Biomedical subjects
Publications and source records attributed to M L Abarca.
Mycotoxins and the fungi that produce them are of increasing importance as causes of human illness, but the diseases produced remain poorly understood at the clinical level. This paper explores four aspects: the increase of interest in ochratoxin A, factors affecting mycotoxin production, toxicology of the major mycotoxins, and the identification of Penicillium species which cause food spoilage and are important in indoor air.
Although Aspergillus fumigatus is the most common etiological agent of invasive aspergillosis, other Aspergillus spp. such as A . flavus, A. niger, A. terreus and A. nidulans (Emericella nidulans) among others, have been also implicated. In this article, the taxonomy of the genus Aspergillus and the characteristics of the species most frequently isolated from patients with nosocomial aspergillosis are reviewed.
Mycotoxins are relatively small molecules characterized by a diversity of chemical structure and a diversity of biological activity. They are often genotypically specific for a group of species, but the same compound can also be formed by fungi belonging to different genera. Most of the mycotoxins known have been recognized as metabolic products of Aspergillus, Penicillium and Fusarium species. This review will be focused on aflatoxins, ochratoxins and fumonisins because of their hazard to animal and human health. The production of these mycotoxins have been usually associated with a small number of species but some recent studies have reported the production of these mycotoxins by some other species. These results show that mycotoxin production is broader than is normally thought, so the possibility can not be ruled out that new species may be a new source of unexpected mycotoxins in their natural substrates.
The DNA that encodes the 5.8S gene of the ribosomal RNA and the two intergenic spacers ITS1 and ITS2 of the two proposed type strains of the Aspergillus niger aggregate (A. niger and Aspergillus tubingensis) have been sequenced. By comparison of sequences we have found that both species could be differentiated by RsaI digestion of the PCR products of the mentioned regions. This method could be a useful tool in the identification of strains of the A. niger aggregate, especially in studies that involve a large number of isolates.
During a survey of the occurrence of Malassezia species in the external ear canals of cats without otitis externa, Malassezia furfur was isolated. This is the first report of the isolation of M. furfur from cats.
A 10-month-old male American Staffordshire terrier was presented to the Autonomous University of Barcelona Veterinary Teaching Hospital because of a 6-month history of a mucopurulent bilateral nasal discharge. The dog had not responded to antibiotics. A follow-up X ray revealed a mixed pattern of osteolysis and increased radiodensity confined to the nasal cavity. Histologic sections of the biopsy specimens revealed the presence of granules containing numerous septate hyphae that were hyaline to pale brown and smooth, one-celled, subspherical-to-elongate conidia that were hyaline to brownish green, and bacteria. Cultures yielded numerous colonies belonging to Scedosporium apiospermum. Susceptibility tests were performed on the isolated strain. The isolate was sensitive to ketoconazole, intermediate to clotrimazole, and resistant to amphotericin B, 5-fluorocytosine, fluconazole, and itraconazole. The dog was treated with oral ketoconazole. During the treatment a general improvement in the lesions was observed. To our knowledge, S. apiospermum has not been implicated previously as an etiologic agent of nasal disease in dogs. This report provides its first description as such.
This retrospective study deals with the main samples studied at the Mycology Diagnostic Service of the Faculty of Veterinary Science of Barcelona: animals with suspected dermatophytosis. Over a ten-year period from 1986 to 1995, 136 dermatophytes were identified from dog and cat cultures submitted for identification and from specimens submitted for mycological examination from a variety of other domestic animals. The most frequent dermatophytes isolated were Microsporum canis (55.9%), Trichophyton mentagrophytes var. mentagrophytes (27.2%), Microsporum gypseum (7.4%) and Trichophyton verrucosum (7.4%). The identity of the dermatophytes from dog and cat cultures submitted for identification was M. canis (77.8%), T. mentagrophytes (13.3%) and M. gypseum (8.9%). Dermatophytes were cultured from 15 of 105 (14.3%) canine specimens and 19 of 56 (33.9%) feline specimens submitted for mycological examination during this period. Microsporum canis was the most common species isolated (73.3% and 94.7% respectively). The percentage of positive microscopic examinations of the specimens of hair in culture positive submissions from dogs and cats was 58.8%. There was a high proportion of positive cultures from both dogs and cats less than 1 year of age, and in some breeds of dogs, but there was no significant difference between the sexes. Although dermatophytes were more frequently isolated in autumn and winter months, no significant difference was detected in the seasonal distribution of the canine and feline dermatophytosis. Trichophyton mentagrophytes was the most prevalent dermatophyte in rabbits and T. verrucosum in ruminants. Other isolated species were T. equinum and M. equinum from horses.
