Molecular diagnosis and characterization of Anaplasma marginale and Ehrlichia ruminantium infecting beef cattle of Maputo Province, Mozambique.

BACKGROUND: Members of the Anaplasmataceae family, such as the Anaplasma and Ehrlichia species, cause economic losses and public health risks. However, the exact economic impact has not been comprehensively assessed in Mozambique due to limited data available on its basic epidemiology. Therefore, we investigated the molecular occurrence and identity of Anaplasma and Ehrlichia spp. infecting beef cattle in Maputo province, Mozambique. METHODS: A total of 200 whole blood samples were collected from apparently healthy beef cattle. Whole blood DNA was extracted and tested for presence of Anaplasma spp. and Ehrlichia ruminantium DNA through amplification of the 16S rRNA and map1 genes. Positive samples to Anaplasma spp. were subject to PCR assay targeting the A. marginale-msp5 gene. Amplicons obtained were purified, sequenced and subject to phylogenetic analyses. RESULTS: Anaplasma spp., A. marginale and E. ruminantium were detected in 153 (76.5%), 142 (71%) and 19 (9.5%) of all the samples analyzed, respectively. On this same sample group, 19 (9.5%) were co-infected with A. marginale and E. ruminantium. The 16S rRNA sequences of Anaplasma spp. obtained were phylogenetically related to A. marginale, A. centrale and A. platys. Phylogenetic analysis revealed that A. marginale-msp5 nucleotide sequences were grouped with sequences from Asia, Africa and Latin America, whereas E. ruminantium-map1 DNA nucleotide sequences were positioned in multiple clusters. CONCLUSION: Cattle in Maputo Province are reservoirs for multiple Anaplasma species. A high positivity rate of infection by A. marginale was observed, as well as high genetic diversity of E. ruminantium. Furthermore, five new genotypes of E. ruminantium-map1 were identified.

Influence of contextual factors on match running performance of starters and non-starters in elite youth male soccer players.

This study investigated the effects of match location, quality of opposition (classified into: strong [1st to 4th position]; intermediate [5th to 15th position]; weak [16th to 20th position]), and match outcome on the match running performance of starters and non-starters from a top elite Brazilian soccer team. Absolute measures were calculated using total distance, high-speed (19.8-25.2 km·h-1), sprinting (≥ 25.2 km·h-1), total distance high-acceleration (> 2m·s2), and deceleration (< -2m·s2) were recorded by GPS units from a sample of young soccer players (N = 25) in a total of 17 matches. Relative measures were calculated by dividing absolute measures by the total duration of the matches. Non-starters covered greater total distance (p = 0.02), sprinting (p = 0.02), high-acceleration and deceleration (p = 0.04), sprinting distance relative per minute played (p = 0.005), and high-acceleration and deceleration relative per minute played (p < 0.001) when the team plays at home, strong opponents, and wins the matches. Starters covered greater total distance in high-speed running (p = 0.04), high-acceleration and deceleration (p = 0.03), and high-speed running relative per minute played (p = 0.04) when the team plays strong opponents and wins the matches. These findings highlight the impact of contextual factors during matches on the locomotor performance of young soccer players.

A New Auto-Regressive Multi-Variable Modified Auto-Encoder for Multivariate Time-Series Prediction: A Case Study with Application to COVID-19 Pandemics.

The SARS-CoV-2 global pandemic prompted governments, institutions, and researchers to investigate its impact, developing strategies based on general indicators to make the most precise predictions possible. Approaches based on epidemiological models were used but the outcomes demonstrated forecasting with uncertainty due to insufficient or missing data. Besides the lack of data, machine-learning models including random forest, support vector regression, LSTM, Auto-encoders, and traditional time-series models such as Prophet and ARIMA were employed in the task, achieving remarkable results with limited effectiveness. Some of these methodologies have precision constraints in dealing with multi-variable inputs, which are important for problems like pandemics that require short and long-term forecasting. Given the under-supply in this scenario, we propose a novel approach for time-series prediction based on stacking auto-encoder structures using three variations of the same model for the training step and weight adjustment to evaluate its forecasting performance. We conducted comparison experiments with previously published data on COVID-19 cases, deaths, temperature, humidity, and air quality index (AQI) in São Paulo City, Brazil. Additionally, we used the percentage of COVID-19 cases from the top ten affected countries worldwide until May 4th, 2020. The results show 80.7% and 10.3% decrease in RMSE to entire and test data over the distribution of 50 trial-trained models, respectively, compared to the first experiment comparison. Also, model type#3 achieved 4th better overall ranking performance, overcoming the NBEATS, Prophet, and Glounts time-series models in the second experiment comparison. This model shows promising forecast capacity and versatility across different input dataset lengths, making it a prominent forecasting model for time-series tasks.

