Pulmonary Telerehabilitation for People With Chronic Obstructive Pulmonary Disease in Brazil: A Mixed-Methods Feasibility Study.

This study assessed the feasibility of pulmonary telerehabilitation‧s (PTR) acceptability, implementation, practicality, and adaptation for people with Chronic Obstructive Pulmonary Disease (COPD) in Brazil. It also explored associations with clinical and socioeconomic features of Brazilians with COPD. This mixed-method study included thirty-one participants with COPD (age 62±10 years; FEV1= 72±14% predicted). Most participants (74.2%) reported good PTR session acceptability on the System Usability Scale and scores of 4.6±0.3 and 4.5±0.6 on a 1-5 Likert-type scale of implementation and practicality, respectively. Participants suggested adaptations for better comfort on the exercise bike and varying exercise modalities. PTR acceptability was associated with participants' younger age (rs=-0.57, p<0.01) and higher education (rs=0.51, p<0.01). PTR is feasible for people with COPD in Brazil regarding acceptability, implementation, practicality, and adaptation. Younger age and higher educational level are associated with greater PTR acceptability.

The Brazilian version of the telehealth usability questionnaire (telehealth usability questionnaire Brazil): translation, cross-cultural adaptation, and psychometric properties.

OBJECTIVE: The objectives of this study were to translate and cross-culturally adapt the telehealth usability questionnaire into Brazilian Portuguese and to evaluate its psychometric properties. METHODS: This was a methodological validation study carried out in two phases. In phase 1, the telehealth usability questionnaire was cross-culturally adapted with 10 participants comprising the expert committee members, including 5 healthcare professionals with theoretical and practical knowledge of telehealth, 1 methodologist, and 4 translators. This phase was performed at Universidade Federal de Juiz de Fora Physiotherapy Clinic School. In phase 2, the psychometric properties of telehealth usability questionnaire Brazil were analyzed. This phase included in-person assessments at Márcio Cunha Hospital, Minas Gerais. The recruitment period for both phases was from April 2020 to February 2021. Content validity, reliability, internal consistency, and criterion validity were analyzed. The criterion validity was evaluated using correlation with a validated instrument: the system usability scale. RESULTS: The telehealth usability questionnaire was adequately translated and cross-culturally adapted. The telehealth usability questionnaire Brazil presented an excellent content validity index of 0.96 with percentages of understanding higher than 90%. The telehealth usability questionnaire Brazil demonstrated great internal consistency (α=0.94 and ω=0.94), excellent intra-rater reliability (intraclass correlation coefficient=0.85, 95%CI 0.75-0.91), no difference between the test and retest [T (0.425), p>0.673], and no proportional bias (p=0.205). There was a moderate correlation between telehealth usability questionnaire Brazil and the system usability scale (r=0.52, p<0.0001). CONCLUSION: The telehealth usability questionnaire was adequately translated and cross-culturally adapted into Brazilian Portuguese and showed adequate psychometric properties for use in telehealth clinical practice and research in Brazilian-Portuguese-speaking individuals.

The Brazilian version of the telehealth usability questionnaire (telehealth usability questionnaire Brazil): translation, cross-cultural adaptation, and psychometric properties

SUMMARY OBJECTIVE: The objectives of this study were to translate and cross-culturally adapt the telehealth usability questionnaire into Brazilian Portuguese and to evaluate its psychometric properties. METHODS: This was a methodological validation study carried out in two phases. In phase 1, the telehealth usability questionnaire was cross-culturally adapted with 10 participants comprising the expert committee members, including 5 healthcare professionals with theoretical and practical knowledge of telehealth, 1 methodologist, and 4 translators. This phase was performed at Universidade Federal de Juiz de Fora Physiotherapy Clinic School. In phase 2, the psychometric properties of telehealth usability questionnaire Brazil were analyzed. This phase included in-person assessments at Márcio Cunha Hospital, Minas Gerais. The recruitment period for both phases was from April 2020 to February 2021. Content validity, reliability, internal consistency, and criterion validity were analyzed. The criterion validity was evaluated using correlation with a validated instrument: the system usability scale. RESULTS: The telehealth usability questionnaire was adequately translated and cross-culturally adapted. The telehealth usability questionnaire Brazil presented an excellent content validity index of 0.96 with percentages of understanding higher than 90%. The telehealth usability questionnaire Brazil demonstrated great internal consistency (α=0.94 and ω=0.94), excellent intra-rater reliability (intraclass correlation coefficient=0.85, 95%CI 0.75-0.91), no difference between the test and retest [T (0.425), p>0.673], and no proportional bias (p=0.205). There was a moderate correlation between telehealth usability questionnaire Brazil and the system usability scale (r=0.52, p<0.0001). CONCLUSION: The telehealth usability questionnaire was adequately translated and cross-culturally adapted into Brazilian Portuguese and showed adequate psychometric properties for use in telehealth clinical practice and research in Brazilian-Portuguese-speaking individuals.

