Comparison of a modified shell vial culture procedure with conventional mouse inoculation for rabies virus isolation

Rabies is a neurotropic disease that is often lethal. The early diagnosis of rabies infection is important and requires methods that allow for the isolation of the virus from animals and humans. The present study compared a modified shell vial (MSV) procedure using 24-well tissue culture plates with the mouse inoculation test (MIT), which is considered the gold standard for rabies virus isolation. Thirty brain samples (25 positive and 5 negative by the fluorescent antibody test) obtained from different animal species at the National Institute of Hygiene Rafael Rangel in Caracas, Venezuela, were studied by the MIT and MSV assays. Nine samples (36%) were positive at 24 h, 10 (40%) were positive at 48 h and six (24%) were positive at 72 h by the MSV assay. With the MIT assay, 76% were positive at six days post inoculation and 12% were positive at 12 and 18 days post inoculation. One sample that was negative according to the MSV assay was positive with MIT on the 12th day. The MSV procedure exhibited a sensitivity of 96.2%, a specificity of 100%, a positive predictive value of 100% and a negative predictive value 80%. This procedure allowed for rapid rabies virus detection. MIT can be employed as an alternative method in laboratories without tissue culture facilities.

Conservación y uso de los recursos genéticos de Annonaceae en México

Se presenta un análisis sobre la conservación y el uso de los recursos genéticos de especies frutales de la familia Annonaceae en México. El diagnóstico realizado en 2010 por la Red de Anonáceas (REMA) del Sistema Nacional de Recursos Fitogenéticos (SINAREFI) sirvió para tal fin y fue complementado por estudios recientes. Partimos de una revisión de registros de herbarios de la Red Mundial de Información sobre Biodiversidad (REMIB) además de recoger información con productores de comunidades rurales y recorridos de campo en estados del país donde las anonáceas se distribuyen. En México se encuentran 14 géneros y 63 especies de Annonaceae distribuidas principalmente en regiones tropicales del Sureste de México. La conservación in situ es esporádica, se está dando en huertos de traspatio para la guanábana (Annona muricata), chirimoya (A. cherimola), chincuya (A. purpurea) y saramuyo (A. squamosa) principalmente. La anona (A. reticulata), ilama (A. diversifolia) y chincuya (A. purpurea) son fomentadas pero no multiplicadas. La conservación ex situ se mantiene en bancos de germoplasma in vivo o colecciones de trabajo para guanábana (30 colectas), chirimoya (70 colectas) e ilama (100 colectas) solamente. Las semillas de estas especies no son ortodoxas y su conservación a mediano y largo plazo no está resuelta en México. El potencial de cultivo para este grupo de frutales es alto pero problemas de falta de variedades, fitosanidad, falta de tecnología y mercados reducidos limitan el cultivo comercial.

Uso combinado de 1-Meticiclopropeno y emulsiones de cera en la conservación de guanábana (Annona muricata)

El objetivo de este trabajo fue evaluar la combinación del 1-metilciclopropeno (1-MCP) y emulsiones de cera sobre la conservación de guanábana. Las condiciones de almacenamiento fueron: 25 ºC y 16 ºC, frutos con emulsiones a base de cera de candelilla o cera de abeja diluídas con agua (10:90 v/v y 15:85 v/v) almacenados a 16 ºC, frutos tratados con 1-MCP (1500 nL L-1, 12 h). En los frutos a 16 ºC con y sin emulsiones se observó daño por frío y no maduraron. En los frutos con la aplicación de 1-MCP solo o combinado con emulsiones, en cualquiera de las dilusiones, no se observaron síntomas daño por frío en la pulpa. La combinación de 1-MCP y emulsión a base de cera de abeja en dilución 15:85 v/v conservaron a los frutos de guanábana por 14-15 días en comparación con los frutos almacenados a 25 ºC (6 días).

