María Teresa Mujica

Interactions between Glomus mosseae and arbuscular mycorrhizal sporocarp-associated saprophytic fungi

The saprophytic fungi Wardomyces inflatus (Marchal) Hennebert, Paecilomyces farinosus (Holm & Gray) A. H. S. Brown & G. Sm., Gliocladium roseum Bain., sterile dark mycelium (SDM-54), Trichoderma pseudokoningii Rifai and Trichoderma harzianum Rifai were isolated from sporocarps of Glomus mosseae. The effect of saprophytic fungi on G. mosseae spore germination was tested on water agar. Wardomyces inflatus decreased the percent germination of G. mosseae spores; G. roseum, T. pseudokoningii and T. harzianum had no effect on germination; and P. farinosus and SDM-54 increased the percentage of spore germination of G. mosseae after 4 d. Wardomyces inflatus significantly decreased hyphal length of spores which germinated, but no other saprophytic fungi affected hyphal growth. Trichoderma pseudokoningii, T. harzianum, P. farinosus and SDM-54 increased the number of auxiliary cells formed by G. mosseae. The effect of saprophytic fungi on arbuscular mycorrhizal colonization of soybean was studied in a greenhouse trial. The percentage of soybean root length colonized was decreased by W. inflatus, unaffected by SDM-54 and T. harzianum, and increased by P. farinosus. Gliocladium roseum decreased root length colonized when plants were 12 wk old, and T. pseudokoningii increased colonization of roots when plants were 4 wk old. Antagonistic, synergistic and neutral actions of G. mosseae upon the saprophytic fungi were observed. The population of T. harzianum decreased and the populations of T. pseudokoningii and SDM-54 increased in the presence of G. mosseae. Our results indicate a complex interaction between G. mosseae and associated saprophytic fungi.

Phenotypic and genotypic characterization of Aspergillus lentulus and Aspergillus fumigatus isolates in a patient with probable invasive aspergillosis.

Characteristic morphological and phylogenetic analyses demonstrated the presence of Aspergillus fumigatus and Aspergillus lentulus as the aetiological agents in a case of probable invasive aspergillosis (IA). This is believed to be the first report of an A. lentulus strain isolated from a patient with probable IA in Argentina.

Phenotypic and genotypic identification of Candida dubliniensis from subgingival sites in immunocompetent subjects in Argentina.

INTRODUCTION It is generally recognized that Candida dubliniensis is commonly found in immunocompromised patients, such as those with advanced human immunodeficiency virus infection, at sites of periodontal disease. Since there are no data available for Argentina, the aim of this study was to determine the prevalence of and to identify C. dubliniensis in periodontal pockets from immunocompetent subjects living in Buenos Aires, Argentina, through a comparison of phenotypic and molecular assays. METHODS Yeasts recovered from subgingival plaque samples were studied for 180 immunocompetent non-smoking patients with periodontal disease. Yeasts were identified by conventional mycological methods and by specific polymerase chain reaction (PCR) assay. Fluconazole and voriconazole susceptibility studies were performed in keeping with the Clinical and Laboratory Standards Institute. RESULTS Among 76 yeasts isolated, C. dubliniensis comprised 10.5% (n = 8; 95% confidence interval 4.7-19.7), which corresponded to 4.4% of patients studied (8/180). C. albicans was the most frequently isolated species of yeast. A great majority of C. dubliniensis isolates was susceptible with only one isolate resistant to both antifungals. CONCLUSION Micromorphology on Staib agar was the phenotypic method that was most concordant with PCR and it was useful for selecting presumptive C. dubliniensis. This is the first report to use PCR to identify C. dubliniensis in subgingival fluid from immunocompetent individuals with periodontal disease in Argentina. On the basis of the findings presented here, we confirm that C. dubliniensis can colonize periodontal pockets of immunocompetent patients with periodontal disease.

