Phitsanu Tulayakul

The toxic effects and fate of intravenously administered zearalenone in goats

To clarify the toxic effects and fate of zearalenone (ZEA) in ruminants, we studied histopathological changes and toxicokinetic profiles in goats administered with a single intravenous (iv) injection of ZEA at doses of 2.4 mg/kg bw and 1.2 mg/kg bw, respectively. The expression of the mRNA of estrogen receptor (ER) alpha and beta in tissues was also investigated. The histopathological study revealed that ZEA caused hepatocellular swelling and lymphocytic infiltration in the liver, kidney, and uterus. The expression of ERalpha mRNA was enhanced by ZEA in association with the histopathological changes, indicating the possible involvement of ERalpha in the toxic effects of ZEA. For toxicokinetic profiles, blood plasma, urine, and feces were collected consecutively after iv injection of ZEA and analyzed for ZEA and its metabolites with high performance liquid chromatography (HPLC). alpha-Zearalenol (ZOL) and beta-ZOL were detected with ZEA, but alpha-zearalanol (ZAL), beta-ZAL, and zearalanone were below the detection limits. The distribution half-life (t(1/2alpha)) and elimination half-life (t(1/2beta)) of ZEA were 3.15 and 28.58h, respectively. ZEA, alpha-ZOL, and beta-ZOL were excreted in urine and feces, with beta-ZOL being the predominant metabolite. The ZEA and ZOL in urine were largely in their glucuronide and/or sulphate conjugated forms, while those in feces were largely in their free forms. This study showed the toxic effect of zearalenone and its metabolites, and their pharmacokinetic characteristics in goats.

Comparative study of heavy metal and pathogenic bacterial contamination in sludge and manure in biogas and non-biogas swine farms

The objective of this study is to determine and compare the heavy metal (Zn, Cu, Cd, Pb) and bacterial (E. coli, coliform and Salmonella spp.) contamination between swine farms utilizing biogas and non-biogas systems in the central part of Thailand. Results showed that average levels of E. coli, coliform, BOD, COD, Zn, Cu and Pb in sludge from the post-biogas pond were higher than the standard limits. Moreover, the levels of E. coli, coliform, Cd and Pb were also higher than the standard limits for dry manure. The levels of E. coli, coliform and BOD on biogas farms were lower than on non-biogas farms. Following isolation of Salmonella spp., it was found that Salmonella serovars Rissen was the most abundant at 18.46% (12/65), followed by Anatum 12.31% (8/65), and Kedougou 9.23% (6/65). The pathogenic strains of Salmonella serovars Paratyphi B var. java and Typhimurium were present in equal amounts at 4.62% (3/65) in samples from all swine farms. This study revealed that significant reduction in E. coli and coliform levels in sludge from covered lagoon biogas systems on swine farms. The presence of Salmonella as well as Cd and Pb, in significant amount in dry manure, suggests that there is a high probability of environmental contamination if it is used for agricultural purposes. Thus, careful waste and manure disposal from swine farms and the regular monitoring of wastewater is strongly recommended to ensure the safety of humans, other animals and the environment.

The effects of subcutaneous administration of T-2 toxin on liver drug metabolizing enzymes in piglets

T-2 toxin is one of trichothecenes, which are a structurally diverse group of toxic secondary metabolites produced by Fusarium and related species of fungi. The toxin usually contaminates cereal grains throughout the world. Although the pig is increasingly being used in pharmacological and toxicological studies, there is not enough information about the effects of T-2 toxin on drug-metabolizing enzymes in pigs. The purpose of this study is to investigate the effects of subcutaneous administration of T-2 toxin on the activities of hepatic Phase I and Phase II metabolizing enzymes in piglet liver. Piglets were administrated 0.3 mg T-2 toxin/Kg BW dissolved in DMSO by single subcutaneous (s.c.) injection. Control animals received only vehicle (DMSO). The activities of Phase I and Phase II enzymes were determined at 24 and 48 h after the last s.c. injection. The activities of cytochrome P-450 (CYP) 1A2 and 2E1 increased slightly at 24 and 48 h (P < 0.05). The CYP3A4 activity increased at 24 (P < 0.01), and tended to decrease at 48 h, but not significantly (P > 0.05). The glutathione S-transferase (GST) activity towards cumene hydroperoxide increased slightly at 24 h (P < 0.05), but decreased slightly at 48 h (P < 0.05). No significant changes were observed in the glutathione S-transferase activity toward 1-Chloro-2,4-dinitrobenzene (CDNB) either at 24 or 48 h. Western blot analyses of the liver fractions revealed increased levels of CYP1A2, 2E1, 3A4, GST , GST M1-1 at 24 h, and that of CYP2E1 at 48 h.The results suggest that T-2 toxin causes modulation of Phase I and Phase II drug metabolizing enzymes in piglets.

