ergoline and roquefortine

In this report, we describe the natural intoxication of 2 dogs that consumed moldy dairy products found in the household garbage and the procedures used to identify and quantify the tremorgenic mycotoxins, roquefortine and penitrem A, in the remaining portions of ingested materials. Following the ingestion of mycotoxins, the dogs of our report developed muscle tremors or seizures that resembled clinical signs of strychnine poisoning. Roquefortine was the predominant mycotoxin in a moldy cream cheese wrapper that was found among scattered garbage consumed by the first dog. Penitrem A was the only mycotoxin detected in discarded moldy macaroni and cheese that was consumed by the second dog. Treatment of dogs with tremorgenic mycotoxin intoxication involves supportive care. Close monitoring is important because the development of aspiration pneumonia is common and has been reported as the cause of death. Clinical signs of intoxication gradually resolve within 24 to 48 hours.

Topics: Animals; Ataxia; Dog Diseases; Dogs; Ergolines; Female; Heterocyclic Compounds, 4 or More Rings; Indoles; Male; Mycotoxicosis; Mycotoxins; Penicillium; Piperazines; Status Epilepticus; Tremor

Mycotoxins are fungal metabolites that induce undesirable effects. The effects of these mycotoxins vary depending on the chemical structure of the toxin and degree of toxicity. Mycotoxins that induce muscle tremors, ataxia, and convulsions are termed tremorgenic mycotoxins. Our report documents the clinical course of 4 dogs from a single household that were simultaneously affected by tremorgenic mycotoxins. Diagnosis of tremorgenic mycotoxicosis was confirmed by stomach content analysis from 1 of the dogs. The mycotoxins identified were penitrem A and roquefortine, which are both produced by Penicillium spp. Treatment goals following tremorgenic mycotoxin ingestion include minimizing absorption, controlling tremors and seizures with methocarbamol and pentobarbital sodium administration, and providing supportive care. Two of the affected dogs required ventilatory support. With early aggressive treatment, prognosis is good and recovery is complete without sequelae. It is helpful for the clinician to be familiar with the typical clinical signs at the time of admission, treatment, and clinical course of dogs with tremorgenic mycotoxicosis.

Topics: Animals; Ataxia; Dog Diseases; Dogs; Ergolines; Gastrointestinal Contents; Heterocyclic Compounds, 4 or More Rings; Indoles; Male; Mycotoxicosis; Mycotoxins; Piperazines; Seizures; Tremor

Two dogs developed alarming tremorgenic nervous stimulation shortly after ingesting discarded rice that had been forgotten in a refrigerator for an undetermined period and that was covered with a grey-green mould. Both dogs exhibited vomition followed by slight salivation, tremors and ataxia and 1 showed such severe agitation and seizures that it necessitated anaesthesia with thiopentone followed, on recovery, by xylazine. The other dog was only sedated with xylazine. They made an uneventful recovery. The rice vomitus yielded a pure culture of Penicillium crustosum. On chemical analysis it was negative for organochlorine, organophosphor and carbamate insecticides, as well as for strychnine, but contained 2.6 microg/g of the mycotoxins penitrem A as well as 34 microg/g of roquefortine as determined by LC-MS and confirmed by MS-MS. This is the 1st South African case of naturally occurring penitrem A toxicosis and also the 1st case where quantification of the levels of mycotoxins in dog vomitus is reported. The tremorgenicity of roquefortine and its contribution towards this syndrome, is questioned.

Topics: Animals; Ataxia; Dog Diseases; Dogs; Ergolines; Female; Food Contamination; Gas Chromatography-Mass Spectrometry; Gastrointestinal Contents; Heterocyclic Compounds, 4 or More Rings; Indoles; Mycotoxicosis; Mycotoxins; Penicillium; Piperazines; Tremor; Vomiting

A short and efficient total synthesis of isoroquefortine C, the 3,12-(Z)-isomer of roquefortine C, from L-tryptophan methyl ester hydrochloride and 4(5)-(hydroxy)methylimidazole hydrochloride is described.

Topics: Ergolines; Heterocyclic Compounds, 4 or More Rings; Indoles; Magnetic Resonance Spectroscopy; Mass Spectrometry; Molecular Structure; Penicillium; Piperazines; Spectrophotometry, Infrared

Penitrem A and roquefortine poisonings were diagnosed in a Laborador retriever following garbage consumption. Clinical signs of mycotoxicosis included polypnea, tachycardia, and ataxia that quickly progressed to lateral recumbency and seizures. Removal of the mycotoxins from the stomach soon after ingestion allowed the dog to recover within 72-96 hours.

