A Contribution towards Sustainable Development in the Amazon Based on a Socioeconomic and Environmental Analysis of Visceral Leishmaniasis in the State of Pará, Brazil.

Human Visceral Leishmaniasis is an endemic public health problem in the Amazon. This article analyzed the spatial distribution of this disease and its relationship with socioeconomic, environmental and public health policy variables in four mesoregions of the state of Pará, from 2011 to 2022. This ecological study used secondary data obtained from official Brazilian agencies. Spatial analysis was performed using the Flow, Kernel and Global Moran bivariate techniques expressed in thematic maps. In the mesoregions studied, 2685 cases of the disease were confirmed, with the highest number of cases in Southeast Pará state. The epidemiological profile followed the national pattern of occurrence of the disease, with a higher number of cases in children below school age. Spatial dependence was observed between the prevalence of the disease and socio-economic indicators. The most intense movement of patients was towards the Belém Metropolitan mesoregion. The disease showed an inhomogeneous pattern of distribution of cases, with a direct relationship between areas with cases and deforestation associated with different anthropic activities. There is a socio-environmental production of the disease that goes beyond the border limits of the mesoregions, and its establishment is related to the unsustainable development model implemented in the region.

New karyotype information for Ctenomys (Rodentia: Ctenomyidae) from Midwest and Northern Brazil.

Its wide karyotypic variation characterizes the genus Ctenomys, and in Brazil, the genus is distributed in the country's southern, Midwest, and Northern regions. Recently, populations of Ctenomys have been found in the Midwest and northern Brazil, with two new lineages named C. sp. "xingu" and C. sp. "central". This work combines classical cytogenetic and molecular analyses to provide new chromosomal information on the boliviensis group distributed in Northern and Midwestern Brazil. This includes the validation of the karyotype of C. bicolor and C. nattereri, and the description of the karyotype of C. sp. "xingu" and C. sp. "central". We found three different karyotypes: 2n=40 for C. bicolor; 2n= 36 for C. nattereri, and specimens from a locality belonging to C. sp. "central"; 2n=34 for the lineage C. sp. "xingu" and specimens from a locality belonging to C. sp. "central". Furthermore, GTG-banding revealed homologous chromosomes between species/lineages and allowed the identification of the rearrangements that occurred, which proved the occurrence of fissions. Considering our results on the variation of 2n in the boliviensis group, we found two possibilities: the first, deduced by parsimony, is that 2n=36 appeared initially, and two fissions produced gave rise to 2n=40, and an independent fusion gave rise to 2n=34 from 2n=36; moreover, the second explanation is that all karyotypes arose independently.

Composition, Antibiofilm, and Antibacterial Potential of Volatile Oils from Geopropolis of Different Stingless Bees' Species.

We aimed to characterize and investigate the antibacterial potential of the native stingless bees geopropolis volatile oils (VO) for the search of potentially new bioactive compounds. Geopropolis samples from Melipona bicolor schencki, M. compressipes manaosensis, M. fasciculata, M. quadrifasciata, M. marginata and M. seminigra merrillae were collected from hives in South Brazil. VO were obtained by hydrodistillation and characterised by gas chromatography coupled to mass spectrometry (GC/MS). Antimicrobial activity was assessed by microplate dilution method. The lowest MIC against cell walled bacteria was 219±0 µg mL-1 from M. quadrifasciata geopropolis VO with Staphylococcus aureus. The M. b. schencki geopropolis VO minimal inhibition concentration (MIC) was 424±0 µg mL-1 against all the mycoplasma strains evaluated. Fractionation resulted in the reduction of 50 % of the MIC value from the original oil. However, its compounds' synergism seems to be essential to this activity. Antibiofilm assays demonstrated 15.25 % eradication activity and 13.20 % inhibition of biofilm formation after 24 h for one subfraction at 2× its MIC as the best results found. This may be one of the essential mechanisms by which geopropolis VOs perform their antimicrobial activity.

Visceral Leishmaniasis and Land Use and Cover in the Carajás Integration Region, Eastern Amazon, Brazil.

