Inhibition of pancreatic carboxypeptidase A: A possible mechanism of interaction between penicillic acid and ochratoxin A

Abstract

Penicillic acid and ochratoxin A are environmentally important toxic fungal metabolites (mycotoxins) that are synergistic in combination. The effects of penicillic acid on the pancreatic enzyme, :arboxypeptidase A were investigated in vitro and in vivo. A broad range of inhibition in vitro of the enzyme by PA was demonstrated with a half‐maximal inhibitory concentration equal to 1.1 x 10^‐4M PA. Inhibition of carboxypeptidase A was time and temperature dependent, and resulted in decreased conversion of parent ochratoxin A to the non‐toxic metabolite, ochratoxin alpha. Studies in vivo demonstrated a penicillic acid‐dependent inhibition of pancreatic carboxypeptidase A activity in the mouse and the chicken following multiple oral exposure. It is postulated that the mode of toxic interaction of the two mycotoxins may be due, in part, to impaired detoxification of ochratoxin A through peni‐cillic acid depletion of carboxypeptidase A activity.     

Multi-bioassay approach for assessing the potency of complex mixtures of polycyclic aromatic hydrocarbons

The chick embryotoxicity screening test (CHEST) and the Salmonella/microsome bioassay were used to evaluate embryotoxic and mutagenic endpoints from crude coal tar (CT) and its fractionated polycyclic aromatic hydrocarbon (PAH) mixtures (designated as A, B, C, D and E). In the CHEST assay, CT and PAH mixtures were injected into the egg yolk. A dose-dependent increase in embryo mortality was observed for all fractions. The E fraction resulted in 47% embryo mortality at a dose of 0.125 mg/kg and was more toxic than CT. At a dose of 1 mg/kg, 85-100% embryonic deaths occurred in fractions C and D and these two fractions were more potent than fractions A and B. The main visual toxic manifestations were liver lesions, discoloration of the liver, and edema. Both CT and fractionated PAH mixtures were also tested in the Salmonella/microsome plate incorporation assay with Salmonella typhimurium strain TA98 and were evaluated with and without metabolic activation at five dose levels. In the presence of S9, the CT and fractions C, D and E induced a dose-dependent positive response. Results from the Salmonella/microsome assay were in good agreement with findings from the CHEST assay suggesting that these two bioassays in combination may facilitate the rapid detection and ranking of complex PAH mixtures.     

Ochratoxin A and Penicillic acid interaction in mice

Abstract

Penicillic Acid (PA) and Ochratoxin A (OA) are toxic fungal metabolites that are synergistic in combination. This interaction was investigated using mice which were dosed orally as follows: control, none; solvent control, 0.2 ml bicarbonate buffer; PA, 40 mg/kg; OA, 10 mg/kg and combination, 40 mg/kg PA + 10 mg/kg OA. The only significant histopathologic change observed was an acute multifocal toxic tubular nephrosis which appeared most severe in the combination‐treated mice killed on day 10. While the combination group had a death rate of 20% (5/25), no deaths occurred in the other treatment groups. The increased death rate and the extensive nephrotoxic findings in the combination group indicate a toxic interaction between OA and PA at sub‐lethal dose levels and is consistent with a renal site of action.     

The combination of zinc compounds and acidic pH limits aerobic growth of a Salmonella typhimurium poultry marker strain in rich and minimal media

The objective of the present study was to examine the combined effects of zinc compounds with different acidic pH levels on the aerobic growth of a S. typhimurium poultry isolate in either rich or minimal media. When overall main effects of pH levels of medium or concentrations of Zn compounds were compared, growth rates of the S. typhimurium poultry isolate were significantly (p < 0.05) decreased by stepwise increase of pH levels of medium (pH 4, 5, 6, and 7) or concentrations (0.67, 3.35, and 6.03%) of Zn compounds (Zn acetate and Zn sulfate). In general growth rates of S. typhimurium poultry isolate appeared to be more reduced by Zn acetate than by Zn sulfate and more reduced in minimal media compared to rich media.     

Behaviour of arsenic in forested catchments following a high-pollution period

Due to high availability of adsorption sites, forested catchments could be net sinks for pollutant arsenic both during the period of increasing and decreasing pollution. We tested this hypothesis along a north-south pollution gradient in spruce die-back affected areas of Central Europe. For two water years (2007-2008), we monitored As fluxes via spruce-canopy throughfall, open-area precipitation, and runoff in four headwater catchments (Czech Republic). Since 1980, atmospheric As inputs decreased 26 times in the north, and 13 times in the south. Arsenic export by runoff was similar to atmospheric inputs at three sites, resulting in a near-zero As mass balance. One site exhibited a net export of As (2.2 g ha⁻¹ yr⁻¹). In contrast, the preceding period (1995-2006) showed much higher As fluxes, and higher As export. Czech catchments do not serve as net sinks of atmospheric As. A considerable proportion of old industrial arsenic is flushed out of the soil.     

Isolation of Salmonella typhimurium Tn5 mutants defective for survival on egg shell surface using transposon footprinting

The objective of this study was to screen the genome of Salmonella typhimurium for genes potentially required for survival on egg shell surfaces using a novel transposon footprinting method based on a previously developed Tn5 mutagenesis technique. Since the amplified DNA fragment length should usually be unique for each mutant, the polymerase chain reaction products separated on an agarose gel generate a transposon footprint with each band in the footprint representing the corresponding Tn5 mutant. By comparing the footprints from the pools of S. typhimurium Tn5 mutants before and after inoculation on egg shell surfaces, Tn5 mutants not recovered after selection were rapidly identified for potential isolation and genetic analysis.