Agar dilution method for susceptibility testing of Neisseria gonorrhoeae

The antibiotic susceptibilities of Neisseria gonorrhoeae isolates obtained from patients attending a clinic for sexually transmitted diseases in Tucumán, Argentina, were determined by the agar dilution method (MIC). 3.5% of the isolates produced ²-lactamase. A total of 96.5% of ²-lactamase negative isolates tested were susceptible to penicillin (MIC < 2 µgml-1); 14.03% of the tested isolates were resistant to tetracycline (MIC < 2 µgml-1), and 98% of the tested isolates were susceptible to spectinomycin (MIC < 64 µgml-1). The MICs for 95% of the isolates, tested for other drugs were: < 2 µgml-1 for cefoxitin, < 0.06 µgml-1 for cefotaxime, < 0.25 µgml-1 for norfloxacin, < 10 µgml-1 for cephaloridine, < 10 µgml-1 for cephalexin, and < 50 µgml-1 for kanamycin. Antibiotic resistance among N. gonorrhoeae isolates from Tucumán, Argentina, appeared to be primarily limited to penicillin and tetracycline, which has been a general use against gonorrhoeae in Tucumán since 1960. Periodic monitoring of the underlying susceptibility profiles of the N. gonorrhoeae strains prevalent in areas of frequent transmission may provide clues regarding treatment options and emerging of drug resistance.

Sensitivity of Lymph Node Aspiration in Localized Cutaneous Leishmaniasis Due to Leishmania (Viannia) braziliensis

Twenty nine patients with localized cutaneous leishmaniasis had lymph node and skin ulcer aspirations for culture of Leishmania with the modified Marzochi´s vacuum aspiratory technique. Sensitivity of lymph node aspiration was 58.6% and 34.5% for skin ulcer aspiration (P=0.06). Combined sensitivity of the two methods was 79.3%. There was no agreement between methods (Kappa Index = -0.084; CI95% -0,45; 0,28) showing the potential complementary roles in diagnostic approach.

Human immunodeficiency virus type 1 Brazilian subtype B variant showed an increasing avidity of the anti-V3 antibodies over time compared to the subtype B US/European strain in São Paulo, Brazil

The Brazilian variant of human immunodeficiency virus type 1 (HIV-1) subtype B, (serotype B"-GWGR), has a tryptophan replacing the proline in position 328 the HIV-1 envelope. A longer median time period from infection to acquired immunodeficiency syndrome (AIDS) for serotype B (B"-GWGR) infected subjects compared to the B-GPGR US/European strain was reported. In a cohort study, in São Paulo city, 10 B"-GWGR patients had a statistically significant increased avidity of the anti-V3 antibodies, from 79% ± 33% to 85% ± 75%, versus from 48% ± 59% to 32% ± 17% for the 10 B-GPGR subjects (p = 0.02). The T CD4+ cells showed a mean increase of + 0.45 cells/month for the B-GPGR subjects and for B"-GWGR the slope was + 1.24 cells/month (p = 0.06), for 62 and 55 months of follow up, respectively. RNA plasma viral load decreased from 3.98 ± 1.75 to 2.16 ± 1.54 log10 in the B"-GWGR group while B-GPGR patients showed one log10 reduction in viral load from 4.09 ± 0.38 to 3.17 ± 1.47 log10 over time (p = 0.23), with a decreasing slope of 0.0042 ± log10,/month and 0.0080 ± log10/month, for B-GPGR and B"-GWGR patients, respectively (p = 0.53). Neither group presented any AIDS defining events during the study, according to Center for Diseases Control criteria. Although the sample size is small, these results may indicate that differences in the pathogenicity of the 2 HIV-1 B serotypes which co-circulate in Brazil may be correlated to the avidity of anti-V3 antibodies.

