Danielle Malta Lima

Técnicas de extrações e procedimentos de clean-up para a determinação de hidrocarbonetos policílicos aromáticos (HPA) em sedimentos da costa do Ceará

Extraction and clean-up are essential points in polycyclic aromatic hydrocarbon (PAHs) analysis in a solid matrix. This work compares extraction techniques and clean-up procedures for PAH analysis. PAH levels, their toxicological significance and source were also evaluated in the waters of the Coco and Ceara rivers. The efficiency of PAH recovery was higher for the soxhlet and ultrasonic techniques. PAH recovery varied from 69.3 to 99.3%. Total PAH concentration (ΣHPA) varied from 720.73 to 2234.76 µg kg-1 (Coco river) and 96.4 to 1859.21 µg kg-1 (Ceara river). The main PAH sources are pyrolytic processes and the levels were classified as medium so that adverse effects are possible.

Hantavirus infection in suspected dengue cases from State of Ceará, Brazil

1. Curso de Medicina, Universidade de Fortaleza, Fortaleza, CE. 2. Departamento de Patologia e Medicina Legal, Universidade Federal do Ceara, Fortaleza, CE. 3. Centro de Pesquisa em Virologia, Faculdade de Medicina, Universidade de Sao Paulo, Ribeirao Preto, SP. 4. Setor de Virologia, Laboratorio Central de Saude Publica do Estado do Ceara, Fortaleza, CE. 5. Nucleo de Vigilância Epidemiologica, Secretaria Estadual de Saude do Estado do Ceara, Fortaleza, CE. 6. Departamento de Saude Publica, Universidade Federal do Ceara, Fortaleza, CE.

Relation factor: A new strategy for quality control in the determination of pesticides in environmental aqueous matrices

The effects promoted by environmental aqueous matrices on pesticide determinations have been assessed, and for the first time, a simple, low-cost and efficient strategy for the correction of analytical results has been determined. This method can be useful as a parameter of quality control in a quality assurance programs. Evaluation of the matrix effect showed that environmental aqueous matrices, e.g., estuarine water, promote a distinctive and significant effect on the determination of pesticides. The picloram, atrazine and methyl parathion pesticides suffered the smallest effects promoted by the estuarine matrix, whereas chlorpyrifos and cypermethrin suffer a significant effect. For picloram, the matrix effect was a function of its physiochemical properties. However, for atrazine, methyl parathion, chlorpyrifos and cypermethrin, the matrix effect was promoted by environmental matrix components. As strategy for analytical quality control, it has been determined that there are relation factors (RFs) between pesticides and the selected surrogates standards. These RFs are not altered by the complexities and compositions of simple and complex aqueous matrices. Predetermined RFs was applied to the picloram, atrazine and methyl parathion assessment in a real sample from the estuary of the Jaguaribe River, and the results showed that when no quality control was applied, the concentration levels would be underestimated, leading to incorrect results and inaccurate conclusions.

Dengue: profile of hematological and biochemical dynamics

Aim The objective of this study was to correlate laboratory tests during the evolution of dengue fever, comparing frequencies between the different clinical forms in order to use test results to predict the severity of the disease. Methods This is an observational, descriptive and retrospective study of 154 patients with clinical and serological diagnoses of dengue fever who, in the period from January to May 2008, were admitted in a tertiary state hospital in the city of Fortaleza that is a referral center for infectious diseases. The patients were allocated to two groups according to age: under 15 years old (n = 66) and 15 years or older (n = 88). The tests analyzed were blood count, platelet count, and serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) concentrations. Results Thrombocytopenia and elevated transaminases were observed in patients with classic dengue fever. The main laboratory abnormalities found in dengue hemorrhagic fever were thrombocytopenia, hemoconcentration and elevated transaminases, similar to severe dengue with the exception of hemoconcentration. Most laboratory abnormalities started on the 3rd day but were more evident on the 5th day with restoration of values by the 11th day; this was more prominent in under 15-year-olds and with the more severe clinical forms. Conclusion These results are relevant in assessing the disease because they can be used as markers for more severe forms and can help by enabling the adaptation of the therapeutic conduct to the needs of individual patients.