Influencia de la deshidratación osmótica y de la adición de cloruro de calcio en la conservación de kivis minimamente procesados

Este trabajo tubo como objetivo estudiar el efecto combinado de la deshidratación osmótica y la influencia del cloruro de calcio en rodajas de kivi sometidos al proceso mínimo. Muestras con y sin deshidratación osmótica fueron almacenadas a 5 °C en embalajes PET. La adición de cloruro de calcio fue realizada durante la deshidratación osmótica. Se evaluaron la pérdida de peso, acidez, sólidos solubles, pH, humedad, coliformes, hongos y levaduras en las muestras hasta 15 días de almacenamiento. La deshidratación osmótica consistió en la inmersión de las rodajas de kivi en solución de sacarosa a 60% y en solución de sacarosa (60%) con adición de cloruro de calcio (0,1 M), ambos tratamientos se realizaron a temperatura ambiente (25 °C) por 24 horas y relación fruta:solución de 1:5. Los resultados mostraron que el pré-tratamiento osmótico con adición de cloruro de calcio aumentó la vida útil hasta 15 días, en cuanto las rodajas tratadas por osmodeshidratación sin adición de sal presentaron vida útil de 12 días. Sensorialmente, los consumidores prefirieron las rodajas de kivi procesadas con pré-tratamiento osmótico y adición de cloruro de calcio.

Madurez fisiológica, germinación y conservación de semillas de guayabito (Psidium cuneatum Camb. - Myrtaceae)

Psidium cuneatum Camb., "guayabito o arasá blanco" es una especie nativa de la provincia de Misiones y norte de Corrientes que tiene importancia ecológica y potencial interés comercial. Actualmente está afectada por el avance de la forestación comercial. El objetivo de este trabajo fue analizar el comportamiento germinativo de sus semillas según la madurez de los frutos, las condiciones de germinación y el almacenamiento en diferente ambientes. En individuos marcados y observados semanalmente durante 2 años, se estableció que la fructificación se produce desde diciembre a julio y que los frutos persisten en el árbol durante un período prolongado antes de dispersarse. La germinación de semillas procedentes de frutos de diferentes tamaños, evaluada a través del índice de velocidad de germinación y del porcentaje de germinación permitió determinar que las semillas procedentes de frutos con diámetro mayor a 1.5 cm exhiben mejor comportamiento germinativo, por lo que el tamaño del fruto es un buen indicador de madurez en esta especie. Por otra parte, se estudió el efecto que el régimen de temperatura tiene sobre la germinación, comprobándose que a 27ºC el porcentaje y la velocidad de germinación fueron significativamente superiores. Para establecer las condiciones para la conservación de las semillas se procedió a almacenarlas durante 30, 90, 180, y 270 días, combinando tipo de recipiente y temperatura. Se comprobó que el envase hermético fue más efectivo para la conservación del poder germinativo cuando se utilizó a baja temperatura, aunque se comprobó una reducción acentuada del poder germinativo, el cual fue de 32% a los 180 días.

Teores de antocianinas, fenóis totais, taninos e ácido ascórbico em uva 'bordô' sobre diferentes porta-enxertos

Este trabalho teve como objetivo avaliar o conteúdo de antocianinas, fenóis totais, taninos e ácido ascórbico, em variedade de uva 'Bordô', cultivada sobre os porta-enxertos '420A', 'IAC-766' e 'Ripária-do-Traviú', em três períodos de colheita distintos. O experimento foi realizado em vinhedo de uva 'Bordô', no município de Missal, no Oeste do Paraná, na safra 2007/2008. O efeito dos porta-enxertos foi avaliado em três períodos de colheita, a partir da época de floração: 90 dias, com bagas em amadurecimento, tendendo ao roxo definitivo; 100 dias, com bagas totalmente roxas; e 110 dias, com bagas em sobrematuração. Foi utilizado o delineamento em blocos casualizados, em parcelas subdivididas no tempo. As variáveis analisadas foram sólidos solúveis, acidez titulável, índice de fenóis totais, antocianinas, taninos e teor de ácido ascórbico. Os resultados mostraram que, nos porta-enxertos '420A' e 'IAC-766', a uva 'Bordô' apresentou respostas similares em acúmulos de antocianinas, taninos, acidez titulável e sólidos solúveis, mas os acúmulos de fenóis totais, tanto na polpa, quanto na epiderme das bagas, foram maiores no '420A'. Ocorreram menores acúmulos de antocianinas, fenóis totais e taninos da epiderme da uva 'Bordô', quando enxertada em 'Ripária-do-Traviú'. Acúmulos superiores de ácido ascórbico foram verificados nas uvas enxertadas em 'IAC-766'. Há evidências que apontam o porta-enxerto 'Ripária-do-Traviú' como inferior aos demais, nos aspectos nutricional e enológico.