Human visceral leishmaniasis is a major public health problem in the Amazon. Thus, we analyzed the spatial distribution of this disease and its relationship with epidemiological, socioeconomic, and environmental variables in the Carajás Integration Region, Pará state, from 2011 to 2020. Epidemiological data for this ecological study were obtained from the State Public Health Secretariat, environmental data were obtained from the National Space Research Institute, and socioeconomic data were obtained from the Brazilian Geography and Statistics Institute. ArcGIS 10.5.1 software was used for classifying land use and cover and for the Kernel and Moran spatial analyses. It was observed in 685 confirmed cases that the epidemiological profile followed the national pattern of the disease occurrence, with a high prevalence in children who were not school-aged. The disease had a non-homogeneous distribution with clusters related to different human activities, such as urbanization, ranching, and mining. A spatial dependence between the disease prevalence and socioeconomic indicators was observed. The municipalities presented gradients of case densities associated with a direct relationship between areas with cases and deforestation. The disease is developing due to risk factors such as establishment and maintenance related to the non-sustainable development model implemented in the region, pointing to the need for its revision.

Antimicrobial Activity of Volatile Oils from Brazilian Stingless Bees Melipona quadrifasciata quadrifasciata and Tetragonisca angustula Propolis.

Few studies were performed on the volatile components' properties of propolis from native Brazilian stingless bees. This work sought to extract, chemically characterize, and evaluate of the antimicrobial properties of volatile oils from propolis of the stingless bees Melipona quadrifasciata quadrifasciata (MQ) and Tetragonisca angustula (TA). The volatile oils from the pulverized propolis samples were obtained in a Clevenger apparatus and characterized by gas chromatography coupled to mass spectrometry (GC/MS) and to flame ionization detector (GC-FID). All samples showed promising activity against Mycoplasma pneumoniae, with MICs from 103 to 224 µg mL-1 , nonetheless, they were less active against cell walled bacteria. Activity against Candida species was moderate, but one MQ sample showed a MIC value of 103 µg mL-1 against C. tropicallis. Oils' fractionation showed no improvement in antibacterial activity. The promising antibacterial effect against this microorganism is likely related to the synergism between the components of the volatile oils.

Mollicute Anti-Adhesive and Growth Inhibition Properties of the Methanolic Extract of Propolis from the Brazilian Native Bee Melipona quadrifasciata.

Hydroalcoholic propolis extracts from the bee species Melipona quadrifasciata have been shown to possess antimicrobial activity against different mollicute strains, but a methanolic extract (ME) could contain an increased diversity of nonpolar bioactive components with a potentially higher antimicrobial activity. The ME obtained by maceration of the propolis sample was fractionated with solvents of different polarities and then, purified by silica gel column chromatography through biomonitoring of its antimicrobial activity against mollicute strains. Analysis by gas chromatography-mass spectrometry (GC/MS) enabled the identification of compounds using the NIST library. Minimum inhibitory concentrations (MICs) of the samples were determined by broth microdilution. Anti-adhesive assays were performed with Mycoplasma pneumoniae cells. The hexane (MIC=62.5 mg/L) and dichloromethane (MIC=125 mg/L) fractions presented the most promising results against M. pneumoniae. They were fractionated into 74 subfractions, and even the best ones did not show better results (MIC>250 mg/L) than their original fractions, likely due to the loss of terpene compounds that seem to act in synergy. The dichloromethane subfraction FD4 was highlighted in the anti-adhesive assay with an inhibitory activity of 21.6 %. A synergistic effect of the nonpolar compounds in M. quadrifasciata propolis may be responsible for its antibacterial activity, but several purified components can improve its anti-adhesive properties.

Coxiella and Bartonella spp. in bats (Chiroptera) captured in the Brazilian Atlantic Forest biome.

BACKGROUND: The role of bats as reservoirs of zoonotic agents, especially pathogenic bacteria such as Bartonella and Coxiella, has been discussed around the world. Recent studies have identified bats as potential hosts of species from the proteobacteria phylum. In Brazil, however, the role of bats in the natural cycle of these agents is poorly investigated and generally neglected. In order to analyze the participation of bats in the epidemiology of diseases caused by Bartonella, Coxiella, Rickettsia, Anaplasma and Ehrlichia, we conducted a descriptive epidemiological study in three biogeographic regions of the Brazilian Atlantic Forest. RESULTS: Tissues of 119 bats captured in preserved areas in the states of Rio de Janeiro, Bahia and Santa Catarina from 2014 to 2015 were submitted to molecular analysis using specific primers. Bartonella spp. was detected in 22 spleen samples (18.5%, 95% CI: 11.9-26.6), whose phylogenetic analysis revealed the generation of at least two independent clusters, suggesting that these may be new unique genotypes of Bartonella species. In addition, four samples (3.4%, 95% CI: 0.9-8.3) were positive for the htpAB gene of C. burnetii [spleen (2), liver (1) and heart (1)]. Rickettsia spp., Anaplasma and Ehrlichia were not identified. This is the first study reporting C. burnetii and Bartonella spp. infections in bats from the Atlantic Forest biome. CONCLUSIONS: These findings shed light on potential host range for these bacteria, which are characterized as important zoonotic pathogens.

