Detection of Plasmodium simium gametocytes in non-human primates from the Brazilian Atlantic Forest.

BACKGROUND: Plasmodium species of non-human primates (NHP) are of great interest because they can naturally infect humans. Plasmodium simium, a parasite restricted to the Brazilian Atlantic Forest, was recently shown to cause a zoonotic outbreak in the state of Rio de Janeiro. The potential of NHP to act as reservoirs of Plasmodium infection presents a challenge for malaria elimination, as NHP will contribute to the persistence of the parasite. The aim of the current study was to identify and quantify gametocytes in NHP naturally-infected by P. simium. METHODS: Whole blood samples from 35 NHP were used in quantitative reverse transcription PCR (RT-qPCR) assays targeting 18S rRNA, Pss25 and Pss48/45 malaria parasite transcripts. Absolute quantification was performed in positive samples for 18S rRNA and Pss25 targets. Linear regression was used to compare the quantification cycle (Cq) and the Spearman's rank correlation coefficient was used to assess the correlation between the copy numbers of 18S rRNA and Pss25 transcripts. The number of gametocytes/µL was calculated by applying a conversion factor of 4.17 Pss25 transcript copies per gametocyte. RESULTS: Overall, 87.5% of the 26 samples, previously diagnosed as P. simium, were positive for 18S rRNA transcript amplification, of which 13 samples (62%) were positive for Pss25 transcript amplification and 7 samples (54%) were also positive for Pss48/45 transcript. A strong positive correlation was identified between the Cq of the 18S rRNA and Pss25 and between the Pss25 and Pss48/45 transcripts. The 18S rRNA and Pss25 transcripts had an average of 1665.88 and 3.07 copies/µL, respectively. A positive correlation was observed between the copy number of Pss25 and 18S rRNA transcripts. Almost all gametocyte carriers exhibited low numbers of gametocytes (< 1/µL), with only one howler monkey having 5.8 gametocytes/µL. CONCLUSIONS: For the first time, a molecular detection of P. simium gametocytes in the blood of naturally-infected brown howler monkeys (Alouatta guariba clamitans) was reported here, providing evidence that they are likely to be infectious and transmit P. simium infection, and, therefore, may act as a reservoir of malaria infection for humans in the Brazilian Atlantic Forest.

Combined computational and experimental study about the incorporation of phosphorus into the structure of graphene oxide.

Phosphorus-containing graphene-based hybrids are materials with outstanding properties for diverse applications. In this work, an easy route to produce phosphorus-graphene oxide hybrid materials is described, involving the use of variable amounts of H3PO4 and H2SO4 during the reaction of oxidation of a graphitic precursor. The physical and chemical features of the hybrids change significantly with the variation in the acid amounts used in the syntheses. XPS and solid-state 13C and 31P NMR results show that the hybrids contain large amounts of oxygen functional groups, with the phosphorus incorporation proceeding mostly through the formation of phosphate-like linkages and other functions with C-O-P bonds. The experimental findings are supported by DFT calculations, which allow the assessment of the energetics and the geometry of the interaction between phosphate groups and graphene-based models; these calculations are also used to predict the chemical shifts in the 31P and 13C NMR spectra of the models, which show good agreement with the experimentally observed solid-state NMR spectra.

Lower Extremity Physical Performance Tests for the Assessment of Athletes via Telehealth are Reliable.