Topics: Animals; Ataxia; Dog Diseases; Dogs; Ergolines; Heterocyclic Compounds, 4 or More Rings; Indoles; Male; Mycotoxicosis; Mycotoxins; Piperazines; Seizures

Several strains of Penicillium are used for the production of mold-ripened cheeses, and some of them are able to produce mycotoxins. The aims of the research were the determination of roquefortine C and PR toxin in domestic and imported blue cheeses, the identification of the penicillia used as starter, and the investigation of their capacity for producing toxins in culture media. Roquefortine C was always found in the cheeses at levels ranging from 0.05 to 1.47 mg/kg, whereas the PR toxin was never found. The identification of the fungal strains present in the domestic cheeses included Penicillium glabrum, Penicillium roqueforti, and Penicillium cyclopium in the Gorgonzola "dolce" and Penicillium roqueforti in the Gorgonzola "naturale"; in one case, the presence of Penicillium crustosum was observed. The strains isolated from the foreign cheeses belonged to P. roqueforti. The strains were able to produce between 0.18 and 8.44 mg/liter of roquefortine in yeast extract sucrose medium and between 0.06 and 3.08 mg/liter and less than 0.05 mg/liter when inoculated in milk at 20 degrees C for 14 days and 4 degrees C for 24 days, respectively. Linear relations between production of roquefortine in culture media and cheeses did not emerge. PR toxin ranged from less than 0.05 to 60.30 mg/liter in yeast extract sucrose medium and was produced in milk at 20 degrees C from only one strain. The low levels and the relatively low toxicity of roquefortine make the consumption of blue cheese safe for the consumer.

Topics: Cheese; Chromatography, High Pressure Liquid; Ergolines; Heterocyclic Compounds, 4 or More Rings; Indoles; Mycotoxins; Naphthols; Penicillium; Piperazines; Time Factors

Seventy-six strains of Penicillium roqueforti used as starter cultures for mould ripened blue cheeses have been analysed for their RAPD genotype by using three different primers. A comparison of the RAPD patterns within each primer group revealed that the genetic constitution of the strains was similar, as most of the strains showed very similar overall patterns. Despite these similarities with each primer, distinct RAPD genotype groups could be identified. With one of the primers, it was possible to detect two heteropolymorphic DNA regions resulting in 13 different groups. With the other two primers, three or four groups could be identified. Between the groups of the different primers marked correspondence with respect to strain distribution could be observed, indicating that the polymorphisms detected by the primers were not independent. The RAPD patterns were compared to the production of secondary metabolites. A correlation was observed between the RAPD patterns of all primers and the production of mycophenolic acid. In addition, one of the primer (ari1) was able to distinguish between P. roqueforti strains producing larger or smaller numbers of metabolites.

Topics: Cheese; DNA Primers; DNA, Fungal; Ergolines; Genes, Fungal; Genotype; Heterocyclic Compounds, 4 or More Rings; Indoles; Mycological Typing Techniques; Mycophenolic Acid; Mycotoxins; Naphthols; Penicillium; Piperazines; Random Amplified Polymorphic DNA Technique

Roquefortine, a cyclopeptide derived from the diketopiperazine cyclo(Trp-dehydroHis), is a secondary metabolite produced by several Penicillium species. It has been reported to cause neurotoxic effect and to inhibit Gram-positive bacteria growth. The mechanisms responsible for its toxicity and metabolism are still unknown. In this study, we investigated the interaction of roquefortine with mammalian cytochromes P450. Roquefortine interaction with rat and human liver cytochromes P450 was monitored by difference UV-vis spectroscopy. It was found to interact with different forms of the cytochromes, giving rise to a type II difference spectrum, characteristic of the binding of an amino function to the heme iron. Roquefortine exhibited high affinity for microsomes from rats treated with various inducers, the K(s) values being in the range 0.2-8 microM. Similar results were observed with human P450 enzymes 1A1, 1A2, 2D6, and 3A4. Roquefortine had no effect on NAPDH cytochrome c reductase. Therefore, inhibition of NADPH consumption was observed using various rat liver microsomes alone or in the presence of 100 microM testosterone in the case of dexamethasone (DEX)-rat microsomes. Enzymatic inhibition was studied in terms of P450 3A activities, i.e., testosterone 6beta-hydroxylase (IC(50) around 10 microM) or bromocriptine metabolism (IC(50) > 50 microM) using DEX-rat liver microsomes or P450 3A4, benzphetamine N-demethylase using phenobarbital-rat liver microsomes (IC(50) > 30 microM), and ethoxyresorufin metabolism using 3-methylcholanthrene-rat liver microsomes (IC(50) 0.1 microM), P450 1A1, and 1A2. Roquefortine was compared with compounds of similar structure: cyclo(Phe-His), cyclo(Phe-dehydroHis), cyclo(Trp-His), phenylahistin. These studies indicate that the =N- imidazole moiety coordinates with the heme iron, and suggest that the dehydroHis moiety and the presence of a fused tetracycle play an important part in roquefortine inhibitory power.