Body fat estimated by equations based on anthropometric parameters correlates with bioelectrical impedance in patients undergoing bariatric surgery / Gordura corporal estimada por equações baseadas em parâmetros antropométricos correlaciona-se com a bioimpedância elétrica em pacientes submetidos à cirurgia bariátrica

Introduction: predictive equations to estimate body fat based on simple anthropometric parameters are easy to use in the clinical practice.Objective: to evaluate the relationship between predictive equations based on anthropometric parameters and bioelectrical impedance to estimate body fat in individuals undergoing bariatric surgery.Methods: a prospective and longitudinal study carried out with individuals undergoing bariatric surgery. Body weight, body mass index, waist circumference and body fat percentage estimated by anthropometric parameters and by impedance were evaluated at three moments, one month before, two and six months after surgery. Data were analyzed by one-way ANOVA for repeated measures with Holm-Sidak ́s post hoc or Friedman test with Tukey ́s post hoc, and Pearson or Spearman correlations, according to data distribution. Significance level adopted 5%.Results: twenty-five subjects composed the final sample. All anthropometric parameters reduced significantly over time (p<0.001). Except for Lean et al equation before surgery, the body fat percentage estimated by other formulas showed a strong correlation with impedance in all moments, with the highest correlation strength observed in Gómez-Ambrosi et al. equation.Conclusion: in the present study, the equations used showed a good correlation with bioelectrical impedance, and the Gómez-Ambrosi et al. equation as a better option to the use of bioimpedance to assess changes in body fat percentage of patients undergoing bariatric surgery for the treatment of severe obesity.

Introdução: Objetivos: analisar o consumo alimentar e os fatores associados ao estado nutricional de crianças menores de dois anos de vida.Método: Estudo de corte transversal realizado com uma amostra de 344 lactentes menores de dois anos de idade e suas respectivas mães, acompanhadas em Unidades de Saúde da Família. As variáveis sociodemográficas, antropométrica das mães e dos lactentes e o consumo alimentar dessas crianças foram avaliadas por meio de questionário estruturado. A força de associação entre as variáveis dependente e as independentes foi avaliada pelo odds ratio, tanto na análise univariada quanto na múltipla, com nível de significância de 5%.Resultados: A prevalência do estado nutricional inadequado foi de 38,08%. Observou que 29,09% das crianças menores de seis meses de idade não chegaram a receber leite materno de forma exclusiva ou o tempo de oferta foi inferior a 30 dias. Notou-se o consumo de alimentos ultraprocessados, principalmente, do suco industrializado no último mês [OR:1,96, IC 95%: 1,05-3,65], baixa ingestão de frutas e o hábito de comer assistindo televisão nos lactentes maiores de seis meses. Após ajuste para variáveis de confusão, permaneceram associadas ao estado nutricional: anemia gestacional [OR: 3,58 IC: 1,46-8,77] e trabalho materno [OR, 0,38, IC 95%: 0,18-0,80].Conclusão: A presença do estado nutricional inadequado, caracterizado pelo baixo ou excesso de peso, associou-se ao fato de a mãe trabalhar e à anemia gestacional. Ademais, constatou-se a participação precoce de alimentos ultraprocessados na alimentação das crianças menores de 24 meses de idade, substituindo alimentos considerados naturais e saudáveis, evidenciando assim práticas alimentares inadequadas frente às recomendações para a faixa etária.

International interlaboratory validation of a nested PCR for molecular detection of Babesia bovis and Babesia bigemina, causative agents of bovine babesiosis.

Babesia bovis and B. bigemina are tick-transmitted parasites causing bovine babesiosis, characterized by significant morbidity and mortality leading to economic losses to the livestock industry in tropical and subtropical regions worldwide. Animals that recover from acute infection remain carriers with low parasitemia acting as a source of transmission, and often escape detection. An improved diagnosis of a B. bovis and/or B. bigemina infection of carrier animals is enabled by the availability of detection methods with high sensitivity. To this end, two nested PCR assays targeting the cytochrome b (cytb) genes of B. bovis and B. bigemina (cytb-nPCR), have been recently developed and an increased sensitivity with respect to reference protocols has been shown (Romero-Salas et al., 2016). In this study, the specificity against a panel of hemoparasites that potentially co-occur with B. bovis and B. bigemina was demonstrated to ensure applicability of the cytb-nPCR assays in a wide range of regions where bovine babesiosis is endemic. Furthermore, we compared both reported cytb-nPCR assays with reference nPCR and qPCR protocols for (i) their capability to detect carrier animals in the field, and (ii) their reproducibility when performed in different laboratories by independent operators. We show that, in a panel of bovine field samples (n = 100), the cytb-nPCR assays detected a considerably higher number of 25% B. bovis and 61% B. bigemina-positive animals compared to 7% and 20% B. bovis and 55% and 49% B. bigemina-positive animals when tested by reference nPCR and qPCR protocols, respectively. Cytb-nPCRs were also found superior in the detection of carrier animals when field samples from Africa were analyzed. In addition, both the B. bovis and B. bigemina cytb-nPCR assays were independently validated in a single blinded study in three laboratories. Importantly, no significant differences in the number/percentage of infected animals was observed using cytb-nPCR assays. In summary, the cytb-nPCR assays detected a considerably higher number of chronically infected B. bovis and B. bigemina carrier animals compared to reference nPCR and qPCR protocols, when applied in different epidemiological field situations. Furthermore, a high reproducibility between laboratories could be demonstrated.

