Stomoxys calcitrans as possible vector of Trypanosoma evansi among camels in an affected area of the Canary Islands, Spain

Introduction Trypanosoma evansi was first identified in the Canary Islands in 1997, and is still present in a small area of the Archipelago. To date, the disease has exclusively affected camel herds, and has not been detected in any other animal hosts. However potential vectors of Trypanosoma evansi must be identified. Methods One Nzi trap was placed on a camel farm located in the infected area for a period of one year. Results Two thousand five hundred and five insects were trapped, of which Stomoxys calcitrans was the sole hematophagous vector captured. Conclusions Stomoxys calcitrans could be exclusively responsible for the transmission of Trypanosoma evansi among camels in the surveyed area, as other species do not seem to be infected by S. calcitrans in the presence of camels.

Larvicidal activity of the methanol extract and fractions of the green fruits of Solanum lycocarpum(Solanaceae) against the vector Culex quinquefasciatus (Diptera: Culicidae)

IntroductionThe larvicidal activity of Solanum lycocarpumagainst Culex quinquefasciatus is unknown.MethodsWe evaluated the larvicidal activity of extracts of the green fruits of Solanum lycocarpum against third and fourth instar larvae of C. quinquefasciatus.ResultsDichloromethane and ethyl acetate fractions showed the greatest larvicidal effect at 200mg/L (83.3% and 86.7%, respectively). The methanol and dichloromethane, ethyl acetate, and hydromethanolic fractions demonstrated larvicidal effects against C. quinquefasciatus, with LC50 values of 126.24, 75.13, 83.15, and 207.05mg/L, respectively.ConclusionsThus, when considering new drugs with larvicidal activity from natural products, S. lycocarpum fruits may be good candidate sources.

Potential for entomopathogenic fungi to control Triatoma dimidiata (Hemiptera: Reduviidae), a vector of Chagas disease in Mexico

Introduction The use of entomopathogenic fungi to control disease vectors has become relevant because traditional chemical control methods have caused damage to the environment and led to the development of resistance among vectors. Thus, this study assessed the pathogenicity of entomopathogenic fungi in Triatoma dimidiata. Methods Preparations of 108 conidia/ml of Gliocladium virens, Talaromyces flavus, Beauveria bassiana and Metarhizium anisopliae were applied topically on T. dimidiata nymphs and adults. Controls were treated with the 0.0001% Tween-80 vehicle. Mortality was evaluated and recorded daily for 30 days. The concentration required to kill 50% of T. dimidiata (LC50) was then calculated for the most pathogenic isolate. Results Pathogenicity in adults was similar among B. bassiana, G. virens and T. flavus (p>0.05) and differed from that in triatomine nymphs (p=0.009). The most entomopathogenic strains in adult triatomines were B. bassiana and G. virens, which both caused 100% mortality. In nymphs, the most entomopathogenic strain was B. bassiana, followed by G. virens. The native strain with the highest pathogenicity was G. virens, for which the LC50 for T. dimidiata nymphs was 1.98 x108 conidia/ml at 13 days after inoculation. Conclusions Beauveria bassiana and G. virens showed entomopathogenic potential in T. dimidiata nymphs and adults. However, the native G. virens strain presents a higher probability of success in the field, and G. virens should thus be considered a potential candidate for the biological control of triatomine Chagas disease vectors.

Ecoepidemiology and biology of Eratyrus mucronatus Stål, 1859 (Hemiptera: Reduviidae: Triatominae), a sylvatic vector of Chagas disease in the Brazilian Amazon

