synthesis of microsomal cholesterol which is transported with LCA and incorporated in the bile canalicular membrane. As the significance of these changes remains unknown the effect of interference with microsomal protein synthesis and/or with the cellular transport of cholesterol was studied. Male Wistar rats were injected i.p. with cycloheximide at the dose of 15 micrograms/100 g BW 3 times over a 24-hour period. After cannulating the common bile duct and collecting bile for one hour, the animals were either injected i.v. with 12 mumoles C14-LCA/100 g BW or with a 7.5% albumin solution and bile was collected for another hour. LCA injection in untreated animals reduced bile flow by more than 90% of control values. In contrast, bile flow in the group treated with cycloheximide and LCA was normal and did not differ from that of animals given cycloheximide alone. Bile salt secretion rate was increased in the cycloheximide-LCA group over the control groups. This was mainly due to the secretion of more than 80% of the injected LCA and was confirmed by the distribution of the radioactivity. By electron microscopy, the liver in the cycloheximide-LCA group did not show any of the well defined changes associated with LCA-induced cholestasis. These data suggest that microsomes play an important role in the pathogenesis of LCA cholestasis and that inhibition of microsomal protein synthesis can prevent its development.     

Quantitative Studies of Total Lipids of Pathogenic Fungi

Ten species of dermatophytes and four of the systemic fungi were assayed for total lipids, acetone-soluble fraction, and phospholipid content in different types of cultures. The yeast phase of each of the systemic fungi grown on solid medium exhibited a higher total lipid content than did the mycelial growth in liquid medium, either shake or still. Shake cultures, in all the fungi tested, produced the least lipids. The yeasts were consistently higher also in the acetone-soluble fraction. Histoplasma duboisii in the yeast phase and Microsporum gypseum produced the greatest amount of phospholipid, and Blastomyces dermatitidis in the yeast phase and M. canis produced the largest acetone-soluble fraction among the systemic fungi and dermatophytes, respectively.     

Contribution of the fluid phase endocytosis to bile flow in cholestasis and choleresis in rats

Using a single iv injection of 1 mg Evans blue dye/100 g body wt in rats, the contribution of the fluid phase endocytosis to bile flow was estimated. In control rats, this pathway contributed 2-4% of the bile flow or 10-20 microliter/hr/100 g body wt. This contribution was not affected by cholestasis induced by taurolithocholic acid or bile duct obstruction or by choleresis induced by dehydrocholic acid. It is concluded that fluid phase endocytosis does not significantly influence bile formation but, rather, may be implicated in the remodeling of liver cell plasma membranes.     

Methods for improved recovery of Listeria monocytogenes from cheese

Method of homogenization (Waring blender versus stomacher), type of diluent (tryptose broth [TB] versus aqueous 2% trisodium citrate), and temperature of diluent (20 versus 40 degrees C) were compared for recovery of Listeria monocytogenes from freshly made and ripened Colby cheese. By using direct plating on McBride listeria agar, significantly higher numbers of L. monocytogenes were recovered when cheese samples were (i) homogenized for 2 min with the blender rather than the stomacher (P less than 0.01), (ii) diluted in trisodium citrate rather than TB (P less than 0.01), and (iii) diluted in diluents at 40 rather than 20 degrees C (P less than 0.05). Based on these results, a new diluent/enrichment medium was developed by adding 2% trisodium citrate to TB (TBC). Despite superior results with the blender, biosafety concerns led to use of the stomacher for homogenization of cheese samples; hence, the stomaching time was increased to 3 min. Results obtained by direct plating indicated that recovery of L. monocytogenes from Colby cheese and from curd samples taken during manufacture of brick cheese increased when samples were diluted 1:10 in TBC at 45 degrees C and stomached for 3 min, as compared with similarly treated samples diluted in TB at 25 degrees C. A similar comparison of both diluents for recovery of L. monocytogenes from cold-pack cheese food yielded bacterial counts which were not significantly different. Recovery of L. monocytogenes from cold-enriched (at 4 degrees C for up to 8 weeks) samples of Colby cheese and cold-pack cheese food was generally similar for samples homogenized in TBC or TB.