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Objective The objective of this study was to evaluate the efficacy of two commercial nucleic acid amplification techniques to identify Mycobacterium tuberculosis in saliva. Subjects and Methods Fifty-two participants were recruited, 32 patients with a clinical and microbiological diagnosis of pulmonary tuberculosis and 20 healthy volunteers as controls. Three sputum samples were collected from each participant and were examined by direct bacilloscopy, cultured in liquid and solid media, and processed using the Mycobacterium tuberculosis direct ( MTD) test for rRNA detection. One saliva sample was collected from each participant using conventional methods and was examined by direct bacilloscopy, cultured, and processed using the MTD test for rRNA detection and the FluoroType Mycobacterium tuberculosis assay for DNA detection. Results In saliva samples, the sensitivity, specificity, and positive and negative predictive values of the MTD test were 71.8%, 95%, 95.8%, and 67.8%, respectively. The values obtained with the FluoroType assay were 56.2%, 90%, 90%, and 56.2%, respectively. Conclusions Our results indicate that when a sufficient volume of sputum cannot be obtained, saliva could be an alternative biological sample for the rapid diagnosis of pulmonary tuberculosis using commercial nucleic acid amplification techniques. [ABSTRACT FROM AUTHOR]

The dynamics of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission are influenced by a variety of factors, including social restrictions and the emergence of distinct variants. In this study, we delve into the origins and dissemination of the Alpha, Delta, and Omicron-BA.1 variants of concern in Galicia, northwest Spain. For this, we leveraged genomic data collected by the EPICOVIGAL Consortium and from the GISAID database, along with mobility information from other Spanish regions and foreign countries. Our analysis indicates that initial introductions during the Alpha phase were predominantly from other Spanish regions and France. However, as the pandemic progressed, introductions from Portugal and the United States became increasingly significant. The number of detected introductions varied from 96 and 101 for Alpha and Delta to 39 for Omicron-BA.1. Most of these introductions left a low number of descendants (<10), suggesting a limited impact on the evolution of the pandemic in Galicia. Notably, Galicia's major coastal cities emerged as critical hubs for viral transmission, highlighting their role in sustaining and spreading the virus. This research emphasizes the critical role of regional connectivity in the spread of SARS-CoV-2 and offers essential insights for enhancing public health strategies and surveillance measures., (© 2024 The Author(s). Journal of Medical Virology published by Wiley Periodicals LLC.)

Perinatal stress experienced by mothers of very premature newborns may influence the mother's milk and the infant's intestinal microbiota. This prospective study of mothers of very preterm infants fed with mother's own milk (MOM) was carried out in a tertiary hospital over a 2-year period. The assessment of maternal stress in 45 mothers of 52 very preterm newborns using the parental stress scale (PSS:NICU) revealed an inverse relationship between stress and MOM production in the first days of life ( p = 0.012). The greatest contributor to stress was the one related to the establishment of a mother-child bond. Maternal stress was lower in mothers in whom the kangaroo method was established early ( p = 0.011) and in those with a higher educational level ( p = 0.032). Levels of fecal calprotectin (FC) decreased with the passage of days and were directly correlated with birthweight ( p = 0.044). FC levels 7 days post-delivery were lower in newborns that received postnatal antibiotics ( p = 0.027). High levels of maternal stress resulted in progressive decreases and increases in the proportions of Firmicutes and Proteobacteria species, respectively, over 15 days post-delivery, both in MOM and in fecal samples from premature newborns. These findings underscore the importance of recognizing and appropriately managing maternal stress in neonatal units, given its marked influence on both the microbiota of maternal milk and the intestinal microbiota of premature newborns.

