Choanoflagellates alongside diverse uncultured predatory protists consume the abundant open-ocean cyanobacterium Prochlorococcus.

Prochlorococcus is a key member of open-ocean primary producer communities. Despite its importance, little is known about the predators that consume this cyanobacterium and make its biomass available to higher trophic levels. We identify potential predators along a gradient wherein Prochlorococcus abundance increased from near detection limits (coastal California) to >200,000 cells mL-1 (subtropical North Pacific Gyre). A replicated RNA-Stable Isotope Probing experiment involving the in situ community, and labeled Prochlorococcus as prey, revealed choanoflagellates as the most active predators of Prochlorococcus, alongside a radiolarian, chrysophytes, dictyochophytes, and specific MAST lineages. These predators were not appropriately highlighted in multiyear conventional 18S rRNA gene amplicon surveys where dinoflagellates and other taxa had highest relative amplicon abundances across the gradient. In identifying direct consumers of Prochlorococcus, we reveal food-web linkages of individual protistan taxa and resolve routes of carbon transfer from the base of marine food webs.

The Bay of Bengal exposes abundant photosynthetic picoplankton and newfound diversity along salinity-driven gradients.

The Bay of Bengal (BoB) is a 2,600,000 km2 expanse in the Indian Ocean upon which many humans rely. However, the primary producers underpinning food chains here remain poorly characterized. We examined phytoplankton abundance and diversity along strong BoB latitudinal and vertical salinity gradients-which have low temperature variation (27-29°C) between the surface and subsurface chlorophyll maximum (SCM). In surface waters, Prochlorococcus averaged 11.7 ± 4.4 × 104 cells ml-1 , predominantly HLII, whereas LLII and 'rare' ecotypes, HLVI and LLVII, dominated in the SCM. Synechococcus averaged 8.4 ± 2.3 × 104 cells ml-1 in the surface, declined rapidly with depth, and population structure of dominant Clade II differed between surface and SCM; Clade X was notable at both depths. Across all sites, Ostreococcus Clade OII dominated SCM eukaryotes whereas communities differentiated strongly moving from Arabian Sea-influenced high salinity (southerly; prasinophytes) to freshwater-influenced low salinity (northerly; stramenopiles, specifically, diatoms, pelagophytes, and dictyochophytes, plus the prasinophyte Micromonas) surface waters. Eukaryotic phytoplankton peaked in the south (1.9 × 104 cells ml-1 , surface) where a novel Ostreococcus was revealed, named here Ostreococcus bengalensis. We expose dominance of a single picoeukaryote and hitherto 'rare' picocyanobacteria at depth in this complex ecosystem where studies suggest picoplankton are replacing larger phytoplankton due to climate change.

No evidence of Phago-mixotropy in Micromonas polaris (Mamiellophyceae), the Dominant Picophytoplankton Species in the Arctic.

In the Arctic Ocean, the small green alga Micromonas polaris dominates picophytoplankton during the summer months but is also present in winter. It has been previously hypothesized to be phago-mixotrophic (capable of bacteria ingestion) based on laboratory and field experiments. Prey uptake was analyzed in several M. polaris strains isolated from different regions and depths of the Arctic Ocean and in Ochromonas triangulata, a known phago-mixotroph used as a control. Measuring ingestion of either fluorescent beads or fluorescently labeled bacteria by flow cytometry, we found no evidence of phago-mixotrophy in any M. polaris strain while O. triangulata was ingesting both beads and bacteria. In addition, in silico predictions revealed that members of the genus Micromonas lack a genetic signature of phagocytotic capacity.

Light-driven synchrony of Prochlorococcus growth and mortality in the subtropical Pacific gyre.

Theoretical studies predict that competition for limited resources reduces biodiversity to the point of ecological instability, whereas strong predator/prey interactions enhance the number of coexisting species and limit fluctuations in abundances. In open ocean ecosystems, competition for low availability of essential nutrients results in relatively few abundant microbial species. The remarkable stability in overall cell abundance of the dominant photosynthetic cyanobacterium Prochlorococcus is assumed to reflect a simple food web structure strongly controlled by grazers and/or viruses. This hypothesized link between stability and ecological interactions, however, has been difficult to test with open ocean microbes because sampling methods commonly have poor temporal and spatial resolution. Here we use continuous techniques on two different winter-time cruises to show that Prochlorococcus cell production and mortality rates are tightly synchronized to the day/night cycle across the subtropical Pacific Ocean. In warmer waters, we observed harmonic oscillations in cell production and mortality rates, with a peak in mortality rate consistently occurring ∼6 h after the peak in cell production. Essentially no cell mortality was observed during daylight. Our results are best explained as a synchronized two-component trophic interaction with the per-capita rates of Prochlorococcus consumption driven either directly by the day/night cycle or indirectly by Prochlorococcus cell production. Light-driven synchrony of food web dynamics in which most of the newly produced Prochlorococcus cells are consumed each night likely enforces ecosystem stability across vast expanses of the open ocean.

