Evaluation of the National Case Report Form for dengue and timely case notification before the co-circulation of multiple arboviruses in Barranquilla, Colombia / Evaluación del formulario nacional de informe de caso para dengue y de la puntualidad en la notificación del caso antes de la co-circulación de múltiples arbovirus en Barranquilla-Colombia

ABSTRACT Objective To evaluate the case report forms and times elapsed between the surveillance steps for dengue virus (DENV) infection in a large Colombian city before the emergence of other arbovirus epidemics. Materials and Methods The descriptive epidemiology of DENV infection cases was analyzed from 2009 to 2013. The completeness of the case report forms filed at the Primary Units of Data Generation (PUDG) were evaluated, as well as the accuracy and suitability of the tests (PPV: positive predictive value). The average time-lags between each step were then calculated. Results There were 7.3, 12.38, 4.66, 6.25 and 29.9 annual cases of dengue infection per 10 000 inhabitants in 2009 to 2013, respectively. In this study, only 57.76% of the cases were classified correctly by the physicians and 26.32% of them were questioned about their home conditions and whether their family/friends had similar symptoms. Patients visited a clinic/hospital on average 4.76 days after developing symptoms and the health system was notified on average 1.75 days later, while 70.6% of them were reported within the one-day target period. There were only minor changes in case reporting times even during a DENV epidemic. Some (12.85%) of the case forms were later modified (average 16.7 days). In the period 2009-2013, the IgM confirmed PPV was 58.60%, while 20 mandatory criteria were absent on more than 25% of the forms. Conclusions The system was accurate, simple, flexible, stable and acceptable, but a number of ways are suggested to improve this case detection and reporting system.(AU)

RESUMEN Objetivo Evaluar los formularios de informe de casos y los tiempos entre los pasos de vigilancia para el dengue en una ciudad colombiana antes de la aparición de otras epidemias de arbovirus. Materiales y Métodos Se analizó la epidemiología descriptiva entre 2009 y 2013. Se evaluó la integridad de los formularios de informes de casos, registrados en las Unidades Primarias de Generación de Datos, así como el valor predictivo (VPP) de las pruebas diagnósticas. Se calcularon los intervalos de tiempo promedio entre cada paso de la vigilancia. Resultados Hubo 7.3, 12.38, 4.66, 6.25 y 29.9 casos anuales por cada 10 000 habitantes en 2009-2013, respectivamente. Solo el 57.76% de los casos fueron clasificados correctamente por los médicos, el 26.32% de ellos fueron interrogados sobre las condiciones de su hogar y si sus familiares/amigos tenían síntomas similares. Los pacientes se presentaron a una clínica/hospital en promedio 4.76 días después de desarrollar síntomas y el sistema de salud fue notificado en promedio 1.75 días más tarde, mientras que el 70.6% de ellos se informaron dentro del período objetivo de un día. Algunos (12.85%) de los formularios de casos se modificaron posteriormente (promedio de 16.7 días). Desde 2009-2013, el VPP confirmado por IgM fue de 58.60%, mientras que veinte criterios obligatorios estuvieron ausentes en más del 25% de los formularios. Conclusiones El sistema fue preciso, simple, flexible, estable y aceptable, pero sugerimos varias formas de mejorar este sistema de detección e informe de casos.(AU)

Evaluation of the National Case Report Form for dengue and timely case notification before the co-circulation of multiple arboviruses in Barranquilla, Colombia.