Complete holography-based system for the identification of microparticles in water samples.

Here, we present a comprehensive holography-based system designed for detecting microparticles through microscopic holographic projections of water samples. This system is designed for researchers who may be unfamiliar with holographic technology but are engaged in microparticle research, particularly in the field of water analysis. Additionally, our innovative system can be deployed for environmental monitoring as a component of an autonomous sailboat robot. Our system's primary application is for large-scale classification of diverse microplastics that are prevalent in water bodies worldwide. This paper provides a step-by-step guide for constructing our system and outlines its entire processing pipeline, including hologram acquisition for image reconstruction.

Molecular and serological detection of Anaplasma spp. in small ruminants in an area of Cerrado Biome in northeastern Brazil.

Anaplasmosis, caused by bacteria of the genus Anaplasma, is an important tick-borne disease that causes economic losses to livestock farms in many countries. Even though Anaplasma spp. have been detected in goats and sheep worldwide, few studies investigate the occurrence and genetic identity of these agents in small ruminants from Brazil. Thus, this work aimed to detect and determine the genetic identity of Anaplasma spp. in small ruminants from the Baixo Parnaíba region, state of Maranhão, northeastern Brazil. For this purpose, blood samples were collected from 161 animals (91 goats; 70 sheep) from 4 municipalities in the Baixo Parnaíba region. Sheep and goat serum samples were subjected to recombinant membrane surface protein (MSP5)-based iELISA. Whole blood samples were subject to DNA extraction and molecular diagnosis using PCR assays for Anaplasma spp. targeting msp1ß, msp1α, 16S rRNA and msp4 genes. Positive samples were sequenced and then subjected to Anaplasma marginale msp1α genetic diversity analysis and phylogenetic inferences based on the 16S rRNA and msp4 genes. The serological survey detected the presence of anti-A. marginale IgG antibodies in 18 animals (11.1%): 2.9% (2/70) sheep and 17.4% (16/91) goats. Anaplasma marginale DNA was detected in 2 goats (1.2%) using qPCR based on the msp1ß gene. Two distinct A. marginale msp1α strains, namely α ß and α ß ΓγΓγΓγΓγ were found in the infected goats, each one found in a different animal, both belonging to the H genotype. Phylogenetic analysis based on the 16S rRNA gene showed the sequences positioned in three different clades and grouped with sequences from 'Candidatus Anaplasma boleense', A. platys and A. marginale. Phylogenetic inferences based on the msp4 gene positioned the sequence variants in the A. marginale clade. The present work represents the first molecular detection of sequence variants phylogenetic associated to 'Candidatus Anaplasma boleense' and A. platys and α ß and α ß ΓγΓγΓγΓγ in goats from Brazil.

Reactivity against Sarcocystis neurona and Sarcocystis falcatula-like in horses from Southeastern and Midwestern Brazil.

Equine protozoal myeloencephalitis (EPM) is a neurological disease caused by Sarcocystis neurona. Immunofluorescence antibody tests (IFATs) have been widely used to identify exposure of horses to S. neurona in Brazil. Here we used IFAT to search for IgG antibodies against Sarcocystis falcatula-like (Dal-CG23) and S. neurona (SN138) in sera from 342 horses sampled in Campo Grande, Mato Grosso do Sul state (Midwestern), and São Paulo, São Paulo state (Southeastern), Brazil. The 1:25 cutoff value was chosen to maximize sensitivity of the test. IgG antibodies against S. neurona were detected in 239 horses (69.88%), whereas IgG antibodies against S. falcatula-like were detected in 177 horses (51.75%). Sera from 132 horses (38.59%) reacted against both isolates. Absence of reactivity was evidenced in 58/342 horses (16.95%). The lower cutoff used, and the presence of opossums infected with S. falcatula-like and Sarcocystis spp. in the regions where the horses were sampled, might justify the high seroprevalence observed here. Owing to the similarity among antigens targeted in immunoassays, reports on S. neurona-seropositive horses in Brazil may also derive from the exposure of horses to other Sarcocystis species. The role of other Sarcocystis species in causing neurological diseases in horses in Brazil remains unclear.