Marine algae and land plants share conserved phytochrome signaling systems.

Phytochrome photosensors control a vast gene network in streptophyte plants, acting as master regulators of diverse growth and developmental processes throughout the life cycle. In contrast with their absence in known chlorophyte algal genomes and most sequenced prasinophyte algal genomes, a phytochrome is found in Micromonas pusilla, a widely distributed marine picoprasinophyte (<2 µm cell diameter). Together with phytochromes identified from other prasinophyte lineages, we establish that prasinophyte and streptophyte phytochromes share core light-input and signaling-output domain architectures except for the loss of C-terminal response regulator receiver domains in the streptophyte phytochrome lineage. Phylogenetic reconstructions robustly support the presence of phytochrome in the common progenitor of green algae and land plants. These analyses reveal a monophyletic clade containing streptophyte, prasinophyte, cryptophyte, and glaucophyte phytochromes implying an origin in the eukaryotic ancestor of the Archaeplastida. Transcriptomic measurements reveal diurnal regulation of phytochrome and bilin chromophore biosynthetic genes in Micromonas. Expression of these genes precedes both light-mediated phytochrome redistribution from the cytoplasm to the nucleus and increased expression of photosynthesis-associated genes. Prasinophyte phytochromes perceive wavelengths of light transmitted farther through seawater than the red/far-red light sensed by land plant phytochromes. Prasinophyte phytochromes also retain light-regulated histidine kinase activity lost in the streptophyte phytochrome lineage. Our studies demonstrate that light-mediated nuclear translocation of phytochrome predates the emergence of land plants and likely represents a widespread signaling mechanism in unicellular algae.

Abundance and Biogeography of Picoprasinophyte Ecotypes and Other Phytoplankton in the Eastern North Pacific Ocean.

Eukaryotic algae within the picoplankton size class (≤2 µm in diameter) are important marine primary producers, but their spatial and ecological distributions are not well characterized. Here, we studied three picoeukaryotic prasinophyte genera and their cyanobacterial counterparts, Prochlorococcus and Synechococcus, during two cruises along a North Pacific transect characterized by different ecological regimes. Picoeukaryotes and Synechococcus reached maximum abundances of 1.44 × 10(5) and 3.37 × 10(5) cells · ml(-1), respectively, in mesotrophic waters, while Prochlorococcus reached 1.95 × 10(5) cells · ml(-1) in the oligotrophic ocean. Of the picoeukaryotes, Bathycoccus was present at all stations in both cruises, reaching 21,368 ± 327 18S rRNA gene copies · ml(-1). Micromonas and Ostreococcus clade OI were detected only in mesotrophic and coastal waters and Ostreococcus clade OII only in the oligotrophic ocean. To resolve proposed Bathycoccus ecotypes, we established genetic distances for 1,104 marker genes using targeted metagenomes and the Bathycoccus prasinos genome. The analysis was anchored in comparative genome analysis of three Ostreococcus species for which physiological and environmental data are available to facilitate data interpretation. We established that two Bathycoccus ecotypes exist, named here BI (represented by coastal isolate Bathycoccus prasinos) and BII. These share 82% ± 6% nucleotide identity across homologs, while the Ostreococcus spp. share 75% ± 8%. We developed and applied an analysis of ecomarkers to metatranscriptomes sequenced here and published -omics data from the same region. The results indicated that the Bathycoccus ecotypes cooccur more often than Ostreococcus clades OI and OII do. Exploratory analyses of relative transcript abundances suggest that Bathycoccus NRT2.1 and AMT2.2 are high-affinity NO3 (-) and low-affinity NH4 (+) transporters, respectively, with close homologs in multiple picoprasinophytes. Additionally, in the open ocean, where dissolved iron concentrations were low (0.08 nM), there appeared to be a shift to the use of nickel superoxide dismutases (SODs) from Mn/Fe/Cu SODs closer inshore. Our study documents the distribution of picophytoplankton along a North Pacific ecological gradient and offers new concepts and techniques for investigating their biogeography.