Background: Hepatitis B virus (HBV) comprises 9 genotypes and multiple subgenotypes that depict differences in geographic distribution, clinical outcome and response to antiviral therapy. However, the molecular epidemiology of HBV geno/subgenotypes is globally scarce. In Spain, HBV genotype D seems to be more prevalent in the northwestern regions compared to the rest of the country for unclear reasons., Methods: HBV genotyping was performed using geno2pheno on a S gene fragment amplified from plasma collected from all chronic hepatitis B individuals attended at one reference hospital in Santiago de Compostela, the Galicia's capital town. Phylogenetic and phylogeographic analyses using a fragment of 345 bp were performed in all viremic specimens. To avoid misleading allocation as consequence of short fragment analysis, several bioinformatic controls were used., Results: A total of 320 individuals with persistent serum HBsAg+ and detectable HBV-DNA were seen between 2000 and 2016 (male 68.4%; median age, 52 years-old; native Spaniards 83.8%). HBV genotype distribution was as follows: A 15.3%; B 1.6%; C 2.5%; D 71.6%; E 3.1%; F 2.2%; G 3.1%; and H 0.6%. HBV genotype D was mostly represented by D4 and D2 subgenotypes (33.4% and 15% of total, respectively). Compared to chronic hepatitis B patients with genotypes B, C, E and G, HBV-D4 carriers tended to be older (54.2% had >50 years-old) and HBeAg-negative (85%). Moreover, 43% were female, 4.7% had cirrhosis, 10.2% hepatitis C and 6.4% HIV coinfection. Phylogenetic analyses could be performed on 82 HBV-D4 specimens; and 79 were confirmed as HBV-D4 using PhyML. Phylogeography using FasTree suggested at least two distinct introductions of HBV-D4 in Galicia, one from the Caribbean and South America, and another from India., Conclusions: HBV subgenotype D4 is the most prevalent HBV variant in chronic hepatitis B patients living in the northwest of Spain, representing 33.4% (107/320) of all chronic hepatitis B infections. This rate of HBV-D4 is among the highest reported worldwide. Epidemiological and phylogenetic analyses suggest a strong association with historical migrant exchanges with Latin America, and especially with the Caribbean basin., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Objective: To evaluate the efficacy of sample pooling compared to the individual analysis for the diagnosis of coronavirus disease 2019 (COVID-19) by using different commercial platforms for nucleic acid extraction and amplification., Methods: A total of 3519 nasopharyngeal samples received at nine Spanish clinical microbiology laboratories were processed individually and in pools (342 pools of ten samples and 11 pools of nine samples) according to the existing methodology in place at each centre., Results: We found that 253 pools (2519 samples) were negative and 99 pools (990 samples) were positive; with 241 positive samples (6.85%), our pooling strategy would have saved 2167 PCR tests. For 29 pools (made out of 290 samples), we found discordant results when compared to their correspondent individual samples, as follows: in 22 of 29 pools (28 samples), minor discordances were found; for seven pools (7 samples), we found major discordances. Sensitivity, specificity and positive and negative predictive values for pooling were 97.10% (95% confidence interval (CI), 94.11-98.82), 100%, 100% and 99.79% (95% CI, 99.56-99.90) respectively; accuracy was 99.80% (95% CI, 99.59-99.92), and the kappa concordant coefficient was 0.984. The dilution of samples in our pooling strategy resulted in a median loss of 2.87 (95% CI, 2.46-3.28) cycle threshold (C t ) for E gene, 3.36 (95% CI, 2.89-3.85) C t for the RdRP gene and 2.99 (95% CI, 2.56-3.43) C t for the N gene., Conclusions: We found a high efficiency of pooling strategies for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA testing across different RNA extraction and amplification platforms, with excellent performance in terms of sensitivity, specificity and positive and negative predictive values., (Copyright © 2020 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Spain has been one of the main global pandemic epicenters for coronavirus disease 2019 (COVID-19). Here, we analyzed >41 000 genomes (including >26 000 high-quality (HQ) genomes) downloaded from the GISAID repository, including 1 245 (922 HQ) sampled in Spain. The aim of this study was to investigate genome variation of novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and reconstruct phylogeographic and transmission patterns in Spain. Phylogeographic analysis suggested at least 34 independent introductions of SARS-CoV-2 to Spain at the beginning of the outbreak. Six lineages spread very successfully in the country, probably favored by super-spreaders, namely, A2a4 (7.8%), A2a5 (38.4%), A2a10 (2.8%), B3a (30.1%), and B9 (8.7%), which accounted for 87.9% of all genomes in the Spanish database. One distinct feature of the Spanish SARS-CoV-2 genomes was the higher frequency of B lineages (39.3%, mainly B3a+B9) than found in any other European country. While B3a, B9, (and an important sub-lineage of A2a5, namely, A2a5c) most likely originated in Spain, the other three haplogroups were imported from other European locations. The B3a strain may have originated in the Basque Country from a B3 ancestor of uncertain geographic origin, whereas B9 likely emerged in Madrid. The time of the most recent common ancestor (TMRCA) of SARS-CoV-2 suggested that the first coronavirus entered the country around 11 February 2020, as estimated from the TMRCA of B3a, the first lineage detected in the country. Moreover, earlier claims that the D614G mutation is associated to higher transmissibility is not consistent with the very high prevalence of COVID-19 in Spain when compared to other countries with lower disease incidence but much higher frequency of this mutation (56.4% in Spain vs. 82.4% in rest of Europe). Instead, the data support a major role of genetic drift in modeling the micro-geographic stratification of virus strains across the country as well as the role of SARS-CoV-2 super-spreaders.

