PubMed Health⌕ Search

Biomedical subjects

M F Dutton

Publications and source records attributed to M F Dutton.

34 records · Page 2Linked to original sources

An enzyme immunoassay for atractyloside, the nephrotoxin of Callilepis laureola (Impila).

Tubers of Callilepis laureola, a traditional remedy, contain an inhibitor of oxidative phosphorylation; atractyloside. A "competitive" ELISA was developed, using the antiserum produced to an atractyloside-protein conjugate. An ovalbumin-atractyloside conjugate was adsorbed to microtitre wells and plates incubated with sample (atractyloside or tuber extract) and antiserum. After successive incubation with secondary antibody-enzyme conjugate and substrate, the absorbance was read at 405 nm. Antibody working dilution was low, but results, confirmed by thin layer chromatography, indicate the immunoassay has diagnostic potential.

Animals↗

T-2 toxin metabolism by ruminal bacteria and its effect on their growth.

The effect of T-2 toxin on the growth rates of different bacteria was used as a measure of its toxicity. Toxin levels of 10 micrograms/ml did not decrease the growth rate of Selenomonas ruminantium and Anaerovibrio lipolytica, whereas the growth rate of Butyrivibrio fibrisolvens was uninhibited at toxin levels as high as 1 mg/ml. There was, however, a noticeable increase in the growth rate of B. fibrisolvens CE46 and CE51 and S. ruminantium in the presence of low concentrations (10 micrograms/ml) of T-2 toxin, which may indicate the assimilation of the toxin as an energy source by these bacteria. Three tributyrin-hydrolyzing bacterial isolates did not grow at all in the presence of T-2 toxin (10 micrograms/ml). The growth rate of a fourth tributyrin-hydrolyzing bacterial isolate was unaffected. B. fibrisolvens CE51 degraded T-2 toxin to HT-2 toxin (22%), T-2 triol (3%), and neosolaniol (10%), whereas A. lipolytica and S. ruminantium degraded the toxin to HT-2 toxin (22 and 18%, respectively) and T-2 triol (7 and 10%, respectively) only. These results have been explained in terms of the presence of two different toxin-hydrolyzing enzyme systems. Studies with B. fibrisolvens showed the presence of a T-2 toxin-degrading enzyme fraction in a bacterial membrane preparation. This fraction had an approximate molecular weight of 65,000 and showed esterase activity (395.6 mumol of p-nitrophenol formed per min per mg of protein with p-nitrophenylacetate as the substrate.

Animals↗

Effects of several mycotoxins on specific growth rate of Butyrivibrio fibrisolvens and toxin degradation in vitro.

Four strains of Butyrivibrio fibrisolvens did not degrade aflatoxin B1. Acetyl T-2 toxin, T-2 toxin, HT-2 toxin, deoxynivalenol, diacetoxyscirpenol, verrucarin A, zearalenone, and ochratoxin A did not affect the specific growth rate of B. fibrisolvens CE51 significantly, but all were degraded to greater or lesser extents. Breakdown products were produced as a result of deacetylation reactions.

Aflatoxin B1↗

Biosynthetic relationship among aflatoxins B1, B2, M1, and M2.

Aflatoxins are a family of toxic, acetate-derived decaketides that arise biosynthetically through polyhydroxyanthraquinone intermediates. Most studies have assumed that aflatoxin B1 is the biosynthetic precursor of the other aflatoxins. We used a strain of Aspergillus flavus which accumulates aflatoxin B2 to investigate the later stages of aflatoxin biosynthesis. This strain produced aflatoxins B2 and M2 but no detectable aflatoxin B1 when grown over 12 days in a low-salt, defined growth medium containing asparagine. Addition of dichlorvos to this growth medium inhibited aflatoxin production with concomitant accumulation of versiconal hemiacetal acetate. When mycelial pellets were grown for 24, 48, and 72 h in growth medium and then transferred to a replacement medium, only aflatoxin B2 and M2 were recovered after 96 h of incubation. Addition of sterigmatocystin to the replacement medium led to the recovery of higher levels of aflatoxins B2 and M2 than were detected in control cultures, as well as to the formation of aflatoxins B1 and M1 and O-methylsterigmatocystin. These results support the hypothesis that aflatoxins B1 and B2 can arise independently via a branched pathway.

Aflatoxin B1↗

The interaction between additives, yeast and patulin production in grass silage.

Both laboratory-prepared and sterile farm silage was found to support growth of Paecilomyces sp. and patulin production. The formation of patulin was affected by the levels of yeast present in the silage, and it was found that there was an inverse relationship between yeast population levels and patulin concentration. The commercial silage additive, "Sylade' had a greater lethal effect on yeast and fungi than "Add F', the latter allowing the formation of patulin by Paecilomyces sp. in the silage.

Animal Feed↗

The conversion of sterigmatocystin to O-methylsterigmatocystin and aflatoxin B1 by a cell-free preparation.