Políticas públicas para o paradesporto: programa bolsa-atleta paralímpico no Mato Grosso do Sul/Brasil / Parasport public policy: paralympic athlete scholarship program in Mato Grosso do Sul/Brazil / Políticas públicas para el paradeporte: programa beca-atleta paralímpico en Mato Grosso do Sul/Brasil

O objetivo foi analisar o Programa Bolsa-Atleta paralímpico no estado de Mato Grosso do Sul, a partir do número de bolsas, modalidades e categorias. Configurou-se como um estudo descritivo-exploratório, com análise de dados na Plataforma de Dados do Inteligência Esportiva, entre os anos 2004-2020. Foram identificados 114 atletas paralímpicos, 77 do gênero masculino e 37 do feminino, contemplados em 12 modalidades paralímpicas, com destaque para o atletismo. Somaram-se 484 bolsas distribuídas entre as categorias do programa, com o predomínio da categoria nacional (61,98%) e estudantil (12,39%). Do total, houve modificação dos atletas entre as categorias, 45,61% mantiveram a bolsa por quatro ou mais anos e 54,39% só tiveram a bolsa entre um e três anos. Conclui-se que o Programa Bolsa-Atleta paralímpico acompanhou os investimentos do governo federal, com o pico de bolsas durante o Plano Brasil Medalhas 2016 e baixa em 2020, em decorrência da pandemia e prioridade governamental. (AU)

The main goal was to analyze the Paralympic Athletes Scholarship Program at Mato Grosso do Sul state, Brazil, based on the number of scholarships, sports modalities and categories. The research was descriptive exploratory, with data from website Sport Intelligence, between the years 2004-2020. We identified 114 Paralympic athletes, 77 male and 37 female, contemplated in 12 Paralympic modalities, with emphasis on athletics. There were 484 scholarships distributed among the categories, predominance the national category (61.98%) and student category (12.39%). From these athletes, there were modifications from categories, 45.61% kept the scholarship for four years or more and 54.39% had the scholarship between one and three years. We concluded that the Paralympic Athletes Scholarship Program followed the Federal Government budget, with high investment with the Brazilian Medal Plan 2016, and the lower investment in 2020, due to the pandemic and government priority. (AU)

El objetivo fue analizar el Programa Beca-Atleta paralímpico en el Estado de Mato Grosso do Sul/Brasil, con base en el número de becas, modalidades y categorías. Se configuró como un estudio descriptivo-exploratorio, con análisis de datos en la Plataforma de Datos de Inteligencia Deportiva, entre los años 2004 y 2020. Se identificaron 114 atletas paralímpicos, 77 del género masculino y 37 del femenino, contemplados en 12 modalidades paralímpicas, donde destaca el atletismo. Se encontraron 484 becas distribuidas entre las categorías del programa, con predominio de la categoría nacional (61,98%) y estudiantil (12,39%). El el total, hubo modificación de los atletas entre las categorías, 45,61% mantuvieron la beca por cuatro o más años y 54,39% solo tuvieron la beca por entre uno y tres años. Se concluyó que el Programa Beca-Atleta paralímpico siguió la inversión del Gobierno Federal, con un pico de becas durante el Plan Brasil Medallas 2016 y el menor número de becas en 2020, como consecuencia del período pandémico y de la prioridad gubernamental. (AU)

Barreiras e facilitadores para o encaminhamento de pacientes com doença pulmonar obstrutiva crônica à reabilitação pulmonar e indicação à prática de atividade física: um estudo qualitativo / Barriers and facilitators for referring patients with chronic obstructive pulmonary disease to pulmonary rehabilitation and physical activity recommendation: a qualitative study

Introdução: A doença pulmonar obstrutiva crônica (DPOC) possui alta prevalência e gera altos custos aos sistemas de saúde. A reabilitação pulmonar (RP) é uma intervenção não-farmacológica que gera benefícios clínicos para os pacientes e economia de gastos para o sistema de saúde devido a redução do número de exacerbações da doença e internações hospitalares. Embora fortemente recomendada, a RP tem sido subutilizada havendo um baixo índice de indicações. Objetivo: Identificar as barreiras e os facilitadores para o encaminhamento de pacientes com DPOC à RP e à prática de atividade física. Métodos: Estudo qualitativo, realizado com médicos atuantes na atenção primária à saúde (APS) e uso de entrevistas semiestruturadas. As entrevistas foram gravadas e transcritas literalmente e submetidas a análise de conteúdo. Itens (trechos) identificados nos discursos e relacionados ao encaminhamento de pacientes com DPOC à RP e à prática de atividade física foram classificados como barreiras ou facilitadores. Tal classificação se baseou entre os quatorze domínios da Theoretical Domains Framework (TDF) e suas definições para identificar determinantes de comportamento entre os participantes relevantes para a mudança na prática clínica. Resultados: Setecentos e cinquenta e seis itens foram identificados nos discursos de vinte e um participantes e classificados como sendo barreiras (n= 279, 37%) ou facilitadores (n= 477, 63%) relacionados principalmente aos domínios da TDF de "Conhecimento" (n= 245, 32%), "Contexto e recursos ambientais" (n= 124, 16%) e "Papel e identidade social/profissional" (n= 107, 14%). Conclusão: As principais barreiras identificadas foram relacionadas aos programas de RP devido ao conhecimento restrito dos profissionais médicos sobre programas de RP, a disponibilidade reduzida de locais que oferecem estes programas e as dificuldades de acesso dos pacientes à RP. Os principais facilitadores identificados foram relacionados a orientação e o conhecimento pelos médicos sobre os benefícios da atividade física e a presença dos grupos de exercícios fornecidos na APS.