Contrasting patterns of RUNX2 repeat variations are associated with palate shape in phyllostomid bats and New World primates.

Establishing the genetic basis that underlies craniofacial variability in natural populations is one of the main topics of evolutionary and developmental studies. One of the genes associated with mammal craniofacial variability is RUNX2, and in the present study we investigated the association between craniofacial length and width and RUNX2 across New World bats (Phyllostomidae) and primates (Catarrhini and Platyrrhini). Our results showed contrasting patterns of association between the glutamate/alanine ratios (Q/A ratio) and palate shape in these highly diverse groups. In phyllostomid bats, we found an association between shorter/broader faces and increase of the Q/A ratio. In New World monkeys (NWM) there was a positive correlation of increasing Q/A ratios to more elongated faces. Our findings reinforced the role of the Q/A ratio as a flexible genetic mechanism that would rapidly change the time of skull ossification throughout development. However, we propose a scenario in which the influence of this genetic adjustment system is indirect. The Q/A ratio would not lead to a specific phenotype, but throughout the history of a lineage, would act along with evolutionary constraints, as well as other genes, as a facilitator for adaptive morphological changes.

Molecular detection of fungi of public health importance in wild animals from Southern Brazil.

Some animals have an important relationship with fungal infections, and searching for pathogens in animal samples may be an opportunity for eco-epidemiological research. Since studies involving wildlife are generally restricted, using samples from road kills is an alternative. The aim of this study was to verify whether pathogenic fungi of public health importance occur in wildlife road kills from Santa Catarina State, Brazil. Organ samples (n = 1063) from 297 animals were analysed according to Polymerase Chain Reaction (PCR) using universal primers to detect fungi in general and, subsequently, using primers specific to Paracoccidioides brasiliensis, Histoplasma capsulatum and Cryptococcus spp. There were 102 samples positive for fungal species. Eight samples were positive for P. brasiliensis, three samples were positive for Cryptococcus spp. and one sample had coinfection by these two fungi. No sample was positive for Histoplasma spp. according to the molecular detection. Genetic sequencing allowed the identification of Fungal sp. in 89 samples, Cryptococcus neoformans in two samples and Aspergillus penicillioides in three samples. This study shows the importance of wild animals in the epidemiology of fungal infections and assists in the mapping of pathogen occurrence in a region that was not previously evaluated.

Pleistocene climatic oscillations in Neotropical open areas: Refuge isolation in the rodent Oxymycterus nasutus endemic to grasslands.

Pleistocene climatic oscillations favoured the expansion of grassland ecosystems and open vegetation landscapes throughout the Neotropics, and influenced the evolutionary history of species adapted to such environments. In this study, we sampled populations of the rodent Oxymycterus nasutus endemic to open areas in the Pampas and Atlantic Forest biomes to assess the tempo and mode of population divergence using an integrative approach, including coalescence theory, ecological niche models, and morphometry. Our results indicated that these O. nasutus populations exhibited high levels of genetic structure. Six major mtDNA clades were found, structuring these biomes into distinct groups. Estimates of their divergence times was indicated to be 0.571 myr. The high degree of genetic structure is reflected in the analyses of geometric morphometric; skull differences between lineages in the two ecoregions were detected. During the last glacial maximum, there was a strong increase in suitable abiotic conditions for O. nasutus. Distinct molecular markers revealed a population expansion over time, with a possible demographic retraction during the post-glacial period. Considering that all clades coalesce with the last interglacial maximum, our results indicated that reduction in suitable conditions during this period may have resulted in a possible vicariance associated with refuge isolation.

New species of Histiotus (Chiroptera: Vespertilionidae) from northeastern Brazil.