Introduction: Influenza A H1N1 community-acquired pneumonia (CAP) is a quite frequent respiratory disease. Despite being considered more serious than other CAPs, there are very few studies comparing its characteristics with noninfluenza CAP. We aim to establish the differences between pneumonia due to H1N1 virus and pneumonia not caused by H1N1 influenza virus and to determine the probability that a pneumonia is due to an H1N1 virus infection based on the most relevant variables., Methods: We used a case-control study where cases were H1N1 CAP patients with confirmed microbiological diagnosis and controls were patients with CAP admitted to hospital. H1N1 and other influenza types were discarded among controls. We calculated the probability of being a case or control using multivariate logistic regression., Results: We included 99 cases and 270 controls. Cases were younger than controls (53 vs 71 years, respectively). Mortality was much higher for H1N1 patients (13% vs 0.3%), and admission to intensive care unit was more frequent for H1N1 cases. The variables most associated with presenting H1N1 CAP were bilateral affectation on chest X-rays (OR: 5.70; 95% CI 2.69-10.40), followed by presence of arthromyalgias, with cases presenting close to three times more arthromyalgias compared to controls. Low leukocytes count and high AST values were also significantly associated with H1N1 CAP. H1N1 CAPs are characterized by bilateral affectation, low leukocyte count, presence of arthromyalgias, and high AST., Conclusions: A few and easy to obtain clinical parameters might be extremely useful to distinguish H1N1 CAP from CAPs of other origin.

Introduction: The American Thoracic Society and the Infectious Diseases Society of America recommend that clinically significant non-tuberculous mycobacteria (NTM) should be identified to the species level in order to determine their clinical significance. The aim of this study was to evaluate identification of rapidly growing NTM (RGM) isolated from clinical samples by using MALDI-TOF MS and a commercial molecular system. The results were compared with identification using a reference method., Methods: We included 46 clinical isolates of RGM and identified them using the commercial molecular system GenoType ® CM/AS (Hain, Lifescience, Germany), MALDI-TOF MS (Bruker) and, as reference method, partial rpoβ gene sequencing followed by BLAST and phylogenetic analysis with the 1093 sequences available in the GeneBank., Results: The degree of agreement between GenoType ® and MALDI-TOF MS and the reference method, partial rpoβ sequencing, was 27/43 (62.8%) and 38/43 cases (88.3%) respectively. For all the samples correctly classified by GenoType ® , we obtained the same result with MALDI-TOF MS (27/27). However, MALDI-TOF MS also correctly identified 68.75% (11/16) of the samples that GenoType ® had misclassified (p=0.005)., Conclusions: MALDI-TOF MS classified significantly better than GenoType ® . When a MALDI-TOF MS score >1.85 was achieved, MALDI-TOF MS and partial rpoβ gene sequencing were equivalent. GenoType ® was not able to distinguish between species belonging to the M. fortuitum complex. MALDI-TOF MS methodology is simple, rapid and associated with lower consumable costs than GenoType ® . The partial rpoβ sequencing methods with BLAST and phylogenetic analysis were not able to identify some RGM unequivocally. Therefore, sequencing of additional regions would be indicated in these cases., (Copyright © 2018 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.)