Introduction: Chronic obstructive pulmonary disease (COPD) has a high prevalence and entails high costs to health systems. Pulmonary rehabilitation (PR) is a non-pharmacological intervention that generates clinical benefits for patients and cost savings for the health system due to a reduced number of exacerbations and hospitalizations. Although strongly recommended, PR has been underutilized, with a low rate of indications. Objective: To identify barriers and facilitators for referring patients with COPD to PR and physical activity. Method: Qualitative study was carried out with medical staff working in primary health care using semi-structured interviews. The interviews were recorded, transcribed literally and underwent content analysis. Items (text excerpts) identified in the speeches and related to the referral of patients with COPD to PR and physical activity were classified according to as barriers or facilitators, and among the fourteen domains of the Theoretical Domains Framework (TDF) and its definitions; aiming to identify the determinants of behaviour which were relevant to change in clinical practice. Results: Seven hundred and fifty-six items were identified in the speeches of twenty-one participants and classified as barriers (n= 279, 37%) or facilitators (n= 477, 63%) mainly related to the TDF domains of "Knowledge" (n= 245, 32%), "Context and environmental resources" (n=124, 16%) and "Role and social/professional identity" (n= 107, 14%). Conclusion: The main barriers identified were related to referring patients PR due to lack of knowledge by medical staff about PR programs, the limited number of programs available, and the restricted access to PR program locations. The main facilitators were related to physical activity, mainly the medical staff's knowledge on physical activity benefits and availability of exercise group activities delivered at primary health care centres.

Expanding our knowledge on African trypanosomes of the subgenus Pycnomonas: A novel Trypanosoma suis-like in tsetse flies, livestock and wild ruminants sympatric with Trypanosoma suis in Mozambique.

Among the subgenera of African tsetse-transmitted trypanosomes pathogenic to livestock, the least known is the subgenus Pycnomonas, which contains a single species, Trypanosoma suis (TSU), a pathogen of domestic pigs first reported in 1905 and recently rediscovered in Tanzania and Mozambique. Analysis by Fluorescent Fragment Length Barcoding (FFLB) revealed an infection rate of 20.3% (108 out of 530 tsetse flies) in a recent study in the Gorongosa and Niassa wildlife reserves in Mozambique, and demonstrated two groups of Pycnomonas trypanosomes: one (14.1%, 75 flies) showing an FFLB profile identical to the reference TSU from Tanzania, and the other (6.2%, 33 flies) differing slightly from reference TSU and designated Trypanosoma suis-like (TSU-L). Phylogenetic analyses tightly clustered TSU and TSU-L from Mozambique with TSU from Tanzania forming the clade Pycnomonas positioned between the subgenera Trypanozoon and Nannomonas. Our preliminarily exploration of host ranges of Pycnomonas trypanosomes revealed TSU exclusively in warthogs while TSU-L was identified, for the first time for a member of the subgenus Pycnomonas, in ruminants (antelopes, Cape buffalo, and in domestic cattle and goats). The preferential blood meal sources of tsetse flies harbouring TSU and TSU-L were wild suids, and most of these flies concomitantly harboured the porcine trypanosomes T. simiae, T. simiae Tsavo, and T. godfreyi. Therefore, our findings support the link of TSU with suids while TSU-L remains to be comprehensively investigated in these hosts. Our results greatly expand our knowledge of the diversity, hosts, vectors, and epidemiology of Pycnomonas trypanosomes. Due to shortcomings of available molecular diagnostic methods, a relevant cohort of trypanosomes transmitted by tsetse flies to ungulates, especially suids, has been neglected or most likely misidentified. The method employed in the present study enables an accurate discrimination of trypanosome species and genotypes and, hence, a re-evaluation of the "lost" subgenus Pycnomonas and of porcine trypanosomes in general, the most neglected group of African trypanosomes pathogenic to ungulates.

Desmascarando o queratocone: efeito do epitélio corneano no queratocone / Unmasking keratoconus: effect of corneal epithelium in keratoconus

Resumo Objetivo: Na presença de uma córnea irregular, como no queratocone, o epitélio tem uma distribuição diferente do normal, "mascarando" a irregularidade do estroma. A avaliação topográfica sem considerar o efeito do epitélio pode induzir uma avaliação menos correta, afetando a precisão de um eventual tratamento guiado por topografia. O objetivo deste trabalho é avaliar o efeito de "máscara" do epitélio no queratocone, comparando a tomografia corneana com e sem epitélio, no decurso da realização de Crosslinking (CXL). Métodos: Cinco olhos de 5 doentes com queratocone em progressão submetidos a CXL segundo o protocolo original de Dresden. Foi realizada tomografia corneana antes e durante o procedimento (imediatamente após a remoção do epitélio), e avaliados os valores da queratometria - K1, K2, K médio (Km), ponto de máxima curvatura (Kmax), valores paquimétricos e astigmatismo corneano. Resultados: Os valores médios de curvatura corneanas antes e após a remoção do epitélio foram de: K1: 43,50 ± 2,54D e 44,32±2,64D (p=0,080); K2: 46,64±2,35D e 49,38±2,86D (p=0,043); Km: 45,48±2,42D e 46,72±2,60D (p=0,042). Observou-se igualmente um aumento estatisticamente significativo do valor de queratometria máxima após a remoção do epitélio (p=0,043). Na paquimetria central observou-se uma redução média de 26,8µm de 524,8±33,0µm para 498,2±37,7µm (p= 0,042). Conclusão: Observou-se um aumento estatisticamente significativo nos valores do Kmax, K2, e Kmédio após remoção do epitélio. Este estudo demonstra o efeito de "máscara" que o epitélio exerce no queratocone.