OBJECTIVE: To evaluate the case report forms and times elapsed between the surveillance steps for dengue virus (DENV) infection in a large Colombian city before the emergence of other arbovirus epidemics. MATERIALS AND METHODS: The descriptive epidemiology of DENV infection cases was analyzed from 2009 to 2013. The completeness of the case report forms filed at the Primary Units of Data Generation (PUDG) were evaluated, as well as the accuracy and suitability of the tests (PPV: positive predictive value). The average time-lags between each step were then calculated. RESULTS: There were 7.3, 12.38, 4.66, 6.25 and 29.9 annual cases of dengue infection per 10 000 inhabitants in 2009 to 2013, respectively. In this study, only 57.76% of the cases were classified correctly by the physicians and 26.32% of them were questioned about their home conditions and whether their family/friends had similar symptoms. Patients visited a clinic/hospital on average 4.76 days after developing symptoms and the health system was notified on average 1.75 days later, while 70.6% of them were reported within the one-day target period. There were only minor changes in case reporting times even during a DENV epidemic. Some (12.85%) of the case forms were later modified (average 16.7 days). In the period 2009-2013, the IgM confirmed PPV was 58.60%, while 20 mandatory criteria were absent on more than 25% of the forms. CONCLUSIONS: The system was accurate, simple, flexible, stable and acceptable, but a number of ways are suggested to improve this case detection and reporting system.

Estimate of the reproduction number of the 2015 Zika virus outbreak in Barranquilla, Colombia, and estimation of the relative role of sexual transmission.

BACKGROUND: In 2015, the Zika arbovirus (ZIKV) began circulating in the Americas, rapidly expanding its global geographic range in explosive outbreaks. Unusual among mosquito-borne diseases, ZIKV has been shown to also be sexually transmitted, although sustained autochthonous transmission due to sexual transmission alone has not been observed, indicating the reproduction number (R0) for sexual transmission alone is less than 1. Critical to the assessment of outbreak risk, estimation of the potential attack rates, and assessment of control measures, are estimates of the basic reproduction number, R0. METHODS: We estimated the R0 of the 2015 ZIKV outbreak in Barranquilla, Colombia, through an analysis of the exponential rise in clinically identified ZIKV cases (n=359 to the end of November, 2015). FINDINGS: The rate of exponential rise in cases was ρ=0.076days-1, with 95% CI [0.066,0.087] days-1. We used a vector-borne disease model with additional direct transmission to estimate the R0; assuming the R0 of sexual transmission alone is less than 1, we estimated the total R0=3.8 [2.4,5.6], and that the fraction of cases due to sexual transmission was 0.23 [0.01,0.47] with 95% confidence. INTERPRETATION: This is among the first estimates of R0 for a ZIKV outbreak in the Americas, and also among the first quantifications of the relative impact of sexual transmission.

A simple, inexpensive, robust and sensitive dot-blot assay for equal detection of the nonstructural-1 glycoprotein of all dengue virus serotypes.

BACKGROUND: Detection of dengue virus (DENV) soluble/excreted (s/e) form of the nonstructural-1 (NS1) glycoprotein in patient acute-phase sera is ideal for diagnosis. The commercially-available detection assays are, however, too expensive for routine use and have low specificity, particularly for the s/e NS1 glycoprotein of DENV-2 and DENV-4, which are important causes of lethal human disease worldwide. METHODS: Mouse monoclonal antibodies (MAbs) were generated and screened against s/e NS1 glycoprotein purified from each DENV serotype to obtain those that reacted equally with each serotype, but not with yellow fever virus (YFV) s/e NS1 glycoprotein or human serum proteins. One MAb, MAb 2C4.6, was further tested against these DENV glycoproteins in human sera using simple, peroxidase-labelled secondary antibody/substrate-developed dot-blot assays. RESULTS: Optimal quenching of endogenous human serum peroxidases was attained using 3% H(2)O(2) in H(2)0 for 5 min. MAb 2C4.6 showed an acceptable detection sensitivity of < 32 ng/ml for the s/e NS1 glycoprotein of each DENV serotype but did not cross-react with the YFV s/e NS1 glycoprotein or human serum proteins. By contrast, the LX1 epitope-specific MAb, 3D1.4, showed similar detection sensitivity against only the DENV-1 NS1 glycoprotein, consistent with results from commercial DENV s/e NS1 glycoprotein detection assays.DENV s/e NS1 glycoproteins were stable in human sera after drying on the nitrocellulose membranes and storage for one month at ambient temperature (28°C) before being processed. The total assay time was reduced to 3 h without any loss of detection sensitivity. This dot-blot format was ideal for the circulating immune complex disruption step, which is required for increased DENV s/e NS1 glycoprotein detection. CONCLUSIONS: This is the first study to determine the detection sensitivity of MAbs against known concentrations of s/e NS1 glycoprotein from each DENV serotype. The preparation of patient serum samples for dot-blot assays can be performed by staff with a basic level of training and storage at low temperatures (e.g., -80°C) is not necessary. These simple, inexpensive (US$ 0.05/sample), robust, sensitive and relatively rapid assays, using improved MAbs such as MAb 2C4.6, should be ideal for the diagnosis of all DENV serotypes in DENV endemic regions.