Bartonella spp. in households with cats: Risk factors for infection in cats and human exposure.

The aim of this study was to estimate the occurrence of Bartonella spp. per household in cats and the risk factors for Bartonella spp. positivity in cats and their owners from Valdivia, Chile. A total of 464 cats (distributed within 324 households) and 326 humans (control group [n = 112] and cat owner [n = 214]) distributed in 262 households were sampled. From the cat owners (n = 214), 128 humans were in households where the cat was also sampled, totaling 84 households with dual sampling. Real-time PCR (qPCR) was used for Bartonella spp. detection in blood from cats and humans, and immunofluorescent immunoassay (IFA) anti-Bartonella henselae was performed in human serum samples. Out of the total of 324 households, 20.43% presented at least one Bartonella positive cat. From the households with dual sampling, 29.7% (25/84) presented at least one qPCR-Bartonella spp. positive cat. However, Bartonella DNA was not amplified in humans, and in 7.3% (6/82) of the households was found at least one of the cat's owners exposed to B. henselae. Cats younger than one year (Odds Ratio (OR) = 5.3), non-neutered (OR 3.46), sampled at home (OR 5.82), and with improper application of tick/flea control products (OR 3.13) showed a higher risk for Bartonella spp. presence. Humans with occupational exposure involving animal contact, were more likely to exhibit B. henselae seropositivity (OR 7.5). Bartonella spp. was present in the cats a moderate number of households, but Bartonella DNA was not detected in owners' blood, inferring that there is a low risk of recent human infection in the studied population.

Gait Training with Functional Electrical Stimulation Improves Mobility in People Post-Stroke.

(1) Background: Stroke is one of the leading causes of disability. To identify the best treatment strategies for people with stroke (PwS), the aim of the current study was to compare the effects of training on a treadmill with functional electrical stimulation (TT-FES) with training on a treadmill (TT), and to analyze the effects of sequence of training on mobility and the parameters of walking ability. (2) Methods: Prospective, longitudinal, randomized and crossover study, in which 28 PwS were distributed into groups, namely the A-B Group (TT-FES followed by TT) and B-A Group (TT followed by TT-FES), using the foot drop stimulator, and were measured with functional tests. (3) Results: We found improved mobility, balance, non-paretic limb coordination, and endurance only in the group that started with TT-FES. However, sensorimotor function improved regardless of the order of training, and paretic limb coordination only improved in the B-A Group, but after TT-FES. These data indicate that the order of the protocols changed the results. (4) Conclusions: Although biomechanical evaluation methods were not used, which can be considered a limitation, our results showed that TT-FES was superior to isolated training on a treadmill with regard to balance, endurance capacity, and coordination of the non-paretic limb.

COVID-19 Patterns in Araraquara, Brazil: A Multimodal Analysis.

The epidemiology of COVID-19 presented major shifts during the pandemic period. Factors such as the most common symptoms and severity of infection, the circulation of different variants, the preparedness of health services, and control efforts based on pharmaceutical and non-pharmaceutical interventions played important roles in the disease incidence. The constant evolution and changes require the continuous mapping and assessing of epidemiological features based on time-series forecasting. Nonetheless, it is necessary to identify the events, patterns, and actions that were potential factors that affected daily COVID-19 cases. In this work, we analyzed several databases, including information on social mobility, epidemiological reports, and mass population testing, to identify patterns of reported cases and events that may indicate changes in COVID-19 behavior in the city of Araraquara, Brazil. In our analysis, we used a mathematical approach with the fast Fourier transform (FFT) to map possible events and machine learning model approaches such as Seasonal Auto-regressive Integrated Moving Average (ARIMA) and neural networks (NNs) for data interpretation and temporal prospecting. Our results showed a root-mean-square error (RMSE) of about 5 (more precisely, a 4.55 error over 71 cases for 20 March 2021 and a 5.57 error over 106 cases for 3 June 2021). These results demonstrated that FFT is a useful tool for supporting the development of the best prevention and control measures for COVID-19.