CONTEXT: Lower extremity physical performance tests (PPTs) have been widely used in sports rehabilitation and are commonly performed in person. However, some situations may disrupt the in-person health care delivery, such as social distancing due to the pandemic, traveling, and living in remote locations. Those situations may require adjustments in planning and applying measurement tests, and telehealth has become an alternative. Nevertheless, the reliability of lower extremity PPT tests via telehealth is still unknown. OBJECTIVES: To verify the test-retest reliability, SEM, and the minimum detectable change (MDC95) of PPTs via telehealth. METHODS: Fifty asymptomatic athletes completed 2 assessment sessions 7 to 14 days apart. The assessment via telehealth consisted of warm-up exercises followed by the single-, triple-, and side-hop tests, and the long jump test, in random order. Intraclass correlation coefficient, SEM, and MDC95 were calculated for each PPT. RESULTS: Single-hop test showed good to excellent reliability, with SEM and MDC95 ranging from 6.06 to 9.24 cm and 16.79 to 25.61 cm, respectively. The triple-hop test showed excellent reliability, with SEM and MDC95 ranging from 13.17 to 28.17 cm and 30.72 to 78.07 cm, respectively. Side-hop tests showed moderate reliability, with SEM and MDC95 ranging from 0.67 to 1.22 seconds and 2.00 to 3.39 seconds, respectively. The long jump test showed excellent reliability, with SEM and MDC95 ranging from 5.34 to 8.34 cm and 14.80 to 23.11 cm, respectively. CONCLUSION: The test-retest reliability of those PPTs via telehealth was acceptable. The SEM and MDC were provided to assist clinicians in interpreting those PPTs.

Physical Performance and Mobility Tests Using Telehealth: A Feasibility Study With Athletes.

CONTEXT: Physical performance tests (PPTs) and mobility tests have been widely used in sports rehabilitation. However, the feasibility of PPTs and mobility tests via telehealth is unknown. OBJECTIVES: To verify the feasibility of PPTs and mobility tests to assess athletes via telehealth. DESIGN: This is a feasibility study. PARTICIPANTS: Athletes enrolled in a sports team or club for at least 2 years and with previous enrollment in a competitive league were recruited through advertisements on social media. The athletes included in this study (mean age = 25.9 y, from different sports) performed a battery of PPTs and mobility tests for the lower-extremity or upper-extremity and trunk, according to their sport modality. MAIN OUTCOME MEASURE: The feasibility was assessed with recruitment, success, and dropout rates. In addition, athletes' perceptions of easiness, satisfaction, and safety during the lower-extremity or upper-extremity and trunk PPTs and mobility tests were assessed. RESULTS: Seventy-three athletes were included, between January and April 2021: 41 were allocated to the lower-extremity and 32 to the upper-extremity and trunk PPTs and mobility test, according to their sports modality. The overall dropout rate was 20.55%; >89% of the athletes agreed that the PPTs and mobility tests via telehealth were easy to perform, >78% were satisfied, and >75% felt safe during the assessments. CONCLUSION: This study indicated that these 2 batteries of performance and mobility tests via telehealth are feasible to assess the lower and upper-extremities, and the trunk of athletes, considering the adherence, athletes' perception of easiness, satisfaction, and safety.

Nanostructured faujasite zeolite as metal ion adsorbent: kinetics, equilibrium adsorption and metal recovery studies.

The hydrothermal synthesis of nano-faujasite has been successfully performed and the effects of some crystallization parameters were investigated, along with the use of this material as a heavy-metal ion adsorbent. X-ray diffraction patterns have shown that the structure of the nano-faujasite is strongly dependent on both the crystallization time and the alkalinity of the synthesis medium. According to N2 physisorption, X-ray fluorescence, SEM/EDS, and solid state 29Si and 27Al NMR data, the produced nano-faujasite consists of a solid with low molar Si/Al ratio (1.7), with high availability of ion exchange sites and high surface area/small particle size, allowing easy diffusion of metal ions to adsorbent active sites. As a consequence, an excellent performance on removal of Cd2+, Zn2+ and Cu2+ ions was found for this solid. The adsorption capacity followed the order Cd2+ (133 mg·g-1) > Zn2+ (115 mg·g-1) > Cu2+ (99 mg·g-1), which agrees with the order of increasing absolute values of the hydration energy of the metal ions. Kinetic studies and adsorption isotherms showed that the metal ion removal takes place by ion exchange on the monolayer surface of the nano-faujasite. The electrochemical recovery of copper in metallic form exhibited an efficiency of 80.2% after 120 min, which suggests that this process can be adequately implemented for full-scale metal removal.