Cross-sectional study of Leptospira seroprevalence in humans, rats, mice, and dogs in a main tropical sea-port city.

Samples were collected from 128 symptomatic humans, 83 dogs, 49 mice, and 20 rats (Rattus rattus: 16; Rattus norvegicus: 4) in neighborhoods where human leptospirosis have been reported within the principal sea-port city of Colombia. Seroprevalences were assessed against 19 pathogenic, 1 intermediate pathogenic, and 1 saprophytic Leptospira serogroups. Pathogenic Leptospira were confirmed using conventional Leptospira-specific polymerase chain-reaction and pulsed-field gel electrophoresis analysis was used for serovar identification. Seroprevalences of 20.4%, 12.5%, 25.0%, 22.9%, and 12.4% were obtained against one to seven different serogroups in mice, R. rattus, R. norvegicus, dogs, and humans, respectively. The DNA was confirmed to be from pathogenic Leptospira by detecting the lipL32 gene in 12.5%, 3.7%, and 0.03% of the R. rattus, dog, and human samples, respectively. The first genetically typed Colombian isolate was obtained from a rat and identified as Leptospira interrogans serovar Icterohaemorrhagiae/Copenhageni.

Simple Prognostic Criteria can Definitively Identify Patients who Develop Severe Versus Non-Severe Dengue Disease, or Have Other Febrile Illnesses.

BACKGROUND: SEVERE DENGUE DISEASE (SDD) (DHF/DSS: dengue hemorrhagic fever/dengue shock syndrome) results from either primary or secondary dengue virus (DENV) infections, which occur 4 - 6 days after the onset of fever. As yet, there are no definitive clinical or hematological criteria that can specifically identify SDD patients during the early acute febrile-phase of disease (day 0 - 3: < 72 hours). This study was performed during a SDD (DHF/DSS) epidemic to: 1) identify the DENV serotypes that caused SDD during primary or secondary DENV infections; 2) identify simple clinical and hematological criteria that could significantly discriminate between patients who subsequently developed SDD versus non-SDD (N-SDD), or had a non-DENV fever of unknown origin (FUO) during day 0 - 3 of fever; 3) assess whether DENV serotype co-infections resulted in SDD. METHODS: First serum samples, with clinical and hematological criteria, were collected from 100 patients during the early acute febrile-phase (day 0 - 3: < 72 hours), assessed for DENV or FUO infections by IgM- and IgG-capture ELISAs on paired serum samples and by DENV isolations, and subsequently graded as SDD, N-SDD or FUO patients. RESULTS: IN THIS STUDY: 1) Thirty-three patients had DENV infections, predominantly secondary DENV-2 infections, including each SDD (DHF/DSS) case; 2) Secondary DENV-2/-3 and DENV-2/-4 serotype co-infections however resulted in N-SDD; 3) Each patient who subsequently developed SDD, but none of the others, displayed three clinical criteria: abdominal pain, conjunctival injection and veni-puncture bleeding, therefore each of these criteria provided definitively significant prognostic (P < 0.001) values; 4) Petechia, positive tourniquet tests and hepatomegaly, and neutrophilia or leukopenia also significantly identified those who: a) subsequently developed SDD versus N-SDD, or had a FUO; b) subsequently developed SDD versus N-SDD; c) subsequently developed N-SDD versus FUOs, respectively. CONCLUSIONS: This is the first report of simple definitively prognostic criteria for SDD patients, including the first assessment and confirmation of conjunctival injection. The three definitive clinical criteria used alone, or supported by the other four criteria, could be essential for specifically identifying those patients needing prompt hospital-based therapies to lessen or avert SDD, without unnecessary hospitalization of the other patients. KEYWORDS: Dengue virus; Severe dengue; Dengue fever; Diagnostic; Criteria; Hemorrhage; Shock.

