MOLECULAR INVESTIGATION OF HEMOTROPIC MYCOPLASMAS IN HUMAN BEINGS, DOGS AND HORSES IN A RURAL SETTLEMENT IN SOUTHERN BRAZIL

SUMMARY The aims of this study were to determine the prevalence of hemoplasmas in a rural Brazilian settlement's population of human beings, their dogs and horses, highly exposed to tick bites; to identify the tick species parasitizing dogs and horses, and analyze factors associated with their infection. Blood samples from 132 dogs, 16 horses and 100 humans were screened using a pan-hemoplasma SYBR green real-time PCR assay followed by a species-specific TaqMan real-time PCR. A total of 59/132 (44.7%) dog samples were positive for hemoplasmas (21 Mycoplasma haemocanisalone, 12 ' Candidatus Mycoplasma haematoparvum' alone and 21 both). Only 1/100 (1.0%) human sample was positive by qPCR SYBR green, with no successful amplification of 16S rRNA or 23 rRNA genes despite multiple attempts. All horse samples were negative. Dogs >1 year of age were more likely to be positive for hemoplasmas ( p= 0.0014). In conclusion, although canine hemoplasma infection was highly prevalent, cross-species hemoplasma transmission was not observed, and therefore may not frequently occur despite overexposure of agents and vectors.

Epidemiological factors related to the transmission risk of Trypanosoma cruzi in a Quilombola community, State of Mato Grosso do Sul, Brazil

INTRODUCTION: This work was an epidemiological investigation of the risk of Trypanosoma cruzi transmission in the rural Quilombola community of Furnas do Dionízio, State of Mato Grosso do Sul, Brazil. METHODS: Of the 71 animals examined, seven were captured (two opossums, Didelphis albiventris; four rats, Rattus rattus; and one nine-banded armadillo, Dasypus novemcinctus) and 64 were domestic (one canine, Canis familiaris; five pigs, Sus scrofa; two bovines, Bos taurus; five caprines, Capra sp.; and 51 ovines, Ovis aries). Parasitological tests were performed to detect parasites in the blood and to identify the morphology of flagellates. These methods included fresh examinations, buffy coat tests and blood cultures. Molecular analysis of DNA for identification of trypanosomatids was performed by polymerase chain reaction (PCR) with primers S35 and S36. RESULTS: The parasitological tests showed flagellates in an opossum and two cattle. The molecular tests showed DNA from T. cruzi in an opossum and a pig. Triatoma sordida was the only triatomine species found in the community, and it colonized households (four specimens) and the surrounding areas (124 specimens). Twenty-three specimens tested positive for flagellates, which were subsequently identified as T. cruzi by PCR. CONCLUSIONS: Data analysis demonstrated that T. cruzi has a peridomestic life cycle that involves both domestic and wild mammals.

Sexually Transmitted Diseases Treatment Guidelines, 2010

These guidelines for the treatment of persons who have or are at risk for sexually transmitted diseases (STDs) were updated by CDC after consultation with a group of professionals knowledgeable in the field of STDs who met in Atlanta on April 18–30, 2009. The information in this report updates the 2006 Guidelines for Treatment of Sexually Transmitted Diseases (MMWR 2006;55[No. RR–11]). Included in these updated guidelines is new information regarding 1) the expanded diagnostic evaluation for cervicitis and trichomoniasis; 2) new treatment recommendations for bacterial vaginosis and genital warts; 3) the clinical efficacy of azithromycin for chlamydial infections in pregnancy; 4) the role of Mycoplasma genitalium and trichomoniasis in urethritis/cervicitis and treatment-related implications; 5) lymphogranuloma venereum proctocolitis among men who have sex with men; 6) the criteria for spinal fluid examination to evaluate for neurosyphilis; 7) the emergence of azithromycin-resistant Treponema pallidum; 8) the increasing prevalence of antimicrobial-resistant Neisseria gonorrhoeae; 9) the sexual transmission of hepatitis C; 10) diagnostic evaluation after sexual assault; and 11) STD prevention approaches.