Introduction Eratyrus mucronatus Stål, 1859 is a wild triatomine vector of Trypanosoma cruzi Chagas, 1909. However, little is known regarding the biology and ecoepidemiology of this triatomine in the Brazilian Amazon. The present study describes the biology of E. mucronatus grown under laboratory conditions and the epidemiological aspects of its natural breeding sites. Methods Five colonies were monitored in the field for 3 years. Temperature and humidity measurements were taken in the mornings and afternoons at the natural breeding sites, and the behavior and distribution of the nymphs and adults were observed in the wild colony. We also monitored the life cycle under controlled laboratory conditions. Results Some factors that were considered decisive for the establishment of these colonies were present at all of the colonies studied in the field. These factors included an active termite nest, a vertebrate for repast, and dry and shaded substrates with temperatures of 24-28°C and with humidity of 80-90%. A generation was developed in 274 days under these microclimatic conditions in the laboratory. Conclusions The climatic variables described in the field indicate that these environmental parameters have a limiting effect on the dispersal and colonization of E. mucronatus to new environments. In addition, the long period of development to adulthood demonstrates that only one generation can develop per year even under the more favorable laboratory conditions.

A rapid and simple method to detect ESBL in Enterobacter cloacae based on MIC of cefepime

INTRODUCTION: The aim of this study was to identify a rapid and simple phenotypic method for extended-spectrum β-lactamase (ESBL) detection in Enterobacter cloacae. METHODS: A total of 79 consecutive, non-repeated samples of E. cloacae were evaluated. Four phenotypic methods were applied for ESBL detection, results were compared to multiplex polymerase chain reaction (PCR) as the gold standard reference method: 1) ceftazidime and cefotaxime disks with and without clavulanate, both with boronic acid added; 2) disk approximation using cefepime and amoxicillin/clavulanate; 3) ESBL screening by minimum inhibitory concentration (MIC) ≥ 16µg/mL and 4) by MIC ≥ 2µg/mL for cefepime. RESULTS: Method 4 showed the best combination of sensitivity (100%) and specificity (94%). CONCLUSIONS: MIC ≥ 2µg/mL for cefepime would be very useful for the phenotypic detection of ESBL in samples of E. cloacae.

Spatial analysis of visceral leishmaniasis in the municipality of Rondonópolis, in the Brazilian State of Mato Grosso, from 2003 to 2012: human, canine and vector distribution in areas of disease transmission

INTRODUCTION: Visceral leishmaniasis (VL) is a zoonosis of great importance to public health and is considered a neglected disease by the World Health Organization. The disease has expanded and become more prevalent in urban areas in Brazil. METHODS: Geospatial analyses were performed and thematic maps of the triad of the disease were produced for the study period (2003-2012) in the urban area of the municipality of Rondonópolis in the midwestern State of Mato Grosso (MT), Brazil, TerraView 4.2.2 software was used for the analyses. RESULTS: A total of 87.9% of the 186 confirmed human cases of VL were cured. Children between the ages of 1 and 4 were the most affected. Registered deaths were predominant among adults aged 60 years or older. The urban area of the municipality consists of eight strata and 12 census districts include 237 neighborhoods. All sectors had confirmed cases of VL. During the study period, human cases of the disease were recorded in 90 neighborhoods. The 23 deaths from the disease were distributed in 21 neighborhoods. Sandflies carrying the parasite were captured in 192 out of 200 neighborhoods evaluated for the presence of the VL vector. The presence of dogs carrying the parasite was confirmed in, 140 out of 154 surveyed neighborhoods. CONCLUSIONS: The data demonstrated the endemic nature of VL, with a high percentage of infected children, a high distribution of canine infection, and a wide adaptation and dispersal of the vectors in the urban environment. These results, illustrate the process of urbanization of VL in the municipality of Rondonópolis, MT, Brazil.

