Polymerase chain reaction (PCR) for the detection of Salmonella in artificially inoculated chicken meat

The aim of this study was to develop a polymerase chain reaction (PCR) protocol for the detection of Salmonella in artificially contaminated chicken meat. Tests were performed with different dilutions of Salmonella Typhimurium or Salmonella Enteritidis cells (10-7, 10-8 or 10-9 CFU/mL) inoculated in chicken meat samples, in order to establish the limits of detection, incubation times (0, 6, 8 and 24 hours of pre-enrichment in PBW 1%) and three DNA extraction protocols (phenol-chloroform, thermal treatment and thermal treatment and Sephaglass). The assay was able to detect until 10-9 CFU/mL of initial dilution of Salmonella cells inoculated in chicken meat, which allows detection of Salmonella within 48 hours, including 24 hours of pre-enrichment and using the phenol-chloroform DNA extraction protocol. As the results are obtained in a shorter time period than that of microbiological culture, this procedure will be useful in the methodology for detection of Salmonella in chicken.

Effects of eucalyptol on house fly (Diptera: Muscidae) and blow fly (Diptera: Calliphoridae)

The effects of eucalyptol were evaluated against the house fly, Musca domestica L., and blow fly, Chrysomya megacephala (F.). The bioassay of adults, using topical application, indicated that M. domestica males were more susceptible than females, with the LD50 being 118 and 177 µg/fly, respectively. A higher LD50 of C. megacephala was obtained; 197 µg/fly for males and 221 µg/fly for females. Living flies of both species yielded a shorter life span after being treated with eucalyptol. The bioassay of larvae, using the dipping method on the third instar, showed that M. domestica was more susceptible than C. megacephala, with their LC50 being 101 and 642 µg/µl, respectively. The emergence of adults, which had been treated with eucalyptol in larvae, decreased only in M. domestica. Having the volatile property, fumigation or impregnated paper test of eucalyptol or the efficacy of repellence or attractiveness merits further investigations to enhance bio-insecticidal efficacy.

New data on Echinococcus spp. in Southern Brazil

40 Echinococcus isolates from sheep and cattle in Southern Brazil were genetically analysed in order to obtain further data on the presence of different taxa of the Echinococcus granulosus complex. Differentiation was done using a PCR technique and sequencing of mitochondrial cytochrome c oxidase subunit 1 (CO1). Most samples (38) could be allocated to the sheep strain (G1) of E. granulosus, while two samples belonged to E. ortleppi, previously known as cattle strain (G5) of E. granulosus. Due to the shorter prepatent period in dogs of the latter taxon, this records have important implications for the design of control measures in this endemic region.

Reproduction of Leishmania (Leishmania ) infantum chagasi in conditioned cell culture growth medium

Leishmanias can be produced by inoculation in conditioned McCoy cell culture growth medium (CGM). Leishmania (Leishmania) infantum chagasi (100 parasites) grown in NNN medium was inoculated in 2.5 mL CGM, kept in plates (24 wells) and its multiplication was observed for five days (120 hours). After day 5, the medium was saturated with the flagellate forms of the parasite (promastigotes). The reproduction of the leishmanias was observed every 24 hours and the number of parasites was calculated by counting the parasites in a drop of 10 µ L and photomicrographied. So the number of Leishmanias was adjusted to 1 mL volume. The advantage of the technique by isolation of Leishmania in CGM demonstrated in this study is its low cost and high efficacy even with a small quantity of parasites (10² promastigotes) used as inoculum. Additionally, isolation of the leishmania can be obtained together with an increase in their density (180 times) as observed by growth kinetics, within a shorter time. These results justify the use of this low-cost technique for the isolation and investigation of the behavior and multiplication of Leishmania both in vertebrates and invertebrates, besides offering means of obtaining antigens, whether whole antigens (leishmanias) or the soluble antigens produced by the parasites which may be useful for the production of new diagnostic kits.

Predictive factors of mortality in burn patients

Burn mortality statistics may be misleading unless they account properly for the many factors that can influence outcome. Such estimates are useful for patients and others making medical and financial decisions concerning their care. This study aimed to define the clinical, microbiological and laboratorial predictors of mortality with a view to focus on better burn care. Data were collected using independent variables, which were analyzed sequentially and cumulatively, employing univariate statistics and a pooled, cross-sectional, multivariate logistic regression to establish which variables better predict the probability of mortality. Survivors and non-survivors among burn patients were compared to define the predictive factors of mortality. Mortality rate was 5.0%. Higher age, larger burn area, presence of fungi in the wound, shorter length of stay and the presence of multi-resistant bacteria in the wound significantly predicted increased mortality. The authors conclude that those patients who are most apt to die are those with age > 50 years, with limited skin donor sites and those with multi-resistant bacteria and fungi in the wound.

