High Prevalence of Prototheca bovis Infection in Dairy Cattle with Chronic Mastitis in Ecuador.

The genus Prototheca, a unicellular, non-photosynthetic, yeast-like microalgae, is a pathogen of concern for the dairy industry. It causes bovine mastitis that currently cannot be cured, and hence generates significant economic losses in milk production. In this study, for the first time in Ecuador, we identify Prototheca bovis as the etiologic agent of chronic mastitis in dairy cattle. Milk samples (n = 458) of cows with chronic mastitis were cultured on Sabouraud Dextrose Agar (SDA). Microscopy and cytB gene sequencing were used to identify Prototheca, whereby Prototheca bovis was isolated from 15.1% (n = 69) of the milk samples, one of the highest infection rates that can be found in the literature in a "non-outbreak" situation. No other Prototheca species were found. We were unable to isolate the alga from environmental samples. We showed that P. bovis was relatively resistant to disinfectants used to sterilize milking equipment on the cattle farms where it was isolated. We discuss how to avoid future infection and also hypothesize that the real prevalence of Prototheca infection in bovine mastitis is probably much higher than what was detected. We recommend a protocol to increase the diagnostic yield in the bacteriology laboratory.

Immunogenicity of a 7-valent pneumococcal conjugate vaccine (PCV7) and impact on carriage in Venezuelan children at risk of invasive pneumococcal diseases.

BACKGROUND AND AIMS: We evaluated the immunogenicity of the 7-valent pneumococcal conjugate vaccine (PCV7), and its impact on pneumococcal carriage in Venezuelan children at high risk for invasive pneumococcal disease (IPD). METHODS: 82 children (age 2-59 months) with sickle cell anemia (n=22), chronic heart disease (n=19), HIV infection (n=12), immune-suppressive therapy (n=11) and other IPD-predisposing conditions (n=18) were vaccinated with PCV7 according to CDC-recommended age-related immunization schedules. Blood samples were taken to determine the concentration of IgG antibody, and nasopharyngeal swabs were obtained to isolate Streptococcus pneumoniae, before the first vaccine dose and 1 month after completion of the vaccination schedule. RESULTS: Pneumococcal carriage prior to the first immunization was 27% (n=22), with the most frequently carried serotypes being vaccine serotypes 6B (22%) and 14 (13%). One month after completion of the vaccination scheme pneumococcal carriage was 22% (n=17), dominated by non-vaccine serotypes 19A (24%) and 7F (12%). Before immunization, 65% of the subjects had IgG antibody titers >0.35 µg/mL for five serotypes tested. Post-vaccination, 100% of the subjects showed titers >1.0 µg/mL for all PCV7 serotypes with geometric mean concentrations (GMC) ranging from 1.75 µg/mL (serotype 23F) to 17.16 µg/mL (serotype 14). Children previously colonized with serotype 6B had a significantly lower GMC to this serotype following immunization than children not carrying 6B prior to the first PCV dose (p<0.05). CONCLUSIONS: PCV7 is highly immunogenic in Venezuelan children at high-risk for IPD. Vaccination was associated with an immediate shift in nasopharyngeal carriage toward non-PCV7 serotypes. Finally, we observed serotype-specific hyporesponsiveness to immunization after natural carriage with the same serotype in high-risk children.

Seguimiento de un brote de infección en tejido blando causado por Mycobacterium abscessus posterior a la mesoterapia en Venezuela / Follow-up on an outbreak in Venezuela of soft-tissue infection due to Mycobacterium abscessus associated with Mesotherapy

Introducción Las infecciones de la piel y los tejidos blandos causadas por micobacterias no tuberculosas (MNT) se han asociado a procedimientos como inyecciones, liposucción, cirugía plástica y acupuntura. Estudiamos un brote de infección en tejidos blandos, debido a MNT posterior a mesoterapia, un procedimiento cosmético que consiste en inyectar una mezcla de sustancias para reducir hipotéticamente el tejido adiposo localizado. Métodos Se entrevistó a pacientes con lesiones en la piel, con antecedentes de mesoterapia, que acudieron al Departamento de Dermatología del Hospital Público de la Ciudad de Barinas, Venezuela, en el período comprendido entre noviembre de 2004 y febrero de 2005. Se tomaron muestras clínicas y ambientales para el aislamiento de micobacterias. Resultados Las entrevistas revelaron que 68 pacientes se infectaron con MNT. Todos recibieron tratamiento en el mismo centro estético, a cargo del mismo terapeuta y con el mismo producto. De las muestras de 5 pacientes se aisló Mycobacterium abscessus. Ninguna de las soluciones utilizadas en la mesoterapia estuvo disponible para el análisis, pero se aisló M. abscessus de una muestra del ambiente tomada en el centro cosmético. La tipificación de las cepas con técnicas basadas en PCR (ERIC-PCR, BOXA1R y RAPD) mostró que los aislados de los pacientes fueron indistinguibles entre sí, pero diferentes del aislado del medio ambiente del centro. Conclusión Lo más probable es que este brote se haya causado por un producto contaminado utilizado en la mesoterapia y no por una micobacteria del ambiente del centro. Hacemos énfasis en la importancia de un mejor control microbiológico de estos productos. Este brote, que afectó al menos a 68 pacientes, es en nuestro conocimiento el más grande descrito en la literatura médica posterior a mesoterapia (AU)

