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F J Cabañes

Publications and source records attributed to F J Cabañes.

At least 19 recordsLinked to original sources

RFLP characterization of Aspergillus niger aggregate species from grapes from Europe and Israel.

In order to characterize by molecular methods the Aspergillus niger aggregate species involved in the ochratoxin A (OTA) contamination of European wine grapes and table grapes from Israel, a total of 173 strains were studied. The ITS-5.8S rDNA fragments of 173 A. niger agreggate strains from grapes included in this study were amplified and their PCR amplicons were RsaI digested in order to classify the strains in the RFLP types, N and T. All of the strains belonging to the A. niger aggregate were classified into the two RFLP types previously defined: type N (43%) and type T (57%). Twenty out of the 173 strains of A. niger aggregate produced OTA (0.1 to 10.5 mug g(-1)). All the OTA producing species belonged to the N-RFLP type.

Aspergillus niger↗

Study of the effect of water activity and temperature on ochratoxin A production by Aspergillus carbonarius.

The effect of water activity (aw) (0.78-0.99) and temperature (15 and 30 degrees C) on growth and production of ochratoxin A (OTA) of six Aspergillus carbonarius strains was studied in two culture media: Czapek yeast autolysate (CYA) agar and yeast extract sucrose (YES) agar, during a period of 30 days. The strains were selected to include different sources and different reported abilities to produce OTA and were characterized by RAPD and ITS-5.8S rDNA sequencing. CYA showed to be better culture medium than YES for OTA production in the isolates tested. OTA concentration was higher at 15 degrees C than at 30 degrees C. At 30 degrees C, ranges for OTA production were more restrictive than those for growth. OTA was produced from 0.86, 0.90 or 0.94 aw depending on the strain. At 15 degrees C, growth and OTA production were detected only in the 0.94-0.99 aw range. The molecular study performed showed that five of the strains were conspecific and no correlation was found between molecular data and the OTA production level or origin. The remaining strain had never been able to produce OTA and will probably represent a new species in the Aspergillus section Nigri. Our results show that A. carbonarius is able to grow and produce OTA in a wide range of water activities at both high and low temperatures.

Aspergillus↗

Effect of water activity on ochratoxin A production by Aspergillus niger aggregate species.

The effect of water activity (a(w)) (0.82-0.99) on growth and ochratoxin A (OTA) production by twelve Aspergillus niger aggregate strains, cultured in Czapek Yeast Autolysate agar (CYA) and Yeast Extract Sucrose agar (YES), was studied for an incubation period of 30 days. The strains were selected to include diverse sources, different reported abilities to produce OTA and different ITS-5.8 S rDNA Restriction Fragment Length Polymorphism (RFLP) pattern. They were characterized by Random Amplification of Polymorphic DNA (RAPD) and ITS-5.8 S rDNA and 28 S rDNA (D1/D2) sequencing. Regardless of the a(w) value tested, YES was a better culture medium than CYA for OTA production. The a(w) range for OTA production was narrower than that for growth. OTA was produced from 0.90, 0.92, 0.94 or 0.96 to 0.99 a(w) depending on the strain and the culture medium. The molecular study differentiated strains into two groups which corresponded to the RFLP types N and T although it did not distinguish them by their source of isolation or OTA producing abilities. Our results show that A. niger aggregate strains are able to grow and produce OTA over a wide a(w) range. These results will lead to a better understanding of the contribution of A. niger aggregate in OTA contamination of food and feed.

Agar↗

Effect of pH on ochratoxin A production by Aspergillus niger aggregate species.

The effect of pH (2-10) on growth and ochratoxin A (OTA) production by 12 Aspergillus niger aggregate strains was studied in two culture media: Czapek yeast autolysate agar (CYA) and yeast extract sucrose agar (YES), over 30 days. The strains were selected to include different sources, different reported abilities to produce OTA and different ITS-5.8S rDNA RFLP patterns. YES was a better culture medium than CYA for OTA production. In this medium, OTA was produced from pH 2 or 3 to 10 depending on the strain. The results show the ability of A. niger aggregate strains not only to grow, but also to produce OTA over a wide pH range. The results will lead to a better understanding of the role of A. niger aggregate strains in the OTA contamination of several food commodities.

Aspergillus niger↗

Ochratoxin A-producing fungi from grapes intended for liqueur wine production.