A cell-free system derived from a versicolorin A-accumulating mutant of Aspergillus parasiticus was found to convert sterigmatocystin to both O-methylsterigmatocystin and aflatoxin B1. It is suggested that the similarity in the chromatographic properties of these two metabolites has caused erroneous conclusions to be made with regards to the biosynthesis of aflatoxin B1.

Aflatoxin B1↗

Role of versicolorin A and its derivatives in aflatoxin biosynthesis.

The involvement of various anthraquinone metabolites in the biosynthesis of aflatoxin B1 was investigated by using a labeled double-substrate technique in a cell-free system. The results showed that both versicolorin A hemiacetal and versicolorin A hemiacetal acetate were converted to aflatoxin B1, whereas versicolorin A was not, even though it was added to the same cell-free system. Thus, versicolorin A hemiacetal, versicolorin A hemiacetal acetate, or both were implicated as key intermediates, whereas versicolorin A and C became side shunt metabolites. These latter compounds reentered the pathway depending on the availability of the appropriate enzymes and suitability of conditions. Dichlorvos, a specific inhibitor of aflatoxin biosynthesis, is considered to have its primary action on either an oxygenase or dehydrogenase involved in the pathway and to act in a secondary capacity as an inhibitor of an esterase which may also be involved in the pathway.

Aflatoxin B1↗

Biosynthesis of versicolorin A.

The incorporation of various potential intermediates into versicolorin A by a versicolorin A-accumulating mutant of Aspergillus parasiticus was studied. Both whole mycelium and cell-free extracts of this mutant were able to convert 14C-labeled versiconal hemiacetal acetate to versicolorin A. By the use of a labeled double substrate technique it was shown that two other compounds, versicolorin A hemiacetal and its acetate derivative, were also converted to versicolorin A. It is concluded that one or both of these compounds are intermediates in the biosynthesis of versicolorin A and therefore may possibly be involved in the biogenesis of the aflatoxins.

Anthraquinones↗

Synthesis of sterigmatocystin derivatives and their biotransformation to aflatoxins by a blocked mutant of Aspergillus parasiticus.

Seven alkyl and aryl homologues of O-methylsterigmatocystin (OMST) were synthesised and fed in separate experiments to a mutant of Aspergillus parasiticus capable of converting sterigmatocystin (ST) to aflatoxin B1 (AFB1). Their conversion to AFB1 was followed over a time period and it was found that O-propylsterigmatocystin (OPRST) was converted to AFB1 more rapidly than O-ethylsterigmatocystin (OEST) or OMST or ST itself. The aryl derivative O-benzoylsterigmatocystin (OBzST) was converted at the slowest rate. These results show that alkyl and aryl homologues of OMST may be converted to AFB1, suggesting that the methylation of ST is not an absolute requirement for its conversion to AFB1. It seems likely that whatever enzyme(s) are involved in this process exhibit relative specificity. As to whether alkylation of ST is an obligatory step in AFB1 biosynthesis is neither supported nor disproved as the fungal cells used are presumably capable of methylating ST. The fact that the propyl derivative showed fastest conversion is not necessarily significant as this may be due to faster diffusion of the least polar of the derivatives through the cell membrane.

Aflatoxins↗

Aflatoxin B1--its effects on an in vitro plant system.

The phytotoxic effects of aflatoxin B1 (AFB1) on in vitro cultures of differentiating calli and regenerating plantlets of Nicotiana tabacum were assessed. Callus appeared more sensitive to the effects of AFB1, with fresh mass accumulation and callus chlorophyll levels affected at low (approximately 0.5 micrograms/ml) aflatoxin concentrations. Transmission electron microscopy revealed early deteriorative alterations in chloroplast morphology. Inhibitory effects of the toxin (up to and including 10 micrograms/ml) on callus fresh mass accumulation were reversed following a 3 week toxin-free recovery period. In tobacco plantlets, root and leaf development, and root and leaf mass were significantly inhibited in a dose-dependent fashion with increasing AFB1 concentration above 0.5 micrograms/ml. Inhibitory effects on plantlet root development were more pronounced that on leaf development.

Aflatoxin B1↗

Occurrence of mycotoxins in cereals and animal feedstuffs in Natal, South Africa.

During the period 1982-1983, just under 800 samples of agricultural commodities, comprising cereals, compound feeds, hay, and silage, were examined for molds and mycotoxins. Aflatoxin B1 showed the highest incidence rate; it occurred in over 27% of all samples analyzed, the highest levels being found in peanut meal at 1500 ppb. Other mycotoxins detected were patulin and a number of trichothecene toxins at incidence rates in all commodities of 5.6 and 3.1%, respectively. The commodities at highest risk were oil seeds, excluding soya bean; the latter was found to be fairly free from contamination with mycotoxins. The most prevalent fungi were Aspergillus flavus and parasiticus, which were found in over 22% of all samples, whereas Penicillium spp. showed the lowest incidence of genera, specifically identified in 8.3% of all samples examined. This latter finding explains in part the low incidence of Penicillium mycotoxins.

Aflatoxin B1↗