Abstract Objective: In the presence of an irregular cornea, like in keratoconus, the corneal epithelium has a different profile "masking" the irregular corneal stroma surface. Topographical analysis without considering the epithelium effect can result in an incorrect assessment, affecting the accuracy of any topography guided treatment. The aim of this study was to evaluate the "masking" effect of the corneal epithelium on corneal curvature in patients with keratoconus, comparing topographical findings before and after removal of the epithelium during Crosslinking (CXL). Methods: Five eyes of 5 patients with progressive keratoconus submitted to CXL according to the original Dresden protocol. Corneal topography was performed before and during the procedure (immediately after epithelium removal) and values of keratometry: K1, K2, mean K (Km), maximum corneal apical curvature (Kmax), corneal thickness and corneal astigmatism were evaluated. Results: The average values of corneal curvature before and after epithelial remove were: K1: 43.50±2.54D and 44.32±2,64D (p=0.080); K2 46.64±2,35D and 49.38±2,86D (p=0.043); Km 45.48±2.42D and 46.72±2.60D (p=0.042). It was also observed a significant increase in the maximum values of corneal apical curvature after removal of the epithelium (p=0.043). In central corneal thickness there was seen a reduction of 26.8µm from 524.8±33.0µm to 498.2±37.7µm (p = 0.042). Conclusion: There was seen a significant increase in the values of Kmax, K2 and mean K. This study demonstrates the masking effect of corneal epithelium on corneal curvature in keratoconus.

Genetic Diversity and Phylogenetic Relationships of Coevolving Symbiont-Harboring Insect Trypanosomatids, and Their Neotropical Dispersal by Invader African Blowflies (Calliphoridae).

This study is about the inter- and intra-specific genetic diversity of trypanosomatids of the genus Angomonas, and their association with Calliphoridae (blowflies) in Neotropical and Afrotropical regions. Microscopic examination of 3,900 flies of various families, mostly Calliphoridae, revealed that 31% of them harbored trypanosomatids. Small subunit rRNA (SSU rRNA) barcoding showed that Angomonas predominated (46%) over the other common trypanosomatids of blowflies of genera Herpetomonas and Wallacemonas. Among Angomonas spp., A. deanei was much more common than the two-other species, A. desouzai and A. ambiguus. Phylogenetic analyses based on SSU rRNA, glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) and internal transcribed spacer rDNA (ITS rDNA) sequences revealed a marked genetic diversity within A. deanei, which comprised four infraspecific genotypes (Dea1-Dea4), and four corresponding symbiont genotypes (Kcr1-Kcr4). Host and symbiont phylogenies were highly congruent corroborating their co-divergence, consistent with host-symbiont interdependent metabolism and symbiont reduced genomes shaped by a long coevolutionary history. We compared the diversity of Angomonas/symbionts from three genera of blowflies, Lucilia, Chrysomya and Cochliomyia. A. deanei, A. desouzai, and A. ambiguus were found in the three genera of blowflies in South America. In Africa, A. deanei and A. ambiguus were identified in Chrysomya. The absence of A. desouzai in Africa and its presence in Neotropical Cochliomyia and Lucilia suggests parasite spillback of A. desouzai into Chrysomya, which was most likely introduced four decades ago from Africa into the Neotropic. The absence of correlation between parasite diversity and geographic and genetic distances, with identical genotypes of A. deanei found in the Neotropic and Afrotropic, is consistent with disjunct distribution due to the recent human-mediated transoceanic dispersal of Angomonas by Chrysomya. This study provides the most comprehensive data gathered so far on the genetic repertoires of a genus of trypanosomatids found in flies from a wide geographical range.

Diagnosis and genetic analysis of the worldwide distributed Rattus-borne Trypanosoma (Herpetosoma) lewisi and its allied species in blood and fleas of rodents.