In vitro susceptibility to antifungal agents of environmental Cryptococcus spp. isolated in the city of Ribeirão Preto, São Paulo, Brazil

Infections by Cryptococcus strains other than C. neoformans have been detected in immunocompromised patients. Of these strains, three are considered human pathogens: C. albidus, C. laurenttii, and C. uniguttulatus. This study deals with the in vitro susceptibility of Cryptococcus to drugs such as amphotericin B, itraconazole, fluconazole, and 5-fluorocytosine. Environmental Cryptococcus isolates (50) distributed as follows: C. neoformans var. neoformans (16), C. albidus (17), C. laurentii (14), and C. uniguttulatus (3) were evaluated by the micro and macrodilution techniques, according to EUCAST and NCCLS recommendations, respectively. Considering both methodologies the respective minimal inhibitory concentrations (MIC) were 0.125 and 2 µg/ml for amphotericin B, 0.06 and 8 µg/ml for itraconazole, and 0.5 and more than 64 µg/ml for fluconazole and 5-fluorocytosine. Agreement percentages for the two methodologies were 100% for amphotericin B and fluconazole for all the strains tested. For itraconazole, the agreement percentage was 81.3% in the C. neoformans strain and 100% for all the others. All species had a agreement percentage of 94.1 to 100% when susceptibility to 5-fluorocytosine was tested. It is concluded that environmental isolates of C. neoformans var. neoformans, C. albidus, C. laurentii, and C. uniguttulatus may show high MICs against certain drugs, suggesting in vitro primary resistance to the antifungals tested.

In vitro and in vivo effects of Enterococcus faecalis CECT7121 on Toxocara canis

The aim of the present paper was to evaluate the larvicidal effect of Enterococcus faecalis CECT7121 (Ef7121) on the Toxocara canis cycle both in vitro and in vivo. For the in vitro experiments, T. canis larvae were incubated with the supernatants of Ef7121 (EI) and mutant Ef7121 (EIm), in a pre-culture of Ef7121 (EII) and in a fresh culture with Ef7121 (EIII) and the Ef7121 mutant strain (EIIIm). The viability of the larvae was calculated after a 48 h incubation. A significant reduction of the viability of T. canis larvae was observed in EI, EII and EIII. A decrease of this inhibitory effect was observed in EIm and EIIIm (p = 0.008). In the in vivo experiments, mice were orally inoculated with three doses of Ef7121. To study the probiotic persistence in the intestine, the animals were sacrificed every four days and their intestines were dissected. The initial average bacterial levels were 9.7 x 10(4) for Ef7121 (colony forming units/g). At the end of the assay the levels were 1.46 x 10(4). No bacterial translocation was detected in mesenteric lymphatic nodules and spleen. Ef7121 interference with the biological cycle was evaluated in mice challenged with T. canis. The interference was significant when the mice were challenged with probiotic and T. canis simultaneously (p = 0.001), but it was not significant when the challenge was performed 15 days after administration of the bacterial inoculum (p = 0.06). In conclusion, Ef7121 possessed in vitro and in vivo larvicidal activity.

Leprosy reactions: the effect of gender and household contacts

Various host-related factors have been reported as relevant risk factors for leprosy reactions. To support a new hypothesis that an antigenic load in local tissues that is sufficient to trigger the immune response may come from an external supply of Mycobacterium leprae organisms, the prevalence of reactional leprosy was assessed against the number of household contacts. The number of contacts was ascertained at diagnosis in leprosy patients coming from an endemic area of Brazil. The prevalence of reactions (patients with reactions/total patients) was fitted by binomial regression and the risk difference (RD) was estimated with a semi-robust estimation of variance as a measure of effect. Five regression models were fitted. Model 1 included only the main exposure variable "number of household contacts"; model 2 included all four explanatory variables ("contacts", "fertile age", "number of skin lesions" and "bacillary index") that were found to be associated with the outcome upon univariate analysis; models 3-5 contained various combinations of three predictors. Male and female patients were analyzed separately. In females, household contacts were a significant predictor for leprosy reactions in model 1 [crude RD = 0.06; 95% confidence interval (CI) = 0.01; 0.12] and model 5 (RD = 0.05; CI = 0.02; 0.09), which included contacts, bacillary index and skin lesions as predictors. Other models were unsatisfactory because the joint presence of fertile age and bacillary index was a likely source of multicollinearity. No significant results were obtained for males. The likely interpretation of our findings might suggest that in female patients, leprosy reactions may be triggered by an external spreading of M. leprae by healthy carrier family members. The small number of observations is an obvious limitation of our study which requires larger confirmatory studies.