Psychological distress and adherence to anti-retroviral therapy or pre-exposure prophylaxis regimens among Urban Black gay and bisexual men (MSM).

BACKGROUND: Urban Black gay, and bisexual men (MSM) bear a disproportionate burden of HIV in the U.S. Mental health is a barrier to adherence to both antiretroviral therapy (ART) and pre-exposure prophylaxis (PrEP). The objective was to determine the association between psychological distress and ART or PrEP adherence among urban Black MSM. METHODS: Using data from a four-year prospective cohort study, adherence to ART was defined as > 95% and PrEP was defined as > 80% of doses taken in the past 30 days. Psychological distress measures included difficulty sleeping; feeling anxious; suicidality; feeling sad or depressed; feeling sick, ill, or not well in the past 3 months; high (vs. low) overall psychological distress was classified as above the median value. Associations were examined using Chi-square, Fisher's exact tests, and logistic regression. RESULTS: Among 165 Black MSM, 44.2% (73) reported high psychological distress. 65.3% (47/72) of participants living with HIV and 39.8% (37/93) of HIV negative participants were ART or PrEP adherent, respectively. Education was significantly associated with PrEP adherence (p = 0.038). Non-injection drug use in the past 3 months (p = 0.008), difficulty sleeping (p = 0.010), feeling anxious (p = 0.003), and feeling sad or depressed (p < 0.001), and overall psychological distress (p < 0.001) were significantly associated with ART adherence. High psychological distress was significantly associated with a reduced odds of ART adherence (aOR 0.23; 95% CI = 0.08-0.70) adjusting for age and non-injection drug use. CONCLUSIONS: Increased psychological distress was significantly associated with ART nonadherence and may represent an important barrier to viral suppression.

Seasonal and Geographical Transitions in Eukaryotic Phytoplankton Community Structure in the Atlantic and Pacific Oceans.

Much is known about how broad eukaryotic phytoplankton groups vary according to nutrient availability in marine ecosystems. However, genus- and species-level dynamics are generally unknown, although important given that adaptation and acclimation processes differentiate at these levels. We examined phytoplankton communities across seasonal cycles in the North Atlantic (BATS) and under different trophic conditions in the eastern North Pacific (ENP), using phylogenetic classification of plastid-encoded 16S rRNA amplicon sequence variants (ASVs) and other methodologies, including flow cytometric cell sorting. Prasinophytes dominated eukaryotic phytoplankton amplicons during the nutrient-rich deep-mixing winter period at BATS. During stratification ('summer') uncultured dictyochophytes formed ∼35 ± 10% of all surface plastid amplicons and dominated those from stramenopile algae, whereas diatoms showed only minor, ephemeral contributions over the entire year. Uncultured dictyochophytes also comprised a major fraction of plastid amplicons in the oligotrophic ENP. Phylogenetic reconstructions of near-full length 16S rRNA sequences established 11 uncultured Dictyochophyte Environmental Clades (DEC). DEC-I and DEC-VI dominated surface dictyochophytes under stratification at BATS and in the ENP, and DEC-IV was also important in the latter. Additionally, although less common at BATS, Florenciella-related clades (FC) were prominent at depth in the ENP. In both ecosystems, pelagophytes contributed notably at depth, with PEC-VIII (Pelagophyte Environmental Clade) and (cultured) Pelagomonas calceolata being most important. Q-PCR confirmed the near absence of P. calceolata at the surface of the same oligotrophic sites where it reached ∼1,500 18S rRNA gene copies ml-1 at the DCM. To further characterize phytoplankton present in our samples, we performed staining and at-sea single-cell sorting experiments. Sequencing results from these indicated several uncultured dictyochophyte clades are comprised of predatory mixotrophs. From an evolutionary perspective, these cells showed both conserved and unique features in the chloroplast genome. In ENP metatranscriptomes we observed high expression of multiple chloroplast genes as well as expression of a selfish element (group II intron) in the psaA gene. Comparative analyses across the Pacific and Atlantic sites support the conclusion that predatory dictyochophytes thrive under low nutrient conditions. The observations that several uncultured dictyochophyte lineages are seemingly capable of photosynthesis and predation, raises questions about potential shifts in phytoplankton trophic roles associated with seasonality and long-term ocean change.

Targeted metagenomic recovery of four divergent viruses reveals shared and distinctive characteristics of giant viruses of marine eukaryotes.