Transient pancytopenia induced by parvovirus B19 in a child with hereditary spherocytosis

OBJECTIVE: To describe the occurrence of transient pancytopenia induced by parvovirus B19 infection in a patient with hereditary hemolytic anemia and to discuss the importance of the diagnosis of this pathology.METHODS: Case report of a child whose diagnosis was made by polymerase chain reaction (PCR) and serology, and review of the literature.CLINICAL REPORT: A twelve year-old male patient with hereditary spherocytosis, presenting non-specific symptoms of an infectious syndrome followed by severe and transient pancytopenia, whose diagnosis was a parvovirus B19 infection.CONCLUSION: The diagnosis of parvovirus infection has a particular importance in hematology, especially on some morbid conditions, among them the hereditary hemolytic anemias. PCR is useful because of its rapidness and sensitivity on the specific diagnosis of this disease.

HIV-1 tropism and CD4 T lymphocyte recovery in a prospective cohort of patients initiating HAART in Ribeirão Preto, Brazil

While human immunodeficiency virus (HIV)-1 chemokine co-receptors 5 tropism and the GWGR motif in the envelope third variable region (V3 loop) have been associated with a slower disease progression, their influence on antiretroviral response remains unclear. The impact of baseline V3 characteristics on treatment response was evaluated in a randomised, double blind, prospective cohort study with patients initiating highly active antiretroviral therapy with lopinavir or efavirenz plus azithothymidine/3TC (1:1) over 48 weeks. Similar virological and immunological responses were observed for both treatment regimens. The 43 individuals had a mean baseline CD4 T cell count of 119 cells/mm(3) [standard deviation (SD) = 99] and a mean viral load of 5.09 log(10) copies/mL (SD = 0.49). The GWGR motif was not associated with a CD4 T cell response, but predicted R5 tropism by the geno2pheno([clinical20%]) algorithm correlated with higher CD4 T cell levels at all monitoring points (p < 0.05). Moreover, higher false-positive rates (FPR) values from this analysis revealed a strong correlation with CD4 T cell recovery (p < 0.0001). Transmitted drug resistance mutations, documented in 3/41 (7.3%) cases, were unrelated to the assigned antiretroviral regimen and had no impact on patient outcomes. In conclusion, naÏve HIV-1 R5 infected patients exhibited higher CD4 T cell counts at baseline; this difference was sustained throughout therapy. The geno2pheno([clinical]) option FPR positively correlated with CD4 T cell gain and may be useful in predicting CD4 T cell recovery.

Performance of rapid diagnostic tests for detection of Hepatitis B and C markers in HIV infected patients

There is little information describing the influence of HIV infection upon the performance of rapid diagnostic tests (RDTs) for hepatitis B and C virus diagnosis. This study aims to evaluate the performance of RDTs for HBsAg and anti-HCV detection among HIV-infected individuals. A total of 362 HIV infected individuals were recruited from clinics between January 2013 to November 2014 in the southeast and northeast of Brazil. HBsAg and anti-HCV were detected using commercial EIAs and four RDTs: HBV (Vikia HBsAg® and Wama Imuno-Rapido HBV®) and HCV (Bioeasy Teste Rápido HCV® and Wama Imuno-Rapido HCV®). Reactive HBsAg and anti-HCV serum samples were tested for HBV DNA and HCV RNA. Sensitivity, specificity and kappa statistic were determined. Using EIA, HBsAg and anti-HCV were detected in 14 (3.9%) and 37 (10.2%) serum samples respectively. Using serum only, HBsAg RDTs demonstrated sensitivities and specificities above 92.0% and Kappa values above 89.0%. Anti-HCV RDTs demonstrated sensitivity and specificities above 82.0% and Kappa higher than 89.0%. Using whole blood samples, Vikia HBsAg® and Wama Imuno-Rapido HCV® showed sensitivity and specificity above 99.0% with Kappa of 66.4% and 100%, respectively. HIV viral load was higher among discordant results for anti-HCV RDT. RDTs demonstrated good performance in HIV infected individuals showing the usefulness of assays in this population.