HTLV- I infection is not associated with a higher risk of death in peruvian HIV-infected patients

Limited and contradictory information exists regarding the prognosis of HIV/HTLV-I co-infection. Our goal was to estimate the effect of HTLV-I infection on mortality in HIV-infected patients at a HIV reference center in Peru. We studied a retrospective cohort of HIV-infected patients, who were exposed or unexposed to HTLV-I. Exposed patients were Western Blot (WB) positive for both retroviruses. Unexposed patients were WB positive for HIV, and had least one negative EIA for HTLV-I. These were selected among patients who entered our Program immediately before and after each exposed patient, between January 1990 and June 2004. Survival time was considered between the diagnosis of exposure to HTLV-I and death or censoring. Confounding variables were age, gender, baseline HIV clinical stage, baseline CD4+ T cell count, and antiretroviral therapy. We studied 50 exposed, and 100 unexposed patients. Exposed patients had a shorter survival compared to unexposed patients [median survival: 47 months (95% CI: 17-77) vs. 85 months (95% CI: 70-100), unadjusted p = 0.06]. Exposed patients had a higher rate of mortality compared to unexposed patients (HIV/HTLV-I (24/50 [48%]) vs. HIV only (37/100 [37%]), univariable p = 0.2]. HTLV-I exposure was not associated to a higher risk of death in the adjusted analysis: HR: 1.2 (0.4-3.5). AIDS clinical stage and lack of antiretroviral therapy were associated to a higher risk of dying. In conclusions, HTLV-I infection was not associated with a higher risk of death in Peruvian HIV-infected patients. Advanced HIV infection and lack of antiretroviral therapy may explain the excess of mortality in this population.

Health problems awareness during travel among faculty members of a large university in Latin America: preliminary report

Health safety during trips is based on previous counseling, vaccination and prevention of infections, previous diseases or specific problems related to the destination. Our aim was to assess two aspects, incidence of health problems related to travel and the traveler's awareness of health safety. To this end we phone-interviewed faculty members of a large public University, randomly selected from humanities, engineering and health schools. Out of 520 attempts, we were able to contact 67 (12.9%) and 46 (68.6%) agreed to participate in the study. There was a large male proportion (37/44, 84.1%), mature adults mostly in their forties and fifties (32/44, 72.7%), all of them with higher education, as you would expect of faculty members. Most described themselves as being sedentary or as taking occasional exercise, with only 15.9% (7/44) taking regular exercise. Preexisting diseases were reported by 15 travelers. Most trips lasted usually one week or less. Duration of the travel was related to the destination, with (12h) or longer trips being taken by 68.2% (30/44) of travelers, and the others taking shorter (3h) domestic trips. Most travelling was made by air (41/44) and only 31.8% (14/44) of the trips were motivated by leisure. Field research trips were not reported. Specific health counseling previous to travel was reported only by two (4.5%). Twenty seven of them (61.4%) reported updated immunization, but 11/30 reported unchecked immunizations. 30% (9/30) reported travel without any health insurance coverage. As a whole group, 6 (13.6%) travelers reported at least one health problem attributed to the trip. All of them were males travelling abroad. Five presented respiratory infections, such as influenza and common cold, one neurological, one orthopedic, one social and one hypertension. There were no gender differences regarding age groups, destination, type of transport, previous health counseling, leisure travel motivation or pre-existing diseases. Interestingly, the two cases of previous health counseling were made by domestic travelers. Our data clearly shows that despite a significant number of travel related health problems, these highly educated faculty members, had a low awareness of those risks, and a significant number of travels are made without prior counseling or health insurance. A counseling program conducted by a tourism and health professional must be implemented for faculty members in order to increase the awareness of travel related health problems.

PARACOCCIDIOIDOMYCOSIS TREATMENT

SUMMARYConsidered to be an emerging endemic mycosis in Latin America, paracoccidioidomycosis is characterized by a chronic course and involvement of multiple organs in immunocompromised hosts. Infection sequelae are mainly related to pulmonary and adrenal insufficiency. The host-parasite interaction results in different expressions of the immune response depending on parasite pathogenicity, fungal load and genetic characteristics of the host. A few controlled and case series reports have shown that azoles and fast-acting sulfa derivatives are useful treatment alternatives in milder forms of the disease. For moderate/severe cases, more prolonged treatments or even parenteral routes are required especially when there is involvement of the digestive tract mucosa, resulting in poor drug absorption. Although comparative studies have reported that shorter treatment regimens with itraconazole are able to induce cure in chronically-infected patients, there are still treatment challenges such as the need for more controlled studies involving acute cases, the search for new drugs and combinations, and the search for compounds capable of modulating the immune response in severe cases as well as the paradoxical reactions.