Polymerase chain reaction-based method for the identification of Leishmania (Viannia) braziliensis and Leishmania (Viannia) guyanensis in mucosal tissues conserved in paraffin

ABSTRACTINTRODUCTION: In the Americas, mucosal leishmaniasis is primarily associated with infection by Leishmania (Viannia) braziliensis. However, Leishmania (Viannia) guyanensis is another important cause of this disease in the Brazilian Amazon. In this study, we aimed at detecting Leishmaniadeoxyribonucleic acid (DNA) within paraffin-embedded fragments of mucosal tissues, and characterizing the infecting parasite species.METHODS: We evaluated samples collected from 114 patients treated at a reference center in the Brazilian Amazon by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses.RESULTS: Direct examination of biopsy imprints detected parasites in 10 of the 114 samples, while evaluation of hematoxylin and eosin-stained slides detected amastigotes in an additional 17 samples. Meanwhile, 31/114 samples (27.2%) were positive for Leishmania spp. kinetoplast deoxyribonucleic acid (kDNA) by PCR analysis. Of these, 17 (54.8%) yielded amplification of the mini-exon PCR target, thereby allowing for PCR-RFLP-based identification. Six of the samples were identified as L. (V.) braziliensis, while the remaining 11 were identified as L. (V.) guyanensis.CONCLUSIONS: The results of this study demonstrate the feasibility of applying molecular techniques for the diagnosis of human parasites within paraffin-embedded tissues. Moreover, our findings confirm that L. (V.) guyanensisis a relevant causative agent of mucosal leishmaniasis in the Brazilian Amazon.

Larvicidal activity of oils, fatty acids, and methyl esters from ripe and unripe fruit of Solanum lycocarpum (Solanaceae) against the vector Culex quinquefasciatus (Diptera: Culicidae)

ABSTRACTINTRODUCTION:The larvicidal activity of oils, fatty acids, and methyl esters of Solanum lycocarpum fruit against Culex quinquefasciatus is unknown.METHODS:The larvicidal activity of samples of ripe and unripe fruit from S. lycocarpum was evaluated against third and fourth instar larvae of C. quinquefasciatus .RESULTS:The oils, fatty acids, and methyl esters of S. lycocarpum showed the greatest larvicidal effect (57.1-95.0%) at a concentration of 100mg/L (LC 50values between 0.70 and 27.54mg/L).CONCLUSIONS:Solanum lycocarpum fruit may be a good source of new natural products with larvicidal activity.

Knowledge and practices related to dengue and its vector: a community-based study from Southeast Brazil

Abstract INTRODUCTION: This study investigated the knowledge of users of primary healthcare services living in Ribeirão Preto, Brazil, about dengue and its vector. METHODS: A cross-sectional survey of 605 people was conducted following a major dengue outbreak in 2013. RESULTS: Participants with higher levels of education were more likely to identify correctly the vector of the disease. CONCLUSIONS: The results emphasize the relevance of health education programs, the continuous promotion of educational campaigns in the media, the role of the television as a source of information, and the importance of motivating the population to control the vector.

The use of a modular titanium endoprosthesis in skeletal reconstructions after bone tumor resections: method presentation and analysis of 37 cases

We analyzed 37 patients who underwent segmental wide resection of bone tumors and reconstruction with a modular titanium endoprosthesis at the Orthopaedic Oncology Group, between 1992 and 1998. Twelve patients were male and 25 were female, with a mean age of 30 years (9 - 81). The mean follow-up was 14 months (2 - 48). The diagnoses were: osteosarcoma (14 cases), metastatic carcinoma (10), Ewing's sarcoma (4), giant cell tumor (4), malignant fibrous histiocytoma (3), chondrosarcoma (1), and aneurysmal bone cyst (1). Eleven articulated total knee, 8 partial proximal femur with bipolar acetabulum, 8 partial proximal humerus, 3 total femur, 2 partial proximal tibia, 2 diaphyseal femur, 2 diaphyseal humerus, and 1 total proximal femur with cementless acetabulum endoprosthesis implant procedures were done. The complications related to the procedure included: infection (5 cases), dislocation (3), module loosening (1), and ulnar nerve paresthesia (1). We used the following criteria for the clinical evaluation: presence of pain, range of motion, reconstruction stability, surgical and oncologic complications, and patient acceptance. The results were good in 56.8% of the cases, regular in 32.4% and poor in 10.8%.