Plasmodium infection and its association with biochemical and haematological parameters in free-living Alouatta guariba clamitans (Cabrera, 1940) (Primates: Atelidae) in Southern Brazil.

BACKGROUND: The influence of Plasmodium spp. infection in the health of Southern brown howler monkey, Alouatta guariba clamitans, the main reservoir of malaria in the Atlantic Forest, is still unknown. OBJECTIVES: The aim of this study was to investigate the positivity rate of Plasmodium infection in free-living howler monkeys in an Atlantic Forest fragment in Joinville/SC and to associate the infection with clinical, morphometrical, haematological and biochemical alterations. METHODS: Molecular diagnosis of Plasmodium infection in the captured monkeys was performed by Nested-polymerase chain reaction (PCR) (18S rRNA and coxI). Haematological and biochemical parameters were compared among infected and uninfected monkeys; clinical and morphometrical parameters were also compared. FINDINGS: The positivity rate of Plasmodium infection was 70% among forty captured animals, the highest reported for neotropical primates. None statistical differences were detected in the clinical parameters, and morphometric measures comparing infected and uninfected groups. The main significant alteration was the higher alanine aminotransferase (ALT) levels in infected compared to uninfected monkeys. MAIN CONCLUSIONS: Therefore, Plasmodium infection in howler monkeys may causes haematological/biochemical alterations which might suggest hepatic impairment. Moreover, infection must be monitored for the eco-epidemiological surveillance of malaria in the Atlantic Forest and during primate conservation program that involves the animal movement, such as translocations.

Hematological and serum biochemistry data on southern brown howler monkeys (Alouatta guariba clamitans) in captivity in Brazil.

BACKGROUND: Physiological values reflect the health condition and responses of individuals to handling in captivity. The aim of this study was to establish hematological and serum biochemistry parameters of clinically healthy animals of the Alouatta guariba clamitans subspecies. METHODS: We collected blood samples from adult males and females kept at the Center for Biological Research of Indaial after chemical containment with 3.9 mg/kg of tiletamine hydrochloride and zolazepam. RESULTS: Significant differences between males and females were found in the levels of erythrocytes, hemoglobin, hematocrit, lymphocytes, neutrophils, platelets, mean corpuscular hemoglobin concentration (MCHC), and gamma-glutamyltranspeptidase (GGT). CONCLUSIONS: Our results suggest the existence of sexual dimorphism in some physiological parameters of A guariba clamitans. The parameters reported herein can be used as reference values for other populations kept under similar conditions.

Therapeutic success and failure in using miltefosine to treat dogs naturally infected with Leishmania infantum.

In urban environments, domestic dogs (Canis familiaris) are a major reservoir for the parasite Leishmania infantum. Miltefosine has been used as the standard treatment for canine visceral leishmaniasis in Brazil. However, therapeutic failures have been reported. In the present study, two dogs (CG03 and CG06) with a diagnosis of infection by L. infantum underwent two cycles of treatment with miltefosine (Milteforan™ - Virbac®). Analyses showed increases in the parasite load of both CG03 and CG06, even after treatment. The clinical score of CG03 dropped from 1 to 0 (after one round of treatment), such that this dog became asymptomatic. CG06 showed clinical worsening, such that its score increased from 1 to 2. After the second therapeutic round, the parasite load in CG03 was found to have decreased, but it was still higher than before drug treatment even though this dog was physically asymptomatic. There was no decrease in the parasite load in CG06 and there was clinical worsening. The clinical response of these dogs to the treatment differed, but the parasite load remained high in both cases, which poses a risk to public health, making it essential take measures to prevent the sandfly vector from accessing the dog.