Giant viruses have remarkable genomic repertoires-blurring the line with cellular life-and act as top-down controls of eukaryotic plankton. However, to date only six cultured giant virus genomes are available from the pelagic ocean. We used at-sea flow cytometry with staining and sorting designed to target wild predatory eukaryotes, followed by DNA sequencing and assembly, to recover novel giant viruses from the Pacific Ocean. We retrieved four 'PacV' partial genomes that range from 421 to 1605 Kb, with 13 contigs on average, including the largest marine viral genomic assembly reported to date. Phylogenetic analyses indicate that three of the new viruses span a clade with deep-branching members of giant Mimiviridae, incorporating the Cafeteria roenbergensis virus, the uncultivated terrestrial Faunusvirus, one PacV from a choanoflagellate and two PacV with unclear hosts. The fourth virus, oPacV-421, is phylogenetically related to viruses that infect haptophyte algae. About half the predicted proteins in each PacV have no matches in NCBI nr (e-value < 10-5), totalling 1735 previously unknown proteins; the closest affiliations of the other proteins were evenly distributed across eukaryotes, prokaryotes and viruses of eukaryotes. The PacVs encode many translational proteins and two encode eukaryotic-like proteins from the Rh family of the ammonium transporter superfamily, likely influencing the uptake of nitrogen during infection. cPacV-1605 encodes a microbial viral rhodopsin (VirR) and the biosynthesis pathway for the required chromophore, the second finding of a choanoflagellate-associated virus that encodes these genes. In co-collected metatranscriptomes, 85% of cPacV-1605 genes were expressed, with capsids, heat shock proteins and proteases among the most highly expressed. Based on orthologue presence-absence patterns across the PacVs and other eukaryotic viruses, we posit the observed viral groupings are connected to host lifestyles as heterotrophs or phototrophs. This article is part of a discussion meeting issue 'Single cell ecology'.

Specialized proteomic responses and an ancient photoprotection mechanism sustain marine green algal growth during phosphate limitation.

Marine algae perform approximately half of global carbon fixation, but their growth is often limited by the availability of phosphate or other nutrients1,2. As oceans warm, the area of phosphate-limited surface waters is predicted to increase, resulting in ocean desertification3,4. Understanding the responses of key eukaryotic phytoplankton to nutrient limitation is therefore critical5,6. We used advanced photo-bioreactors to investigate how the widespread marine green alga Micromonas commoda grows under transitions from replete nutrients to chronic phosphate limitation and subsequent relief, analysing photosystem changes and broad cellular responses using proteomics, transcriptomics and biophysical measurements. We find that physiological and protein expression responses previously attributed to stress are critical to supporting stable exponential growth when phosphate is limiting. Unexpectedly, the abundance of most proteins involved in light harvesting does not change, but an ancient light-harvesting-related protein, LHCSR, is induced and dissipates damaging excess absorbed light as heat throughout phosphate limitation. Concurrently, a suite of uncharacterized proteins with narrow phylogenetic distributions increase multifold. Notably, of the proteins that exhibit significant changes, 70% are not differentially expressed at the mRNA transcript level, highlighting the importance of post-transcriptional processes in microbial eukaryotes. Nevertheless, transcript-protein pairs with concordant changes were identified that will enable more robust interpretation of eukaryotic phytoplankton responses in the field from metatranscriptomic studies. Our results show that P-limited Micromonas responds quickly to a fresh pulse of phosphate by rapidly increasing replication, and that the protein network associated with this ability is composed of both conserved and phylogenetically recent proteome systems that promote dynamic phosphate homeostasis. That an ancient mechanism for mitigating light stress is central to sustaining growth during extended phosphate limitation highlights the possibility of interactive effects arising from combined stressors under ocean change, which could reduce the efficacy of algal strategies for optimizing marine photosynthesis.

Red de vigilancia de resistencia antimicrobiana PRONARES: informe primer semestre 2001 / PRONARES antimicrobial surveillance network on antimicrobial agents resistance: report of the first semester 2001