Histiotus are vespertilionid bats endemic to South America, easily recognized by its very long ears. During a twelve-month bat inventory in northeastern Brazil, eleven specimens of Histiotus were collected with a unique combination of characters that did not match those of any known species. In this paper, we describe these specimens as a new species. Histiotus sp. nov is distinguished from its congeners by its pale transparent wings and translucent ears, a triangular-shaped ear with a prominent lobe in the inner border connected by a band (~4 mm) across the forehead; its general golden-brownish body color and well-marked bicolor dorsal hairs. Its geographic distribution is unique among vespertilionids, arranged in a northeast-southwest diagonal across South America, includes the Caatinga and Cerrado of Brazil and Chaco of Bolivia. The available data suggest a seasonal reproductive pattern, with births occurring in the mid to late rainy season.

A new species of swamp rat of the genus Scapteromys waterhouse, 1837 (Rodentia: Sigmodontinae) endemic to Araucaria angustifolia forest in Southern Brazil.

A new species of swamp rat of the genus Scapteromys from the Meridional Plateau of Southern Brazil is described. Morphological, molecular, and karyological analysis support the recognition of the new species, distinct from S. aquaticus and S. tumidus. Scapteromys sp. nov. is significantly smaller than the congeneric taxa considering most of the external and craniometric measurements and the pelage is conspicuously grayer and darker. It can be distinguished from S. tumidus by the laterally extended thenar pad of the manus and the parallel edges of the hamular process of the pterygoid, and from S. aquaticus by a grayer and darker pelage and smaller values of most external and craniometric measurements. Karyological analysis indicated a difference in chromosome numbers across the distributional range: 2n=34 and 2n=36. A total of 11 haplotypes were found along the range of the new species within the biogeographic province of Araucaria angustifolia Forest. Strongly supported substructure was found within the new taxon, resulting in two reciprocally monophyletic clades.

DNA barcoding of sigmodontine rodents: identifying wildlife reservoirs of zoonoses.

Species identification through DNA barcoding is a tool to be added to taxonomic procedures, once it has been validated. Applying barcoding techniques in public health would aid in the identification and correct delimitation of the distribution of rodents from the subfamily Sigmodontinae. These rodents are reservoirs of etiological agents of zoonoses including arenaviruses, hantaviruses, Chagas disease and leishmaniasis. In this study we compared distance-based and probabilistic phylogenetic inference methods to evaluate the performance of cytochrome c oxidase subunit I (COI) in sigmodontine identification. A total of 130 sequences from 21 field-trapped species (13 genera), mainly from southern Brazil, were generated and analyzed, together with 58 GenBank sequences (24 species; 10 genera). Preliminary analysis revealed a 9.5% rate of misidentifications in the field, mainly of juveniles, which were reclassified after examination of external morphological characters and chromosome numbers. Distance and model-based methods of tree reconstruction retrieved similar topologies and monophyly for most species. Kernel density estimation of the distance distribution showed a clear barcoding gap with overlapping of intraspecific and interspecific densities < 1% and 21 species with mean intraspecific distance < 2%. Five species that are reservoirs of hantaviruses could be identified through DNA barcodes. Additionally, we provide information for the description of a putative new species, as well as the first COI sequence of the recently described genus Drymoreomys. The data also indicated an expansion of the distribution of Calomys tener. We emphasize that DNA barcoding should be used in combination with other taxonomic and systematic procedures in an integrative framework and based on properly identified museum collections, to improve identification procedures, especially in epidemiological surveillance and ecological assessments.

Description of the karyotype of Rhagomys rufescens Thomas, 1886 (Rodentia, Sigmodontinae) from Southern Brazil Atlantic forest.

Rhagomys rufescens (Rodentia: Sigmodontinae) is an endemic species of the Atlantic forest from Southern and Southeastern Brazil. Some authors consider Rhagomys as part of the tribe Thomasomyini; but its phylogenetic relationships remain unclear. Chromosomal studies on eight specimens of Rhagomys rufescens revealed a diploid number of 2n = 36 and a number of autosome arms FN = 50. GTG, CBG and Ag-NOR banding and CMA(3) /DAPI staining were performed on metaphase chromosomes. Eight biarmed and nine acrocentric pairs were found in the karyotype of this species. The X and Y chromosomes were both acrocentric. Most of the autosomes and the sex chromosomes showed positive C-bands in the pericentromeric region. The X chromosome showed an additional heterochromatic block in the proximal region of the long arm. Nucleolus organizer regions (NORs) were located in the pericentromeric region of three biarmed autosomes (pairs 4, 6 and 8) and in the telomeric region of the short arm of three acrocentrics (pairs 10, 12 and 17). CMA (3) /DAPI staining produced fluorescent signals in many autosomes, especially in pairs 4, 6, and 8. This study presents cytogenetic data of Rhagomys rufescens for the first time.