PCA and logistic regression in 2-[18F]FDG PET neuroimaging as an interpretable and diagnostic tool for Alzheimer's disease.

Objective.to develop an optimization and training pipeline for a classification model based on principal component analysis and logistic regression using neuroimages from PET with 2-[18F]fluoro-2-deoxy-D-glucose (FDG PET) for the diagnosis of Alzheimer's disease (AD).Approach.as training data, 200 FDG PET neuroimages were used, 100 from the group of patients with AD and 100 from the group of cognitively normal subjects (CN), downloaded from the repository of the Alzheimer's Disease Neuroimaging Initiative (ADNI). Regularization methods L1 and L2 were tested and their respective strength varied by the hyperparameter C. Once the best combination of hyperparameters was determined, it was used to train the final classification model, which was then applied to test data, consisting of 192 FDG PET neuroimages, 100 from subjects with no evidence of AD (nAD) and 92 from the AD group, obtained at the Centro de Diagnóstico por Imagem (CDI).Main results.the best combination of hyperparameters was L1 regularization andC≈ 0.316. The final results on test data were accuracy = 88.54%, recall = 90.22%, precision = 86.46% and AUC = 94.75%, indicating that there was a good generalization to neuroimages outside the training set. Adjusting each principal component by its respective weight, an interpretable image was obtained that represents the regions of greater or lesser probability for AD given high voxel intensities. The resulting image matches what is expected by the pathophysiology of AD.Significance.our classification model was trained on publicly available and robust data and tested, with good results, on clinical routine data. Our study shows that it serves as a powerful and interpretable tool capable of assisting in the diagnosis of AD in the possession of FDG PET neuroimages. The relationship between classification model output scores and AD progression can and should be explored in future studies.

Fungicides from rice cultivation (tebuconazole and azoxystrobin) alters biochemical and histological markers of hammertoad tadpoles (Boanafaber).

Tebuconazole (TBZ) and azoxystrobin (AZX) are fungicides frequently used in rice cultivation. Despite protecting crops against fungal diseases, these compounds can contaminate the natural environments close to the crops, exerting negative effects on non-target organisms, the present study aimed to characterize the contamination by fungicides of a river that flows in an area dominated by rice cultivation in the north of the state of Santa Catarina, SC, Brazil. Concentrations of TBZ and AZX found in the field were used to evaluate their negative effects on development, biochemical biomarkers and histopatology of the liver of a native tadpole species, the hammerfrog (Boana faber). Tadpoles were exposed for 16 days to the lowest (1.20 µg/L) and highest (2.60 µg/L) concentration of TBZ, lowest (0.70 µg/L) and highest (1.60 µg/L) concentration of AZX, and the mix of both fungicides at lowest and highest concentration of each found in field analyses. Exposure to the lower TBZ concentration and both concentrations of the Mix accelerated the development of tadpoles. AZX caused an increase in the activities of glutathione S-transferase (GST), carboxylesterase (CbE) and glucose-6-phosphate dehydrogenase (G6PDH) in the liver, an increase in the levels of protein carbonyls (PC) in the liver and an increase in the activity of acetylcholinesterase (AChE) in muscle of tadpoles. TBZ, on the other hand, generated an increase in GST, G6PDH, PC and histopathological severity scores in liver and in muscle AChE activity. The effects were more intense in the groups exposed to the Mix of contaminants. No treatment altered brain AChE. The data showed that the fungicides from in rice cultivation found in natural aquatic environments around the crops pose risks to the health of the animals, compromising their metabolism and development.

Exposure to pesticides used in rice farming (bentazone, chlorantraniliprole and tebuconazole) affects biochemical biomarkers and hepatic histopathological parameters of hammertoad tadpoles (Boana faber).