Topics: Animals; Anti-Inflammatory Agents; Cytochrome P-450 Enzyme System; Dexamethasone; Ergolines; Heme; Heterocyclic Compounds, 4 or More Rings; Humans; Indoles; Liver; Male; Piperazines; Rats; Rats, Sprague-Dawley; Spectrum Analysis; Testosterone

Topics: Animals; Cytochrome P-450 CYP3A; Cytochrome P-450 Enzyme Inhibitors; Cytochrome P-450 Enzyme System; Ergolines; Heterocyclic Compounds, 4 or More Rings; Indoles; Liver; Male; Microsomes, Liver; Mixed Function Oxygenases; Molecular Structure; NADP; Piperazines; Rats; Testosterone

A method is described for the determination of roquefortine C in (blue) cheese. After liquid-liquid extraction with a mixture of hydrochloric acid and methanol, and filtration, an aliquot is analysed using column-switching reversed-phase liquid chromatography. The recovery of roquefortine C in Fetta cheese is about 85%, the calibration curve is linear from 10 to 2500 ng g(-1) (r2 = 0.998), and the detection limit is about 10 ng g(-1). In different batches of Danish Blue concentrations of 1000-2000 ng g(-1) of roquefortine C are found. As regards the stability of roquefortine C its half-life in diffuse daylight is ca. 50 min, while after irradiation with ultraviolet light, it is about 10 min.

Topics: Acetates; Buffers; Calibration; Cheese; Chromatography, Liquid; Drug Stability; Ergolines; Heterocyclic Compounds, 4 or More Rings; Hydrochloric Acid; Indoles; Methanol; Mycotoxins; Piperazines; Regression Analysis; Reproducibility of Results; Sensitivity and Specificity; Sunlight; Ultraviolet Rays

A method is described for the determination of roquefortine C in (blue) cheese. After liquid liquid extraction with a mixture of hydrochloric acid and methanol, and filtration, an aliquot is analysed using column-switching reversed-phase liquid chromatography. The recovery of roquefortine C in Fetta cheese is about 85%, the calibration curve is linear from 10 to 2500 ng g(-1) (r2 = 0.998), and the detection limit is about 10 ng g(-1). In different batches of Danish Blue concentrations of 1000-2000 ng g(-1) of roquefortine C are found. As regards the stability of roquelfortine C its half-life in diffuse daylight is ca. 50 min, while after irradiation with ultraviolet light, it is about 10 min.

Topics: Cheese; Chromatography, Liquid; Drug Stability; Ergolines; Heterocyclic Compounds, 4 or More Rings; Indoles; Light; Mycotoxins; Piperazines; Sensitivity and Specificity; Ultraviolet Rays

Topics: Animals; Dog Diseases; Dogs; Ergolines; Gas Chromatography-Mass Spectrometry; Heterocyclic Compounds, 4 or More Rings; Indoles; Mass Spectrometry; Mycotoxicosis; Mycotoxins; Piperazines

The uptake of exogeneous 14C-roquefortine by Penicillium farinosum cells and its metabolism was followed. Growth and roquefortine biosynthesis of three Penicillium species during a cultivation period of 14 d were investigated. Furthermore, the effect of cultivation conditions viz. various culture media on roquefortine uptake and biosynthesis as well as biomass production were studied. Some general problems of physiology and metabolic regulation of alkaloid formation in Penicillium species were discussed.

