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There is no suitable vaccine against human visceral leishmaniasis (VL) and available drugs are toxic and/or present high cost. In this context, diagnostic tools should be improved for clinical management and epidemiological evaluation of disease. However, the variable sensitivity and/or specificity of the used antigens are limitations, showing the necessity to identify new molecules to be tested in a more sensitive and specific serology. In the present study, an immunoproteomics approach was performed in Leishmania infantum promastigotes and amastigotes employing sera samples from VL patients. Aiming to avoid undesired cross-reactivity in the serological assays, sera from Chagas disease patients and healthy subjects living in the endemic region of disease were also used in immunoblottings. The most reactive spots for VL samples were selected, and 29 and 21 proteins were identified in the promastigote and amastigote extracts, respectively. Two of them, endonuclease III and GTP-binding protein, were cloned, expressed, purified and tested in ELISA experiments against a large serological panel, and results showed high sensitivity and specificity values for the diagnosis of disease. In conclusion, the identified proteins could be considered in future studies as candidate antigens for the serodiagnosis of human VL.
Tuberculosis (TB) is the leading cause of death among infectious diseases worldwide. Among the estimated cases of drug-resistant TB, approximately 60% occur in the BRICS countries (Brazil, Russia, India, China and South Africa). Among Brazilian states, primary and acquired multidrug-resistant TB (MDR-TB) rates were the highest in Rio Grande do Sul (RS). This study aimed to perform molecular characterisation of MDR-TB in the State of RS, a high-burden Brazilian state. We performed molecular characterisation of MDR-TB cases in RS, defined by drug susceptibility testing, using 131 Mycobacterium tuberculosis (M.tb) DNA samples from the Central Laboratory. We carried out MIRU-VNTR 24loci, spoligotyping, sequencing of the katG, inhA and rpoB genes and RDRio sublineage identification. The most frequent families found were LAM (65.6%) and Haarlem (22.1%). RDRio deletion was observed in 42 (32%) of the M.tb isolates. Among MDR-TB cases, eight (6.1%) did not present mutations in the studied genes. In 116 (88.5%) M.tb isolates, we found mutations associated with rifampicin (RIF) resistance in rpoB gene, and in 112 isolates (85.5%), we observed mutations related to isoniazid resistance in katG and inhA genes. An insertion of 12 nucleotides (CCAGAACAACCC) at the 516 codon in the rpoB gene, possibly responsible for a decreased interaction of RIF and RNA polymerase, was found in 19/131 of the isolates, belonging mostly to LAM and Haarlem families. These results enable a better understanding of the dynamics of transmission and evolution of MDR-TB in the region.
Cadillo is an invasive species in Florida pastures and natural areas. Despite its invasiveness, relatively few studies have evaluated cadillo management. Thus, the objective of this research was to determine effective POST herbicides for cadillo control in Florida. Greenhouse and field studies were conducted at the Range Cattle Research and Education Center near Ona, FL, in 2015 and 2016. In the greenhouse study, triclopyr-ester, aminopyralid, metsulfuron, 2,4-D amine, aminopyralid+metsulfuron, aminocyclopyrachlor+metsulfuron, and imazapyr+aminocyclopyrachlor+metsulfuron provided ≥80% control of cadillo 28 d after treatment (DAT). Aminocyclopyrachlor at 17 and 35 g ha–1 were the only treatments with <80% control, with 70% and 75% control, respectively. Similar results were reflected in cadillo dry biomass reduction. The herbicide treatments used in the field study were triclopyr-ester, aminopyralid, 2,4-D amine, aminocyclopyrachlor, and triclopyr+fluroxypyr. Most treatments provided excellent control in the field (≥90% control) 30 DAT, and by 60 DAT all treatments provided 100% control. Results from these studies suggest that cadillo is susceptible to many of the common POST herbicides utilized in pastures and natural areas in Florida.