New method for evaluation of cutaneous sensibility in diabetic feet: preliminary report

Diabetic neuropathy is an important complication of the disease, responsible for ulceration and amputation of the foot. Prevention of these problems is difficult mainly because there is no method to correctly access sensibility on the skin of the foot. The introduction of the Pressure-Specified Sensory Device (PSSD TM) in the last decade made possible the measurement of pressure thresholds sensed by the patient, such as touch, both static and in movement, on a continuous scale. This paper is the first in Brazil to report the use of this device to measure cutaneous sensibility in 3 areas of the foot: the hallux pulp, the calcaneus, and the dorsum, which are territories of the tibial and fibular nerves. METHOD: Non-diabetic patients were measured as controls, and 2 groups of diabetic patients - with and without ulcers - were compared. The PSSD TM was used to test the 3 areas described above. The following were evaluated: 1 PS (1-point static), 1 PD (1-point dynamic), 2 PS (2-points static), 2 PD (2-points dynamic). RESULTS: The diabetic group had poorer sensibility compared to controls and diabetics with ulcers had poorer sensibility when compared to diabetics without ulcers. The differences were statistically significant (P <.001). CONCLUSION: Due to the small number of patients compared, the results should be taken as a preliminary report.

PKO: alternative method for isolating mycobacteria from sputum

We elaborated an alternative culture method, which we denominated PKO (initials in tribute of respect to Petroff, Kudoh and Ogawa), for isolating Mycobacterium tuberculosis from sputum for diagnosis of pulmonary tuberculosis (TB), and to compare its performance with the Swab and Petroff methods. For the technique validation, sputum samples from patients suspected of pulmonary TB cases were examined by acid-fast microscopy (direct and concentrated smear), PKO, Swab and Petroff methods. We found that Petroff and PKO methods have parity in the effectiveness of M. tuberculosis isolation. However, by the PKO method, 65% of isolated strains were detected in a period of £15 days, while by the Petroff method the best detection was in an interval of 16-29 days (71%). In positive smear samples, the average time of PKO isolation is only superior to the one related for Bactec 460TB. In conclusion, the exclusion of the neutralization stage of pH in the PKO reduces the manipulation of the samples, diminishes the execution time of the culture according to the Petroff method and facilitates the qualification of professionals involved in the laboratorial diagnosis of Tuberculosis.

A new species of Cerqueirellum Py-Daniel, 1983 (Diptera: Simuliidae) and proven new vector of mansonelliasis from the Ituxi River, Amazon basin, Brazil

A new species of the genus Cerqueirellum Py-Daniel, 1983 (Diptera: Simuliidae) is described. The adults are similar to the species C. oyapockense (Floch & Abonnenc, 1946) and C. roraimense (Nunes de Mello, 1974), of which the females are similar, and the males present discrete differences. The main differences of this new species to others of the genus Cerqueirellum are the integument of the larva recovered from stout spines and long cephalic trichomes in the pupa. Some females were infected with Mansonella ozzardii (Manson, 1897) (Nematoda, Onchocercidae) and probably transmit mansonelliasis in the Ituxi river, state of Amazonas, Brazil.

Reliability of the interpretation of coronary angiography by the simple visual method

OBJECTIVE: Evaluation of inter and intraobserver reproducibility of by the visual method interpretation of cineangiogram in a clinically based context. METHODS: Five interventional cardiologists analyzed 11 segments of 8 coronary cineangiograms at a two month apart sessions. The percent luminal reduction by the lesions were analyzed by two different classifications: in one (A) the lesions were graded in 0% = absent, 1-50% = mild, 51 - 69 = moderate, and > or = 70% = severe; the other classification (B) was a dichotomic one : <70% = nonsignificant and > or = 70%=significant lesions. The agreement were measured by the kappa (k) index. RESULTS: Interobserver agreement was moderate for classification A (1st measurement, k = 0.36 -- 0.63, k m = 0.49; 2nd measurement, k = 0.39-0.68, k m = 0.52) and good for classification B (1st measurement, k = 0.55-0.73, k m = 0.63; 2nd measurement, k = 0.37-0.82, k m = 0.61). Intraobserver levels of agreement were k = 0.57-0.95 for classification A and 0.62-1.0 for classification B. CONCLUSION: The higher level of reproducibility obtained by adopting the dichotomous criteria usually considered for ischemic limits demonstrates that in the present clinical context, the reliability of the simple visual method is adequate for the identification of patients with clinically significant lesions and candidates for myocardial revascularization procedures.