Pathogens that infect the gastrointestinal and respiratory tracts are subjected to intense pressure due to the environmental conditions of the surroundings. This pressure has led to the development of mechanisms of bacterial tolerance or persistence which enable microorganisms to survive in these locations. In this review, we analyze the general stress response (RpoS mediated), reactive oxygen species (ROS) tolerance, energy metabolism, drug efflux pumps, SOS response, quorum sensing (QS) bacterial communication, (p)ppGpp signaling, and toxin-antitoxin (TA) systems of pathogens, such as Escherichia coli , Salmonella spp., Vibrio spp., Helicobacter spp., Campylobacter jejuni , Enterococcus spp., Shigella spp., Yersinia spp., and Clostridium difficile , all of which inhabit the gastrointestinal tract. The following respiratory tract pathogens are also considered: Staphylococcus aureus , Pseudomonas aeruginosa , Acinetobacter baumannii , Burkholderia cenocepacia , and Mycobacterium tuberculosis Knowledge of the molecular mechanisms regulating the bacterial tolerance and persistence phenotypes is essential in the fight against multiresistant pathogens, as it will enable the identification of new targets for developing innovative anti-infective treatments., (Copyright © 2018 American Society for Microbiology.)

Objective: Pulmonary nocardiosis is an uncommon pulmonary infection caused by aerobic gram-positive bacteria of the genus Nocardia. Nocardia sp. are environmental organisms spread worldwide. Approximately 50 Nocardia species have been described to date, about 30 of which are known to cause human disease. Nocardia cyriacigeorgica was first reported in 2001., Case Report: We report a case of infection caused by N. cyriacigeorgica in a patient with B-cells non-Hodgkin lymphoma and diabetes mellitus. The microbiological findings reflect a possible co-infection by N. cyriacigeorgica and Aspergillus fumigatus., Conclusions: Patient's background and information related to risk factors are essential to detect the growth of Nocardia sp. in the laboratory. Furthermore, diagnosis of invasive pulmonary aspergillosis is particularly controversial, especially in intensive care units patients. Taking everything into account, we will discuss a possible co-infection by N. cyriacigeorgica and A. fumigatus in a critically ill patient.

Incorporation of rapid detection systems to identify mutations in M. tuberculosis complex that confer resistance to isoniazid and rifampicin has potentiated the knowledge of their distribution, given the geographical variability. We performed antibiograms of the 2,993 strains isolated in Galicia, Spain (2008-2013). In the strains resistant to isoniazid, a concentration of 0.4 mg/mL and MTBDRplus Genotype test (Hain Lifescience, Germany) were used. We found that 3.64 % of strains were resistant to isoniazid, while 0.43 % were resistant to isoniazid and rifampicin (multidrug resistant, MDR). The MTBDRplus test showed an overall sensitivity of 72.48 %, with 62.5 % sensitivity for non MDR isoniazid-resistant strains and 100 % sensitivity for MDR strains. The katG gene mutation was detected at codon 315 in 38.53 % of strains. The S315T mutation appeared in 61.54 % of MDR strains and 34.38 % of non-MDR strains. The 28.44 % had mutations in inhA, (93.55 % in C15T), and 38.46 % of MDR strains were mutated. In non-MDR strains, 37.50 % were wild-type, 35.42 % and 27.08 % had mutations in katG and inhA, respectively. The most frequent mutation in rpoβ was S531L (46.15 %). The 38.71 % and 41.9 % of strains with resistance to isoniazid and streptomycin had mutations in katG and inhA, respectively (2 strains with mutations in T8C and T8A). The distribution pattern of resistance among strains with high and low concentrations of isoniazid showed statistically significant differences in relation to the mutation in katG and wild-type. The sensitivity of the Genotype MTBDRplus test for non-MDR strains in our area was at the lower threshold described.

Objective: Phylogenetically, hepatitis B virus (HBV) is classified into genotypes and subgenotypes used for epidemiological studies. The aim of this study is to know the distribution of HBV subgenotypes D in our environment., Methods: From 401 patients HBV surface antigen positive, HBV DNA-positive, partial HBV-DNA S gene was amplified, sequenced and analysed using geno2pheno (hbv) (Max-Planck Institute) on line application., Results: We found 259 (64.6%) patients with HBV genotype D: 53 not subgenotypable, 9 (4%) D1, 61 (30%) D2, 15 (7%) D3 and 121 (59%) D4. Patients with D1 subgenotype were, on average, 23 years younger (p = 0.0001), with a higher proportion of women (p < 0.05)., Conclusions: HBV subgenotype D4 was the most prevalent in our area. Patients with D1 subgenotype came from abroad were younger than the other subgenotypes and mostly women. These results show the interest of conducting studies at HBV subgenotype level.

