Detection of Candida dubliniensis in patients with candidiasis in Caracas, Venezuela

Candida species are responsible for 80% of all nosocomial fungal infections. In 1995 a new yeast species was described, Candida dubliniensis which shares with Candida albicans characteristics. We have studied 109 yeast isolates identified as C. albicans to investigate the presence of C. dubliniensis by microbiological studies and PCR using DUBR/DUBF primers. Positive results using microbiological tools were between 90 and 98%. Two morphological and physiological of the 80 DNA examined samples (2.5%) showed a PCR product of 288 bp which allow the identification of C. dubliniensis. This is the first report in Venezuela of identification of this species using a PCR approach.     

Admixture estimates for Caracas, Venezuela, based on autosomal, Y-chromosome, and mtDNA markers

The present Venezuelan population is the product of admixture of Amerindians, Europeans, and Africans, a process that was not homogeneous throughout the country. Blood groups, short tandem repeats (STRs), mtDNA, and Y-chromosome markers have been used successfully in admixture studies, but few such studies have been conducted in Venezuela. In this study we aim to estimate the admixture components of samples from two different socioeconomic levels from Caracas, Venezuela's capital city, compare their differences, and infer sexual asymmetry in the European Amerindian union patterns. Gene frequencies for blood groups ABO and Rh (CDE) and for the STRs VWA, F13A01, and FES/FPS and mtDNA and Y-chromosome haplogroups were studied in a sample of 60 individuals living in Caracas, taken from a private clinic (high socioeconomic level), and 50 individuals, also living in Caracas, drawn from a public maternity clinic (low socioeconomic level). The admixture analysis for the five autosomal markers gives a high European component (0.78) and an almost negligible African sub-Saharan component (0.06) for the high socioeconomic level, whereas for the low socioeconomic level the sub-Saharan, European, and Amerindian components were 0.21, 0.42, and 0.36, respectively. Estimates of admixture based on mtDNA and Y-chromosome markers reveal that the Amerindian contribution to these Caracas samples is almost entirely through females, because the Y-chromosome Amerindian and African sub-Saharan chromosomes found in this study were scarce. Our study reveals that the identification of the grandparents' geographic origin is an important methodological aspect to take into account in genetic studies related to the reconstruction of historical events.     

Plant reproductive systems in a secondary deciduous forest remnant in the Caracas valley,Venezuela

Plant Ecology - Plant sexual and breeding systems, temporal variations in sex expression, and herkogamy were investigated. Of the 294 plant species surveyed, 73.8% were hermaphroditic, 18.7%...     

Datos sobre Tinea capitis en Venezuela

El Dr.Borelli es Professor Asociado de la Fac. de Med., U.C.V., en las Cátedras de Dermatología (Dr.Carlos, J. Alarcón) y en la de Microbiología (Dr.Leopoldo Briceño I.). La Dra.Coretti es Primer Adjunto al Servicio de Medicina III, Hospital de Niños J. M. de los Ríos (Dr.Pedro J. Alvarez).Trabajo presentado en ocasión de las Primeras Jornadas Pediátricas, Caracas 18–24 Febrero 1962.     

Molecular Characterization of Sewage-Borne Pathogens and Detection of Sewage Markers in an Urban Stream in Caracas,Venezuela