In Brazil, rabies surveillance is based on monitoring domestic and wild animals, although the most prevalent lineage of the rabies virus (RABV) currently diagnosed in Brazil is associated with bats, particularly non-haematophagous bats. Disease control is based on the mass vaccination of dogs and cats. We used data collected by the passive surveillance system of the city of Campinas from 2011 to 2015, to describe the temporal and geographic distributions of the bat specimens and RABV and discuss the current rabies surveillance with the advent of the declaration of canine and feline rabies-free areas in Brazil. We described the species, locations and health statuses of the collected bat specimens. Moreover, all samples were submitted for RABV diagnosis. Then, we performed a time series decomposition for each bat family. Additionally, we determined the spatiotemporal relative risk for RABV infection using the ratio of the kernel-smoothed estimates of spatiotemporal densities of RABV-positive and RABV-negative bats. From the 2537 bat specimens, the most numerous family was Molossidae (72%), followed by Vespertilionidae (14%) and Phyllostomidae (13%). The bat families behaved differently in terms of seasonal and spatial patterns. The distribution of bats varied geographically in the urban environment, with Molossidae and Phyllostomidae being observed downtown and Vespertilionidae being observed in peripheral zones. Concurrently, a significant relative risk of RABV infection was observed downtown for Vespertilionidae and in peripheral zones for Molossidae. No RABV-positive sample clusters were observed. As a result of the official declaration of RABV-free areas in southern Brazil, mass dog and cat vaccinations are expected to halt in the near future. This stoppage would make most dog and cat populations susceptible to other RABV lineages, such as those maintained by non-haematophagous bats. In this scenario, all information available on bats and RABV distribution in urban areas is essential. Currently, few studies have been conducted. Some local health authorities, such as that in Campinas, are spontaneously basing their surveillance efforts on bat rabies, which is the alternative in reality scenario of increased susceptibility to bat-associated RABV that is developing in Brazil.
The fraction of organic matter present affects the fragmentation behavior of sialoliths; thus, pretherapeutic information on the degree of mineralization is relevant for a correct selection of lithotripsy procedures. This work proposes a methodology for in vivo characterization of salivary calculi in the pretherapeutic context. Sialoliths were characterized in detail by X-ray computed microtomography (μCT) in combination with atomic emission spectroscopy, Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, and transmission electron microscopy. Correlative analysis of the same specimens was performed by in vivo and ex vivo helical computed tomography (HCT) and ex vivo μCT. The mineral matter in the sialoliths consisted essentially of apatite (89 vol%) and whitlockite (11 vol%) with average density of 1.8 g/cm3. In hydrated conditions, the mineral mass prevailed with 53 ± 13 wt%, whereas the organic matter, with a density of 1.2 g/cm3, occupied 65 ± 10% of the sialoliths’ volume. A quantitative relation between sialoliths mineral density and X-ray attenuation is proposed for both HCT and μCT.
Gluten is only partially digested by intestinal enzymes and can generate peptides that can alter intestinal permeability, facilitating bacterial translocation, thus affecting the immune system. Few studies addressed the role of diet with gluten in the development of colitis. Therefore, we investigate the effects of wheat gluten-containing diet on the evolution of sodium dextran sulphate (DSS)-induced colitis. Mice were fed a standard diet without (colitis group) or with 4·5 % wheat gluten (colitis + gluten) for 15 d and received DSS solution (1·5 %, w/v) instead of water during the last 7 d. Compared with the colitis group, colitis + gluten mice presented a worse clinical score, a larger extension of colonic injury area, and increased mucosal inflammation. Both intestinal permeability and bacterial translocation were increased, propitiating bacteria migration for peripheral organs. The mechanism by which diet with gluten exacerbates colitis appears to be related to changes in protein production and organisation in adhesion junctions and desmosomes. The protein α-E-catenin was especially reduced in mice fed gluten, which compromised the localisation of E-cadherin and β-catenin proteins, weakening the structure of desmosomes. The epithelial damage caused by gluten included shortening of microvilli, a high number of digestive vacuoles, and changes in the endosome/lysosome system. In conclusion, our results show that wheat gluten-containing diet exacerbates the mucosal damage caused by colitis, reducing intestinal barrier function and increasing bacterial translocation. These effects are related to the induction of weakness and disorganisation of adhesion junctions and desmosomes as well as shortening of microvilli and modification of the endocytic vesicle route.