Introduction: Candida infection has become a major health problem worldwide. The epidemiology of Candidaemia has substantially changed by the emergence of the species Candida non-albicans. This variation is particularly important in the choice of prophylaxis and empirical treatment. The methods based on biochemical and molecular biology have limitations for the correct identification of Candida species. The aim of this study is to demonstrate the ability of the MALDI-TOF mass spectrometry for the identification of these species and compare it with the technology used today., Methods: We included all isolates collected over 2 years (n=73) of Candida non-albicans from non-invasive samples. The identification was carried out by Vitek-2 systems YST and API CAUX. The MALDI-TOF identifications were made with Confidence Axima system (Shimadzu Corporation) using the Shimadzu Launchpad software and database SARAMIS (AnagnosTec GmbH). Discrepancies were resolved by SeptiFast LightCycler multiplex PCR, specific PCR C. glabrata and enzymatic digestion with BanI SADH fragment in isolates of C. parapsilosis., Results: Of the 73 isolates of Candida non-albicans, the biochemical methods conclusively identified 67 to species level and 6 at the genus level. The MALDI-TOF system obtained identifications at the species level in all cases. The correlation in the species of all isolates studied was 85.07%, reaching 94.52% when the correlation was made between the identification obtained by biochemical methods and the methods for the analysis of the discrepancies. In isolates of C. parapsilosis, MALDI-TOF system obtained an identification of C. orthopsilosis. In 3 of them it was confirmed by digestion with BanI SADH fragment., Conclusion: This study has demonstrated the use of mass spectrometry (MALDI-TOF system) to provide the microbiology laboratory with greater efficiency and reliability to identify isolates of Candida non-albicans to species level. It also shows its potential usefulness in identifying related species, such as C. parapsilosis, metapsilosis and orthopsilosis., (Copyright © 2010 Elsevier España, S.L. All rights reserved.)

In the Health Care Area of Santiago de Compostela, during three years (2004-2006), we studied 218 Streptococcus pneumoniae isolates from invasive disease (10 CSF, 133 blood culture, 7 pleural fluid, 9 corneal ulcer and 59 lower respiratory tract), to determine the predominant serotypes and antimicrobial susceptibility. 77.1% of the isolates were from adults and 22.9% from pediatric patients. There were 33 different serotypes, six of them occurring in more than 5% of cases, in decreasing order: 19 (16.97%), 3 (11.46%), 1 (10.55%), 14 (10.55%), 23 (6.88%) and 6 (6.88%). The predominant serogroup in CSF was 19 and in blood culture predominant serogroups were 19, 1, 14 and 3. 63.76% of the isolates were susceptible to all antibiotics tested. The most frequent resistance phenotypes were to erythromycin (24.31%), followed by penicillin (18.34%). Both phenotypes were associated with serotypes 19, 14 and 6. The resistance to levofloxacin was 0.9% and 0.5% to cefotaxime. We did not find any vancomycin resistance.

Objective: To describe the characteristics of patients with multidrug-resistant tuberculosis (MDR-TB), a descriptive prospective study was carried out applying a combination of exhaustive conventional epidemiology with molecular genotyping., Setting: All patients diagnosed with MDR-TB in Galicia, Spain, between 1998 and 2004 were included in the study., Design: Of 9895 diagnosed cases of TB, 58 were MDR-TB (0.59%). The site of disease was pulmonary in 56 cases and 46 were smear-positive. Only two cases were co-infected with the human immunodeficiency virus (HIV) and seven were immigrants. Twenty-five (43%) had received previous TB treatment. These cases presented more risk factors for treatment default and a lower frequency of contact with cases of MDR-TB., Results: Genotyping analysis was performed in 57 patients, showing evidence of four clusters (30 patients, 52.6%), each with identical genetic patterns. The patients included in the clusters were younger, and most had primary forms or had had contact with another case of MDR-TB, especially in hospital. Neither the Beijing/W nor the B strain was identified., Conclusion: There is a low prevalence of MDR-TB in Galicia. Unlike previous studies, there was a high rate of transmissibility, including nosocomial transmission. Transmission is not associated with HIV or previously reported strains with a high capacity for transmission.