Influence of soil impoverishment on the interaction between Glomus mosseae and saprobe fungi

Abstract The effect of the saprobe fungi Wardomyces inflatus (Marchal) Hennebert, Paecilomyces farinosus (Holm & Gray) A. H. S. Brown & G. Sm., Gliocladium roseum Bain., Trichoderma pseudokoningii Rifai and T. harzianum Rifai, isolated from sporocarps of Glomus mosseae, on arbuscular mycorrhizal (AM) colonisation and plant dry matter of soybean was studied in 2/3 and 1/5 diluted soils in a greenhouse trial. Soil dilution to 1/5 had no effect on shoot dry matter of soybean but decreased AM colonisation and root dry weight of plants. CFU of saprobe fungi, except T. harzianum, were higher in 1/5 than in 2/3 diluted soils. W. inflatus and Gliocladium roseum decreased the shoot dry weight of soybean plant when inoculated together with Glomus mosseae. The saprobe fungi P. farinosus and T. pseudokoningii increased the shoot dry weights of plants grown in 1/5 diluted soil. The shoot dry weight and AM colonisation in 1/5 diluted soil were also increased when T. harzianum was inoculated together with Glomus mosseae. Thus, saprobe fungi increased AM colonisation of soybean plants by indigenous endophytes. The AM colonisation of plants at both soil dilutions was increased by Glomus mosseae. The highest level of AM colonisation was observed when P. farinosus and T. pseudokoningii were inoculated together Glomus mosseae. The dilution of soils influenced the interaction between inoculated microorganisms and their effect on plant growth.

Saksenaea erythrospora infection following a serious sailing accident

Saksenaea erythrospora is a species of the order Mucorales recently described and reported as a cause of human mucormycosis. We report a case of S. erythrospora in a man involved in a serious sailing accident causing deep skin and soft tissue contamination with soil and water. Direct microscopic examination of the clinical sample with Giemsa stains showed hyaline and non-septate hyphae belonging to the order Mucorales. Fungal identification was performed by culture of biopsy material on SDA, and identification of species by floating an agar block containing the fungus in a nutritionally deficient medium consisting of sterile distilled water supplemented with 0.05 % yeast extract; and by sequencing the ITS region of the rDNA. This is the first report to our knowledge of infection with S. erythrospora in Argentina, confirming the presence of this fungus in this country.

A prospective observational study of vulvovagintis in pregnant women in Argentina, with special reference to candidiasis

To evaluate the frequency of yeast, bacteria or protozoa in pregnant women and to correlate the possible associations of these microorganisms and their relationships with vulvovaginitis (VV) and cervicitis. Vaginal specimens were collected and prepared for smears in microscope slides for the evaluation of yeast, Trichomonas vaginalis and bacteria. Samples were cultured in specific culture medium. Cervical specimens were used to investigate the presence of Neisseria gonorrhoeae, Chlamydia trachomatis, Ureaplasma spp. and Mycoplasma hominis. We enrolled 210 pregnant women, aged 10–42 years old. Of them, 38.1% were symptomatic. Symptoms were most prevalent in the second and third trimesters of pregnancy coincident with a major prevalence of microorganisms. In this study, 39.5% of pregnant women had normal microbial biota and symptoms of VV due to non‐infectious causes were observed (6.2%). The occurrence of vulvovaginal candidiasis was 25% and Candida albicans with a prevalence of 80.7% was the dominant species (P = 0.005) while non‐albicans Candida species and other yeast were more common in asymptomatic ones (P = 0.0038). The frequency of bacterial vaginosis, T. vaginalis, C. trachomatis and N. gonorrhoeae were 18.1%, 1.4, 1.4% and 0.5% respectively.

Prevalence of "Candida albicans", "Candida dubliniensis" and "Candida africana" in pregnant women suffering from vulvovaginal candidiasis in Argentina

BACKGROUND Vulvovaginal candidiasis (VVC) is a vulvovaginitis commonly diagnosed in gynecology care. In recent years, the taxonomy of the most important pathogenic Candida species, such as Candida albicans have undergone significant changes. AIMS This study examined the prevalence of C. albicans, Candida africana, and Candida dubliniensis in vaginal specimens from 210 pregnant women suffering from vulvovaginitis or having asymptomatic colonization. METHODS Phenotypic and molecular methods were used for the identification of the species. RESULTS During the studied period, 55 isolates of Candida or other yeasts were obtained from specimens collected from 52 patients suffering from vulvovaginitis (24.8%). C. albicans was the predominant Candida species in 42 isolates (80.7%), either alone or in combination with other species of the genus (5.7%, n=3). Additionally, nine isolates of C. albicans (50%) were obtained from asymptomatic patients (n=18). C. dubliniensis was the causative agent in 2 (3.8%) cases of VVC, and was also isolated in one asymptomatic patient. Molecular assays were carried out using specific PCR to amplify the ACT1-associated intron sequence of C. dubliniensis. The amplification of the HWP1 gene also correctly identified isolates of the species C. albicans and C. dubliniensis. No C. africana was isolated in this work. Some C. albicans isolates were either homozygous or heterozygous at the HWP1 locus. The distribution of heterozygous and homozygous C. albicans isolates at the HWP1 locus was very similar among patients suffering from VVC and asymptomatic patients (p=0.897). CONCLUSIONS The presence of C. albicans and C. dubliniensis, and the absence of C. africana in pregnant is noteworthy.