Primary resistance of HIV to antiretrovirals among individuals recently diagnosed at voluntary counselling and testing centres in the metropolitan region of Recife, Pernambuco

Determining the prevalence and type of antiretroviral (ARV) resistance among ARV-naïve individuals is important to assess the potential responses of these individuals to first-line regimens. The prevalence of primary resistance and the occurrence of recent infections among individuals with human immunodeficiency virus (HIV)/acquired immune deficiency syndrome (AIDS) were identified among recently diagnosed patients at five sexually transmitted disease/AIDS testing and counselling centres in the metropolitan region of Recife (RMR), Pernambuco, Brazil, between 2007-2009. One-hundred and eight samples were analysed using the Calypte® BED assay. Males predominated (56%), as did patients aged 31-50 years. Twenty-three percent presented evidence of a recent HIV infection. The median CD4+ T lymphocyte count was 408 cells/mm³ and the median viral load was 3.683 copies/mL. The prevalence of primary resistance was 4.6% (confidence interval 95% = 1-8.2%) based on criteria that excluded common polymorphisms in accordance with the surveillance drug resistance mutation criteria. The prevalence of resistance to non-nucleoside reverse transcriptase, nucleoside/nucleotide reverse transcriptase and protease inhibitors were 3.8%, 1.5% and 0.8%, respectively. Fifty-seven percent of strains were from clade B, 37.7% were clade F and 3.1% were clade C; there were no statistically significant differences with respect to resistance between clades. Recent infection tended to be more common in men (p = 0.06) and in municipalities in the south of the RMR (Jaboatão dos Guararapes and Cabo de Santo Agostinho) (p = 0.046). The high prevalence of recent infection and the high prevalence of non-B strains in this poor Brazilian region merit further attention.

Comparative evaluation of the microplate nitrate reductase assay and the rezasurin microtitre assay for the rapid detection of multidrug resistant Mycobacterium tuberculosis clinical isolates

The microplate nitrate reductase assay (MNRA) and the rezasurin microtitre assay (REMA) were used for the susceptibility testing of 73 clinical isolates and the results were compared with those that were obtained using the Bactec 460 TB and Bactec MGIT 960 systems. The REMA and the MNRA were performed in 96-well plates. For the REMA, the concentrations of isoniazid (INH) and rifampicin (RIF) ranged from 1.0-0.01 µg/mL and 2.0-0.03 µg/mL, respectively. For the MNRA, the INH concentration was between 1.0-0.03 µg/mL and the RIF concentration was between 2.0-0.06 µg/mL. For the MNRA, the sensitivity, specificity, positive predictive value, negative predictive value and INH/RIF agreement were 100/95.6, 97.6/100, 96.8/100, 100/98 and 98.6/98.6, respectively, and for the REMA, they were 100/91.3, 90.4/100, 88.5/100, 100/96.1 and 94.5/97.2, respectively. Our data suggest that these two rapid, low-cost methods may be inexpensive, alternative assays for the rapid detection of multidrug resistant tuberculosis in low-income countries.

Variation in haematological parameters in children less than five years of age with asymptomatic Plasmodium infection: implication for malaria field studies