Trypanosoma (Herpetosoma) lewisi is a cosmopolitan parasite of rodents strongly linked to the human dispersal of Rattus spp. from Asia to the rest of the world. This species is highly phylogenetically related to trypanosomes from other rodents (T. lewisi-like), and sporadically infects other mammals. T. lewisi may opportunistically infect humans, and has been considered an emergent rat-borne zoonosis associated to poverty. We developed the THeCATL-PCR based on Cathepsin L (CATL) sequences to specifically detect T. (Herpetosoma) spp., and assess their genetic diversity. This method exhibited high sensitivity using blood samples, and is the first molecular method employed to search for T. lewisi in its flea vectors. THeCATL-PCR surveys using simple DNA preparation from blood preserved in ethanol or filter paper detected T. lewisi in Rattus spp. from human dwellings in South America (Brazil and Venezuela), East Africa (Mozambique), and Southeast Asia (Thailand, Cambodia and Lao PDR). In addition, native rodents captured in anthropogenic and nearby human settlements in natural habitats harbored T. (Herpetosoma) spp. PCR-amplified CATL gene fragments (253bp) distinguish T. lewisi and T. lewisi-like from other trypanosomes, and allow for assessment of genetic diversity and relationships among T. (Herpetosoma) spp. Our molecular surveys corroborated worldwide high prevalence of T. lewisi, incriminating Mastomys natalensis as an important carrier of this species in Africa, and supported its spillover from invader Rattus spp. to native rodents in Brazil and Mozambique. THeCATL-PCR provided new insights on the accurate diagnosis and genetic repertoire of T. (Herpetosoma) spp. in rodent and non-rodent hosts, revealing a novel species of this subgenus in an African gerbil. Phylogenetic analysis based on CATL sequences from T. (Herpetosoma) spp. and other trypanosomes (amplified using pan-trypanosome primers) uncovered rodents harboring, beyond mammal trypanosomes of different subgenera, some species that clustered in the lizard-snake clade of trypanosomes.

New insights from Gorongosa National Park and Niassa National Reserve of Mozambique increasing the genetic diversity of Trypanosoma vivax and Trypanosoma vivax-like in tsetse flies, wild ungulates and livestock from East Africa.

BACKGROUND: Trypanosoma (Duttonella) vivax is a major pathogen of livestock in Africa and South America (SA), and genetic studies limited to small sampling suggest greater diversity in East Africa (EA) compared to both West Africa (WA) and SA. METHODS: Multidimensional scaling and phylogenetic analyses of 112 sequences of the glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) gene and 263 sequences of the internal transcribed spacer of rDNA (ITS rDNA) were performed to compare trypanosomes from tsetse flies from Gorongosa National Park and Niassa National Reserve of Mozambique (MZ), wild ungulates and livestock from EA, and livestock isolates from WA and SA. RESULTS: Multidimensional scaling (MDS) supported Tvv (T. vivax) and TvL (T. vivax-like) evolutionary lineages: 1) Tvv comprises two main groups, TvvA/B (all SA and WA isolates plus some isolates from EA) and TvvC/D (exclusively from EA). The network revealed five ITS-genotypes within Tvv: Tvv1 (WA/EA isolates), Tvv2 (SA) and Tvv3-5 (EA). EA genotypes of Tvv ranged from highly related to largely different from WA/SA genotypes. 2) TvL comprises two gGAPDH-groups formed exclusively by EA sequences, TvLA (Tanzania/Kenya) and TvLB-D (MZ). This lineage contains more than 11 ITS-genotypes, seven forming the lineage TvL-Gorongosa that diverged from T. vivax Y486 enough to be identified as another species of the subgenus Duttonella. While gGAPDH sequences were fundamental for classification at the subgenus, major evolutionary lineages and species levels, ITS rDNA sequences permitted identification of known and novel genotypes. CONCLUSIONS: Our results corroborate a remarkable diversity of Duttonella trypanosomes in EA, especially in wildlife conservation areas, compared to the moderate diversity in WA. Surveys in wilderness areas in WA may reveal greater diversity. Biogeographical and phylogenetic data point to EA as the place of origin, diversification and spread of Duttonella trypanosomes across Africa, providing relevant insights towards the understanding of T. vivax evolutionary history.

Microsatellite analysis supports clonal propagation and reduced divergence of Trypanosoma vivax from asymptomatic to fatally infected livestock in South America compared to West Africa.

BACKGROUND: Mechanical transmission of the major livestock pathogen Trypanosoma vivax by other biting flies than tsetse allows its spread from Africa to the New World. Genetic studies are restricted to a small number of isolates and based on molecular markers that evolve too slowly to resolve the relationships between American and West African populations and, thus, unable us to uncover the recent history of T. vivax in the New World. METHODS: T. vivax genetic diversity, population structure and the source of outbreaks was investigated through the microsatellite multiloci (7 loci) genotype (MLGs) analysis in South America (47isolates from Brazil, Venezuela and French Guiana) and West Africa (12 isolates from The Gambia, Burkina Faso, Ghana, Benin and Nigeria). Relationships among MLGs were explored using phylogenetic, principal component and STRUCTURE analyses. RESULTS: Although closely phylogenetically related, for the first time, genetic differences were detected between T. vivax isolates from South America (11 genotypes/47 isolates) and West Africa (12 genotypes/12 isolates) with no MLGs in common. Diversity was far greater across West Africa than in South America, where genotypes from Brazil (MLG1-6), Venezuela (MLG7-10) and French Guiana (MLG11) shared similar but not identical allele composition. No MLG was exclusive to asymptomatic (endemic areas) or sick (outbreaks in non-endemic areas) animals, but only MLGs1, 2 and 3 were responsible for severe haematological and neurological disorders. CONCLUSIONS: Our results revealed closely related genotypes of T. vivax in Brazil and Venezuela, regardless of endemicity and clinical conditions of the infected livestock. The MLGs analysis from T. vivax across SA and WA support clonal propagation, and is consistent with the hypothesis that the SA populations examined here derived from common ancestors recently introduced from West Africa. The molecular markers defined here are valuable to assess the genetic diversity, to track the source and dispersion of outbreaks, and to explore the epidemiological and pathological significance of T. vivax genotypes.