The ochratoxigenic mycobiota of grapes intended for liqueur wines from four Spanish vineyards were studied. The specific wine-making technology of these wines requires overripening of the grapes on the vine or extended post-harvest exposure of the grapes in the sun. In every vineyard, samples were taken at three different developmental stages: veraison, harvesting time and after over-ripening. With the maturation of the berries there was a clear increase of Aspergillus spp. In the last sampling time studied, they were isolated from the 90.3% of the plated berries. Black aspergilli (mainly A. niger aggregate and A. carbonarius) were predominant among the different Aspergillus spp. isolated and constituted 98.5% of the total Aspergillus strains isolated. At harvesting time and after over-ripening, the percentage of colonized berries with A. carbonarius exceeded that of Aspergillus niger aggregate. Due to their low frequency of isolation, Penicillium spp. and Aspergillus spp. outside black aspergilli are not an important source of ochratoxin A in grapes for liqueur wine production. On the contrary, 98.5% of the A. carbonarius isolates screened were able to produce ochratoxin A. Although the possible participation of different ochratoxin A-producing species may occur, our results confirm that A. carbonarius is the most important source of ochratoxin A in liqueur wines, increasing its occurrence along the ripening of grapes.

Aspergillus↗

Genetic typing of Malassezia pachydermatis from different domestic animals.

In the present study, random amplification of polymorphic DNA, which detects DNA polymorphism in fungal genomic DNA, was applied for genetic typing of Malassezia pachydermatis isolates. Fifty-five isolates from different domestic animals and body sites and the neotype strain CBS 1879 were characterized. Primers M13 and OPT-20 were used to analyse their genetic relatedness and similarity. This technique allowed us to distinguish four different genetic types. The predominant genetic type was observed in isolates recovered from different anatomical locations in all animals. It was the only genetic type found in cats, horse, goat and pig. The other three genetic types were observed only in isolates from external ear canals of dogs. Types II and IV were only recovered from external otitic ears and type III from healthy ears. An animal was colonised by more than one type of M. pachydermatis and different genetic types were detected in the same body site. Some genetic types were only isolated from diseased skin.

Animals↗

Ochratoxigenic species from Spanish wine grapes.

The ochratoxigenic mycobiota of grapes belonging to representative wine regions located along the Mediterranean coast of Spain at different developmental stages was identified. During the development of the berries, the occurrence of Aspergillus spp. increased while the percentage of berries contaminated by non-ochratoxin A (OTA) producing species such as Alternaria spp. and Cladosporium spp. decreased. Penicillium verrucosum, the only confirmed Penicillium spp. that is able to produce OTA, was not isolated. The contamination by OTA-producing species comes from the surface of the berries and not from the inner fruit. Black aspergilli were predominant among the different Aspergillus spp. isolated. All the Aspergillus carbonarius isolates were able to produce OTA at different concentrations. None of the isolates belonging to Aspergillus niger aggregate and to Aspergillus japonicus var. aculeatus were able to produce OTA. These results are a strong evidence of the contribution of A. carbonarius in the OTA contamination in wine grapes, mainly at the last developmental stages of the berries.

Aspergillus↗

Comparison of disk diffusion method and broth microdilution method for antifungal susceptibility testing of dermatophytes.

The use of the agar diffusion Neo-Sensitabs method to determine antifungal susceptibility of 59 isolates of dermatophytes, namely Epidermophyton floccosum, Microsporum canis, M. gypseum, Trichophyton mentagrophytes, T. rubrum and T. tonsurans to Clotrimazole (CLZ), Itraconazole (ITZ) and Terbinafine (TBF) is described. Results obtained are compared to the minimum inhibitory concentrations (MIC) determined by an adaptation of the NCCLS-M38-A procedure. Using the diffusion method, all strains showed a broad zone of inhibition at the first available reading time (3 or 7 days). Using the broth microdilution method, the geometric mean MIC (microg/ml) with regard to all isolates was < or = 0.03 for TBF, < or = 0.069 for CLZ and < or = 0.919 for ITZ. In both methods, TBF was the most active antifungal agent against all isolates tested. The two methods evaluated were able to detect the resistance of the quality control strains of Aspergillus fumigatus to ITZ. Even though a reference method for testing dermatophytes still has not been developed, our data suggest that the Neo-Sensitabs diffusion method could provide a simple procedure for the antifungal susceptibility testing of dermatophytes in the routine clinical laboratory.

Antifungal Agents↗

Molecular analysis of Malassezia sympodialis-related strains from domestic animals.