Influence of Compound 16.842 on hookworm infections

Compound 16.842 was tried with three different groups of patients in order to evaluate its tolerancy, and efficacy as well, with a view of using the drug in mass campaigns against hookworm. Group I, used for a preliminary trial, consisted of 38 patients attending an out-patient clinic, and living either in the out-skirts or in the various urban areas of the city of Rio de Janeiro, including some inmates of an orphanage. Group II, a field trial, was carried out in two farms, where the drug was administered both to the positive cases (124) and to the rest of the population (nearly 90%). Group III, a field trial was also carried out in a small town where nearly 40% of the total population was treated with the Compound. Tolerability was considered rather satisfactory, mainly among the patients receiving two single doses (50-150mg), according to the age, 4 - 6 weeks apart). These results suggest that 2-4 courses of therapy within a shorter span of time should be the ideal for a mass treatment campaign. Efficacy varied from 26.6% to 76.2% parasitological cure in the various groups, with a wide range of variation also in the percent of mean reduction of eggs for hookworm. The drug showed also some effect against Ascaris lumbricoides giving cure rates between 10,5% and 35.7% in the various groups, with a percentage reduction in mean egg counts of 27% to 83.3% according to the various groups. It was concluded that Compound 16.842 possesses a marked effect on hookworm and a mild effect on A. lumbricoides. The findings indicate the need for more accurate studies to determine the most efficient schedules of treatment and the real value of the drug, as compared to other antihelminthics against the two parasites under study.

Evolution of sarcoma 180 (ascitic tumor) in mice infected with Schistosoma mansoni

Mice infected with 60 cercariae of Schistosoma mansoni were more resistant to the sarcoma 180 ascites tumor. Tumor inoculation was performed 50 days after schistosoma infection and the animals were observed and weighed at 48 hours intervals for development and progression of malignancy. In infected mice the weight gain (ascites formation) started later and was shorter than in uninfected Controls. Also, the number of tumor cells into the peritoneal cavity 72h after tumor implantation was shorter in infected group than incontrols. This in creased resistance against a transplantable tumor probably is related to the effect of endotoxin on tumoricidal activity of macrophages activated by the infection. The immunodepression induced by Schistosoma mansoni infection enhances the proliferation of endogenous bacteria increasing the amount of endotoxin absorbed from the gut.

Comparison of feeding and defecation patterns between fifth-instar nymphs of Triatoma patagonica (Del Ponte, 1929) and Triatoma infestans (Klug, 1934) under laboratory conditions

Among the vectors of Chagas disease, Triatoma patagonica is a species in the process of adaptation to the human environment. However, its vector competence is not well known. This study had the aim of evaluating and comparing feeding and defecation patterns among fifth-instar nymphs of Triatoma patagonica and Triatoma infestans that were fed ad libitum. The results showed that nymphs of Triatoma patagonica had a feeding pattern similar to that of Triatoma infestans. Sixty nine percent and 58% of nymphs of Triatoma patagonica and Triatoma infestans, respectively, produced their first defecation within five minutes after being fed. Triatoma patagonica defecated during feeding, with an average time until first defecation that was shorter than that of Triatoma infestans (3.4 and 6.2 min, respectively). The nymphs of Triatoma patagonica were capable of defecating during or immediately after feeding.

Evaluation of laboratory markers of progression of HIV disease to death

INTRODUCTION: One of the important current problems in HIV/AIDS infection is the establishment of epidemiological and laboratorial prognostic parameters during patient follow-up. This study aimed at analyzing the evolution of laboratory tests: CD4 lymphocyte count, viral load, hemoglobin (Hb), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and the epidemiological variables sex and age as prognostic factors for survival in progression to death among AIDS patients. METHODS: A retrospective study was conducted using analysis of medical records, and prospective 24-month follow-up of patients with HIV/ AIDS attended at the President Vargas Hospital Outpatient Clinic, a reference center in HIV/ AIDS attendance in the State of Maranhão, Brazil. The study analyzed patients aged 10 to 60 years old, who manifested AIDS and who were not using antiretroviral therapy or had used it for less than 5 years. The Chi-square test was used for statistical analysis. RESULTS: The sample included 100 patients - 57 were current outpatients, and 43 had died. The variables viral load (p=0.726), ALT (p=0.314), sex (p=0.687), and age (p=0.742) were analyzed, and no evidence of association between them and worst prognosis was observed. CONCLUSIONS: A significant relation was verified between low Hb levels (p=0.000) and CD4 (p=0.000) and shorter survival.