Congopain genes diverged to become specific to Savannah, Forest and Kilifi subgroups of Trypanosoma congolense, and are valuable for diagnosis, genotyping and phylogenetic inferences.

Trypanosoma congolense is the most important agent of nagana, a wasting livestock trypanosomosis in sub-Saharan Africa. This species is a complex of three subgroups (Savannah, Forest and Kilifi) that differ in virulence, pathogenicity, drug resistance, vectors, and geographical distribution. Congopain, the major Cathepsin L-like cysteine protease (CP2) of T. congolense, has been extensively investigated as a pathogenic factor and target for drugs and vaccines, but knowledge about this enzyme is mostly restricted to the reference strain IL3000, which belongs to the Savannah subgroup. In this work we compared sequences of congopain genes from IL3000 genome database and isolates of the three subgroups of T. congolense. Results demonstrated that the congopain genes diverged into three subclades consistent with the three subgroups within T. congolense. Laboratory and field isolates of Savannah exhibited a highly polymorphic repertoire both inter- and intra-isolates: sequences sharing the archetypical catalytic triad clustered into SAV1-SAV3 groups, whereas polymorphic sequences that, in general, exhibited unusual catalytic triad (variants) assigned to SAV4 or not assigned to any group. Congopain homologous genes from Forest and Kilifi isolates showed, respectively, moderate and limited diversity. In the phylogenetic tree based on congopain and homologues, Savannah was closer to Forest than to Kilifi. All T. congolense subgroup nested into a single clade, which together with the sister clade formed by homologues from Trypanosoma simiae and Trypanosoma godfreyi formed a clade supporting the subgenus Nannomonas. A single PCR targeting congopain sequences was developed for the diagnosis of T. congolense isolates of the three subgroups. Our findings demonstrated that congopain genes are valuable targets for the diagnosis, genotyping, and phylogenetic and taxonomic inferences among T. congolense isolates and other members of the subgenus Nannomonas.

Trypanosoma livingstonei: a new species from African bats supports the bat seeding hypothesis for the Trypanosoma cruzi clade.

BACKGROUND: Bat trypanosomes have been implicated in the evolutionary history of the T. cruzi clade, which comprises species from a wide geographic and host range in South America, Africa and Europe, including bat-restricted species and the generalist agents of human American trypanosomosis T. cruzi and T. rangeli. METHODS: Trypanosomes from bats (Rhinolophus landeri and Hipposideros caffer) captured in Mozambique, southeast Africa, were isolated by hemoculture. Barcoding was carried out through the V7V8 region of Small Subunit (SSU) rRNA and Fluorescent Fragment Length barcoding (FFLB). Phylogenetic inferences were based on SSU rRNA, glyceraldehyde phosphate dehydrogenase (gGAPDH) and Spliced Leader (SL) genes. Morphological characterization included light, scanning and transmission electron microscopy. RESULTS: New trypanosomes from bats clustered together forming a clade basal to a larger assemblage called the T. cruzi clade. Barcoding, phylogenetic analyses and genetic distances based on SSU rRNA and gGAPDH supported these trypanosomes as a new species, which we named Trypanosoma livingstonei n. sp. The large and highly polymorphic SL gene repeats of this species showed a copy of the 5S ribosomal RNA into the intergenic region. Unique morphological (large and broad blood trypomastigotes compatible to species of the subgenus Megatrypanum and cultures showing highly pleomorphic epimastigotes and long and slender trypomastigotes) and ultrastructural (cytostome and reservosomes) features and growth behaviour (when co-cultivated with HeLa cells at 37°C differentiated into trypomastigotes resembling the blood forms and do not invaded the cells) complemented the description of this species. CONCLUSION: Phylogenetic inferences supported the hypothesis that Trypanosoma livingstonei n. sp. diverged from a common ancestral bat trypanosome that evolved exclusively in Chiroptera or switched at independent opportunities to mammals of several orders forming the clade T. cruzi, hence, providing further support for the bat seeding hypothesis to explain the origin of T. cruzi and T. rangeli.

Evolutionary insights from bat trypanosomes: morphological, developmental and phylogenetic evidence of a new species, Trypanosoma (Schizotrypanum) erneyi sp. nov., in African bats closely related to Trypanosoma (Schizotrypanum) cruzi and allied species.

Parasites of the genus Trypanosoma are common in bats and those of the subgenus Schizotrypanum are restricted to bats throughout the world, with the exception of Trypanosoma (Schizotrypanum) cruzi that also infects other mammals and is restricted to the American Continent. We have characterized trypanosome isolates from Molossidae bats captured in Mozambique, Africa. Morphology and behaviour in culture, supported by phylogenetic inferences using SSU (small subunit) rRNA, gGAPDH (glycosomal glyceraldehyde 3-phosphate dehydrogenase) and Cyt b (cytochrome b) genes, allowed to classify the isolates as a new Schizotrypanum species named Trypanosoma (Schizotrypanum) erneyi sp. nov. This is the first report of a Schizotrypanum species from African bats cultured, characterized morphologically and biologically, and positioned in phylogenetic trees. The unprecedented finding of a new species of the subgenus Schizotrypanum from Africa that is closest related to the America-restricted Trypanosoma (Schizotrypanum) cruzi marinkellei and T. cruzi provides new insights into the origin and evolutionary history of T. cruzi and closely related bat trypanosomes. Altogether, data from our study support the hypothesis of an ancestor trypanosome parasite of bats evolving to infect other mammals, even humans, and adapted to transmission by triatomine bugs in the evolutionary history of T. cruzi in the New World.