En el transcurso de los años hemos sido testigos de un creciente problema de resistencia antimicrobiana, fenómeno que involucra cada día mayor número de cepas, nuevas especies y nuevos mecanismos. En noviembre de 1997 iniciamos una red de vigilancia de resistencia antimicrobiana PRONARES (Programa Nacional de Resistencia), en diferentes hospitales del país, que trabajando un protocolo común (20 cepas por síndrome clínico por mes) y utilizando un programa computacional WHONET (diseñado para vigilancia), nos permitiera detectar y monitorear el problema de la resistencia bacteriana en Chile. Los resultados del primer semestre de este año reportan 5.251 cepas de diferentes síndromes clínicos. En ITU, Escherichia coli (1.088 cepas) demostró alta susceptibilidad a todos los antimicrobianos, Klebsiella (1.000 cepas) demostró un perfil de resistencia más elevado, en cepas de Enterococcus spp se observó 30 por ciento de resistencia a ciprofloxacina y 2 por ciento a nitrofurantoína. Entre 899 cepas procedentes de infecciones invasoras, Staphylococcus aureus (555 cepas), mostró elevado perfil de resistencia a cloxacilina 40 por ciento superior -40 por ciento (21 por ciento) al observado en cepas aisladas de piel y tejido blandos (550). Shigella spp, (137 cepas) presentó 80 por ciento de resistencia ampicilina y 32 por ciento a cloranfenicol; ciprofloxacina y furazolidona demostraron muy buena actividad in vitro frente a este enteropatógeno. Al comparar cepas nosocomiales y de la comunidad, las primeras mostraron un perfil de mayor resistencia. Mantener una red nacional de vigilancia de resistencia se hace cada vez más necesario para orientar el uso adecuado de antibacterianos y evitar así que el fenómeno aumente

Escherichia coli enterotoxigena y campylobacter jejuni en el sindrome diarreico agudo en lactantes Chilenos / Enterotoxigenic escherichia coli and campylobacter jejuni in acute diarrheal syndrome in Chilean infants

An investigation carried out between the months of November 1982 and November 1983 looked at the frequency of isolation of enterotoxigenic Escherichia coli and Campylobacter jejuni compared with other enteropathogenic bacteria in 144 children under 2 years of age who had acute diarrhea and in 66 controls, all of them from lower socioeconomic strata, in a hospital and an outpatient office in Santiago, Chile. In addition, seasonal variations in frequency of the bacteria were determined. During the summer enteropathogenic bacteria were isolated in 64.7 per cent of the patients with diarrhea, while in winter this figure fell to 45.2 per cent. In the control group these proportions were 37.8 and 23.8 per cent, respectively (p0.01). Enteropathogenic E. coli was the bacterium that was isolated most frequently from sick infants (37.3 per cent in summer and 19.0 per cent in winter), followed by enterotoxigenic E. coli (21.6 and 19.0 per cent), several species of Shigella (12.7 and 4.8 per cent), C. jejuni (7.8 and 9.5

), enteroinvasive E. coli (1.9 and 2.4 per cent), and various species of Salmonella (3.9 and 0 per cent); no strains of Yersinia were isolated. Infections caused by more than one enteropathogenic bacterium were observed in 19.6 per cent of the summer cases and 9.5 per cent of the winter ones

Emergencia de resistencia a macrólidos en streptococcus pyogenes / Emergence of macrolide resistant streptococcus pyogenes strains

Background: Diseases produced by Streptoccocus pyogenes are still a problem in Chile, as in the rest of the world. It exhibits in vitro susceptibility to different antimicrobials, but penicillin continues to be the treatment of choice. Alternative drugs have been developed for allergic patients, such as erythromycin, new macrolides and cephalosporins. Nevertheless, resistant strains are appearing due to the indiscriminate use of macrolides. Aim: To assess present antimicrobial susceptibility of S Pyogenes strains isolated from chilean patients. Material and Methods: The susceptibility to penicillin, macrolides, clindamycin, cephalotin and vancomycin of 153 S Pyogenes strains, obtained from different health centers of the Metropolitan Region and isolated between 1996 and 1998, was assessed using the Kirby-Bauer method. Agar dilution minimal inhibitory concentration was then determined to macrolide resistant strains. Results: All strains were susceptible to penicillin. There was a 7.2 percent cross-resistance to macrolides. Conclusions: These results confirm that S Pyogenes resistance to macrolides has increased considerably in the Metropolitan Region of Chile during the last years

Actividad comparativa in vitro de moxifloxacina y otros antimicrobianos frente a patógenos respiratorios / Comparative in vitro activity of moxifloxacin and other antimicrobials against respiratory pathogens