Molecular diversity of Sarcocystis spp. in opossums (Didelphis spp.) from Southeastern and Midwestern Brazil.

South American opossums (Didelphis spp.) are definitive hosts of Sarcocystis neurona, Sarcocystis speeri, Sarcocystis lindsayi and Sarcocystis falcatula. In Brazil, diverse studies have demonstrated a high frequency of Sarcocystis falcatula-like in sporocysts derived from opossums, and high genetic diversity has been observed in surface antigen-encoding genes (SAGs). In this study, genetic diversity of Sarcocystis spp. derived from Didelphis albiventris and Didelphis aurita from the cities of Campo Grande and São Paulo, was accessed by sequencing SAG2, SAG3, SAG4, the first internal transcribed spacer (ITS-1) and cytochrome c oxidase subunit I (cox1). Molecular identification was performed for 16 DNA samples obtained from sporocyst or culture-derived merozoites. The ITS-1, cox1, and SAG3 fragments were cloned, whereas SAG2 and SAG4 were sequenced directly from PCR products. Four alleles variants were found for SAG2, 13 for SAG3 and seven for SAG4, from which four, 13 and four, respectively, were novel. Twenty-seven allele variants were found for ITS-1, all phylogenetically related to S. falcatula-like previously described in Brazil. Sarcocystis sp. phylogenetically related to Sarcocystis rileyi was evidenced by cox1 in three opossums. Further studies are needed to clarify the role of Didelphis spp. as definitive hosts of Sarcocystis spp. other than that previous described.

Bovine anaplasmosis as a risk factor for retained placenta, mastitis, and abomasal displacement in dairy cattle.

This study aimed to evaluate the frequency of IgG antibodies against A. marginale, the occurrence of this bacterium by qPCR, and the effect of bovine anaplasmosis as a risk factor for clinical cases of retained placenta, mastitis, and abomasal displacement in dairy cattle. For that 179 Holstein cows out of three dairy herds, in the municipality of Sertão, Rio Grande do Sul, Brazil. These cows were on farms that were vulnerable to risk factors that are crucial to susceptibility among these animals to this intracellular hemoparasite. The mean seropositivity for A. marginale from the periods evaluated was 54% on farm A, 69.4% on farm B, and 27.3% on farm C. Molecular diagnosis was performed with qPCR and the mean positivity for A. marginale among the cows on farms A, B, and C in December 2017 was 34.6% (67/179). Infected animals showed clinical cases of retained placenta (6.1%), mastitis (6.1%), and abomasal displacement (0.5%). The association between positivity for anaplasmosis and these clinical cases was assessed through the odds ratio. Our results show that females with a positive qPCR assay for A. marginale had 52.48 times increased probability (OR) to develop clinical cases of retained placenta and mastitis (P < 0.001). These clinical cases negatively impact the productivity of positive females. Thus, implementing preventive and prophylactic control measures to ensure the sanitary quality of the herds is needed to avoid losses due to morbidity and mortality and diminish the economic losses suffered by farmers.

Bartonella spp. and Typhus Group Rickettsiae among Persons Experiencing Homelessness, São Paulo, Brazil.

Persons experiencing homelessness in São Paulo, Brazil, were seropositive for Bartonella spp. (79/109, 72.5%) and typhus group rickettsiae (40/109, 36.7%). Bartonella quintana DNA was detected in 17.1% (14/82) body louse pools and 0.9% (1/114) blood samples. Clinicians should consider vectorborne agents as potential causes of febrile syndromes in this population.

Serological evidence of exposure to Bartonella sp. in dogs with suspected vector-borne diseases, toxoplasmosis and neosporosis.