Pesticides used in rice cultivation can cause negative health effects to non-target organisms representative of natural biodiversity. In this context, the present study aimed to investigate the occurrence of pesticides in surface waters from a river that flows in the middle of a rice farming-dominated area. We were also interested in evaluate biochemical and histological effects caused by exposure (16 d) to the lower and higher concentrations of the main found herbicide (bentazone, BTZ), insecticide (chlorantraniliprole, CTP) and fungicide (tebuconazole, TBZ), isolated or mixed, in Boana faber tadpoles. No significant differences were observed in the development of the animals. Tadpoles exposed to the herbicide BTZ showed higher hepatic levels of malondialdehyde (MDA). In animals exposed to CTP, MDA levels were lower than controls. Animals exposed to the fungicide TBZ showed higher hepatic activity of glutathione S-transferase and carboxylesterase (CbE), as well as higher levels of carbonyl proteins and MDA. Animals exposed to Mix showed higher activity in CbE and glucose-6-phosphate dehydrogenase activity in the liver, as well as higher levels of MDA. In the brain and muscle of tadpoles exposed to Mix, acetylcholinesterase activity was higher. Histological changes were also observed in pesticide-exposed animals, such as increased occurrence of melanomacrophages, inflammatory infiltrates and congestion. Our data evidences the contamination of natural aquatic environments by rice pesticides, and the adverse effects of main ones in B. faber tadpoles, which suggests the contribution of pesticides derived from rice cultivation to the degradation of local biodiversity health.

Diversity of gastrointestinal parasites and molecular characterization of Giardia duodenalis in free-living and captive howler monkeys (Alouatta guariba clamitans) in southern Brazil.

Non-human primates (NHPs) are the group that most share infectious agents with humans due to their close taxonomic relationship. The southern brown howler monkeys (Alouatta guariba clamitans) are endemic primates from Brazil and Argentina's Atlantic Forest. This study aimed to investigate the presence of intestinal parasites in free-living (FL) and captive (CA) southern brown howler monkeys. Thirty-nine stool samples were collected in two areas in southern Brazil, 15 FL and 24 CA. Stool sediments obtained by centrifugal sedimentation technique were used for microscopic analysis and direct immunofluorescence assay and evaluated by molecular analysis through amplification and sequencing of TPI fragments. Intestinal parasites Giardia duodenalis, Cryptosporidium spp., and Trypanoxyuris minutus were detected at coproparasitological analysis. This is the first report of the presence of Cryptosporidium spp. in free-living howlers. The molecular characterization of G. duodenalis isolates indicated assemblage B for the first time found in free-living A. guariba clamitans. The high prevalence of G. duodenalis transmission in CA howler monkeys can be explained by direct contact with humans and frequent soil contact. The presence of a potentially zoonotic assemblage in these animals indicates that the process of fragmentation and cohabitation with humans and livestock affects the wildlife, thus indicating a need for eco-health measures.

Serosurvey of Trypanosoma cruzi in persons experiencing homelessness and shelter workers of Brazil.

Although Chagas disease, caused by Trypanosoma cruzi, has been associated with social vulnerability worldwide, producing disability and mortality, no study to date has assessed this protozoal infection in persons experiencing homelessness. Accordingly, the present study aimed to assess anti-T. cruzi antibodies by Wiener Chagatest ELISA recombinant v.3.0 in serum samples of persons experiencing homelessness and related shelter workers in São Paulo, a city with reported vectors but no recent autochthonous case report. Overall, seropositivity to T. cruzi resulted in three of 203 (1.5%) persons experiencing homelessness and two of 87 (2.3%) shelter workers, with similar seroprevalence likely associated with their past social vulnerability. Although the seropositivity in persons experiencing homelessness and shelter workers was within 0 to 25.1% seroprevalence for chronic Chagas disease in the general Brazilian population, the disease has almost decreased 2-fold from the 1980s to 2000s, and such a wide range may not reflect the local disease status. In addition, the authors hypothesized that the similar seroprevalence and exposure between homeless persons and shelter workers herein may be more associated with shared past and present low-income social vulnerability than migratory movements, which may also include infection by sharing injecting drugs, vertical transmission, or blood transfusion. Thus, future studies are needed to confirm the active transmission of Chagas disease in São Paulo city. Moreover, Chagas disease should be considered as differential diagnosis in homeless persons and shelter workers, even in major disease-free Brazilian or other worldwide cities, mostly due to early exposure and vulnerable living conditions.