Leishmania infantum Infection in Blood Donors, Northeastern Brazil

To the Editor: Leishmania infantum is endemic to northeastern Brazil. It is responsible for visceral leishmaniasis (VL), a major emerging health problem in urban areas. Transmission occurs predominantly by the Lutzomyia longipalpis sand fly, but transfusion-associated VL, caused by L. infantum, has been reported from southern Europe and, by L. donovani, on the Indian subcontinent (1,2). Most L. infantum infections are asymptomatic (3), raising concern that the parasite could be present in donated blood from otherwise healthy residents in areas to which it is endemic (4). n nVL caused by L. infantum is endemic to 20 of Brazil’s 27 states; an annual average of 3,553 cases occur nationwide, with 54% of all cases reported from Brazil’s northeastern region. The state of Ceara historically ranks first or second in number of cases; an annual average of 467 cases were reported during the last decade (5). Thirty-eight percent of cases were reported from Fortaleza, the capital, where 28.4% of the state’s population resides. Over a 10-year-period, 277 (7.8%) persons with VL have died, and 109 (39%) of VL-related deaths have occurred in Fortaleza. n nSixty-nine percent of blood donors for Ceara reside in Fortaleza. To determine the prevalence of Leishmania infection among healthy blood donors, we tested blood donated to the State of Ceara Public Blood Bank. Compulsory serologic testing was also done for Trypanosoma cruzi (Chagas disease), hepatitis B and C, Treponema pallidum (syphilis), human T-cell lymphotropic virus types 1 and 2, and HIV-1 and -2. In the blood bank, 60% of units are centrifuged to separate the buffy coat in preparation for platelet separation. During May–November 2011, we randomly selected 431 buffy coats and tested them for Leishmania spp. by ELISA and PCR. To separate plasma from cells, 10 mL of buffy coat was centrifuged in Ficoll-Hypaque (Histopaque −1077, Sigma-Aldrich, Sao Paulo, Brazil). We tested plasma for leishmanial IgG by ELISA using a modified protocol of Evans et al. (6). IOC/L2906 L. infantum strain (MHOM/BR/2002/LPC-RPV) was used as the source of promastigote antigens. In addition, DNA was extracted from the mononuclear cell preparation, and PCR was performed with primers 150 (5′-GGG[G/T]AGGGGCGTTCT[G/C]CGAA-3′) and 152 (5′-[G/C][G/C][G/C][A/T]CTAT[A/T]TTACACCAACCCC-3′) that target the 120-bp conserved region of the Leishmania kDNA minicircle present in all Leishmania spp (7). As a positive control, kDNA was extracted from L. amazonensis promastigotes, strain BA-125 (MHOM/BR/87), characterized by PCR and isoenzymes (8). All PCR-positive samples were purified and sent to Ludwig Biotec (Alvorada, Brazil) for sequencing by ACTGENE-Molecular Analysis. The Federal University of Ceara Ethics Committee approved this study. n nBuffy coats from 57 (13.2%) serum samples from 431 donors were positive for leishmanial IgG, and 20 (4.6%) were positive for Leishmania spp. DNA. Sequencing of all PCR-positive samples confirmed the Leishmania genus. Three donors tested positive by both ELISA and PCR. Overall, the prevalence of leishmanial infection was 17.1% of blood donors. Eighty of the 431 units tested positive for >1 of compulsorily screened infections and were rejected. Of the remaining 351 that were negative for co-infection, 43 (12.2%) were positive for leishmanial IgG and 15 (4.3%) for Leishmania spp. DNA. Two donors were positive for both by ELISA and PCR. The prevalence of Leishmania infection among blood units accepted for transfusion was 16%. n nThe results demonstrate a surprisingly high prevalence of Leishmania infection in blood donors in Fortaleza, several times higher than that other diseases for which blood is screened (Figure). In a recent study in Salvador, Brazil (9), 5.4% of blood donors had leishmanial antibodies, of which 68% were positive by its PCR targeting kDNA amplification. n n n nFigure n nComparison of the prevalence of Leishmania infantum as tested by PCR and ELISA and of other infections compulsorily tested in 431 blood donors in Fortaleza, state of Ceara, northeastern Brazil. HBV, hepatitis B virus; HCV, hepatitis C virus; HTLV, ... n n n nThe percentages of antibody- or PCR-positive units capable of transmitting Leishmania and the outcomes are unknown. Viable Leishmania might not be in the blood of all PCR-positive donors, and even when present, the inoculum might be reduced by removal of infected circulating mononuclear phagocytes in the buffy coat, or parasites might be affected by steps involved in preparation or storage. However, if we consider units that test positive by PCR as being potentially infectious, the number of recipients at risk is of substantial concern. For example, in 2011, there were 99,933 blood donations to the State of Ceara Public Blood Bank. After compulsory screening for the other bloodborne pathogens, 93,238 units were accepted for transfusion. Extrapolating from the PCR-positive rate of 4.3%, a total of 4,009 recipients possibly were exposed to infection. Further studies are needed to determine whether recipients of blood from donors who are PCR positive and/or leishmanial antibody positive become infected with L. infantum. Persons with advanced AIDS or other immunosuppressive conditions seemingly would be at greatest risk for VL. n nIn Brazil, legislation requires that all blood for transfusion be tested for T. cruzi ., hepatitis B and C, T. pallidum, human T-cell lymphotropic virus types 1 and 2, and HIV-1 and -2. As additional information becomes available, screening for L. infantum also might be advisable to reduce the possibility of the recipient becoming infected, developing VL, and possibly being a reservoir of infection in the community (10), particularly in Ceara and other regions where the prevalence of L. infantum infection is high.