Caracterización microecologica de nemátodos parásitos presentes en caprinos de zonas áridas de Venezuela

Se determinó que los caprinos de las zonas áridas de Venezuela son parasitados por lassiguientes especies de nemátodos: Haemonchus contortus, Trichostrongylus axei, T. colubriformis, Cooperia curticei, Oesophagostomun columbianum, Skrjabinema ovis y Trichuris globulosa. Los valores del índice de diversidad de Shannon-Weaver oscilaron entre 0,045 y 1,73 bits (X = 1,15 ± 0,24 bits). La diversidad máxima por mes presenta valores que van desde 1 a 2,80 bits (X = 2,49 ± 0,28 bits) y la equitabilidad varió de 0,045 a 0,67 (X = 0,44 ± 0,09). Los valores obtenidos con los datos agrupados en un solo bloque son de 1,25 bits, 2,80 bits y 0,40 para el índice de diversidad de Shannon-Weaver, la diversidad máxima y la equitabilidad respectivamente. En lo referente a las asociaciones parasitarias, se encontró un primer grupo conformado por T. axei, T. colubriformis y H. contortus, un segundo grupo por T. colubriformis, H. contortus y O. columbianum, y un tercero integrado por T. colubriformis, S. ovis y T. globulosa. Los resultados obtenidos permiten además, sugerir em torno al control químico que en los meses en los cuales el valor del índice de diversidad de Shannon-Weaver se aproxime al de la diversidad máxima y el del índice de equitabilidad se aproxime a la unidad, se pueden utilizar antihelmínticos de amplio espectro.

Etiology of community-acquired pneumonia in hospitalized children based on WHO clinical guidelines

Rapport de synthèse: Enjeux de la recherche : La pneumonie communautaire chez l'enfant est un problème de santé publique considérable. Elle est responsable de 2 millions de mort par année, 70% survenant dans les pays en voie de développement. Sous nos latitudes son incidence est de 40/1000 enfants par année, ce qui représente une morbidité importante. Deux difficultés surviennent lorsqu'on cherche à diagnostiquer une pneumonie. La première est de distinguer une pneumonie bactérienne d'une virale, particulièrement chez les petits enfants où les infections virales des voies respiratoires inférieures sont fréquentes. L'OMS a définit la pneumonie selon des critères exclusivement cliniques et une étude effectuée à Lausanne en 2000 a montré que ces critères peuvent être utilisés dans nos contrées. La seconde difficulté est de définir l'agent causal de la pneumonie, ceci pour plusieurs raisons : L'aspiration endotrachéale, seul examen fiable, ne peut être obtenue de routine chez l'enfant vu son caractère invasif, la culture des secrétions nasopharyngées reflète la flore physiologique de la sphère ORL et une bactériémie n'est présente que dans moins de 10% des pneumonies. L'étiologie de la pneumonie reste souvent inconnue, et de ce fait plusieurs enfants reçoivent des antibiotiques pour une infection non bactérienne ce qui contribue au développement de résistances. L'objectif de cette étude était d'effectuer une recherche extensive de l'agent causal de la pneumonie et de déterminer quels facteurs pourraient aider le clinicien à différencier une pneumonie virale de bactérienne, en corrélant l'étiologie avec la sévérité clinique et les marqueurs de l'inflammation. Contexte de la recherche : II s'agissait d'une étude prospective, multicentrique, incluant les enfants âgés de 2 mois à 5 ans hospitalisés pour une pneumonie, selon les critères de l'OMS, dans le service de pédiatrie de Lausanne et Genève entre mars 2003 et Décembre 2005, avant l'implantation de la vaccination antipneumococcique de routine. Chaque enfant, en plus des examens usuels, bénéficiait d'une recherche étiologique extensive : Culture virale et bactérienne, PCR (Mycoplasma Pneumoniae, Chlamydia Pneumoniae, Virus Influenza A et B, RSV A et B, Rhinovirus, Parainfluenza 1-3, enterovirus, human metapneumovirus, coronavirus OC43, E229 ; et NL 63) et détection d'AG viraux dans les sécrétions nasopharyngées ; sérologies virales et bactériennes à l'entrée et 3 semaines après la sortie (AG Influenza A et B, Parainfluenza 1,2 et 3, RSV, Adenovirus, M.Pneumoniae et S.Pneumoniae). Conclusions : Un agent pathogène a été découvert chez 86% des 99 patients retenus confirmant le fait que plus la recherche étiologique est étendue plus le pourcentage d'agent causal trouvé est élevé. Une infection bactérienne a été découverte chez 53% des patients dont 45% avaient une infection à S. Pneumoniae confirmant l'importance d'une vaccination antipneumococcique de routine. La déshydratation et les marqueurs de l'inflammation tels que la C-Reactive Protein et la Procalcitonine étaient significativement plus élevés dans les pneumonies bactériennes. Aucune corrélation n'a été trouvée entre le degré de sévérité de la pneumonie et l'étiologie. L'étude a confirmé la haute prévalence d'infections virales (67%) et de co-infection (33%) dans la pneumonie de l'enfant sans que l'on connaisse le rôle réel du virus dans la pathogenèse de la pneumonie. Perspectives : d'autres études à la suite de celle-ci devraient être effectuées en incluant les patients ambulatoires afin de déterminer, avec un collectif plus large de patient, une éventuelle corrélation entre sévérité clinique et étiologie. Abstract : Community-acquired pneumonia (CAP) is a major cause of death in developing countries and of morbidity in developed countries. The objective of the study was to define the causative agents among children hospitalized for CAP defined by WHO guidelines and to correlate etiology with clinical severity and surrogate markers. Investigations included an extensive etiological workup. A potential causative agent was detected in 86% of the 99 enrolled patients, with evidence of bacterial (53%), viral (67%), and mixed (33%) infections. Streptococcus pneumoniae was accounted for in 46% of CAP. Dehydration was the only clinical sign associated with bacterial pneumonia. CRP and PCT were significantly higher in bacterial infections. Increasing the number of diagnostic tests identifies potential causes of CAP in up to 86% of children, indicating a high prevalence of viruses and frequent co-infections. The high proportion of pneumococcal infections re-emphasizes the importance of pneumococcal immunization.