Recently, several new lipid-dependent species belonging to the genus Malassezia have been described. Some of them, such as Malassezia dermatis, Malassezia nana, and the tentatively named "Malassezia equi," have similar phenotypes and are genetically close to Malassezia sympodialis Simmons et Gueho 1990. DNA characterization by D1/D2 26S rRNA gene and internal transcribed spacer (ITS)-5.8S rRNA gene sequencing analysis of lipid-dependent strains from different animal species close to M. sympodialis is described and illustrated. Phylogenetic analysis of both the D1/D2 regions of 26S rRNA gene and ITS-5.8S rRNA gene sequences showed four distinct clusters. Cluster I included isolates from different animal species (horse, pig, and lamb) and the type culture of M. sympodialis. Cluster II included isolates from horses grouping close to the "M. equi" AJ305330 sequence. Cluster III comprised isolates mainly from goats. Cluster IV contained isolates mainly from cats grouping together with the M. nana AB075224 sequence. This last cluster included isolates from healthy and external otitic ears. All of these strains had identical 26S rRNA gene and ITS regions. It is not clear whether the value of these genetic differences is for the definition of species or whether they only demonstrate genetic variation among strains from different origins within M. sympodialis, which are in the course of differentiation and probably adaptation to specific animal hosts.

Animals↗

Influence of pH and incubation time on ochratoxin A production by Aspergillus carbonarius in culture media.

The effect of pH (2 to 10) and temperature (15 and 30 degrees C) on growth and production of ochratoxin A (OTA) of six strains of Aspergillus carbonarius was studied in two culture media: Czapek yeast autolysate agar and yeast extract sucrose agar. Isolates were selected by their different source and different reported ability to produce OTA. Regardless of the initial pH or the temperature tested, Czapek yeast autolysate agar has been shown to be the best culture medium for OTA production by A. carbonarius. In this medium, OTA was produced from pH 2 to 10 at the two incubation temperatures tested. The results obtained show the ability of A. carbonarius to not only grow but also produce OTA over a wide pH range at high or low temperatures. This may help explain why this species is considered the main OTA source in some substrata.

Agar↗

Selective efficacy of culture media recommended for isolation and enumeration of Fusarium spp.

Selective culture media, such as Nash and Snyder medium (NS), dichloran-chloramphenicol peptone agar (DCPA), modified Czapek-Dox agar (MCz), Czapek Dox iprodione dichloran agar (CZID), potato dextrose iprodione dichloran agar (PDID), or malachite green agar (MGA 2.5), have been developed for isolating and enumerating Fusarium spp. from natural samples. However, some of these culture media are not very selective because they allow the growth of many other fungal species. In this study, a comparison of the selective efficacy of these culture media, using different strains of Fusarium spp. (F. anthophilum, F. culmorum, F. dlamini, F. graminearum, F. napiforme, F. nygamai, F. oxysporum, F. proliferatum, F. semitectum, F. solani, F. subglutinans, and F. verticillioides) and natural samples has been carried out. Among the six recommended selective culture media assayed, no statistical differences were detected in colony counts of the Fusarium spp. strains tested, although the colony diameters in MGA 2.5 were significantly lower than in NS, MCz, DCPA, CZID, and PDID media. With natural samples, MGA 2.5 performs as a potent selective medium for Fusarium spp., whereas the other recommended selective media allow the growth of many other different fungal species including Zygomycetes and yeasts.

Colony Count, Microbial↗

Aspergillus carbonarius as the main source of ochratoxin A contamination in dried vine fruits from the Spanish market.

Ochratoxin A (OTA) can occur in a wide range of foods, but unexpectedly high concentrations have been detected in dried vine fruits of various origins. The European Union has recently established a maximum OTA limit of 10 microg/kg for these foodstuffs. In order to determine the likely origin of OTA, a mycological study of 50 dried fruit samples (currants, raisins, and sultanas) representative of the Spanish market was conducted. Fungal contamination was detected in 49 of 50 (98%) samples. Black aspergilli were isolated from all of the positive samples. Aspergillus niger var. niger was isolated from 98% of the samples, and Aspergillus carbonarius was found in 58% of the samples. One hundred sixty-eight A. niger var. niger isolates and 91 A. carbonarius isolates were screened for their ability to produce OTA. Eighty-eight (96.7%) A. carbonarius isolates and one (0.6%) A. niger var. niger isolate were found to be OTA producers. Black aspergilli were the dominant fungi. Among black aspergilli, A. carbonarius has shown a consistent ability to produce OTA and is the most probable source of this mycotoxin in these substrates.

Aspergillus↗

What is the source of ochratoxin A in wine?