Sôbre o Trypanosoma conorrhini, hemoparasito do rato transmitido pelo Triatoma rubrofasciata: presença do vector infectado na cidade do Rio de Janeiro

Os autores, após historiar a desccberta do Trypanosoma conorrhini (DONOVAN, 1909) (sinonimia Crithidia conorrhini DONOVAN, 1909, Trypanosoma boy lei LAFONT, 1912) no inseto transmissor e no hospedeiro vertebrado, o Rattus rattus diardi, fazem um estudo sumário do seu vector intermediário, o barbeiro cosmopolita Triatoma rubrofasciata (DE GEER, 1773) . Em seguida, referem a captura, no centro da cidade do Rio de Janeiro, Brasil, de um exemplar adulto dêste barbeiro parasitado por flagelados que foram identificados ao Trypanosoma conorrhini, fato êste pela primeira vez verificado no Novo Mundo. Certas formas evolutivas dêste parasito no inseto são muito semelhantes às do Schizotrypanum cruzi, mas os tripanosomas metacíclicos apresentam alguns caractéres morfológicos que permitem seu reconhecimento: os mais notáveis são o pequeno tamanho e a colocação do núcleo muito para trás, bem junto ao blefaroplasto. São dadas as medidas de 100 tripanosomas metaciclicos do T. conorrhini e de Schizotrypanum, cujo comprimento total médio foi, respectivamente, 13.88 u e 19.85 u. Nas seguintes espécies de barbeiro foi facilmente obtida a evolução do Trypanosoma conorrhini: Triatoma infestans, Triatoma vitticeps, Rhodnius prolixus e Panstrongylus megistus. Por inoculação de triatomas infectados foi obtida a transmissão do T. conorrhini ao camondongo branco, ao rato e ao macaco rhesus. As infecções foram muito fracas, custando-se a ver o tripanosoma no sangue, a fresco. Em gotas espêssas êle é encontrado com mais facilidade, mas o método de escôlha para verificação da sua presença no sangue dos animais é o xenodiagnóstico. Nesta prova o Triatoma infestans foi empregado com os resultados mais favoráveis. Por êste processo foi observada a infecção inaparente de um camondongo até 53 dias depois da inoculação. A forma sanguícola do T. conorrhini é um tripanosoma ttpico, de grande dimensões (40-60u), de extremidade posterior muito alongada e pontuda, membrana ondulante ampla e com largas pregas, blefaroplasto subcentral; em muitos indivíduos é notável uma estrutura particular, junto e à frente do blefaroplasto. Poucas horas depois de inoculados com as dejeções, os flagelados passam para a circulação, transformando-se em 3-4 dias nos grandes tripanosomas que acabam de ser descritos. O número de parasitos depende do número de flagelados inoculados. Não se conhece o processo de multiplicação do T. conorrhini no vertebrado, nunca tendo sido observados tripanosomas em via de divisão no sangue nem formas proliferativas nos tecidos. As sub-inoculações (de animal a animal) em geral não passam da primeira (MORISHITA, 1935). Tentativas até agora realizadas por outros pesquisadores no sentido de cultivar o T. conorrhini a partir do sangue de animais infectados, têm sido negativas. Por enquanto as pesquisas tendentes a determinar o hospedeiro vertebrado do parasito no Rio de Janeiro limitam-se a aplicação do xenodiagnós-tico em cinco ratos do Morro de Santo Antônio, cujo resultado...