Parasitological stool sample exam by spontaneous sedimentation method using conical tubes: effectiveness, practice, and biosafety

INTRODUCTION: Spontaneous sedimentation is an important procedure for stool examination. A modification of this technique using conical tubes was performed and evaluated. METHODS: Fifty fecal samples were processed in sedimentation glass and in polypropylene conical tubes. Another 50 samples were used for quantitative evaluation of protozoan cysts. RESULTS: Although no significant differences occurred in the frequency of protozoa and helminths detected, significant differences in protozoan cyst counts did occur. CONCLUSIONS: The use of tube predicts a shorter path in the sedimentation of the sample, increases concentration of parasites for microscopy analysis, minimizes the risks of contamination, reduces the odor, and optimizes the workspace.

Artesunate + amodiaquine versus artemether-lumefantrine for the treatment of uncomplicated Plasmodium falciparum malaria in the Colombian Pacific region: a noninferiority trial

INTRODUCTION: In Colombia, there are no published studies for the treatment of uncomplicated Plasmodium falciparum malaria comparing artemisinin combination therapies. Hence, it is intended to demonstrate the non-inferior efficacy/safety profiles of artesunate + amodiaquine versus artemether-lumefantrine treatments. METHODS: A randomized, controlled, open-label, noninferiority (Δ≤5%) clinical trial was performed in adults with uncomplicated P. falciparum malaria using the 28‑day World Health Organization validated design/definitions. Patients were randomized 1:1 to either oral artesunate + amodiaquine or artemether-lumefantrine. The primary efficacy endpoint: adequate clinical and parasitological response; secondary endpoints: - treatment failures defined per the World Health Organization. Safety: assessed through adverse events. RESULTS: A total of 105 patients was included in each group: zero censored observations. Mean (95%CI - Confidence interval) adequate clinical and parasitological response rates: 100% for artesunate + amodiaquine and 99% for artemether-lumefantrine; the noninferiority criteria was met (Δ=1.7%). There was one late parasitological therapeutic failure (1%; artemether-lumefantrine group), typified by polymerase chain reaction as the MAD20 MSP1 allele. The fever clearance time (artesunate + amodiaquine group) was significantly shorter (p=0.002). Respectively, abdominal pain for artesunate + amodiaquine and artemether-lumefantrine was 1.9% and 3.8% at baseline (p=0.68) and 1% and 13.3% after treatment (p<0.001). CONCLUSIONS: Uncomplicated P. falciparum malaria treatment with artesunate + amodiaquine is noninferior to the artemether-lumefantrine standard treatment. The efficacy/safety profiles grant further studies in this and similar populations.

Risk factors associated with hantavirosis fatality: a regional analysis from a case-control study in Brazil

Introduction: In Brazil, hantavirus cardiopulmonary syndrome (HCPS) has a high lethality rate that varies by region. This study aimed to identify the risk factors associated with fatal hantavirosis. Methods: This study was a case-control study that included all laboratory confirmed cases of hantavirosis. The cases were stratified by the different Brazilian regions using data from the Notifiable Diseases Information System. “Cases” were patients who progressed to death, whereas “controls” were patients who were cured. The odds ratio (OR) and the adjusted OR were calculated. Results: Overall, 158 cases and 281 controls were included in this study. In the Midwest region, the cases were 60% less likely to present with flank pain, and the time between the beginning of symptoms and death was shorter than the time between the beginning of symptoms and a cure. In the Southeast region, the cases were 60% less likely to present with thrombocytopenia or reside in rural areas compared to those who progressed to a cure. Additionally, the cases sought medical assistance, notification and investigation more quickly than the controls. In the Southern region, the cases that died were 70% less likely to be male compared to the controls. Conclusions: HCPS manifests with nonspecific symptoms, and there are few published studies related to the condition, so determining a patient's therapeutic strategy is difficult. This study presents findings from different Brazilian regions and highlights the need for further investigations to improve comprehension about regional risk factors associated with hantavirosis and to reduce morbimortality.