Introduction Skin and soft tissue infections caused by nontuberculous mycobacteria (NMT) are reported to be associated with injections, liposuction, plastic surgery, and acupuncture. Herein, we describe an outbreak of soft tissue infection due to NMT following mesotherapy, a cosmetic procedure involving injection of poorly defined mixtures alleged to reduce local adiposity. Methods Patients with skin lesions and a history of mesotherapy treatment, who visited the dermatology department of the public hospital in Barinas, Venezuela, from November 2004 to February 2005 were interviewed. Clinical and environmental samples were taken for mycobacteria isolation. Results The interviews revealed that 68 patients who had been treated for cosmetic purposes at the same clinic by the same therapist had received injections with the same product and were infected with NMT. Clinical specimens from 5 patients grew Mycobacterium abscessus. No mesotherapy solution was available for analysis but M. abscessus was isolated from an environmental sample in the clinic. PCR-based strain typing techniques (ERIC-PCR, BOXA1R and RAPD) showed that the patient's isolates were undistinguishable from each other but different from the environmental isolate. Conclusions This outbreak was likely caused by a contaminated injectable mesotherapy product and not by mycobacteria from the clinic environment. We emphasize the importance of better microbiological control of these products. To our knowledge, this outbreak, which affected at least 68 patients, appears to be the largest ever associated with mesotherapy and described in the literature (AU)

[Persistent positive baciloscopy after several months of tuberculosis therapy: not always it represents a treatment failure]. / Baciloscopia positiva persistente en fase avanzada de la terapia anti-tuberculosa: no siempre indica fracaso del tratamiento.

International agencies for tuberculosis (TB) control (WHO, IUATLD, KNCV) consider the presence of acid-fast bacilli (AFB) in the sputum smear after completion of TB therapy as treatment failure. We describe a case of a pulmonary TB patient who received and correctly completed 6 months of antituberculous treatment with a positive AFB sputum smear. Treatment was continued and sputum cultures were performed for resistance determination. After 9 months, despite the persistence of AFB in the sputum smear, the treatment was suspended because none of the sputum samples yielded a positive culture, indicating no viable bacilli. In addition, the patient was asymptomatic and had recovered his body weight. An exhaustive review of medical literature allowed us to conclude that a positive sputum smear after therapy has been detected in up to 5% of cases and does not always represent a treatment failure as defined by international guidelines. For scenarios where culture methods are not available we propose a scheme to evaluate these patients. This includes compliance with the treatment, severity of the disease at the moment of diagnosis, clinical symptoms after specific therapy and rate of in vitro resistance of M. tuberculosis in the community.

Evaluación de productos tuberculicidas comerciales: normalización. Respuesta de los autores / Assessment of commercial tuberculocidal products: normalization. Authors’ reply

No disponible

Levels of complement C3 and C4 components in Amerindians living in an area with high prevalence of tuberculosis

The levels of complement C3 and C4 components were determined in non-indigenous (creoles) and indigenous (Warao) populations, the latter with an extremely high tuberculosis (TB) rate. Serum samples from 209 adults were studied and classified in 4 groups taking into account tuberculin skin tests (TST): (1) the group of Warao patients (58 positive for the TST, WP TST+ and 9 negative for the TST, WP TST-), (2) the group of creole patients (34 positive for the TST, CP TST+ and 9 negative for the TST, CP TST-), (3) the group of healthy Warao controls (38 positive and 14 negative for TST, WC TST+ and WC TST-, respectively), (4) the creole controls (26 positive and 21 negative for the TST, CC TST+ and CC TST-, respectively). With respect to the results concerning the measurement of both complement C3 and C4 components with the exception of the WC TST and the CC groups, the WP TST+ and WP TST- as well as WC TST+ groups showed a significant frequency of individuals with decreased levels of complement C3 component (20.6, 33.3, and 26.3 percent, respectively) and also C4 component (12.0, 11.1, and 13.3 percent, respectively) in comparison to both creole patients (CP TST+, 8.82 percent and CP TST-, 0 percent and CP TST+, 5.88 percent and CP TST-, 0 percent) for C3 and C4, respectively. The study of these parameters carried out in 15 Warao subjects with active infection, before and after anti-TB chemotherapy,statisticallyconfirmedthat the effective chemotherapy did not restore normal levels of the complement C3 and C4 components among Warao patients. Aditional tests for hepatitis B or hepatitis C infection, and the profile of the hepatic proteins were not associated to the deficiency in production of the complement components.In conclusion, the results show that within the Warao population, a high percentage of subjects exhibit decreased levels of both complement C3 and C4 components independent of latent or active infection and the status of TST.