Experiencia de enseñanza-aprendizaje: cambio de creencias y conductas en el manejo del agua de consumo

Se propuso modificar las creencias y conductas de los niños con respecto al manejo del agua de consumo, utilizando una experiencia de enseñanza-aprendizaje, con mostración al microscopio del agua contaminada. Se administró una pre-encuesta estructurada y una post-encuesta (resolución de problemas) a 63 niños (28 varones y 35 mujeres) de sexto y séptimo grado de la escuela primaria. Estos niños provenían, en su totalidad, de una población marginal del norte del Gran Buenos Aires, Argentina. El promedio de edad fue 12 años 8 meses y el ED = 1 año 1 mes. Cada conducta adecuada fue evaluada con un punto: higiene y protección de los recipientes de recolección y almacenamiento, tratamiento del agua (hervido o clorado) y periodo de conservación y cada creencia óptima, dos puntos (evitar la contaminación que produce enfermedad). Se determinó el número de alumnos que modificaron sus conductas y creencias en la post-encuesta y se halló si era significativa la diferencia por la prueba de Chi cuadrado (X²). Se comprobó que los niños, en general, no mejoraron sus conductas y creencias. Se mantuvieron las conductas adecuadas de lavado; no se modificaron las de protección del recipiente en la recolección del agua y se mantuvieron comportamientos inadecuados en el almacenamiento. Sólo se adquirieron conductas correctas en el tratamiento, a través del hervido y más específicamente del clorado (P < 0,01). Las creencias no se modificaron. El 30% de alumnos señaló todos los pasos correctos en el manejo del agua y el 18% sólo le faltó señalar la protección del envase en la post-encuesta. En próximas acciones educativas se deberá reforzar las conductas adecuadas de lavabo y clorado y revertir las erróneas del periodo de conservación y el momento de clorado y a su vez, sustentar estos procedimientos correctos con sólidos conocimientos de los procesos de contaminación, haciéndolos extensivos a otros aspectos del saneamiento básico.

Outbreak of persistent cutaneous abscesses due to Mycobacterium chelonae after mesotherapy sessions, Lima, Peru

Outbreaks of rapidly growing mycobacteria have been occasionally described. The article reports an outbreak of cutaneous abscesses due to Mycobacterium chelonae following mesotherapy in Lima, Peru. From December 2004 through January 2005, 35 subjects who had participated in mesotherapy training sessions presented with persistent cutaneous abscesses. Thirteen (37%) of these suspected cases consented to underwent clinical examination. Skin punch-biopsies were collected from suspicious lesions and substances injected during mesotherapy were analyzed. Suspected cases were mainly young women and lesions included subcutaneous nodules, abscesses and ulcers. Mycobacterium chelonae was isolated from four patients and from a procaine vial. In conclusion, it is important to consider mesotherapy as a potential source of rapidly growing mycobacteria infections.