A case of canine visceral leishmaniasis of unknown origin in Curitiba (state of Paraná, Brazil) treated successfully with miltefosine.

There are no records of autochthonous cases of canine visceral leishmaniasis in the city of Curitiba, Paraná state, Brazil. In 2020, a male French bulldog (CW01), approximately 2 years old was taken by its owners to a private veterinarian clinic. The suspicion of CVL was confirmed by means of a serology test (ELISA/IFAT reagent), rapid chromatographic immunoassay (DPP®) (ELISA - Biomanguinhos®), parasitological culture and quantitative polymerase chain reaction (qPCR). The animal routinely frequented parks in Curitiba and was taken on several trips to the municipalities of Bombinhas and Balneário Camboriú (Santa Catarina) and to Matinhos (Paraná) where CVL had not previously been reported. Treatment was initiated orally with Milteforan™ which resulted in a significant reduction in the parasitic load. The suspicion of autochthony was investigated through entomological research. A total of 10 traps were installed, one at the animal's home, seven in adjacent city blocks and two in a forest edge. No sandflies were trapped in the dog's home and adjacent houses. The traps in the forest edge caught one Migonemyia migonei female and five Brumptomyia spp. females. This case serves as a warning of the possible introduction of CVL in the city of Curitiba.

IgM antibody responses against Plasmodium antigens in neotropical primates in the Brazilian Atlantic Forest.

Introduction: Zoonotic transmission is a challenge for the control and elimination of malaria. It has been recorded in the Atlantic Forest, outside the Amazon which is the endemic region in Brazil. However, only very few studies have assessed the antibody response, especially of IgM antibodies, in Neotropical primates (NP). Therefore, in order to contribute to a better understanding of the immune response in different hosts and facilitate the identification of potential reservoirs, in this study, naturally acquired IgM antibody responses against Plasmodium antigens were evaluated, for the first time, in NP from the Atlantic Forest. Methods: The study was carried out using 154 NP samples from three different areas of the Atlantic Forest. IgM antibodies against peptides of the circumsporozoite protein (CSP) from different Plasmodium species and different erythrocytic stage antigens were detected by ELISA. Results: Fifty-nine percent of NP had IgM antibodies against at least one CSP peptide and 87% against at least one Plasmodium vivax erythrocytic stage antigen. Levels of antibodies against PvAMA-1 were the highest compared to the other antigens. All families of NP showed IgM antibodies against CSP peptides, and, most strikingly, against erythrocytic stage antigens. Generalized linear models demonstrated that IgM positivity against PvCSP and PvAMA-1 was associated with PCR-detectable blood-stage malaria infection and the host being free-living. Interestingly, animals with IgM against both PvCSP and PvAMA-1 were 4.7 times more likely to be PCR positive than animals that did not have IgM for these two antigens simultaneously. Discussion: IgM antibodies against different Plasmodium spp. antigens are present in NP from the Atlantic Forest. High seroprevalence and antibody levels against blood-stage antigens were observed, which had a significant association with molecular evidence of infection. IgM antibodies against CSP and AMA-1 may be used as a potential marker for the identification of NP infected with Plasmodium, which are reservoirs of malaria in the Brazilian Atlantic Forest.

Comparing human to electronic observers to monitor hand hygiene compliance in an intensive care unit.