Cross-Sectional Study to Determine the Prevalence of Hepatitis B and C Virus Infection in High Risk Groups in the Northeast Region of Brazil

Background: HBV (Hepatitis B Virus) and HCV (Hepatitis C Virus) infections are more prevalent in vulnerable populations than the general population. The objective of this study was to investigate the prevalence of HBV and HCV infection in HIV-positive patients (GI), chronic renal failure (CRF) patients (GII) and coagulation disorder individuals (GIII). Methods: A cross-sectional study was conducted from June 2014 to March 2015. Serum samples were tested for markers of hepatitis B and C by enzyme-linked immunosorbent assay (ELISA). Sociodemographic, epidemiological, clinical and laboratory data and accompanying statistical analyses were performed using Epi Info™ 7. Results: A total of 348 individuals were recruited, i.e., 154 HIV-positive, 143 CRF and 51 coagulopathy patients. Among them, more than 66% were men, and the predominant age group was 26–35 years in GI and 56–65 years in GIII. Most patients had more than 8 years of education (66.2% in GI, 60.6% in GIII and 46.1% in GII), with a family income between 100–400 dollars in more than 48% of patients. The prevalence of the HBsAg marker was 3.9%, 7% and 3.9%, total anti-HBc was 28.6%, 55.9% and 31.4%, and anti-HCV was 1.3%, 12.6% and 47% for GI, GII and GIII, respectively. However, the prevalence of anti-HBs was greater than 70% in all groups. Conclusions: This study shows a high prevalence of HBV and HCV among specific groups compared to the general population. Factors such as age, income, number of sexual partners, sexually transmitted disease burden, blood transfusion history or blood products and blood transfusions before 1994 were associated with a higher prevalence for these infections.

A NEW POSSIBILITY FOR SURVEILLANCE: DO WE IDENTIFY ALL CASES OF LEPTOSPIROSIS?

SUMMARY Leptospirosis is a febrile disease with a typically underestimated global incidence, especially in regions where dengue is endemic. Therefore, it is difficult to accurately determine the number of leptospirosis cases in these areas, which contributes to significant under-reporting this disease. In this study, we estimated the number of possible leptospirosis cases among dengue-like cases that were reported during 2008, 2010, and 2012 in the city of Fortaleza, northeast Brazil. Patients were evaluated for dengue and leptospirosis using immunoenzymatic tests for IgM antibodies that were specific to each pathogen. Among the suspected cases of dengue that resulted as negative in laboratory tests, 10.8% (2008), 19.2% (2010), and 30.8% (2012) were confirmed to be leptospirosis. Considering the cases reported by the surveillance authority as dengue that were subsequently discarded based on the laboratory test results, we estimate that the number of actual leptospirosis cases may be 26 to 49 times higher than those diagnosed and reported by the Health Services. Furthermore, we believe that approximately 20% of dengue-like cases may be leptospirosis cases in areas where the two diseases are endemic.