Radiometric studies of mycobacterium tuberculosis

An in vitro assay system that included automated radiometric quantification of 14CO2 released as a result of oxidation of 14C- substrates was applied for studying the metabolic activity of M. tuberculosis under various experimental conditions. These experiments included the study of a) mtabolic pathways, b) detection times for various inoculum sizes, c) effect of filtration on reproducibility of results, d) influence of stress environment e) minimal inhibitory concentrations for isoniazid, streptomycin, ethambutol and rifampin, and f) generation times of M. tuberculosis and M. bovis. These organisms were found to metabolize 14C-for-mate, (U-14C) acetate, (U-14C) glycerol, (1-14C) palmitic acid, 1-14C) lauric acid, (U-14C) L-malic acid, (U-14C) D-glucose, and (U-14C) D-glucose, but not (1-14C) L-glucose, (U-14C) glycine, or (U-14C) pyruvate to 14CO2. By using either 14C-for-mate, (1-14C) palmitic acid, or (1-14C) lauric acid, 10(7) organisms/vial could be detected within 24 48 hours and as few as 10 organisms/vial within 16-20 days. Reproducible results could be obtained without filtering the bacterial suspension, provided that the organisms were grown in liquid 7H9 medium with 0.05% polysorbate 80 and homogenized prior to the study. Drugs that block protein synthesis were found to have lower minimal inhibitory concentrations with the radiometric method when compared to the conventional agar dilution method. The mean generation time obtained for M. bovis and different strains of M. tuberculosis with various substrates was 9 ± 1 hours.

Successful control program to implement the appropriate antibiotic prophylaxis for cesarean section

OBJECTIVE: Describe implementation of a successful program to reduce doses (cefazolin 2 to 1 g) used for antimicrobial prophylaxis. METHODS: Evaluation of an intervention program to reduce prophylactic antimicrobial doses. The intervention included weekly staff discussions, automatic dispensation of 1g-vial of cefazolin by the pharmacy unless expressly requested by surgeon and increase in post-discharge surveillance as a strategy to reassure surgeons of the safety of the reduction. In the pre and post intervention periods, a prospective study of antimicrobial consumption and surgical site infections were measured. RESULTS: There were 5,164 and 5,204 deliveries in 2001-2002 and 2003-2004, respectively; 1,524 (29.5%) and 1,363 (26%) were cesarean sections. There was a 45% decrease in cefazolin vials used on average per cesarean section (2.29 to 1.25). Patients evaluated increased from 16% to 67% and the SSI rates in both periods were 3.34% to 2.42%, respectively. CONCLUSION: An ample intervention, including administrative and educational measures, led to high compliance with dose reduction and saved more than US$4,000 in cefazolin, considered important because government reimbursement in Brazil for cesarean section is $80.

POLYCLONAL OUTBREAK OF BLOODSTREAM INFECTIONS CAUSED BY Burkholderia cepacia COMPLEX IN HEMATOLOGY AND BONE MARROW TRANSPLANT OUTPATIENT UNITS

Aim: The objective was to describe an outbreak of bloodstream infections by Burkholderia cepacia complex (Bcc) in bone marrow transplant and hematology outpatients.Methods: On February 15, 2008 a Bcc outbreak was suspected. 24 cases were identified. Demographic and clinical data were evaluated. Environment and healthcare workers' (HCW) hands were cultured. Species were determined and typed. Reinforcement of hand hygiene, central venous catheter (CVC) care, infusion therapy, and maintenance of laminar flow cabinet were undertaken. 16 different HCWs had cared for the CVCs. Multi-dose heparin and saline were prepared on counter common to both units.Findings: 14 patients had B. multivorans(one patient had also B. cenopacia), six non-multivorans Bcc and one did not belong to Bcc. Clone A B. multivorans occurred in 12 patients (from Hematology); in 10 their CVC had been used on February 11/12. Environmental and HCW cultures were negative. All patients were treated with meropenem, and ceftazidime lock-therapy. Eight patients (30%) were hospitalized. No deaths occurred. After control measures (multidose vial for single patient; CVC lock with ceftazidime; cleaning of laminar flow cabinet; hand hygiene improvement; use of cabinet to store prepared medication), no new cases occurred.Conclusions: This polyclonal outbreak may be explained by a common source containing multiple species of Bcc, maybe the laminar flow cabinet common to both units. There may have been contamination by B. multivorans (clone A) of multi-dose vials.