[Cutaneous myiasis by Cochliomyia hominivorax (Coquerel) (Diptera Calliphoridae) in Hospital Universidad del Norte, Soledad, Atlántico]. / Miasis cutánea por Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae) en el Hospital Universidad del Norte, Soledad, Atlántico.

Human myiasis is the parasitism of human tissues by fly larvae. Diagnoses are based on clinical pattern of tissue damage and presence of insect stages. Herein, a case myiasis is described in a seven-year-old female child. She presented with fever associated with abscessed scalp lesions containing exposed larvae. Severe pediculosis was also observed. The patient was hospitalized and treated with clindamycin, gentamicin (for bacterial secondary infections) and ivermectin (treatment for lice) after which the patient showed clinical improvement and was discharged four days later. Since human myiasis can be caused by a number of different species, larvae were collected from the patient and identified as those of Cochliomyia hominivorax (Diptera: Calliphoridae). Because other cases of coinfestation of flies and lice are on record, health workers are to be alerted about the possible pediculosis-myasis risk.

Miasis cutánea por Cochliomyia hominivorax (Coquerel) (Díptera: Calliphoridae) en el Hospital Universidad del Norte, Soledad, Atlántico / Cutaneous myiasis by Cochliomyia hominivorax (Coquerel) (Díptera Calliphoridae) in Hospital Universidad del Norte, Soledad, Atlántico

La miasis humana es el parasitismo de órganos y tejidos producido por especies de larvas del orden Díptera. El diagnóstico se realiza con base en hallazgos clínicos y se confirma con estudios entomológicos. Se presenta el caso de una niña de siete años de edad que fue llevada por su padre al servicio de urgencias por presentar fiebre asociada a una lesión abscedada en el cuero cabelludo, con salida espontánea de larvas. Como hallazgo en el examen físico se reportó pediculosis grave. La paciente fue hospitalizada y tratada con clindamicina, gentamicina e ivermectina, y mostró mejoría de sus condiciones clínicas. Se dio alta médica al cuarto día de estancia hospitalaria. Se recolectaron larvas en estadio dos de Cochliomyia hominivorax (Diptera: Calliphoridae) directamente del área lesionada, observándose la asociación miasis-pediculosis; por lo tanto, se alerta a los trabajadores del área de la salud del riesgo potencial que representa la pediculosis para el desarrollo de la miasis.

Human myiasis is the parasitism of human tissues by fly larvae. Diagnoses are based on clinical pattern of tissue damage and presence of insect stages. Herein, a case myiasis is described in a seven-year-old female child. She presented with fever associated with abscessed scalp lesions containing exposed larvae. Severe pediculosis was also observed. The patient was hospitalized and treated with clindamycin, gentamicin (for bacterial secondary infections) and ivermectin (treatment for lice) after which the patient showed clinical improvement and was discharged four days later. Since human myiasis can be caused by a number of different species, larvae were collected from the patient and identified as those of Cochliomyia hominivorax (Diptera: Calliphoridae). Because other cases of coinfestation of flies and lice are on record, health workers are to be alerted about the possible pediculosis-myasis risk.

Altered enzyme-linked immunosorbent assay immunoglobulin M (IgM)/IgG optical density ratios can correctly classify all primary or secondary dengue virus infections 1 day after the onset of symptoms, when all of the viruses can be isolated.