During a one year period, 944 dogs from the Municipal kennel of Barcelona were examined to detect animals with suspected dermatophytosis. Only a few animals (1.8%) presented skin lesions but none of them had dermatophytosis. A representative number of dogs without visible skin lesions (n = 172), selected at random, were used to carry out a seasonal study of the mycobiota of their fur. Fifteen isolates belonging to the genera Microsporum and Trichophyton were isolated from 14 of the 172 (8.1%) dogs without lesions. The identity of these fungi was Microsporum gypseum (6/15), Trichophyton terrestre (4/15), M. canis (2/15), M. cookei (2/15) and Trichophyton ajelloi (1/15) (one strain each of M. gypseum and T. ajelloi were isolated from one dog). Species of Penicillium (% prevalence = 89.5%), Alternaria (86.6%), Cladosporium (84.9%), Aspergillus (77.3%), Scopulariopsis (65.7%) and Chrysosporium (64.5%) were the most prevalent. No significant differences in the fungal biota were observed with respect to age, gender, hair length or between mixed and pure breed dogs. A large number of isolates, including species belonging to the genera Beauveria, Chrysosporium, Malbranchea and Scopulariopsis, that macroscopically and/or microscopically resemble dermatophytes and may be mistaken for them, produced a red color change in Dermatophyte Test Medium. No significant seasonal difference was detected among the isolates belonging to the most frequently encountered genera, with the exception of Scopulariopsis (higher in summer and autumn) and Chrysosporium (higher in summer). Species from other genera, with lower occurrence also presented significant differences in their seasonal distribution. Arthrinium, Aureobasidium, Chaetomium and Phoma spp. presented maximum prevalence peaks in spring, Fusarium, Paecilomyces, Phoma and Rhizopus spp. in summer and Geotrichum and Mucor spp. in autumn. The Microsporum and Trichophyton species were more frequently isolated in summer.
The effects of three dyes on the colony enumeration of nine fungal strains (including members of the Deuteromycetes and Zygomycetes) in pure and mixed cultures were investigated. Using malt extract agar as basal and control medium, the following dyes and concentrations were assayed: auramine (25 ppm), gentian violet (5 ppm) and malachite green (1 ppm). The chemicals commonly used in commercial media dichloran (2 ppm) and rose bengal (50 ppm) were included in the study as reference mould-spreading inhibitors. Higher counts were usually obtained in the media containing dichloran, rose bengal or auramine, including the control medium in the absence of chemical when the mixed-conidium inocula did not include a spreading mould. Nevertheless in most cases no significant differences were observed between them. Malachite green (1 ppm) performed mainly as a strong inhibitor of spreading moulds, only allowing adequate colony development and recoveries of both Fusarium and Aspergillus strains tested.
In a survey of the occurrence of ochratoxin A (OA)-positive strains isolated from feedstuffs, two of the 19 isolates of Aspergillus niger var. niger that were studied produced OA in 2% yeast extract-15% sucrose broth and in corn cultures. This is the first report of production of OA by this species.
The ability of strains of genus Epidermophyton to grow at different concentrations of NaCl (ranged from 0 to 10%) was studied. A reduction in the diameter of colonies was observed as the concentration of NaCl increased. A nearly complete absence of macroconidia were shown even at the lowest concentration assayed. The geophilic species E. stockdaleae tolerated higher concentrations of NaCl (> 7%) than E. floccosum (< 3%).