Molecular characterization of two sewage-borne pathogens identified hepatitis A virus (HAV) subgenotype IA and Giardia duodenalis assemblages A and B as predominant genotypes circulating in an urban area of Venezuela. This study reveals epidemiological features of human pathogens of worldwide distribution and the efficacy of molecular methods for accurate assessment of sewage pollution.Multiple microbial pathogens may frequently be found in surface waters that receive uncontrolled municipal sewage discharges. The range and diversity of sewage-borne pathogens in surface waters are geographically specific and strongly dependent on the burden of infectious diseases in the population, the seasonal patterns of infectious diseases in the community, and the availability of sewage treatment processing (5, 7). The metropolitan city of Caracas, the capital and largest city of Venezuela, is located in northern South America near the Caribbean coast. Water pollution is a big issue in Caracas, like in many other cities in South America, where most of the human sewage (∼97%) from overpopulated urbanized areas is discharged without any treatment into nearby rivers and coastal environments. Despite these facts, the seriousness of sewage-related health issues is not at the forefront of public concern in this country.Giardia is the protozoan parasite most frequently detected in human fecal samples submitted to diagnostic laboratories from major cities in Venezuela. The frequency of giardiasis reported in the population varies between 21% and 45% but may increase up to 75% among school age children (8). Notwithstanding, the epidemiology of giardiasis in Venezuela remains unknown, and no previous studies have documented the distribution of species and genotype assemblages associated with human infections.Hepatitis A virus (HAV), the etiological agent of hepatitis A in humans, has distinguishable epidemiological patterns of distribution and endemicity closely related to socioeconomic development (9, 12). Water- and food-borne outbreaks of HAV have been well documented worldwide (6, 19). Seroepidemiological studies conducted in selected populations in Venezuela have demonstrated high endemicity of hepatitis A infection among low socioeconomic population strata, with seroprevalences between 48% and 98% (13). Nevertheless, studies on HAV genotype circulation in major urban areas of the country are scarce, as is research on predominant exposure routes and potential transmission patterns through the environment.The analysis of nucleic acid sequences of sewage-borne pathogens may provide relevant information on predominant species and genotypes of human-pathogenic viruses and parasites circulating in specific geographical areas (11, 12, 15). The molecular approach may be of relevance for countries lacking reliable disease surveillance programs and proper understanding of the potential transmission of specific human pathogens through the environment. In this research, Giardia cysts and HAV recovered from an urban stream were characterized by multiple molecular methods along with nucleotide sequence analysis to identify predominant genotypes circulating in a major urban area of Venezuela''s capital. The strength and efficacy of multiple molecular methods for accurate assessment of human sewage pollution and risks of exposure to sewage-borne pathogens were also investigated.Dry season sampling (October through March) was conducted in a heavily polluted urban stream (>10⁶ fecal coliforms/100 ml) that flows in a southeast direction through the metropolitan city of Caracas (16). Water sample volumes of 100 ml were collected in three sterile centrifuge tubes two to four times per month. Giardia cysts were concentrated by centrifugation (100 ml at 1,500 × g for 15 min) followed by DNA extraction by the freeze-thaw method in the presence of Chelex-100 (3) for sucrose-purified cysts. Human-pathogenic assemblage occurrence was determined by nested PCR amplification and sequence analysis of the triosephosphate isomerase (tpi) gene (18). Multiple sequence alignments were performed with ClustalW (21), and phylogenetic analyses were conducted using MEGA4 software (20). The genetic diversity of Giardia isolates was inferred by the neighbor-joining method (17) using a bootstrap test of 1,000 replicates. Giardia cysts counts were obtained by fluorescence microscopy using BTF EasyStain monoclonal antibody stain (BTF Precise Microbiology, Inc.) and 4′6-diamidino-2-phenylindole (DAPI). The recovery efficiency of cysts was determined in five experiments using ColorSeed C&G spike suspensions as internal quality controls (14).HAV particles were concentrated from 35 ml by ultracentrifugation and elution with 0.25 N glycine buffer following procedures previously described (16). Viral RNA was extracted from sample concentrates with Trizol (Invitrogen, Inc., Carlsbad, CA) following the manufacturer''s instructions. General detection of HAV was based on amplification of the 5′ nontranslated region (5′ NTR), while analysis of genetic diversity involved sequencing and phylogenetic analysis of the VP1 amino terminus and the full VP1 gene (2, 10). Sequence alignment was conducted with the DNAman software 5.2.2 (Lynnon BioSoft, Quebec, Canada) followed by phylogenetic analysis.Molecular detection of sewage pollution was accomplished by PCR amplification of Bacteroidales human-specific 16S rRNA genes, Bacteroides thetaiotamicron 16S rRNA genes, and the nifH gene of Methanobrevibacter smithii using primers and PCR conditions originally described by Field et al. (4), Carson et al. (1), and Ufnar et al. (22), respectively.Giardia duodenalis tpi nucleotide sequences amplified directly from urban stream waters were included after phylogenetic analysis into two well-defined clusters of assemblages A and B. These results were supported by high bootstrap values, as indicated in Fig. ​Fig.1.1. The level of cysts recovered from these samples ranged from 10,400 to 62,000 cysts/liter; however, mean percent recoveries varied from 20% to 50%, which suggests that the urban stream may harbor and receive much higher loads of cysts.Open in a separate windowFIG. 1.Phylogenetic tree of G. duodenalis assemblages A (VPW2, VPW3, and VPW7) and B (VPW1, VPW4, VPW5, and VPW6) from urban stream samples forming two clusters in a neighbor-joining analysis of tpi nucleotide sequences. Only bootstrap values >80% are shown in the tree.Three genomic regions used for detection and characterization of HAV revealed the predominance of HAV strains belonging to subgenotype IA, the most frequent genotype associated with human disease worldwide (9). A neighbor-joining tree constructed from the alignment of nucleotide sequences from urban stream samples and sequences of HAV strains from case patients (unpublished data) was used to investigate the relationship between genotypes present in environmental and clinical samples. The comparative analysis indicated a high degree of identity (98 to 99%) between nucleotide sequences from the urban stream and the strains from sporadic HAV cases. The phylogenetic analysis grouped all of these sequences into two unique clades within subgenotype IA, strongly supported by significant bootstrap values (Fig. 2A, B, and C).Open in a separate windowFIG. 2.Phylogenetic analysis of the HAV 5′ NTR (A; 284 nucleotides [nt]), VP1 amino terminus (B; 172 nt), and complete VP1 (C; 820 nt) regions. Nucleotide sequences of HAV reference strains are designated by their GenBank accession number, including the name of the country of origin, except for Venezuelan isolates, which are shown in bold. S, isolates derived from human sporadic cases; W, urban stream isolates. Phylogenetic analysis was performed by neighbor joining, and phylogenetic distances were calculated by the Kimura two-parameter test. Bootstrap values ≥90% are shown in the trees. Letters in bold indicate the subtype.Three reliable published assays for detection of human-specific markers of fecal pollution identified and confirmed the predominant point source of water pollution. Sequence analysis of three randomly selected PCR products from each marker revealed ≥99% sequence identity with published sequences (GenBank) derived from different geographical areas, thus indicating the validity and specificity of the molecular markers as reliable indicators of human sewage pollution in Venezuela.The results of this research demonstrate that the molecular assays applied for detection and characterization of sewage-borne pathogens in surface waters may have practical applications for epidemiological investigations on distribution of predominant human-specific genotypes circulating in urban populations. Previous studies identified the most predominant waterborne gastroenteritis viruses circulating in Metropolitan Caracas (16).The molecular-based monitoring approach for rapid and precise identification of sewage-borne pathogens and sewage markers in surface waters has important implications for sewage-related health issues that require special attention in Venezuela and South America. Deficient sewerage coverage and lack of municipal wastewater treatment, commonly associated with informal settlements around densely populated urban areas, are responsible for many of the environmental degradation and public health problems that occur in these countries. The precise identification of human pathogens in the environment offers an appropriate and alternative approach for initial assessment of risks of exposure to waterborne pathogens. Current bacterial indicators of fecal pollution (fecal coliforms, Escherichia coli, and enterococci) do not allow identification of the relative sources of impacts (i.e., sewage, urban runoff, and agricultural waste) on surface waters. Thus, the molecular detection of sewage-borne pathogens and sewage markers in surface waters may be more effective than the bacterial indicator approach for forecasting pathogen distribution and for managing and reducing risks associated with inappropriate sewage disposal into natural waters in Venezuela and South America.     