Objective: The purpose of this study was to determine the epidemiological characteristics of tuberculosis (TB) in the Public Health System District of Santiago de Compostela (population : 386125) from 1999 to 2002., Methods: Inclusion criteria were: 1). microbiological and/or pathological diagnosis of TB in any specimen, and 2). patient younger 35 years old with recent medical history of TB. Mantoux test positive, and pleural effusion with linfocitosis and adenosine deaminase >47 IU/ml., Results: 946 patients were included (568 men and 378 women), with ages ranging from 2 months to 96 years. The incidence of TB was 60.9/100000 in 1999, 67.6/100000 in 2000, 61.9/100000 in 2001 and 54.6/100000 in 2002. The incidence rate of tuberculous meningitis was 1.03/100000 in 1999 and 2000, 0.77/100000 in 2001 and 0.51/100000 in 2002. The percentage of cases associated with HIV was 3.4% in 1999, 1.9% in 2000, 2.4% in 2001 and 2002. We found an increase in the rate of males over 55 years of age; with incidence per 100000 inhabitants of 122.4 in 1999, 142.8 in 2000, 115 in 2001 and 119 in 2002, whereas in females the incidence was 40.6 in 1999, 60.9 in 2000, 54.1 in 2001 and 39.1 in 2002., Conclusions: In last four years the incidence of tuberculosis has decreased but remains high in males over 55 years old.

Among all infectious diseases, tuberculosis has probably been the most frequent cause of death and morbidity in the history of humanity. On a world-wide basis, the tuberculosis (TB) represents a serious public health problem, existing great differences between developed and developing countries in terms of controlling the disease. One of the problems inherent in this disease is the resistance of Mycobacterium tuberculosis to drugs, specially the multi-resistance (defined as resistance to isoniziade and rifampicine), it represents a problem of global health. Faced with clinical suspect, its diagnosis must be immediately done and treatment must be started as soon as possible. For this purpose, we have at our disposal such effective diagnosis and therapeutic methods. However, the most important the most important action is that all these patients must be subjected to a close evolved control carried by specialized units.

Ligase chain reaction amplification (LCx Abbott Laboratories) was used to detect the presence of M. tuberculosis in 101 adenopathy specimens obtained from 98 patients. A total of 30 cases of lymph node tuberculosis were diagnosed, and the data were compared with results obtained using conventional techniques. The sensitivity of auramine staining and culture were 50.0% and 66.7%, respectively. The sensitivity, specificity, positive predictive value and negative predictive value of the LCx assay were 80.0, 98.6, 96.0, and 92.1% respectively. The results confirm the reliability of ligase chain reaction amplification for the detection of lymph node tuberculosis.

Objective: To analyse the distribution of the forms of presentation of tuberculosis (TPF) in elderly patients., Material and Methods: The medical records of patients diagnosed with tuberculosis attending the Tuberculosis Prevention and Control Unit of the Santiago Health District were reviewed over of six years period. The classification of TPF was: pulmonary forms (P), disease confined to the lung; extrapulmonary forms (EF), disease outside the lung; mixed forms (MF), the presence of both pulmonary and extrapulmonary tuberculosis; disseminated forms (DF), the presence of two or more extrapulmonary locations; and miliary TB, which was defined by a diffuse pulmonary radiographic pattern or diagnosis was undertaken by necropsy., Results: A total of 278 tuberculosis infected patients were observed, 156 (56.2%) were men and 122 (43.8%) women, their mean age was 75.3 years (range 65-95). The distribution of TPF was: 155 (55.8%) P forms; 66 (23.7%) EF, of which 27 (41.0%) were ganglionary location, 12 (18.2%) bone and joint, 8 (12.0%) intestinal, 6 (9.1%) peritoneal, 5 (7.6%) meningeal, and other locations 8 (12.1%); MF 47 cases (16.9%); miliary TB 7 cases (2.5%) and. DF 3 cases (1.1%). None case was observed of HIV infected patient., Conclusions: Our findings confirm high incidence of extrapulmonary TB in elderly patients. Our experience shows a modification to the classical presentation of the disease, and thus the need for sensitivity in locating the disease.