We compared dengue virus (DV) isolation rates and tested whether acute primary (P) and acute/probable acute secondary (S/PS) DV infections could be correctly classified serologically when the patients' first serum (S1) samples were obtained 1 to 3 days after the onset of symptoms (AOS). DV envelope/membrane protein-specific immunoglobulin M (IgM) capture and IgG capture enzyme-linked immunosorbent assay (ELISA) titrations (1/log(10) 1.7 to 1 log(10) 6.6 dilutions) were performed on 100 paired S1 and S2 samples from suspected DV infections. The serologically confirmed S/PS infections were divided into six subgroups based on their different IgM and IgG responses. Because of their much greater dynamic ranges, IgG/IgM ELISA titer ratios were more accurate and reliable than IgM/IgG optical density (OD) ratios recorded at a single cutoff dilution for discriminating between P and S/PS infections. However, 62% of these patients' S1 samples were DV IgM and IgG titer negative (or=2.60 and <2.60) discriminatory IgM/IgG OD (DOD) ratios on these S1 samples than those published previously to correctly classify the highest percentage of these P and S/PS infections. The DV isolation rate was highest (12/12; 100%) using IgG and IgM titer-negative S1 samples collected 1 day AOS, when 100% of them were correctly classified as P or S/PS infections using these higher DOD ratios.

Investigation of relationships between Aedes aegypti egg, larvae, pupae, and adult density indices where their main breeding sites were located indoors.

Aedes aegypti (L.) density indices obtained in a dengue fever (DF) endemic area were compared. One hundred and twenty premises, in an urban area of Colombia where dengue type-1 and type-2 virus cocirculated, were randomly selected and sampled for 7 months. The geometric mean monthly numbers (density index, DI) of Ae. aegypti eggs (ODI), 4th instar larvae (LDI), pupae (PDI), and adults (ADI) were calculated based on the use of ovitraps, nets, and manual aspirators, respectively. A negative temporal correlation was observed between the LDI and the ODI (r = -0.83, df = 5, and P < 0.01). Positive temporal correlations were only observed between the LDI and the PDI (r = 0.90, df = 5, and P < 00.5) and the Breteau and House indices (r = 0.86, df = 5, and P < 0.01). No other correlations were found between these indices and any of the other density indices or the incidence of suspected DF cases in residents, the temperature, the rainfall, or seasonal fluctuations. Our results were, therefore, probably due to the most productive Ae. aegypti breeding sites (large water containers) being located indoors within this study area. The number of adult female Ae. aegypti/person (n = 0.5) and pupae/person (n = 11) in our study area were lower and dramatically higher than the transmission thresholds previously reported for adult and pupae, respectively. Because there were confirmed DF cases during the study period, the transmission threshold based on the Ae. aegypti pupae was clearly more reliable. We found that the mean ovitrap premise index (OPI) was 98.2% during this study and that the mean larval (L-4th instars) premise index (LPI) was 59.2%, and therefore we suggest that the OPI and LPI would be more sensitive methods to gauge the effectiveness of A. aegypti control programs.

An inexpensive intervention for the control of larval Aedes aegypti assessed by an improved method of surveillance and analysis.

A sampling method coupled with statistical calibration factors was developed to accurately assess the numbers of larvae and pupae of Aedes aegypti in large water-storage containers of variable capacities and water levels. Aedes aegypti productivity in different types of breeding sites found in an urban study area in central Colombia was assessed and compared. In this study, water-storage tanks and drums were found to comprise 79% of the containers positive for larval Ae. aegypti, which contributed to 93 and 92% of the total production of populations of 4th-stage larvae and pupae, respectively. These main breeding sites of Ae. aegypti were found at an indoor to outdoor ratio of 2.4:1 and no correlation was found between temporal fluctuation of populations of larval Ae. aegypti and monthly rainfall. Netted lids that used inexpensive local materials were designed to prevent oviposition by Ae. aegypti. During a 6-month trial period, 56% of inspected containers had netted lids correctly in place. Of these, 78% had no mosquito larvae. Because only 37% of uncovered containers were free of mosquito larvae, a significant difference was demonstrated when these inexpensive mechanical barriers were used (chi2 = 138.7; P < 0.001). These netted lids and the improved methods described to assess the productivity of larval and pupal Ae. aegypti in this study are now being used in combination with other strategies to assess and control these populations of dengue virus vectors in the main port city on the Atlantic Coast of Colombia.