Bartonellosis is a vector-borne zoonotic disease with worldwide distribution that infect a broad spectrum of mammalian species. Despite the recent studies carried out in Brazil, information regarding Bartonella in dogs are scarce. Therefore, we performed a retrospective study to investigate the exposure to Bartonella sp. in dogs by indirect immunofluorescence assay (IFA). Three hundred and thirty-five archived serum samples from dogs previously tested for vector-borne pathogens, Toxoplasma gondii, and Neospora caninum were screened for the presence of IgG antibodies to Bartonella sp. All dogs originated from the Metropolitan region of Ribeirão Preto, northeast of the State of São Paulo. Twenty-eight samples (8.3%) were positive for Bartonella sp. at the cut-off of 64. Among the 28 seropositive samples for Bartonella sp., 16 (57.1%) were also seropositive for Ehrlichia canis, 12 (42.8%) for Babesia vogeli, five (17.8%) for T. gondii and three (10.7%) for L. infantum and N. caninum. Our results demonstrated that dogs sampled were exposed to Bartonella sp. Since all the animals sampled in the present study were from private owners, our findings demonstrate that these people may also be exposed to Bartonella sp. Further studies designed to assess whether the infection by other arthropod-borne pathogens such as B. vogeli and E. canis are risk factors for Bartonella infection are needed.

Novel Ehrlichia and Hepatozoon genotypes in white-eared opossums (Didelphis albiventris) and associated ticks from Brazil.

White-eared opossums (Didelphis albiventris) are well adapted to anthropized areas. The increased contact with domestic animals and humans mediates the transmission of arthropod-borne pathogens. Despite the worldwide occurrence of tick-borne Anaplasmataceae and Hepatozoidae species in a variety of vertebrates, few studies reported serological evidence or molecular detection of theses agentes in marsupials. Up to now, while Ehrlichia/Anaplasma spp. have only been detected in marsupials from Brazil, Hepatozoon spp. have been reported in marsupials from Chile, Australia and Brazil. The present work aimed to investigate, using molecular techniques and blood smear analysis, the presence of Ehrlichia spp., Anaplasma spp., and Hepatozoon sp. in the blood and ticks collected from D. albiventris in urban forest fragments from midwestern Brazil. Between May and December 2017, 43 D. albiventris (27 males and 16 females) were captured for blood and tick collection in the city of Campo Grande, state of Mato Grosso do Sul, midwestern Brazil. Ticks (46 Amblyomma dubitatum nymphs and 24 Amblyomma spp. larvae) were collected from 14 out 43 (32.5%) of the white-eared opossums. Panoptic-stained blood smears were performed using peripheral blood (tail tip) of the captured opossums. DNA extracted from blood and tick samples were subjected to PCR/qPCR assays for Anaplasmataceae agents (rrs, gltA, groEL, sodB, and dsb genes, and 23S-5S intergenic region) and Hepatozoon spp. (18S rRNA gene), followed by Sanger sequencing, BLASTn and phylogenetic analyses. An inclusion resembling Ehrlichia morulae was found in a white-eared opossum's monocyte from a blood smear stained with Panoptic. Five (11.63% [5/43]) white-eared opossums' blood samples and 7 (25% [7/28]) tick samples (2 pools of Amblyomma spp. larvae and 5 pools of A. dubitatum nymphs) were positive for Anaplasmataceae via a PCR assay targeting the conserved rrs gene. Phylogenetic analysis based on the rrs gene positioned three sequences obtained from opossums and ticks together as a subclade within the Ehrlichia canis clade. However, all samples were negative in a qPCR assay specific for E. canis based on the dsb gene. Phylogenetic analyses positioned the gltA and 23S-5S ITS sequences obtained from opossums' blood samples in a separate clade from the other validated Ehrlichia species. One (2.3% [1/43]) opossum blood sample was positive for the 18S rRNA gene of Hepatozoon sp. The phylogenetic analysis positioned the Hepatozoon sp. sequence obtained from a D. albiventris specimen in a clade with a sequence previously detected in a black storm petrel (Oceanodroma melania) from Mexico. All the other sequences of Hepatozoon sp. previously detected in marsupials from Brazil were positioned in a separated clade. The present work showed the occurrence of putative novel genotypes of Ehrlichia sp. and Hepatozoon sp. in white-eared opossums and associated A. dubitatum ticks from midwestern Brazil.

Micro aerial vehicle with basic risk of operation.