During a microvinification trial using natural mouldy grapes from a research experimental vineyard, ochratoxin A (OTA) contaminated white wine was obtained. Potential OTA-producing mycobiota of grape samples used in this microvinification process was assessed. Only Aspergillus carbonarius isolates were detected as producers of OTA. Our report is a strong evidence of the contribution of A. carbonarius in the OTA contamination in wine.

Aspergillus↗

Occurrence of Malassezia spp. in horses and domestic ruminants.

During a study on the occurrence of Malassezia spp. in 112 animals (50 horses, 25 goats, 25 sheep and 12 cows), Malassezia spp. were isolated from 60% of horses, 28% of sheep, 44% of goats and 58% of cows. In these animals, the occurrence of lipid-dependent species (42%) was much greater than the occurrence observed for M. pachydermatis (3%). Among the results obtained, we point out the first isolation of M. sympodialis, M. globosa and M. restricta from sheep, M. pachydermatis, M. furfur, M. sympodialis, M. obtusa, M. globosa and M. restricta from goats and M. furfur, M. slooffiae, M. obtusa, M. globosa and M. restricta from horses. To the authors' knowledge, this survey also states the first description of the presence of M. restricta in animals.

Animals↗

Occurrence of Malassezia spp. in the external ear canals of dogs and cats with and without otitis externa.

We studied the lipophilic microbiota of the external ear canals of 332 animals (264 dogs and 68 cats), with and without otitis externa, over an 11-year period from 1988 to 1999. Malassezia pachydermatis was isolated from 62.2% and 50% of dogs with and without otitis externa, respectively, and from 41.2% and 17.6% of cats with and without otitis externa, respectively. In the group of animals studied for lipid-dependent species, these yeasts were isolated from 4.5% of dogs with otitis externa and from 23.1% and 8.9% of cats with and without otitis externa, respectively. M. sympodialis and M. furfur were isolated from cats and M. furfur and M. obtusa from dogs. Our findings show that lipid-dependent Malassezia species may contribute to the etiology of otitis externa in dogs and cats.

Animals↗

An easy screening method for fungi producing ochratoxin A in pure culture.

A simple screening method has been developed for detecting ochratoxin production by fungi, based on high-performance liquid chromatographic determinations on extracts obtained from agar plugs cut from pure Petri dish cultures. Two culture media. Yeast Extract Sucrose agar and Czapek Yeast Extract agar, and three extraction solvents (methanol, methylene chloride/formic acid, and methanol/formic acid) were compared. All of the isolates tested produced ochratoxin A in one or both culture media after 7 or 14 days of incubation. Based on the results obtained, the use of both culture media is recommended. As extraction solvent, either methanol or methanol-formic acid could be used. This method also provides quantitative information on the level of ochratoxin produced by the cultures. The simplicity of the method makes it very useful when many fungal isolates need to be screened.

Cells, Cultured↗

Distribution of ochratoxin A producing strains in the A. niger aggregate.

Ochratoxin A (OA) production of 92 isolates belonging to the A. niger aggregate was tested. All these isolates were grouped into the two proposed species A. niger and A. tubingensis, according to their ITS-5.8S rDNA RFLP patterns. The distribution of the isolates into the two species was very similar since 52.2% were classified as pattern T (corresponding to A. tubingensis), and 47.8% were classified as pattern N (corresponding to A. niger). Six out of the 92 isolates studied produced OA. All the OA producing strains were classified as pattern N while none of the isolates classified as pattern T produced OA.

Animals↗

Current importance of ochratoxin A-producing Aspergillus spp.

Ochratoxin A (OA) is receiving attention worldwide because of the hazard it poses to human and animal health. OA contamination of commodities, such as cereals or pork and poultry meat, is well recognized. Nevertheless, there is an increasing number of articles reporting OA contamination in other food commodities, such as coffee, beer, wine, grape juice, and milk, in the last few years. This continuous and increasing exposure to OA that humans experience is reflected in the high incidence of OA in both human blood and milk in several countries. OA was believed to be produced only by Aspergillus ochraceus and closely related species of section Circumdati and by Penicillium verrucosum; however, in the genus Aspergillus, the production of OA has been recently reported by species outside the section Circumdati. Thus, it has been clearly established as a metabolite of different species of the section Nigri, such as Aspergillus niger and Aspergillus carbonarius. OA production ability by Aspergillus spp. is more widespread than previously thought; therefore, there is the possibility that unexpected species can be new sources of this mycotoxin in their natural substrates.

Animals↗