One hundred twenty-one paraffin-embedded cervical biopsy specimens were tested for the presence of human papillomavirus (HPV) DNA by in situ hybridization and polymerase chain reaction. By in situ hybridization using probes for HPV types 6/11, 16/18, 31/33/35, 42/43/44, 51/52, and 45/56, HPV DNA was found in none of 20 normal/squamous metaplasia biopsy specimens, in one of 76 HPV equivocal biopsy specimens, in seven of 12 condyloma/mild dysplasia biopsy specimens, and in 12 of 13 moderate/severe dysplasia biopsy specimens. Polymerase chain reaction using HPV L1 consensus sequence primers followed by filter hybridization of the amplification products was positive for HPV DNA in two of 20 normal/squamous metaplasia biopsy specimens, in 23 of 76 HPV equivocal biopsy specimens, in eight of 12 condyloma/mild dysplasia biopsy specimens, and in 12 of 13 moderate/severe dysplasia biopsy specimens. Among biopsies that tested positive by polymerase chain reaction but that were negative by in situ hybridization, the most commonly identified HPV was type 16. We conclude that although HPV equivocal biopsy specimens contain HPV DNA more frequently than histologically normal tissue, the majority of biopsy specimens in this category test negative for HPV DNA. The clinical significance of a positive test for HPV, in the absence of unequivocal histologic changes, remains to be determined.
The influence of dilution plating technique, nature of diluent, culture media and incubation period on the enumeration of moulds have been studied. Three new culture media containing Auramine, Gentian Violet and Malachite Green respectively have been induced in this study. No significant differences were observed between results obtained after 3, 5 and 7 days of incubation. Significantly higher recoveries were obtained using the surface-spread method than pour plate method. Using the first technique no effect of diluent was observed, and among the different culture media studied higher counts were obtained with medium containing Auramine.
The effects of a wide range of concentrations of 13 dyes on the colony diameters of nine fungal strains (including members of the Deuteromycetes and Zygomycetes) were evaluated. Auramine at a concentration of 50 ppm (50 micrograms/ml), methylene blue at a concentration of 500 ppm, gentian violet at a concentration of 5 ppm, and phenol red at a concentration of 50 ppm performed as well as the commonly used dyes dichloran at a concentration of 2 ppm and rose bengal at a concentration of 50 ppm in that they allowed adequate colony development of the Deuteromycetes strains tested and controlled rapidly spreading fungi.
The occurrence of saprophytic fungi on hair and feathers samples taken from apparently healthy domestic animals (cows, pigs, rabbits, and chickens) has been studied. A total of 221 strains classified in 24 genera were isolated. The most frequent genera were Aspergillus, Penicillium, Mucor, Absidia and Alternaria, and some statistically significant differences of contamination were detected among the animal species.
Thirty seven strains of the Aspergillus flavus group isolated from animal mixed feeds have been screened for their ability to produce aflatoxins in yeast extract and sucrose (YES), aflatoxin producing ability (APA), and coconut agar medium (CAM) media. The concentration and detection of the aflatoxins by different methods is compared. Five known aflatoxin-positive and one aflatoxin-negative strains have been used as controls. Only 5 out of the 37 strains (13.5%) were aflatoxin-producers in YES medium. Of these five strains and the five known aflatoxin-positive strains, only three showed blue fluorescence in APA medium and four in CAM medium. Generally, the aflatoxin concentration in CAM medium was higher than in YES and APA media. Using the 'agar-plug method' and by direct spotting of the YES broth on TLC plates, some aflatoxin-producing strains were not detected.
A 5-year-old female Siamese cat was presented to the veterinary teaching hospital with a history of bronchopneumonia for 20 days. The cat had not responded to antibacterial chemotherapy and had developed a pronounced submandibular lymphadenopathy. Characteristically encapsulated yeast cells with narrow-necked buds were clearly seen in a fine-needle aspirate of the lymph node with an India ink preparation. Cryptococcus neoformans var. neoformans was identified. Susceptibility tests on the isolated strain were performed using antifungal tablets. The strain was sensitive to amphotericin B, fluconazole, itraconazole and ketoconazole and was resistant to 5-fluorocytosine. The cat was positive for feline immunodeficiency virus. Nevertheless, the cat was treated with ketoconazole for 3 months and apparently recovered. Three months later the animal was presented in a precomatose state. The owners refused to treat the animal and the cat was destroyed.