Infecciones en tejidos blandos por micobacterias no tuberculosas secundarias a mesoterapia. ¿Cuánto vale la belleza? / Soft tissue infections due to non-tuberculous mycobacteria following mesotherapy. What is the price of beauty?

Introducción. La mesoterapia se utiliza mucho en Latinoamérica con fines cosméticos, especialmente en pacientes con obesidad. En este estudio se describen las características clínicas y epidemiológicas, el diagnóstico microbiológico, el tratamiento y el seguimiento de infecciones por micobacterias no tuberculosas en un grupo de pacientes en Caracas (Venezuela) con antecedentes de mesoterapia. Metodología. Entre marzo de 2002 y diciembre de 2003 se evaluaron 49 pacientes con infección en la piel y tejidos blandos secundaria a mesoterapia. Se tomaron muestras de las lesiones para el aislamiento de micobacterias y se elaboró una ficha clínica. Además, se analizaron 15 productos utilizados en mesoterapia. Las micobacterias aisladas fueron identificadas a través del polimorfismo de fragmentos de restricción del gen hsp65. Resultados. De los 49 pacientes evaluados, en el 81,6% se confirmó una infección por micobacterias no tuberculosas. Las especies más comunes fueron Mycobacterium abscessus y M. fortuitum pero también se aislaron M. chelonae, M. peregrinum, M. simiae y una nueva especie que fue designada M. cosmeticum. Los pacientes recibieron tratamiento específico para cada especie durante un período de 3 hasta 18 meses. La investigación de la fuente de infección reveló que 21 pacientes estaban agrupados en tres brotes y se encontraron dos productos contaminados, uno con M. fortuitum y otro con M. abscessus. Conclusiones. Los médicos deben estar atentos ante aquellos pacientes con antecedentes de mesoterapia que desarrollen tardíamente lesiones en piel y tejidos blandos, que no respondan al tratamiento antimicrobiano convencional, ya que éstas podrían ser causadas por micobacterias no tuberculosas (AU)

Introduction. Mesotherapy is widely used In Latin America for cosmetic purposes, particularly in obese individuals. We describe the clinical and epidemiological characteristics, microbiological diagnosis, treatment and follow-up of patients from Caracas (Venezuela) with soft tissue infection caused by non-tuberculous mycobacteria following mesotherapy. Methods. Between March 2002 and December 2003, we evaluated 49 cases of skin and soft tissue infection following mesotherapy. Specimens obtained from the lesions and 15 products used in the mesotherapy procedure were cultured for the presence of non-tuberculous mycobacteria. Isolated mycobacteria were identified by PCR restriction fragment length polymorphism analysis of the hsp65 gene. Results. Infection by non-tuberculous mycobacteria was confirmed in 81.6% of the 49 cases. Mycobacterium abscessus and M. fortuitum were the most common species, but M. chelonae, M. peregrinum, M. simiae and a new species that was designated "M. cosmeticum" were also isolated. Patients were treated with species-specific antibiotic agents for 3 to 18 months. Investigation into the source of the infection revealed that 21 patients were clustered within 3 different outbreaks and two products were found to be contaminated with M. fortuitum and M. abscessus, respectively. Conclusions. Physicians should be alerted to the possibility of infection by non-tuberculous mycobacteria in patients with a history of mesotherapy who develop late-onset skin and soft tissue infection, particularly if they do not respond to conventional antibiotic treatment (AU)

Inactivación de micobacterias con desinfectantes registrados como tuberculicidas / Inactivation of mycobacteria by disinfectants with a tuberculocidal label