Objective: A retrospective study of the correlation between CD4 lymphocytes and the viral load in 16 HIV-patients with tuberculosis., Material and Methods: The clinical forms of presentation of tuberculosis were classified according to the location/s of the disease into: pulmonary, distinguishing between typical pulmonary and atypical according to the radiological pattern; extrapulmonary; mixed forms: pulmonary and extrapulmonary; and miliary tuberculosis., Results: Tuberculosis was exclusively pulmonary in 7 cases (44%), all were atypical; extrapulmonary 6 (38%); mixed 2 (12%); and miliary 1 (6%). The mean CD4 lymphocyte count was 111.1 (range 5-360), in 11 (69%) the counts were below 200 cells/mm3. The lowest CD4 count was in the mixed forms with a mean of 45 cells/mm3 whilst the highest was obtained in pulmonary forms with a mean of 128.3 cells/mm3. The mean viral load was 4.82 log (range, 0-5.93), the highest load was for mixed forms with a mean 5.69 log, whereas the lowest load was for pulmonary forms with a mean of 4.19 log. No significant correlation was observed between CD4 lymphocytes and viral load (correlation coefficient--0.1163)., Conclusions: Though no significant correlation was observed, a high CD4 was associated to a low viral load and inversely a low CD4 with a high viral load.

Background: The aim of this work was to evaluate the applicability of polymerase chain reaction (PCR) in the microbiological diagnostic of meningococcal meningitis as compared with the conventional methods (Gram stain and culture)., Methods: One hundred and fifteen cerebrospinalis fluid samples from 115 patients with suspicious symptoms of meningitis were studied. 47 of them belonged to patients suspicious for meningococcal disease; 28 to patients with bacterial meningitis of other infectious etiologies; 10 to patients with meningitis showing lymphocytic pleocytosis and 30 to patients with an unconfirmed meningitis. The cerebrospinalis fluid samples were processed for culture by standard procedures and by PCR according to the method described by Newcombe et al for peripheral blood samples., Results: Thirty five out of 39 patients suspicious of meningococcal meningitis were microbiologically confirmed, being 22 culture and PCR positive, 3 microscopically and PCR positive, 1 only microscopically positive, and 9 positive only by PCR. By using PCR methodology, the number of confirmed diagnostics of meningococcal meningitis increased in a 23% as compared to those obtained by microscopic observation and culture. Sensitivity, specificity, predictive positive value and predictive negative value were 87.1, 98.7, 97.1 and 94.1 respectively for the PCR method., Conclusions: Our results indicate that PCR can be used on routine basis as a complementary technique to the standard laboratory procedures for diagnosis of meningococcal meningitis.

Objective: To analyze the clinical characteristics and diagnose of tuberculosis in patients infected by human immunodeficiency virus in our setting., Methods: We have revised the clinical, microbiological and histological characteristics of 92 cases diagnosed tuberculosis in seropositive patients, during a span time of 128 months., Results: Of them, 71 were male and most of them (81.5%) were intravenous drugs users. The most (93.4%) were sintomatic when tuberculosis was diagnosed mainly fever and general and respiratory sintomatology. The most common signs were the presence of lymphadenopathies and hepatomegaly. The tuberculosis affected mainly pulmonary and ganglionary system. 34 cases had only pulmonary pathology, 24 extrapulmonary, 25 pulmonary and extrapulmonary and 9 miliary. The tuberculosis diagnose was based in microbiologic criteris. The most frequent source, as histologic critery, has been the ganglionary tissue., Conclusions: The only or associated pulmonary forms are more frequent. The most commonly extrapulmonary form is the ganglionary localization. The most of diagnose methods were obtained from respiratory tract and lymphadenopathy samples.

Background: The aim of this study was to determine the clinico-epidemiologic characteristics of meningitis caused by Neisseria meningitidis., Methods: A retrospective study was performed of the bacterial meningitis with LCR positive cultures for Neisseria meningitidis from January 1990 to 31 March, 1997. To calculate the rate of incidence data from the 1990 population census were used corresponding to a population of 465,786 inhabitants per year attended in our hospital., Results: A growth was observed in the strains of N. meningitidis in 61 LCR, representing 30% of all positive LCRs. Thirty-three strains belonged to serogroup B (54%) and 28 of serogroup C (46%). Ninety-one point nine percent of the cases were found in patients under the age of 20. The annual rates of incidence ranged from 2.3 cases/100,000 inhabitants in 1990 to 3.4 cases/100,000 inhabitants in 1996 with a slight decrease between these two dates. In patients under the age of 15 years, the rates of incidence ranged from 12.3/100,000 in 1990 to 13.4 per 100,000 inhabitants in 1996. In the remaining years the rates decreased with a minimum of 2.2 cases/100,000 inhabitants. The incidence for N. meningitis serogroup C ranged from 0 to 0.9 cases/100,000 inhabitants between 1990 and 1995. In 1996 the rate increased up to 2.6 cases/100,000 inhabitants. The rate of mortality was 6.6% and sequelae 8.7%. Since 1995 strains with decreased sensitivity to penicillin have been isolated, with percentages ranging from 20% to 56.25%. All strains remained sensitive to third generation cephalosporins and rifampicin., Conclusions: Neisseria meningitidis remains the most frequent etiologic agent of acute bacterial meningitis. The increase in serogroup C strains and the ever more frequent appearance of strains with decreased resistance to penicillin are confirmed, as is the persistence of high levels of endemia in our medium.