BirdLife International´s Important Bird and Biodiversity Areas (IBA) Programme has identified, documented and mapped over 13,000 sites of international importance for birds. IBAs have been influential with governments, multilateral agreements, businesses and others in: (1) informing governments’ efforts to expand protected area networks (in particular to meet their commitments through the Convention on Biological Diversity); (2) supporting the identification of Ecologically or Biologically Significant Areas (EBSAs) in the marine realm, (3) identifying Wetlands of International Importance under the Ramsar Convention; (4) identifying sites of importance for species under the Convention on Migratory Species and its sister agreements; (5) identifying Special Protected Areas under the EU Birds Directive; (6) applying the environmental safeguards of international finance institutions such as the International Finance Corporation; (7) supporting the private sector to manage environmental risk in its operations; and (8) helping donor organisations like the Critical Ecosystems Partnership Fund (CEPF) to prioritise investment in site-based conservation. The identification of IBAs (and IBAs in Danger: the most threatened of these) has also triggered conservation and management actions at site level, most notably by civil society organisations and local conservation groups. IBA data have therefore been widely used by stakeholders at different levels to help conserve a network of sites essential to maintaining the populations and habitats of birds as well as other biodiversity. The experience of IBA identification and conservation is shaping the design and implementation of the recently launched Key Biodiversity Areas (KBA) Partnership and programme, as IBAs form a core part of the KBA network.
Important Bird and Biodiversity Areas (IBAs) are sites identified as being globally important for the conservation of bird populations on the basis of an internationally agreed set of criteria. We present the first review of the development and spread of the IBA concept since it was launched by BirdLife International (then ICBP) in 1979 and examine some of the characteristics of the resulting inventory. Over 13,000 global and regional IBAs have so far been identified and documented in terrestrial, freshwater and marine ecosystems in almost all of the world’s countries and territories, making this the largest global network of sites of significance for biodiversity. IBAs have been identified using standardised, data-driven criteria that have been developed and applied at global and regional levels. These criteria capture multiple dimensions of a site’s significance for avian biodiversity and relate to populations of globally threatened species (68.6% of the 10,746 IBAs that meet global criteria), restricted-range species (25.4%), biome-restricted species (27.5%) and congregatory species (50.3%); many global IBAs (52.7%) trigger two or more of these criteria. IBAs range in size from < 1 km2 to over 300,000 km2 and have an approximately log-normal size distribution (median = 125.0 km2, mean = 1,202.6 km2). They cover approximately 6.7% of the terrestrial, 1.6% of the marine and 3.1% of the total surface area of the Earth. The launch in 2016 of the KBA Global Standard, which aims to identify, document and conserve sites that contribute to the global persistence of wider biodiversity, and whose criteria for site identification build on those developed for IBAs, is a logical evolution of the IBA concept. The role of IBAs in conservation planning, policy and practice is reviewed elsewhere. Future technical priorities for the IBA initiative include completion of the global inventory, particularly in the marine environment, keeping the dataset up to date, and improving the systematic monitoring of these sites.
Diarrhoeagenic Escherichia coli (DEC) is a leading cause of infectious diarrhoea worldwide. In recent years, Escherichia albertii has also been implicated as a cause of human enteric diseases. This study describes the occurrence of E. coli pathotypes and serotypes associated with enteric illness and haemolytic uremic syndrome (HUS) isolated in Brazil from 2011 to 2016. Pathotypes isolated included enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC) and Shiga toxin-producing E. coli (STEC). PCR of stool enrichments for DEC pathotypes was employed, and E. albertii was also sought. O:H serotyping was performed on all DEC isolates. A total of 683 DEC and 10 E. albertii strains were isolated from 5047 clinical samples. The frequencies of DEC pathotypes were 52.6% (359/683) for EPEC, 32.5% for EAEC, 6.3% for ETEC, 4.4% for EIEC and 4.2% for STEC. DEC strains occurred in patients from 3 months to 96 years old, but EPEC, EAEC and STEC were most prevalent among children. Both typical and atypical isolates of EPEC and EAEC were recovered and presented great serotype heterogeneity. HUS cases were only associated with STEC serotype O157:H7. Two E. albertii isolates belonged to serogroup O113 and one had the stx2f gene. The higher prevalence of atypical EPEC in relation to EAEC in community-acquired diarrhoea in Brazil suggests a shift in the trend of DEC pathotypes circulation as previously EAEC predominated. This is the first report of E. albertii isolation from active surveillance. These results highlight the need of continuing DEC and E. albertii surveillance, as a mean to detect changes in the pattern of pathotypes and serotypes circulation and provide useful information for intervention and control strategies.