Establishment and Characterization of a Cell Line from the Mosquito Anopheles albimanus (Diptera: Culicidae)

A new cell line designated LSB-AA695BB, was established from embryos of the mosquito Anopheles albimanus. The primary culture was initiated in April, 1995, and the first passage was made 48 days later. Serial subcultures of the cells have been carried through 90 passages from Abril 1995 to February 1996. The cells were grown at 28°C in MK/VP12 medium, supplemented with 20% fetal bovine serum; the pH tolerance ranged between 6.8 to 7.0. The cells have also been adapted to MM/VP12 medium under the same pH, temperature and serum concentration. The majority of the cells were a fibroblast-type. Isozyme characterization showed a pattern similar to that of An. albimanus pupae and adults but distinct from Ae. taeniorhynchus and Ae. albopictus (C6/36) mosquito cell lines. The culture was shown to be free of mycoplasma, bacteria and fungi. Microsporidia contamination of transovarial transmission was controlled with 6.0 mg/ml of albendazole

A new continuous cell line from the mosquito Psorophora confinnis (Diptera: Culicidae) and its susceptibility to infections with some arboviruses

A new cell line, PC-0199-BR, was established from embryonated eggs of the mosquito Psorophora confinnis. To date (September 2000) it has had 62 continuous passages. This is the first report of a cell line of mosquitoes belonging to the genus Psorophora. Cell growth initially was achieved in the MM/VP12 medium, supplemented with 20% fetal bovine serum; however, the subcultures were later adapted to Grace's medium with 10% fetal bovine serum. Cell morphology in the primary cultures was heterogeneous; but later in the established cell line, the predominant cell type was epithelioid. Cultured cells were predominantly diploid (2n=6); however, chromosome abnormalities were observed in a small proportion of the cells in later passages. C and G band patterns were also determined in the karyotype. The cell line isozyme profiles coincided with pupae and adult samples of the species taken from the same colony. A preliminary arbovirus susceptibility study for the cell line was undertaken. No evidence was observed of contamination of the cell line with bacteria, fungi or mycoplasma.

Do Archaea and bacteria co-infection have a role in the pathogenesis of chronic chagasic cardiopathy?

Chronic cardiopathy (CC) in Chagas disease is a fibrotic myocarditis with C5b-9 complement deposition. Mycoplasma and Chlamydia may interfere with the complement response. Proteolytic enzymes and archaeal genes that have been described in Trypanosoma cruzi may increase its virulence. Here we tested the hypothesis that different ratios of Mycoplasma, Chlamydia and archaeal organisms, which are frequent symbionts, may be associated with chagasic clinical forms. MATERIALS AND METHODS: eight indeterminate form (IF) and 20 CC chagasic endomyocardial biopsies were submitted to in situ hybridization, electron and immunoelectron microscopy and PCR techniques for detection of Mycoplasma pneumoniae (MP), Chlamydia pneumoniae(CP), C5b-9 and archaeal-like bodies. RESULTS: MP and CP-DNA were always present at lower levels in CC than in IF (p < 0.001) and were correlated with each other only in CC. Electron microscopy revealed Mycoplasma, Chlamydia and two types of archaeal-like bodies. One had electron dense lipid content (EDL) and was mainly present in IF. The other had electron lucent content (ELC) and was mainly present in CC. In this group, ELC correlated negatively with the other microbes and EDL and positively with C5b-9. The CC group was positive for Archaea and T. cruzi DNA. In conclusion, different amounts of Mycoplasma, Chlamydia and archaeal organisms may be implicated in complement activation and may have a role in Chagas disease outcome.