Topics: Ergolines; Heterocyclic Compounds, 4 or More Rings; Indoles; Mycotoxins; Penicillium; Piperazines; Time Factors

Auxotrophic, spore color, and alkaloid biosynthetic mutants of Penicillium roquefortii were selected after N-methyl-N'-nitro-N-nitrosoguanidine treatment. Diploids were obtained via protoplast fusion techniques, and the segregants from a diploid were genetically analyzed. The data demonstrated the potential of parasexual recombination in this organism. Evidence was obtained which suggests that the his and sts (sensitivity to Sulfatase) genes may be linked. The genetic information obtained in this study can serve as a starting point for further mapping of genes in P. roquefortii, and indications are that this organism may serve as a promising vehicle for the genetic study of the formation of ergoline alkaloids.

Topics: DNA, Fungal; Ergolines; Genes, Fungal; Haploidy; Heterocyclic Compounds, 4 or More Rings; Indoles; Mutation; Penicillium; Piperazines; Protoplasts; Recombination, Genetic; Spores, Fungal

We tested the mutagenic potential of crude extracts of 18 strains of Penicillium camemberti and 6 strains of P. roqueforti, which are used commercially in the production of mould ripened cheese in Switzerland. No mutagenic activity could be detected in any of the extracts. Roquefortine, a mycotoxin of P. roqueforti, often found in Blue cheese, was negative in the Amestest. The results obtained do not lead reasons for experting undesired long term effects from the consumption of mould ripened cheese.

Topics: Animals; Ergolines; Heterocyclic Compounds, 4 or More Rings; Indoles; Mutagenicity Tests; Mutagens; Mycotoxins; Penicillium; Piperazines; Rats

Roquefortine and the penitrems were biosynthesised concurrently at an approximately equimolar rate by Penicillium crustosum after growth and sporulation. [14C]mevalonic acid was incorporated (15% efficiency) into the isoprenoid regions of the penitrem and roquefortine molecules to an extent consistent with their 6:1 molar ratio of isoprenoid components. [14C]penitrem A (specific activity, 3.4 X 10(2) mu Ci mmol-1) and 14C-penitrems B, C, and E readministered to young cultures were metabolically interconverted, indicating considerable metabolic flux, though generally directed towards penitrem A as the end product and suggesting a metabolic grid for the penitrem metabolites. Addition of bromide to the medium preferentially favored the production of bromo-analogs rather than the usual chloropenitrems.

Topics: Ergolines; Fermentation; Heterocyclic Compounds, 4 or More Rings; Indoles; Kinetics; Mycotoxins; Penicillium; Piperazines; Structure-Activity Relationship

Two new indolic metabolites were isolated from Penicillium crustosum and separated from other penitrem mycotoxins by high-performance liquid chromatography. Penitrem D is a deoxy-penitrem A. Penitrem E is dechloro-penitrem A and was shown to be tremorgenic in mice, although it has only one-third of the activity of penitrem A. Roquefortine was also shown, for the first time, to be an important metabolic product of P. crustosum.

Topics: Animals; Chemical Phenomena; Chemistry; Chromatography, Liquid; Ergolines; Heterocyclic Compounds, 4 or More Rings; Indoles; Mice; Mycotoxins; Penicillium; Piperazines; Tremor

A method is described for the determination of roquefortine in blue cheese and blue cheese dressing. The method involves sample extraction with ethyl acetate, cleanup by liquid-liquid partition, and determination by high pressure liquid chromatography with ultraviolet and electrochemical detectors connected in series. Recoveries of roquefortine added to cheese at levels of from 16 to 320 ng/g averaged 74.9%. This method was applied to the analysis of 12 samples of blue cheese and 2 samples of blue cheese dressing, all of which were produced in the United States; roquefortine was found in all of the samples at average levels of 424 ng/g for the blue cheese and 45 ng/g for the blue cheese dressing.

Topics: Cheese; Chromatography, High Pressure Liquid; Electrochemistry; Ergolines; Heterocyclic Compounds, 4 or More Rings; Indoles; Mycotoxins; Piperazines; Ultraviolet Rays

Topics: Chromatography, Thin Layer; Densitometry; Ergolines; Heterocyclic Compounds, 4 or More Rings; Indoles; Mycotoxins; Naphthols; Penicillium; Piperazines; Spectrophotometry, Ultraviolet

Topics: Administration, Oral; Animals; Epoxy Compounds; Ergolines; Female; Heterocyclic Compounds, 4 or More Rings; Indoles; Injections, Intraperitoneal; Lethal Dose 50; Male; Mice; Mycotoxins; Naphthols; Penicillium; Piperazines