Use of oxfendazole to control porcine cysticercosis in a high-endemic area of Mozambique

A randomized controlled field trial to evaluate the effectiveness of a single oral dose of 30 mg/kg of oxfendazole (OFZ) treatment for control of porcine cysticercosis was conducted in 4 rural villages of Angónia district, north-western Mozambique. Two hundred and sixteen piglets aged 4 months were selected and assigned randomly to OFZ treatment or control groups. Fifty-four piglets were treated at 4 months of age (T1), while another 54 piglets were treated at 9 months of age (T2) and these were matched with 108 control pigs from the same litters and raised under the same conditions. Baseline data were collected on the prevalence of porcine cysticercosis using antigen ELISA (Ag-ELISA), as well as knowledge and practices related to Taenia solium transmission based on questionnaire interviews and observations. All animals were followed and re-tested for porcine cysticercosis by Ag-ELISA at 9 and 12 months of age when the study was terminated. Overall prevalence at baseline was 5.1% with no significant difference between groups. At the end of the study, 66.7% of the controls were found positive, whereas 21.4% of the T1 and 9.1% of the T2 pigs were positive, respectively. Incidence rates of porcine cysticercosis were lower in treated pigs as compared to controls. Necropsy of 30 randomly selected animals revealed that viable cysts were present in none (0/8) of T2 pigs, 12.5% (1/8) of T1 pigs and 42.8% (6/14) of control pigs. There was a significant reduction in the risk of T. solium cysticercosis if pigs were treated with OFZ either at 4 months (OR = 0.14; 95% CI: 0.05-0.36) or at 9 months of age (OR = 0.05; 95% CI: 0.02-0.16). Strategic treatment of pigs in endemic areas should be further explored as a means to control T. solium cysticercosis/taeniosis.

Prevalence and risk factors of porcine cysticercosis in Angónia district, Mozambique

Taenia solium is an important zoonosis in many developing countries. Cysticercosis poses a serious public health risk and incurs sizeable economic losses to pig production. Because data on the epidemiology of porcine cysticercosis in Mozambique are scarce, the present study was conducted to determine the prevalence and risk factors for porcine cysticercosis. A cross-sectional survey was carried out in 11 villages in Angónia district, Tete province in northwestern Mozambique. Between September and November, 2007, a total of 661 pigs were tested serologically and examined by tongue inspection. Serum samples were tested for the presence of circulating parasite antigen using a monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (Ag-ELISA). In addition, a questionnaire survey to collect information on pig production, occurrence and transmission of porcine cysticercosis, risk factors and awareness of porcine cysticercosis was conducted in the selected households from which pigs were sampled. Two hundred thirty-one samples (34.9%) were found positive by the Ag-ELISA, while by tongue inspection on the same animals cysticerci were detected in 84 pigs (12.7%). Increasing age (OR = 1.63; 95% CI = 1.13-2.37) and free-range pig husbandry system (OR = 3.81; 95% CI = 2.08-7.06) were important risk factors for porcine cysticercosis in the district. The present findings indicate that porcine cysticercosis is endemic in the region, and that increasing pig age and pig husbandry practices contribute significantly to porcine cysticercosis transmission. Further epidemiological studies on the prevalence and transmission of porcine cysticercosis in rural communities in Mozambique are needed to enable collection of more baseline data and implementation of effective control strategies within the country.

Cathepsin L-like genes of Trypanosoma vivax from Africa and South America--characterization, relationships and diagnostic implications.

We characterized sequences from genes encoding cathepsin L-like (CatL-like) cysteine proteases from African and South American isolates of Trypanosoma vivax and T. vivax-like organisms, and evaluated their suitability as genetic markers for population structure analysis and diagnosis. Phylogenetic analysis of sequences corresponding to CatL-like catalytic domains revealed substantial polymorphism, and clades of sequences (TviCatL1-9) were separated by large genetic distances. TviCatL1-4 sequences were from cattle isolates from West Africa (Nigeria and Burkina Faso) and South America (Brazil and Venezuela), which belonged to the same T. vivax genotype. T. vivax-like genotypes from East Africa showed divergent sequences, including TviCatL5-7 for isolates from Mozambique and TviCatL8-9 for an isolate from Kenya. Phylogenetic analysis of CatL-like gene data supported the relationships among trypanosome species reflected in the phylogenies based on the analysis of small subunit (SSU) of ribosomal RNA gene sequence data. The discovery of different CatL-like sequences for each genotype, defined previously by ribosomal DNA data, indicate that these sequences provide useful targets for epidemiological and population genetic studies. Regions in CatL-like sequences shared by all T. vivax genotypes but not by other trypanosomes allowed the establishment of a specific and sensitive diagnostic PCR for epidemiological studies in South America and Africa.