Association of human papillomavirus and Chlamydia trachomatis with intraepithelial alterations in cervix samples

The influence of different infectious agents and their association with human papillomavirus (HPV) in cervical carcinogenesis have not been completely elucidated. This study describes the association between cytological changes in cervical epithelium and the detection of the most relevant aetiological agents of sexually transmitted diseases. Samples collected from 169 patients were evaluated by conventional cytology followed by molecular analysis to detect HPV DNA, Chlamydia trachomatis, herpes simplex virus 1 and 2,Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, andTreponema pallidum, besides genotyping for most common high-risk HPV. An association between cytological lesions and different behavioural habits such as smoking and sedentariness was observed. Intraepithelial lesions were also associated with HPV and C. trachomatis detection. An association was also found between both simple and multiple genotype infection and cytological changes. The investigation of HPV and C. trachomatisproved its importance and may be considered in the future for including in screening programs, since these factors are linked to the early diagnosis of patients with precursor lesions of cervical cancer.

Intra-puparial development of the females of Chrysomya albiceps (Wiedemann) (Diptera, Calliphoridae)

Intra-puparial development of the females of Chrysomya albiceps (Wiedemann) (Diptera, Calliphoridae). The chronology and morphological changes that take place during intra-puparial development of Chrysomya albiceps is described based on 254 specimens reared in the laboratory. Larvae were obtained from the eggs laid by a single female. The pre-pupae were separated according to the reduction of larval length and the degree of pigmentation and sclerotization of the cuticle. After pupation, 10 individuals were fixed in Carnoy's solution and preserved in 70% ethanol, 10 individuals were fixed every 3 hours up to complete the first 24 hours (n = 80), the remaining individuals were fixed every six hours up to the 90th hour (n = 110) when 54 females emerged. The pupae were immersed in 5% formic acid for 48 hours and maintained in 70% ethanol, and then dissected and analyzed. C. albiceps shows four intra-puparial stages, each of which were described and compared with those described for Musca domestica, Calliphora erythrocephala, Sarcophaga bullata, Cuterebra tenebrosa, Oestrus ovis and Dermatobia hominis. Four developmental stages may be described: (1) the larva-pupa apolysis, after three hours; (2) the criptocephalic pupa, after six hours; (3) the phanerocephalic pupa, after nine hours; (4) the pharate pupa, after nine hours. The pharate adult is completely formed after 81 hours.

Parachlamydia and rhabdochlamydia: emerging agents of community-acquired respiratory infections in children.

T O THE E DITOR-Besides viruses, Mycoplasma pneumoniae and Chlamydia pneumoniae are common causes of community-acquired respiratory infections (CARI) in children. However, the causal agent of CARI remains unknown in many cases [ 1]. Growing evidence suggests that Chlamydia-related bacteria might have a pathogenic role in humans [ 2, 3]. Parachlamydia acanthamoebae and Protochlamydia naegleriophila have been detected in respiratory clinical samples [ 4, 5], and the role of Parachlamydia acanthamoebae in pneumonia is supported by in vitro studies and animal models [ 6]. Rhabdochlamydia crassificans and Rhabdochlamydia porcellionis are intracellular pathogens of arthropods that also belong to the Chlamydiales order [ 7, 8]. A recent analysis suggests that Rhabdochlamydia species might affect morbidity and mortality in premature newborns [ 9], but their role ...