Genetic diversity of Histoplasma capsulatum strains isolated from Argentina based on nucleotide sequence variations in the internal transcribed spacer regions of rDNA.

The internal transcribed spacer (ITS) regions of rDNA genes of 49 Histoplasma capsulatum (48 from clinical samples and one from soil) isolates were examined. Nucleotide sequence heterogeneity within this region was useful for phylogenetic classification of H. capsulatum and species identification. Thus, in 45 of 49 isolates we observed higher percentages of identity in the nucleotide sequences of ITS regions when the isolates studied herein were compared with those reported in our country in the South America B clade. Phylogenetic analyses of rDNA sequences corresponding to the 537 bp of the ITS region obtained from H. capsulatum isolates assigned South America type B clade (45 isolates), North America type 1 and Asia clade (2 isolates each one). H. capsulatum strains isolated from soil and from patients living in Argentina (45 of 49) clustered together with the H. capsulatum isolates of the South America B clade. The high level of genetic similarity among our isolates suggests that almost one genetic population is present in the microenvironment. Isolates described as H. capsulatum var. capsulatum or var. farciminosum (2 isolates) did not form a monophyletic group and were found in the Asia clade. Subsequent studies are needed to properly identify these isolates.

Phenotypic and genotypic characterisation of Candida dubliniensis isolate in a patient with disseminated candidiasis

Analia Fernandez, Virginia M. Jewtuchowicz, Maria Celina Malzone, Gabriela Lopez-Daneri, Gladys Saa, Cristina A. Iovannitti, Marta Tokumoto and Maria T. Mujica Laboratorio Central Servicio de Microbiologı́a, Hospital Universitario Fundación Favaloro, Buenos Aires, Argentina, Unidad de Cuidados Intensivos Pediátricos, Hospital Universitario Fundación Favaloro, Buenos Aires, Argentina and Departamento de Microbiologı́a, Parasitologı́a e Inmunologı́a de la Facultad de Medicina de la Universidad de Buenos Aires, Buenos Aires, Argentina

Estudio de la variabilidad genética entre aislamientos clínicos de Candida albicans formadores de biopelículas

Resumen Las biopeliculas estan constituidas por microcolonias incluidas dentro de una matriz polimerica y representan una forma de crecimiento microbiano. Candida albicans puede colonizar las superficies de cateteres, protesis y epitelios formando biopeliculas que son resistentes a las drogas antifungicas. El objetivo de este trabajo fue la caracterizacion genotipica de aislamientos clinicos de C. albicans formadores de biopeliculas usando la tecnica RAPD (Random Amplified Polymorphic DNA) . Fueron estudiados 25 aislamientos clinicos de C. albicans de fauces, sangre, piel, unas, materia fecal, biopsia de esofago, y flujo vaginal de pacientes con candidiasis. Cada cepa fue previamente analizada en su capacidad de crecer y adherirse a la superficie de poliestireno, y la cuantificacion de la biopelicula formada fue realizada mediante el ensayo de reduccion de XTT 2,3-bis (2-metoxi-4 nitro-5 sulfofenil) -2 H tetrazolio-5 carboxanilida.. Los coeficientes de similitud generados por RAPD variaron entre el 49 y el 91% con los cuatro iniciadores usados, revelando un alto nivel de variabilidad genetica. El dendrograma agrupo los aislamientos en cuatro grupos, incluyendo todos ellos cepas con muy diferente capacidad para formar biopeliculas. Aislamientos con genotipos similares mostraron diferente capacidad de formacion de biopeliculas. Las cepas fueron agrupadas independientemente del origen de la muestra. Nuestros resultados sugieren que en poblaciones naturales de C. albicans no existe correlacion entre la capacidad de formar biopeliculas y el genotipo determinado por PCR-RAPD.