During the season of high malaria transmission, most children are infected by Plasmodium, which targets red blood cells (RBCs), affecting haematological parameters. To describe these variations, we examined the haematological profiles of two groups of children living in a malaria-endemic area. A cross-sectional survey was conducted at the peak of the malaria transmission season in a rural area of Burkina Faso. After informed consent and clinical examination, blood samples were obtained from the participants for malaria diagnosis and a full blood count. Of the 414 children included in the analysis, 192 were not infected with Plasmodium, whereas 222 were asymptomatic carriers of Plasmodium infection. The mean age of the infected children was 41.8 months (range of 26.4-57.2) compared to 38.8 months (range of 22.4-55.2) for the control group (p = 0.06). The asymptomatic infected children tended to have a significantly lower mean haemoglobin level (10.8 g/dL vs. 10.4 g/dL; p < 0.001), mean lymphocyte count (4592/µL vs. 5141/µL; p = 0.004), mean platelet count (266 x 103/µL vs. 385 x 103/µL; p < 0.001) and mean RBC count (4.388 x 106/µL vs. 4.158 x 106/µL; p < 0.001) and a higher mean monocyte count (1403/µL vs. 1192/µL; p < 0.001) compared to the control group. Special attention should be applied when interpreting haematological parameters and evaluating immune responses in asymptomatic infected children living in malaria-endemic areas and enrolled in vaccine trials.

Increased frequency of micronuclei in the lymphocytes of patients chronically infected with hepatitis B or hepatitis C virus

In this study, we analysed the frequency of micronuclei (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) and evaluated mutagen-induced sensitivity in the lymphocytes of patients chronically infected with hepatitis B virus (HBV) or hepatitis C virus (HCV). In total, 49 patients with chronic viral hepatitis (28 HBV-infected and 21 HCV-infected patients) and 33 healthy, non-infected blood donor controls were investigated. The frequencies (‰) of MN, NPBs and NBUDs in the controls were 4.41 ± 2.15, 1.15 ± 0.97 and 2.98 ± 1.31, respectively. The frequencies of MN and NPBs were significantly increased (p < 0.0001) in the patient group (7.01 ± 3.23 and 2.76 ± 2.08, respectively) compared with the control group. When considered separately, the HBV-infected patients (7.18 ± 3.57) and HCV-infected patients (3.27 ± 2.40) each had greater numbers of MN than did the controls (p < 0.0001). The HCV-infected patients displayed high numbers of NPBs (2.09 ± 1.33) and NBUDs (4.38 ± 3.28), but only the HBV-infected patients exhibited a significant difference (NPBs = 3.27 ± 2.40, p < 0.0001 and NBUDs = 4.71 ± 2.79, p = 0.03) in comparison with the controls. Similar results were obtained for males, but not for females, when all patients or the HBV-infected group was compared with the controls. The lymphocytes of the infected patients did not exhibit sensitivity to mutagen in comparison with the lymphocytes of the controls (p = 0.06). These results showed that the lymphocytes of patients who were chronically infected with HBV or HCV presented greater chromosomal instability.

Infusão de insulina em terapia intensiva: ensaio controlado randomizado

Ensaio clínico controlado e aleatorizado que comparou o uso de protocolo de insulina intensivo e convencional na evolução clínica de pacientes em sepse grave e choque séptico, nas primeiras 72 h. Foi conduzido em um hospital universitário na cidade de São Paulo. Os pacientes (n=46) foram alocados em dois grupos: glicêmico intensivo (glicemia entre 80-110mg/dl) e convencional (180-220mg/dl). Utilizaram-se testes t-Student e Qui-Quadrado na análise dos dados. Observou-se diferença estatisticamente significativa (p<0,001) na média glicêmica, mas não houve diferença para as variáveis pressão arterial média mínima (p=0,06) e máxima (p=0,11), creatinina sérica (p=0,33) e na mortalidade (p=0,11). Apesar de não haver diferença entre os grupos quanto à mortalidade, a instabilidade hemodinâmica no grupo convencional foi mais duradoura e somente nele ocorreram óbitos.

Adherence to statin treatment and associated factors in female users from the Unified Health System (SUS)

Objective: To identify the adherence rate of a statin treatment and possible related factors in female users from the Unified Health System. Method: Seventy-one women were evaluated (64.2 ± 11.0 years) regarding the socio-economic level, comorbidities, current medications, level of physical activity, self-report of muscular pain, adherence to the medical prescription, body composition and biochemical profile. The data were analyzed as frequencies, Chi-Squared test, and Mann Whitney test (p<0.05). Results: 15.5% of women did not adhere to the medical prescription for the statin treatment, whose had less comorbidities (p=0.01), consumed less quantities of medications (p=0.00), and tended to be younger (p=0.06). Those patients also presented higher values of lipid profile (CT: p=0.01; LDL-c: p=0.02). Musculoskeletal complains were not associated to the adherence rate to the medication. Conclusion: The associated factors to adherence of dyslipidemic women to statin medical prescription were age, quantity of comorbidities and quantity of current medication.