Airborne dispersal ofCecropia peltata L. (Moraceae) pollen,a neotropical species,in the vicinity of Caracas,Venezuela

In this paper, airborne dispersion of the pollen ofCecropia peltata L. (Moraceae) in a location close to the Equator is described. The Cecropias are fast-growing pioneer trees unique to the Neotropics, where they grow within a large altitude range, from Mexico to Brazil.Cecropia pollen was the most abundant grain throughout the year. The highest recovery occurred during April–May. As widely distributed members of the Moraceae family and because of their small pollen size, the Cecropias are prime suspects for being the source of inhalant allergens. The numbers ofCecropia airborne pollen grains that were recorded are well above those deemed necessary for human sensitization.     

habitat de Nocardia asteroides,Phialophora pedrosoi y Cryptococcus neoformans en Venezuela

Resumen Se estudiaron 336 muestras de tierra, madera y plantas provenientes de diferentes zonas del Edo. Mérida/Venezuela y Estados vecinos. Fueron inoculados cobayos y hamsters intratesticularmente y ratones por via endovenosa. Testículos y pulmones respectivamente fueron examinados por medio de cultivos e histológicamente.De 11 muestras se aislaronNocardia asteroides, de 6Phialophora pedrosoi y de unaCryptococcus neoformans.Se discuten las probables causas de porqué no se logró aislarParacoccidioides brasiliensis.

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Summary 336 samples of soil, wood and plants from several aereas of the State of Mérida/Venezuela and adjacent regions were studied.The samples were injected into the testes of guinea pigs and into mice intravenously. The testes and lungs of the animals respectively were cultured and examined histologically. From eleven samples were isolatedNocardia asteroides, from sixPhialophora pedrosoi and from oneCryptococcus neoformans.It is discussed, why it was not possible to isolateParacoccidioides brasiliensis.

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Zusammenfassung Es wurden 336 Erd-, Holz- und Pflanzenproben aus verschiedenen Gebieten des Staates Mérida/Venezuela und benachbarten Zonen untersucht. Das Material wurde Meerschweinchen intratestikulär und weissen Mäusen intravenös injiziert. Hoden und Lungen der Tiere wurden auf Nährböden verbracht und histologisch untersucht.Elfmal wurdeNocardia asteroides, sechsmalPhialophora pedrosoi und einmalCryptococcus neoformans isoliert und histologisch nachgewiesen. Es werden die möglichen Gründe diskutiert, warum es nicht gelangParacoccidioides brasiliensis zu isolieren.

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Del Instituto de Anatomía Patológica y Laboratorio de Micología de la Universidad de Los Andes, Mérida/Venezuela y del Laboratorio de Micología, Escuela de Medicina de la Universidad de Cincinnati/Ohio EEUU.

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Este trabajo pudo realizarse con la ayuda económica proporcionada por el Sr. Rector de la U.L.A., Dr.P. Rincón Gutiérrez a quien damos nuestras gracias más sinceras.A Wm. Bridge Cooke, Ph. D. y al Dr.G. A. de Vries agradecemos la identificación de las cepas dePhialophora pedrosoi.

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A los Dres.R. Kohler yP. N. Tablante G., y al Sr.Oswaldo Juergenson nuestro agradecimiento por la ayuda prestada en la recolección de las muestras.