In the current study, phage-exposed mimotopes as targets against tegumentary leishmaniasis (TL) were selected by means of bio-panning cycles employing sera of TL patients and healthy subjects, besides the immune stimulation of peripheral blood mononuclear cells (PBMCs) collected from untreated and treated TL patients and healthy subjects. The clones were evaluated regarding their specific interferon-γ (IFN-γ) and interleukin-4 (IL-4) production in the in vitro cultures, and selectivity and specificity values were calculated, and those presenting the best results were selected for the in vivo experiments. Two clones, namely A4 and A8, were identified and used in immunization protocols from BALB/c mice to protect against Leishmania amazonensis infection. Results showed a polarized Th1 response generated after vaccination, being based on significantly higher levels of IFN-γ, IL-2, IL-12, tumour necrosis factor-α (TNF-α) and granulocyte-macrophage colony-stimulating factor (GM-CSF); which were associated with lower production of specific IL-4, IL-10 and immunoglobulin G1 (IgG1) antibodies. Vaccinated mice presented significant reductions in the parasite load in the infected tissue and distinct organs, when compared with controls. In conclusion, we presented a strategy to identify new mimotopes able to induce Th1 response in PBMCs from TL patients and healthy subjects, and that were successfully used to protect against L. amazonensis infection.
Knowing the burden of influenza is helpful for policy decisions. Here we estimated the contribution of influenza-like illness (ILI) visits associated with laboratory-confirmed influenza among all clinic visits in a Senegal sentinel network. ILI data from ten sentinel sites were collected from January 2013 to December 2015. ILI was defined as an axillary measured fever of more than 37.5 °C with a cough or a sore throat. Collected nasopharyngeal swabs were tested for influenza viruses by rRT-PCR. Influenza-associated ILI was defined as ILI with laboratory-confirmed influenza. For the influenza disease burden estimation, we used all-case outpatient visits during the study period who sought care at selected sites. Of 4030 ILI outpatients tested, 1022 were influenza positive. The estimated proportional contribution of influenza-associated ILI was, per 100 outpatients, 1.2 (95% CI 1.1–1.3), 0.32 (95% CI 0.28–0.35), 1.11 (95% CI 1.05–1.16) during 2013, 2014, 2015, respectively. The age-specific outpatient visits proportions of influenza-associated ILI were higher among children under 5 years (0.68%, 95% CI: 0.62–0.70). The predominant virus during years 2013 and 2015 was influenza B while A/H3N2 subtype was predominant during 2014. Influenza viruses cause a substantial burden of outpatient visits particularly among children under 5 of age in Senegal and highlight the need of vaccination in risk groups.
A study was conducted over eight consecutive days in February 2010 in which daily variations in the vertical distributions of heterotrophic bacteria, mesozooplankton and ichthyoplankton at 1–1200 m in the South-western Atlantic Ocean were investigated. Diurnal and nocturnal samples were collected at an oceanographic station at four regional depths: Tropical Water (TW) (1 m), South Atlantic Central Water (SACW) (250 m), Antarctic Intermediate Water (AAIW) (800 m) and Upper Circumpolar Deep Water (UCDW) (1200 m). Bacterial, mesozooplankton and larval fish densities significantly differed between sample depths but not between sampling tow times. In total, 154 zooplankton species and 18 larval fish species were identified. The highest number of taxa was obtained from the night-time TW trawls. This depth zone had the highest densities of mesozooplankton, larval fish and bacterioplankton (auto and heterotrophic), associated with the highest temperature and salinity and the lowest inorganic nutrient concentrations. Two sample groups were identified based on their mesozooplankton and larval fish compositions: night-time TW and other water masses (daytime TW, SACW, AAIW and UCDW). Thirty-two indicator species were detected in night-time TW. The copepod Nullosetigera impar was, to the best of our knowledge, identified for the first time on the Brazilian coast. Our results showed significant variability in the abundance and vertical distribution of mesozooplankton, bacterioplankton and larval fish along the water column in an oceanic area. We have provided new data and insights on the composition and vertical distribution of mesozooplankton, larval fish and bacterioplankton in deep waters in the South-western Atlantic Ocean.