Crescimento e acúmulo de crômio em alface cultivada em dois latossolos tratados com CrCl3 e resíduos de curtume

Resíduos sólidos de curtume e CrCl3 foram aplicados em dois solos, Latossolo Roxo eutrófico (LRe) unidade Ribeirão Preto e Latossolo Vermelho-Amarelo (LVA) unidade Laranja Azeda, que se diferenciaram, dentre outros atributos, pelo teor de manganês facilmente redutível. Os resíduos utilizados foram lodo do efluente de caleiro com concentração 0,06 g kg-1 de crômio (LCL) e um lodo do decantador primário (LCR), contendo 17,4 g kg-1 de crômio, ambos na matéria seca, aplicados em doses correspondentes a 10, 20 e 30 Mg ha-1 e 19, 38 e 57 Mg ha-1 (base seca), respectivamente, de acordo com o teor de nitrogênio total de cada um. O CrCl3 foi aplicado nas doses de 330, 660 e 990 kg ha-1 de Cr, equivalentes às doses do metal aplicadas na forma de lodo (LCR). Realizou-se o experimento em vasos alocados em casa de vegetação (blocos ao acaso), que foram monitorados quanto à formação de Cr6+, aos 1, 6, 14, 28, 54 e 86 dias da instalação. Após o 56° dia de incubação, foi transplantada uma muda de alface (Lactuca sativa L.) para cada vaso, cultivada por um período de trinta dias. A oxidação do Cr3+ a Cr6+ foi verificada apenas para o LRe nos tratamentos que receberam doses crescentes de CrCl3. A formação de Cr6+ teve máximo entre 0,72 e 1,16% do Cr3+ aplicado, após um dia de incubação, decrescendo com o tempo, não sendo detectada a sua presença, para nenhuma das doses, após o 54° dia. A aplicação dos resíduos elevou a condutividade elétrica do extrato de saturação (2:1) de 1,40 a 5,07 Ds m-1 e a RAS de 3,05 a 14,12, afetando o desenvolvimento da alface e causando a morte das plantas nas doses mais altas, sendo tais efeitos mais pronunciados no LVA. A concentração de crômio na parte aérea das plantas aumentou, nem sempre de forma proporcional, com o aumento das doses aplicadas na forma de lodo ou sal, com efeito mais acentuado para o LVA do que para o LRe. A aplicação de resíduos de curtume no experimento, para ambos os solos, mostrou-se mais limitante pelo seu conteúdo de sais do que pela presença de crômio.