An exhaustive search for the clinical records of patients diagnosed with tuberculous disease was done in the hospitals of the area under study, which involves 392,000 population. During the years 1992, 1993 and 1994. There were included: 1) patients who had positive bacilloscopy and/or positive Lowenstein's culture in any specimen: 2) patients younger than 35-years-old who had pleural effusion, significant Mantoux and adenosine deaminase (ADA) over 47 U/I in the pleural effusion. In total 814 patients remained in the study with an average age of 38.39(19.39 DE) in 1992, 39.02 (20.04 DE) in 1993, and 34.1 years-old (19.2 DE) in 1994, with extreme ages of 2 months and 87 years-old. The incidence/100,000 H was: in 1992: 67.86, in 1993: 66.58 and in 1994: 73.2. The contagious forms incidence/100,000 H was: 1.5 in 1992 and 1993; and 1.79 in 1994. The hospital mortality incidence/100,000 H was 2.04 in 1992, 2.30 in 1993 and 2.6 in 1994. We conclude that tuberculosis is endemic in our area with moderately high and stationary incidence.

In order to assess the incidence of tuberculous disease in our health area, we reviewed the clinical records of patients from the three hospitals of the area under study during the years 1989, 1990 and 1991, who had positive bacilloscopy, positive Lowenstein's culture in any specimen and/or compatible anatomopathologic report. After excluding 26 patients because they belonged to other health areas, 885 patients remained in the study, out of which 251 (64% men and 36% women) were from the year 1989, 270 (64% men and 36% women) from the year 1990 and 364 (62% men and 38% women) from the year 1991. The mean age was 38.4 (SD 20.5). Fifty-one percent of the patients were between 20 and 35 years old. The rate of new cases was 65.87 per 100.000 population in 1989, 71.05 in 1990 and 95.53 in 1991. Seventy-four cases were HIV-positive (8%). Tuberculous meningitis was present in 12 patients. The highest mortality was 1.79 per 100.000 population in 1990. We conclude that tuberculosis presents a medium-high incidence in our health area.

We studied the antibiotic sensitivity of 130 Streptococcus pneumonia strains isolated from respiratory samples (96 cases), blood (13 cases), CSF (6 cases), ocular secretions (5 cases) and other origins (10 cases). High-level penicillin-resistance was found in 13.08% of strains, and intermediate-level penicillin resistance was also found in 22.3% of strains. A multiple-antibiotic resistance pattern was shown by 80.44% of all penicillin-resistant strains. Resistance rates found against antibiotics were as follows: tetracycline, 41.53%; chloramphenicol 17.69%; cotrimoxazole, 16.92% and erythromycin 6.92%. All isolated strains were sensitive to rifampin and vancomycin.

To evaluate the epidemiology, etiology, mortality and prognostic factors of bacteremia in a district hospital, we carried out a retrospective study of all blood cultures (Bc) performed in 1987 in our institution. The criteria for contamination community or hospital acquired disease, and terminal or rapidly fatal disease were those commonly accepted. Two patients were excluded owing to incomplete data. Overall 1,693 Bcs were performed; 257 of these were positive, 189 were considered as true positive (11.1%) and corresponded to 70 episodes of bacteremia (9.1/1000 admitted patients). Sixty-eight patients were included, 48 from the community and 20 of hospital origin. The male/female ratio was 43/25. There was shock in 27% of the community acquired and in 55% of the hospital acquired cases (p less than 0.1). There were no mortality differences between both groups, but there were differences in the spectrum of causative organisms. A comparison with a previous series from our institution was made. Its results shown some peculiarities of the disease in our area.