FACTORS ASSOCIATED TO ADHERENCE TO DIFFERENT TREATMENT SCHEMES WITH MEGLUMINE ANTIMONIATE IN A CLINICAL TRIAL FOR CUTANEOUS LEISHMANIASIS

The favorable outcome of the treatment of a disease is influenced by the adherence to therapy. Our objective was to assess factors associated with adherence to treatment of patients included in a clinical trial of equivalence between the standard and alternative treatment schemes with meglumine antimoniate (MA) in the treatment of cutaneous leishmaniasis (CL), in the state of Rio de Janeiro. Between 2008 and 2011, 57 patients with CL were interviewed using a questionnaire to collect socioeconomic data. The following methods were used for adherence monitoring: counting of vial surplus, monitoring card, Morisky test and modified Morisky test (without the question regarding the schedule); we observed 82.1% (vial return), 86.0% (monitoring card), 66.7% (Morisky test) and 86.0% (modified Morisky test) adherence. There was a strong correlation between the method of vial counting and the monitoring card and modified Morisky test. A significant association was observed between greater adherence to treatment and low dose of MA, as well as with a lower number of people sleeping in the same room. We recommend the use of the modified Morisky test to assess adherence to treatment of CL with MA, because it is a simple method and with a good performance, when compared to other methods.

Influence of light conditions on adult mortality and egg-laying of Triatoma brasiliensis neiva, 1911 (Hemiptera: Reduviidae: Triatominae)

We compared, for Triatoma brasiliensis, the egg-laying process and the mortality of adults under conditions of almost permanent darkness and with normal laboratory luminosity. Mortality did not differ between groups. The egg-laying per vial and per female was significantly greater in the group of normal luminosity. We consider that it is not recommendable to keep the adults of this species under complete darkness. Other biological aspects should be analysed in relation to luminosity.

Retrospective study of 151 patients with cutaneous leishmaniasis treated with meglumine antimoniate

We retrospectively analyzed a series of 151 cases of cutaneous leishmaniasis treated between 1967 and 1982. One-hundred-and-thirty-nine (92%) patients presented with active lesions and were treated with daily doses of meglumine antimoniate: 81 adults received a 5-ml vial IM and 58 children received 1 to 5ml. Forty-five (32.4%) patients underwent continuous treatment with meglumine antimoniate for 25 to 116 days without rest intervals, and 94 (67.6%) intermittent treatment with 2 to 5 series of meglumine antimoniate. Intermittent series could include schedules of daily IM applications for 10 to 25 days each and intervals varying from 10 to 60 days. Antimony dose was calculated for 66 (47.5%) patients and ranged from 3.9 to 28.7 Sb5+/kg/day. Of these, 35 patients received >10mg and 31 patients <10mg Sb5+/kg/day. Median time of healing was longer for lesions on the legs and feet - 67.5 days versus 48.7 days (p < 0.001) for other sites. However, there were no significant differences in the median time of healing between adults and children, intermittent and continuous regimens or high and low antimony doses. Fifty-one patients were reassessed 5 to 14 years after treatment and showed no evidence of disease. These results support further investigation (clinical trials) on treatment using low doses of antimony.

Oropharyngeal histoplasmosis: report of eleven cases and review of the literature

INTRODUCTION: Histoplasmosis is a systemic mycosis endemic in Brazil, especially in the State of Rio Grande do Sul, where Histoplasma capsulatum was isolated from the soil. H. capsulatum may compromise unusual areas, including the oropharynx, particularly in patients presenting disseminated histoplasmosis; which is associated with a state of immunosuppression, such as AIDS. METHODS: During database analysis of a total of 265 cases of histoplasmosis, the medical records of 11 patients with histological or microbiological diagnoses of oral histoplasmosis (OH) between 1987 and 2008 were retrospectively reviewed. RESULTS: This work reports 11 cases of OH, the majority presenting histopathological or microbiological evidence of disseminated histoplasmosis (DH). In the patients with DH, OH was the first manifestation of histoplasmosis. Five of the 11 patients discussed were HIV-seropositive with clinical and laboratory findings of AIDS. Four patients presented active pulmonary tuberculosis concomitant with histoplasmosis. Treatment was based on the use of itraconazole and amphotericin B deoxycholate. Eight patients responded successfully to therapy after one year, two did not come back for reevaluation and one died despite adequate therapy. CONCLUSIONS: Oral histoplasmosis is closely associated with immunosuppression status, especially in patients presenting AIDS; moreover, in many cases, OH is the first sign of disseminated histoplasmosis.