We draw current efforts towards proposing a wing-type micro UAV with characteristics of being a basic operation risk self handled (Micro-Brosh) platform. Its micro-sized wingspan and weight, which are less than 0.30 m and 0.150 kg, respectively, guarantee the low risk to the operator and installations in case of crashing. It can be launched manually without using an appropriate runway, besides using a soft grass field for landing is recommended. Its associated costs for construction and maintenance are very low (below US$ 500) if compared to traditional aircraft. The main contribution here is the architectural design, besides we provide detailed documentation including techniques for determining lift, thrust, drag, minimum flight velocity, maximum time of flight and distance (autonomy), and other issues that we have solved related to the aircraft development. We validate our prototype with experiments including several tests done with the UAV with results that have proven its flight ability. To the end, we understand that this work provides a nice starting document to researchers that intend to develop a UAV or enter this field.

Diabetic foot infection caused by bacteria of the Burkholderia cepacia complex: report of an unusual case and a scoping literature review.

Burkholderia cepacia complex (BCC) is group of widespread gram-negative bacillus organized in over 20 phylogenetically distinct bacterial species. According to previous studies, BCC species pathogens are widely reported in patients with cystic fibrosis (CF), but not in individuals with diabetes mellitus (DM). In this case report, a 42-year-old male patient with DM and a foot infection caused by BCC is presented. The patient was hospitalized after antibiotic treatment failure and improved after two surgical debridement procedures and a high-dose extended infusion (EI) of meropenem. The team of vascular surgeons and the infectious disease specialists worked fervently to solve the case. Finally, a scoping review was conducted to map BCC infections in patients with DM.

Threat under cats' claws: Molecular detection and risk factors for zoonotic Bartonella species in blood and claw samples from cats in Brazil.

Bartonelloses are zoonoses widely dispersed throughout the world caused by bacteria of the genus Bartonella. Domestic cats play an important role in the epidemiology of bartonelloses, since these animals are considered natural hosts of B. henselae, B. koehlerae and B. clarridgeiae. This study aimed to determine the occurrence of Bartonella spp. in domestic cats' blood and claw samples in the southern region of Bahia, northeastern Brazil. Additionally, the main clinical and hematological changes in Bartonella-positive animals were investigated, as well as the risk factors associated with the infection. For this purpose, 188 indoor house domestic cats were clinically evaluated and submitted to claw and blood sample collection. Additionally, data regarding the clinical history of the animals were recorded. Out of 188 cats' blood samples, 20.7% (39/188) were positive in the qPCR for Bartonella spp. based on the nuoG gene. Out of 39 claw samples collected, 23.9% (9/39) were positive for Bartonella spp. The parameters of the blood and claw samples ranged from 1.42 to 395,000 and 4.32 - 108,000 copies/µL of a fragment of Bartonella nuoG gene, respectively. The amplified sequences shared identity ranging from 99% to 100% with the three main cat-related Bartonella species. Higher platelet values (p = 0.0082) were observed in animals positive for Bartonella spp. Young and unsterilized cats with outdoor access were more prone to infection by Bartonella spp. The data reported here demonstrated the occurrence of Bartonella spp. in blood and claw samples from cats in northeastern Brazil showing no significant clinical and hematological disorders.

Sustainable Solutions for Sea Monitoring With Robotic Sailboats: N-Boat and F-Boat Twins.

Strategic management and production of internal energy in autonomous robots is becoming a research topic with growing importance, especially for platforms that target long-endurance missions, with long-range and duration. It is fundamental for autonomous vehicles to have energy self-generation capability to improve energy autonomy, especially in situations where refueling is not viable, such as an autonomous sailboat in ocean traversing. Hence, the development of energy estimation and management solutions is an important research topic to better optimize the use of available energy supply and generation potential. In this work, we revisit the challenges behind the project design and construction for two fully autonomous sailboats and propose a methodology based on the Restricted Boltzmann Machine (RBM) in order to find the best way to manage the supplementary energy generated by solar panels. To verify the approach, we introduce a case study with our two developed sailboats that have planned payload with electric and electronics, and one of them is equipped with an electrical engine that may eventually help with the sailboat propulsion. Our current results show that it is possible to augment the system confidence level for the potential energy that can be harvested from the environment and the remaining energy stored, optimizing the energy usage of autonomous vehicles and improving their energy robustness.