Composição corporal e exigências nutricionais em magnésio, potássio e sódio de cordeiros Santa Inês

O trabalho objetivou determinar a composição corporal de cordeiros Santa Inês e estimar suas exigências de magnésio, potássio e sódio, para ganho de peso. Foram conduzidos dois experimentos com 18 cordeiros machos em cada um, com peso médio inicial de 25 e 15 kg no primeiro e no segundo experimentos, respectivamente. Em cada experimento, seis animais foram abatidos, para determinação das quantidades de cada mineral retido no corpo, servindo como animais-referência para a técnica do abate comparativo. Os doze animais remanescentes em cada experimento foram divididos em dois grupos: seis animais receberam alimentação ad libitum e seis receberam alimentação restrita. Os cordeiros do grupo ad libitum e restrito foram abatidos quando os do grupo ad libitum atingiram 35 e 25 kg de peso vivo no primeiro e segundo experimentos, respectivamente. A composição corporal em Ca, P, Mg, K e Na foi estimada a partir de equações de regressão do logaritmo da quantidade desses minerais presentes no corpo vazio dos animais, em razão do peso corporal vazio. As exigências líquidas desses minerais por kg de ganho de peso vivo, obtidas a partir da derivação das equações de predição da composição corporal foram: 0,47 e 0,41 g de Mg, 2,32 e 2,05 g de K e 1,33 e 0,55 g de Na, respectivamente, em animais com 15 e 35 kg.

Composição corporal e exigências nutricionais de cálcio e fósforo de cordeiros Santa Inês em crescimento

Dois experimentos foram conduzidos com o objetivo de determinar a composição corporal e estimar as exigências de Ca e P, para ganho em peso, de cordeiros Santa Inês em crescimento. Em cada experimento foram usados 18 animais com 25 a 35 kg de peso vivo (PV), no primeiro, e com 15 a 25 kg no segundo experimento. Seis animais foram abatidos no início de cada experimento para avaliar o conteúdo de Ca e P corporal, servindo como animais referência para o método do abate comparativo, seis animais receberam alimentação ad libitum e seis, alimentação restrita. As exigências líquidas de Ca e P para o ganho em peso foram estimadas a partir da derivação de equações de regressão do logaritmo da quantidade desses minerais presentes no corpo vazio, em função do peso do corpo vazio. As exigências líquidas por quilo de ganho de PV para animais com 15, 25 e 35 kg de PV foram, respectivamente: 11,63, 10,52 e 9,82 g de Ca e 5,82, 4,99 e 4,28 g de P.

Genetic variability of the Brazilian hair sheep breeds

The objectives of this work were to investigate the genetic structure of the Brazilian hair sheep breeds and to determine the origin of the Santa Inês breed. Molecular similarity was determined using Randomly Amplified Polymorphic DNA - Polymerase Chain Reaction markers in 238 individuals from five naturalized sheep breeds: Santa Inês (48 animals), Rabo Largo (48), Somali (48), Morada Nova (48) and Bergamasca (46), collected in Goiás, Sergipe, Bahia, and Ceará States as well as in the Federal District. Fifty-four loci were selected from 19 primers, after a pilot test using 140 primers. Qualitative analyses indicate diagnostic markers for all breeds. All breeds were significantly different from each other. Interbreed differences were explained by 14.92% of the total variation. Santa Inês clustered with Bergamasca (97% bootstrap) and with Rabo Largo, composing the third member of the group (81% bootstrap) while Morada Nova and Somali breeds clustered separately. Each breed should be considered as a separate management and conservation unit, and special care should be taken with Rabo Largo, Morada Nova and Somali breeds, represented by small herds in Brazil.

Gene Ontology Function prediction in Mollicutes using Protein-Protein Association Networks

Abstract Background: Many complex systems can be represented and analysed as networks. The recent availability of large-scale datasets, has made it possible to elucidate some of the organisational principles and rules that govern their function, robustness and evolution. However, one of the main limitations in using protein-protein interactions for function prediction is the availability of interaction data, especially for Mollicutes. If we could harness predicted interactions, such as those from a Protein-Protein Association Networks (PPAN), combining several protein-protein network function-inference methods with semantic similarity calculations, the use of protein-protein interactions for functional inference in this species would become more potentially useful. Results: In this work we show that using PPAN data combined with other approximations, such as functional module detection, orthology exploitation methods and Gene Ontology (GO)-based information measures helps to predict protein function in Mycoplasma genitalium. Conclusions: To our knowledge, the proposed method is the first that combines functional module detection among species, exploiting an orthology procedure and using information theory-based GO semantic similarity in PPAN of the Mycoplasma species. The results of an evaluation show a higher recall than previously reported methods that focused on only one organism network.