Introducción. Se evaluó la actividad micobactericida de desinfectantes disponibles en el mercado venezolano, registrados como tuberculicidas y que son utilizados en la clínica y el laboratorio para la desinfección de materiales críticos y semicríticos. Materiales y métodos. Se utilizó la Prueba de Suspensión Cuantitativa de la Comunidad Europea prEN 14348 bajo circunstancias limpias y sucias, y se evaluó la actividad de Gerdex® y K-ller® (bromuro de lauril dimetil bencil amonio, en concentraciones de 10 y 0,16%, respectivamente), Microplus-Action® (5% polimetilen diurea), Cidex® (2% glutaraldehído) y Cidex® OPA (0,55% orthophthaldialdehído). La actividad micobactericida se evaluó con las cepas de referencia Mycobacterium smegmatis ATCC 19420, y con cepas de M. fortuitum, M. chelonae y M. abscessus aisladas de muestras clínicas. La actividad tuberculocida se evaluó con M. tuberculosis H37Rv. Resultados. Cidex y Cidex OPA fueron efectivos frente a todas las micobacterias evaluadas bajo condiciones limpias y sucias con un > 5-Log10 de reducción de viabilidad después de 5 min de exposición. Gerdex y K-ller lograron solamente un 2-Log10 de reducción con M. tuberculosis, M. abscessus y M. chelonae, aun después de 60 min de exposición. El Microplus-Action mostró un 4-Log10 de reducción de viabilidad con M. tuberculosis, pero menos de 1-Log10 de reducción con M. abscessus y M. chelonae después 60 min de exposición. Conclusiones. La actividad tuberculicida y micobactericida de varios desinfectantes disponibles en Venezuela, Gerdex®, K-ller® y Microplus-Action®, que indican sus fabricantes resulta inadecuada cuando es evaluada según las normas europeas. El uso de estos compuestos para la desinfección de materiales críticos y semicríticos podría dar como resultado infecciones iatrogénicas (AU)

Introduction. Disinfectants available in Venezuela, registered as tuberculocidal, and used in clinical and laboratory settings were tested for their mycobactericidal activity. Methods. The European Standard Quantitative Suspension test prEN14348 was used in simulated clean and dirty conditions. The disinfectants tested included Gerdex® and K-ller® (both containing the quaternary ammonium compound dimethyl benzyl lauryl ammonium bromide in concentrations of 10% and 0.16% respectively), Microplus-Action® (5% polymethylene urea), Cidexì (2% glutaraldehyde) and Cidex® OPA (0.55% ortho-phthalaldehyde). The mycobactericidal activity was evaluated with M. smegmatis ATCC 19420 and in clinical isolates of M. fortuitum, M. chelonae and M. abscessus. The tuberculocidal activity was assessed with M. tuberculosis H37Rv. Results. Cidex and Cidex OPA were effective against all the test organisms, showing a > 5-log 10 reduction in CFU after 5 minutes of exposure under both clean and dirty conditions. Gerdex and K-ller only achieved a 2-log reduction in M. tuberculosis, M. abscessus and M. chelonae CFU, even after 60 minutes of exposure. Micro Plus Action showed a 4-log reduction in M. tuberculosis viability but less than a 1-log reduction in M. abscessus and M. chelonae CFUs after 60 minutes of incubation. Conclusion. The tuberculocidal activity claimed by the manufacturers of several disinfectants sold in Venezuela, Gerdex®, K-ller® and Microplus-Action® is inaccurate when tested by European norms. The use of these products for disinfecting material used in critical and semi-critical patients could result in iatrogenic infections (AU)

Tuberculosis en menores de 15 años en la población Warao de Venezuela / Tuberculosis in subjects under 15 years of age in the population of Warao in Venezuela

It is difficult to establish a definitive diagnosis of tuberculosis in rural areas where there is no access to a large hospital. The Warao people of the Delta Amacuro State in Venezuela, have a very high prevalence of adult TB, and we suspected that the Warao children would also have a high prevalence of the disease, almost entirely undiagnosed. We applied a simple methodology to select children suspicious for tuberculosis that is based on a rating system using clinical criteria, reactivity to tuberculin and intradomicilliary contacts. Of the 502 children under the age of 15 that were evaluated with this rating system, 27 were determined to be suspicious of TB and were further evaluated by a chest X-ray. Radiologic confirmation of TB was found in 16 (60) of the 27 suspicious children. Of these 16 patients, 13 (81) were PPD positive and 3 were PPD negative. Additionally, 7 of the 16 children with pathologic x-ray changes had one or more confirmatory findings: 3 were positive by culture or smear examination and 5 had a positive serologic B diagnostic test. In conclusion this methodology proved to be highly efficient in diagnosing childhood tuberculosis in this population, and should also be useful in other rural populations with a high prevalence of adult TB.