Efeito do pH na adsorção-dessorção de chumbo em Latossolos brasileiros

Reações de adsorção-dessorção de chumbo em solos são influenciadas por atributos de superfície dos colóides dos solos e pela composição da solução do meio. Este estudo avaliou o efeito do pH sobre a adsorção-dessorção de chumbo em Latossolos brasileiros. Amostras do horizonte A de cada solo, suspensas em Ca(NO3)2 5 mmol L-1, foram tituladas com HNO3 7 mmol L-1 ou solução saturada de Ca(OH)2, para que fosse atingido o valor de pH estipulado em cada experimento (4,5, 5,5 e 6,5). Atingido o pH de equilíbrio, as amostras foram equilibradas com Pb(NO3)2, com vistas em obter uma concentração final de 0,15 mmol L-1 (relação solo:solução 1:100; força iônica 15 mmol L-1), por um período de 72 h. A dessorção foi realizada em Ca(NO3)2 5 mmol L-1, pH 5,5. O aumento do pH de 4,5 para 6,5 causou aumento da adsorção de até 16,7 vezes, um aumento médio de 2,9 vezes de pH 4,5 para 5,5; 1,4 vez de pH 5,5 para 6,5 e 4,2 vezes de pH 4,5 para 6,5. A relação Pb adsorvido/Pb adicionado foi, em média, de 0,33 para pH 4,5; 0,75 para pH 5,5 e 0,94 para pH 6,5. A fração média de Pb dessorvido (Pb dessorvido/Pb adsorvido) decresceu de 0,36 a pH 4,5, para < 0,06 a pH 6,5. O efeito dos atributos do solo sobre a adsorção-dessorção de chumbo decresceu quando o pH aumentou, evidenciado por uma maior diferenciação na quantidade adsorvida pelos solos em valores mais baixos de pH. A adsorção de Pb foi positivamente correlacionada (e geralmente a fração dessorvida foi negativamente correlacionada) com área superficial específica, CTC a pH 7,0, teores de caulinita, hematita, Fe2O3 extraídos pelo ditionito-citrato-bicarbonato de sódio e oxalato ácido de amônio e SiO2 e Fe2O3 extraídos pelo ataque sulfúrico. O fato de considerável fração de chumbo permanecer adsorvida em pH 4,5 mostra a reduzida disponibilidade deste metal em Latossolos, mesmo em baixos valores de pH.

Reconsolidation of the soil surface after tillage discontinuity, with and without cultivation, related to erosion and its prediction with RUSLE

Site-specific regression coefficient values are essential for erosion prediction with empirical models. With the objective to investigate the surface-soilconsolidation factor, Cf, linked to the RUSLE's prior-land-use subfactor, PLU, an erosion experiment using simulated rainfall on a 0.075 m m-1 slope, sandy loam Paleudult soil, was conducted at the Agriculture Experimental Station of the Federal University of Rio Grande do Sul (EEA/UFRGS), in Eldorado do Sul, State of Rio Grande do Sul, Brazil. Firstly, a row-cropped area was excluded from cultivation (March 1995), the existing crop residue removed from the field, and the soil kept clean-tilled the rest of the year (to get a degraded soil condition for the intended purpose of this research). The soil was then conventional-tilled for the last time (except for a standard plot which was kept continuously cleantilled for comparison purposes), in January 1996, and the following treatments were established and evaluated for soil reconsolidation and soil erosion until May 1998, on duplicated 3.5 x 11.0 m erosion plots: (a) fresh-tilled soil, continuously in clean-tilled fallow (unit plot); (b) reconsolidating soil without cultivation; and (c) reconsolidating soil with cultivation (a crop sequence of three corn- and two black oats cycles, continuously in no-till, removing the crop residues after each harvest for rainfall application and redistributing them on the site after that). Simulated rainfall was applied with a Swanson's type, rotating-boom rainfall simulator, at 63.5 mm h-1 intensity and 90 min duration, six times during the two-and-half years of experimental period (at the beginning of the study and after each crop harvest, with the soil in the unit plot being retilled before each rainfall test). The soil-surface-consolidation factor, Cf, was calculated by dividing soil loss values from the reconsolidating soil treatments by the average value from the fresh-tilled soil treatment (unit plot). Non-linear regression was used to fit the Cf = e b.t model through the calculated Cf-data, where t is time in days since last tillage. Values for b were -0.0020 for the reconsolidating soil without cultivation and -0.0031 for the one with cultivation, yielding Cf-values equal to 0.16 and 0.06, respectively, after two-and-half years of tillage discontinuation, compared to 1.0 for fresh-tilled soil. These estimated Cf-values correspond, respectively, to soil loss reductions of 84 and 94 %, in relation to soil loss from the fresh-tilled soil, showing that the soil surface reconsolidated intenser with cultivation than without it. Two distinct treatmentinherent soil surface conditions probably influenced the rapid decay-rate of Cf values in this study, but, as a matter of a fact, they were part of the real environmental field conditions. Cf-factor curves presented in this paper are therefore useful for predicting erosion with RUSLE, but their application is restricted to situations where both soil type and particular soil surface condition are similar to the ones investigate in this study.