Organ differences in microsomes and cytosol metabolism of Aflatoxin B1 in piglets

In order to delineate the features of aflatoxin B1 (AFB1) metabolism in various organs of piglets, in vitro metabolism of AFB1 by microsomes and cytosol of the various piglet organs was studied. The AFB1 was converted efficiently to AFP1 by the kidney microsomes. A less efficient metabolism was noted from the AFB1 to AFQ1, AFM1, and aflatoxicol (AFL) in the various organs. The microsomal ability to form AFB1-DNA adduct was higher in liver when compared to the other organs. The cytosolic glutathione-S-transferase activity to convert AFB1-epoxide to AFB1-glutathione conjugate product was relatively higher in the liver and the small intestine. The reductase activity to convert AFB1-dialdehyde to AFB1-dialcohol was similar in all the organs. The results suggest that the variation in susceptibility to the aflatoxin among different organs is attributable mainly to the organ differences in cytochrome P450 activity to form AFB1-epoxide.

Occurrence of Penicillium brocae and Penicillium citreonigrum, which Produce a Mutagenic Metabolite and a Mycotoxin Citreoviridin, Respectively, in Selected Commercially Available Rice Grains in Thailand

Commercially available rice grains in Thailand were examined to isolate the monoverticillate Penicillium species responsible for toxic yellowed rice. Penicillium species were obtained from seven out of 10 rice samples tested. Among them, one Penicillium citreonigrum isolate and six Penicillium brocae isolates were morphologically identified. The P. citreonigrum isolate produced the mycotoxin citreoviridin on a yeast extract sucrose broth medium. Mycotoxin surveys showed that citreoviridin was not detected in any samples, but one out of 10 rice samples tested was positive for aflatoxin B1 at a level of 5.9 μg/kg. An Ames test revealed that methanol extracts from rice grains inoculated with selected P. brocae isolates were positive for strains TA100 and YG7108 of Salmonella typhimurium, suggesting the presence of base-pair substitution and DNA alkylation mutagens. Our data obtained here demonstrated that aflatoxin B1 and toxic P. citreonigrum were present on domestic rice grains in Thailand, although limited samples were tested. Penicillium brocae, which may produce mutagenic metabolites, was isolated for the first time from the surface of Thai rice grains.

Comparison of zinc, lead, cadmium, cobalt, manganese, iron, chromium and copper in duck eggs from three duck farm systems in Central and Western, Thailand

This was a comparative study of the heavy metal levels (Zn, Pb, Cd, Co, Mn, Fe, Cr and Cu) in eggs from free grazing duck, small-scale, and large-scale farms in central and western regions of Thailand. A questionnaire was used to gather demographic data for the analysis of heavy metal contamination in feed, drinking water and wastewater. The correlation between the amounts of heavy metal contamination in eggs was studied against the heavy metals found in feed, drinking water and wastewater. The levels of Pb, Cd, Cr and Cu in eggs from large-scale farms were significantly higher than small farms and free grazing farms at P < 0.001. Zn in eggs from free grazing farms was higher than in the small farms and large-scale farms sampled. The contamination of Pb in eggs from all types of farms exceeded the standard limits of ACFS 6703-2005. The average levels of Pb in the eggs from small-scale farms correlated significantly with the level of Pb found in the feed at P < 0.05, while the average levels of Pb in eggs from free grazing duck farms correlated significantly with the levels of Pb found in the drinking water at P < 0.05. Additionally, the average level of Cu in duck egg from large-scale farms correlated significantly with the level of Cu found in the feeds at P < 0.001. Furthermore, from a calculation of the provisional tolerable daily intake (WHO-FAO) of heavy metals in this study, it was concluded that consumers face health risks from Cd contamination. Thus, heavy metal contamination, especially Pb and Cd in duck egg, must be of concern due to the health risks and the route of crucial heavy metals contamination should be elucidated and long - term monitoring of heavy metals posing health effects in farm systems should be carried out.