Background: Respiratory pathogens are becoming increasingly resistant to antimicrobials. A new group of drugs, called respiratory quinolones have been synthesized to overcome this problem. Aim: To study the in vitro susceptibility of respiratory pathogens to old and new antimicrobials. Material and methods: Forty five strains of S pneumoniae, 44 strains of H influenzae, 21 strains of M catarrhalis, 10 strains of methicillin susceptible S aureus and 20 strains of methicillin resistant S aureus were studied. All were isolated from community acquired respiratory infections during 1999. Minimal inhibitory concentrations of moxifloxacin, amoxicilin, amoxicilin/clavulanic acid, clarithromycin, azithromycin, ciprofloxacin and levofloxacin were determined using the Etest method. Betalactamase production by H influenzae and M catarrhalis was also studied. Results: S pneumoniae strains were 100 percent susceptible to quinolones and cotrimoxazole, 2 percent were resistant to macrolides, 11 percent were resistant to amoxicilin/clavulanic acid and 47 percent were resistant to cefuroxime. H influenzae was 100 percent susceptible to quinolones, azithromycin and amoxicilin/clavulanic acid. There was a 53 percent resistance to cotrimoxazole, 21 percent to amoxicilin, 9 percent to clarithromycin and 7 percent to cefuroxime. M catarrhalis was 100 percent susceptible to quinolones and 100 percent resistant to amoxicilin, 5 percent resistant to macrolides, 14 percent resistant to amoxicilin/clavulanic acid, 20 percent to cefuroxime and 30 percent to cotrimoxazole. Methicilline susceptible S aureus was susceptible to all antimicrobials and methicillin resistant S aureus was resistant to all. Conclusions: Moxifloxacin and the new respiratory quinolones can be useful in the treatment of respiratory infections

Prevalencia de chlamdia trachomatis en conjuntivitis neonatal determinada mediante las técnicas de inmuno-fluorescencia y amplificación génica / Indirect fluorescence and genic amplification for the determination of chlamydia trachomatis prevalence in neonatal conjunctivitis

Backgrund: Chlamydia trachomatis is one of the most common identifiable infectious agents in neonatal conjunctivitis. It also causes pneumonitis, that is preceded by conjunctivitis in one third of cases. Aim: To asses the prevalence of Chlamydia trachomatis in newborns with conjunctivitis. Patients and methods: In 162 newborns, coming from 14 Primary Health Centers from Santiago de Chile, C. trachomatis was detected by indirect fluorescence and two polymerase chain reaction (PCR 1 and 2), wich amplified different sequences from the common endogenous plasmid. Those patients with positive indirect fluorescence and PCR 2 were definedas infected: Results: The prevalence of C. trachomatis was 8 percent, and the distribution of the positive cases was similar in the different Health Centers. Other isolates were: S. aureus (9.8 percent), S. pneumoniae (8 percent), S. viridans (6.2 percent) y H. influenzae (5.5 percent). Conclusions: The prevalence of C. trachomatis in neonatal conjunctivitis in Chile is similar to that of developed countries. Therefore, C. trachomatis should be considered in the election of antimicrobials for the treatment of neonatal conjunctivitis, to avoid ocular and respiratory complications

Prevalencia de streptococcus pneumoniae resistente a penicilina en niños que asisten a jardines infantiles en Santiago / Penicillin resistant streptococcus pneumoniae among children attending day care centers

Las infecciones neumocócicas causan enfermedades graves en los niños. Se han observado cambios epidemiológicos en las infecciones por Streptococcus pneumoniae, como aumento de la incidencia, desarrollo creciente de resistencia antimicrobiana y brotes infecciosos epidérmicos.El estudio de portadores es útil para vigilar la aparición y desarrollo de la resistencia antimicrobiana y de los serotipos prevalentes. Con el propósito de determinar la prevalencia de cepas resistentes de S. pneumoniae en niños que asisten a jardines infantiles de Santiago y los serotipos más frecuentes en la nasofaringe, se efectuó cultivo de hisopado faríngeo y nasal obtenido entre noviembre de 1994 y agosto de 1995 de 200 de ellos, cuya edad era entre 3 meses y 4 años (97 < de 2 años y 103 > de 2 años). Las muestras fueron sembradas en agar sangre con 5 por ciento de sangre de cordero, con o sin gentamicina (5 µg/ml) e incubadas en ambiente con dióxido de carbono. La resistencia antimicrobiana se evaluó mediante disco de oxacilina,microdilución en caldo y E-test.En 60,2 por ciento de los casos se aisló S. pneumoniae de la nasofaringe; 28,4 por ciento de las cepas eran resistentes a penicilina (20,3 por ciento con resistencia moderada y 2,1 por ciento elevada), 2 por ciento eran resistentes a cloranfenicol. No se encontraron cepas resistentes a eritromicina, cefotaxima o vancomicina. La frecuencia de cepas resistentes a la penicilina fue similar en los niños menores y mayores de 2 años. E serotipo más frecuente fue el 6A