Objective: We sought to determine whether an electronic hand hygiene (HH) system could monitor HH compliance at similar rates to direct human observation. Methods: This 4-year proof-of-concept study was conducted in an intensive care unit (ICU) of a private tertiary-care hospital in São Paulo, Brazil, where electronic HH systems were installed in 2 rooms. HH compliance was reported respectively using direct observation and electronic counter devices with an infrared system for detecting HH opportunities. Results: In phase 1, HH compliance by human observers was 56.3% (564 of 1,001 opportunities), while HH compliance detected by the electronic observer was 51.0% (515 of 1,010 opportunities). In phase 2, human observers registered 484 HH opportunities with a HH compliance rate of 64.7% (313 of 484) versus 70.6% (346 of 490) simultaneously detected by the electronic system. In addition, an enhanced HH electronic system monitored activity 24 hours per day and HH compliance without the presence of a human observer was 40.3% (10,642 of 26,421 opportunities), providing evidence for the Hawthorne effect. Conclusions: The electronic HH monitoring system had good correlation with human HH observation, but compliance was remarkably lower when human observers were not present due to the Hawthorne effect (25%-30% absolute difference). Electronic monitoring systems can replace direct observation and can markedly reduce the Hawthorne effect.

Comparison of two electronic hand hygiene systems using real-time feedback via wireless technology to improve hand hygiene compliance in an intensive care unit.

Background: Most hand hygiene (HH) intervention studies use a quasi-experimental design, are primarily uncontrolled before-and-after studies, or are controlled before-and-after studies with a nonequivalent control group. Well-funded studies with improved designs and HH interventions are needed. Objectives: To evaluate healthcare worker (HCW) HH compliance with alcohol-based hand rub (ABHR) through direct observation (human observer), 2 electronic technologies, a radio frequency identification (RFID) badge system, and an invasive device sensor. Methods: In our controlled experimental study, 2,269 observations were made over a 6-month period from July 1 to December 30, 2020, in a 4-bed intensive care unit. We compared HH compliance between a basic feedback loop system with RFID badges and an enhanced feedback loop system that utilized sensors on invasive devices. Results: Real-time feedback by wireless technology connected to a patient's invasive device (enhanced feedback loop) resulted in a significant increase in HH compliance (69.5% in the enhanced group vs 59.1% in the basic group; P = .0001). Conclusion: An enhanced feedback loop system connected to invasive devices, providing real-time alerts to HCWs, is effective in improving HH compliance.

Increased Leishmania infantum resistance to miltefosine and amphotericin B after treatment of a dog with miltefosine and allopurinol.

BACKGROUND: Leishmania infantum is the most important etiological agent of visceral leishmaniasis in the Americas and Mediterranean region, and the dog is the main host. Miltefosine was authorized to treat canine leishmaniasis (CanL) in Brazil in 2017, but there is a persistent fear of the emergence of parasites resistant not only to this drug but, through cross-resistance mechanisms, also to meglumine antimoniate and amphotericin B. Additionally, the literature shows that acquisition of resistance is followed by increased parasite fitness, with higher rates of proliferation, infectivity and metacyclogenesis, which are drivers of parasite virulence. In this context, the aim of this study was to analyze the impact of treating a dog with miltefosine and allopurinol on the generation of parasites resistant to miltefosine, amphotericin B and meglumine antimoniate. METHODS: In vitro susceptibility tests were conducted against miltefosine, amphotericin B and meglumine antimoniate with T0 (parasites isolated from a dog before treatment with miltefosine plus allopurinol), T1 (after 1 course of treatment) and T2 (after 2 courses of treatment) isolates. The rates of cell proliferation, infectivity and metacyclogenesis of the isolates were also evaluated. RESULTS: The results indicate a gradual increase in parasite resistance to miltefosine and amphotericin B with increasing the number of treatment courses. An increasing trend in the metacyclogenesis rate of the parasites was also observed as drug resistance increased. CONCLUSION: The data indicates an increased L. infantum resistance to miltefosine and amphotericin B after the treatment of a dog with miltefosine plus allopurinol. Further studies with a larger number of L. infantum strains isolated from dogs with varied immune response profiles and undergoing different treatment regimes, are advocated.