This study aimed to evaluate the risk factors for tuberculosis (TB) treatment default in a priority city for disease control in Brazil. A cohort of TB cases diagnosed from 2008 to 2009 was followed up from patients’ entry into three outpatient sites, in Juiz de Fora, Minas Gerais (Brazil), until the recording of the outcomes. Drug addiction, alcoholism and treatment site appeared to be independently associated with default. Current users of crack as the hardest drug (odds ratio (OR) 12·25, 95% confidence interval (CI) 3·04–49·26) were more likely to default than other hard drug users (OR 5·67, 95% CI 1·34–24·03), former users (OR 4·12, 95% CI 1·11–15·20) and those not known to use drugs (reference group). Consumers at high risk of alcoholism (OR 2·94, 95% CI 1·08–7·99) and those treated in an outpatient hospital unit (OR 8·22, 95% CI 2·79–24·21%) also were more likely to default. Our results establish that substance abuse was independently associated with default. National TB programmes might be more likely to achieve their control targets if they include interventions aimed at improving adherence and cure rates, by diagnosing and treating substance abuse concurrently with standard TB therapy.
Phosphorus and calcium deficiency in horses represents an important factor responsible for the low equine production in Brazil. The basic mechanisms of P and Ca metabolism differ substantially among species. Regulation of P and Ca metabolism is less well understood in horses than in others species. With the use of the isotopic dilution technique is possible to evaluate the metabolism for this mineral. The aim of the present experiment was to study the effect of different Ca levels in the diet on P and Ca metabolism in horses.
Anti nutritional factors in tropical legumes are very common and the chemical analysis for tannins has become an important tool to evaluate alternative ruminant feeds in the tropics. However, frequently results from tannin chemical analysis are not in agreement with biological response when animals are fed those feeds. There is a lack of information concerning the biological effects (activity or reactivity) of tannins on ruminants. Usually the effects of tannins are tested in vitro by adding compounds with capacity of biding tannins. The aim of this work was to compare two binding compounds to evaluate the effect of tannins of tanniniferous feeds for ruminants.
In vitro and in situ techniques for research on ruminants are currently much in focus. Since they have good correlations with in vivo data, they are feasible alternatives to predict the nutrition rates of feeds and may be applied in equine research on in vivo apparent digestibility. On the other hand, the disadvantage of these methods is due to the fact that fistulated animals are required to obtain the inoculum. Theodorou et al., (1994) developed an extremely promising gas production technique to assess feeds for ruminants, but still require rumen inoculum obtained from operated animals. Faecal microorganisms function similarly to those in the rumen and in the large intestine of equines. The objective this experiment was to compare rumen liquor and equine faeces as inoculum to determine in vitro digestibility of equine feeds.
Concentrate mixtures fed ruminants generally are composed by cereals where phosphorus is present mainly in phytate form or phytin (Maga, 1982). Phytate phosphorus is thought to be completely available to ruminants due to the presence of phytase enzyme that hydrolyzes phytate phosphorus making it available for absorption. Researches have shown that this fact is not always true, depending on different conditions (Park et al., 2000). The aim of this paper was to study the influence of phytate phosphorus on calcium availability.
It is a general believe that the local sheep breed Churra da Terra Quente (CTQ), reared in the Northeast of Portugal is well adapted to the conditions of its production system. However, there are large gaps on our knowledge to allow understanding of such claimed adaptation. Several researchers have found differences in digestibility between sheep breeds (e.g. Givens and Moss, 1994; López et al., 2001), suggesting that those fed natural resources can have an improved ability to digest fibre. This experiment was designed to compare digestibility coefficients of a high– and a low–roughage diet measured on sheep from CTQ and Ile-de-France (IF) breeds at the same degree of maturity.
In Northeast of Portugal sheep is reared under extensive systems. These systems frequently involve expressive body composition changes due to the storing and mobilization of body reserves, mainly fat. Body condition score (BCS) is the most common way to assess these reserves and the nutritional status of ewes. However due to the subjective nature of BCS, their quality has been questioned and other alternatives has been studied. For cattle there are some studies that use the real time ultrasonography (RTU) to evaluate the BCS (Schwager-Suter et al., 2000; Broring et al., 2003), but this approach was not tested in ewes. Therefore the aim of the present study was to establish a relationship between the BCS and ultrasound subcutaneous fat (SF) and Longissimus thoracis et lumborum muscle (LM) measurements.