TRANSLATION AND TRANSCULTURAL ADAPTATION OF THE INJURY REPORT FORM FOR RUGBY UNION.

OBJECTIVE: To translate into Brazilian Portuguese and conduct the cross-cultural adaptation of the "Injury Report Form for Rugby Union" questionnaire. METHODS: This is a cross-sectional study, level of evidence II, with following the steps: translation; synthesis; back-translation; review by a committee of experts and pre-final version; pre-test to verify comprehension; elaboration of the final version of the instrument and clinical application. RESULTS: The two versions resulting from the translation and adaptation process did not show great differences. The pre-final version was filled by 23 male rugby players; three questions were not understood by 40%, 27% and 82.5% of the players, respectively, which required a new meeting with a multidisciplinary committee of experts. The modifications were made, requiring then a new application. The new version was filled by 25 male rugby players, aged 29.44 ± 5.90 years; of which 56% had complete higher education; finalizing the process of cross-cultural adaptation. CONCLUSION: The questionnaire "Formulário para Notificação/Avaliação de Lesão em Jogadores de Rugby" was translated and transculturally adapted into Brazilian Portuguese. Level of Evidence II, Diagnostic Studies - Investigating a Diagnostic Test.

OBJETIVO: Traduzir para o português brasileiro e realizar adaptação transcultural do questionário "Injury Report Form for Rugby Union". MÉTODOS: Estudo transversal, nível de evidência II, etapas seguidas: tradução; síntese; retrotradução; revisão pelo comitê de especialistas e versão pré-final; pré-teste para verificar a compreensão; elaboração da versão final do instrumento e aplicação clínica. RESULTADOS: No processo de tradução e adaptação as duas versões não apresentaram grandes diferenças entre si. A versão pré-final foi preenchida por 23 jogadores de rugby do sexo masculino, três questões não foram compreendidas por 40%, 27% e 82,5% dos jogadores, respectivamente, convocando-se nova reunião com o comitê multidisciplinar de especialistas. As modificações foram realizadas, sendo necessária nova aplicação. A nova versão foi preenchida por 25 jogadores de rugby do sexo masculino, com idade entre 29,44± 5,90 anos; sendo que 56% possuíam nível de escolaridade superior completo; finalizando o processo de adaptação transcultural. CONCLUSÃO: O questionário "Formulário para Notificação/Avaliação de Lesão em Jogadores de Rugby" foi traduzido e adaptado transculturalmente para o português brasileiro. Nível de Evidência II, Estudos diagnósticos - Investigação de um exame para diagnóstico.

Evaluation of schistosomula crude antigen (SCA) as a diagnostic tool for Schistosoma mansoni in low endemic human population.

The study aimed to determine the potential of schistosomula crude antigen (SCA) as a diagnostic target for anti-S. mansoni antibody detection. Cercariae were transformed into schistosomula, homogenized through sonication, and then centrifuged to obtain the SCA. SCA was evaluated using ELISA and dot blots immunoassays on 30 S. mansoni infected sera samples obtained from chronic patients and 30 non-infected humans' sera samples. Either Kato-Katz or saline gradient method or both were employed as the diagnostic reference. Dot blots immunoassay was further performed on protein eluted from 10 to 12 kDa immunoreactive band identified by Western blot analysis. The area under the ROC curve was 0.95 (AUC 0.95, CI 0.88-1.01, p < 0.0001). The sensitivity and specificity of SCA-ELISA and dot blots assays were 96.67% and 86.67% respectively. The human IgG-specific response against SCA was significantly higher in S. mansoni infected individuals (OD = 0.678 ± 0.249) compared to the non-infected population (OD = 0.235 ± 0.136) (p < 0.0001). Our study showed that SCA and its 10-12 kDa component could be useful as diagnostic tools for chronic schistosomiasis.