Description of the karyotype of Rhagomys rufescens Thomas, 1886 (Rodentia, Sigmodontinae) from Southern Brazil Atlantic forest

Rhagomys rufescens (Rodentia: Sigmodontinae) is an endemic species of the Atlantic forest from Southern and Southeastern Brazil. Some authors consider Rhagomys as part of the tribe Thomasomyini; but its phylogenetic relationships remain unclear. Chromosomal studies on eight specimens of Rhagomys rufescens revealed a diploid number of 2n = 36 and a number of autosome arms FN = 50. GTG, CBG and Ag-NOR banding and CMA3/DAPI staining were performed on metaphase chromosomes. Eight biarmed and nine acrocentric pairs were found in the karyotype of this species. The X and Y chromosomes were both acrocentric. Most of the autosomes and the sex chromosomes showed positive C-bands in the pericentromeric region. The X chromosome showed an additional heterochromatic block in the proximal region of the long arm. Nucleolus organizer regions (NORs) were located in the pericentromeric region of three biarmed autosomes (pairs 4, 6 and 8) and in the telomeric region of the short arm of three acrocentrics (pairs 10, 12 and 17). CMA3/DAPI staining produced fluorescent signals in many autosomes, especially in pairs 4, 6, and 8. This study presents cytogenetic data of Rhagomys rufescens for the first time.

Antioxidant defenses, longevity and ecophysiology of South American bats.

Microchiropteran bats sustain very high oxygen consumption rates when active, but they also exhibit drastic daily drops in oxygen consumption when torpid. In addition, bats are also characterized by an extraordinary longevity considering their body mass and high specific metabolic rate when compared to other mammals of related size. Therefore, they consist of a very interesting group regarding the free radical theory of aging. The present study was carried out to measure the antioxidant defenses of several tissues of five South American bat species, attempting to correlate the antioxidant status, ecophysiology and longevity. Superoxide dismutase (SOD) and catalase (CAT) activities in blood, liver and kidney were higher compared to other tissues. The contents of alpha-tocopherol and beta-carotene found in liver, heart, kidneys, and pectoral muscles were one to two orders of magnitude higher than those usually found in rat and mouse liver. Also, these contents in liver were generally inversely related to lipoperoxidation measured as TBARS contents. Blood GSH contents and the activities of SOD and CAT were higher in torpid Sturnira lillium compared to active ones, thus suggesting that the elevation of such antioxidants might be daily modulated to minimize the oxidative stress related to the transition from torpid to active state in bats. The lower ROS generation reported in the literature for other bat species, their high constitutive antioxidant defenses, and the daily energy sparing associated with torpor appear to be closely related to their ecophysiological adaptations and to their extended longevity.

Variações na coloração de iraras (Eira barbara Linnaeus, 1758 - Carnivora, Mustelidae) da Reserva Biológica Estadual do Sassafrás, Santa Catarina, sul do Brasil / Variation in coat color of tayras (Eira barbara Linnaeus, 1758 - Carnivora, Mustelidae) in the State Biological Reserve of Sassafrás, Santa Catarina, South Brazil

Iraras apresentam variações de coloração por toda sua área de distribuição, sendo o objetivo deste estudo contribuir para o conhecimento destas variações. Na Reserva Biológica Estadual do Sassafrás, Santa Catarina, Brasil, foram obtidos 21 registros fotográficos de iraras, abrangendo diferentes colorações de pelagem. Duas classes de coloração foram observadas entre os indivíduos fotografados: corpo escuro com cabeça e pescoço cinzas e corpo, cabeça e pescoço branco-amarelados. A freqüência dos registros não variou significativamente entre as duas classes de coloração, indicando que a coloração de pelagem branco-amarelada é comum para esta população local.

Tayras present color variations along their range of distribution. The objective of this study is to contribute to the understanding of these variations. In the State Biological Reserve of Sassafrás, Santa Catarina, Brazil, 21 photographic records of tayras were obtained, comprising different coat colors. Two coat color classes were observed among photographed individuals: dark body with gray head and neck, and body, head and neck homogeneously white-yellowish. The frequency of records did not varied significantly between the two color classes, indicating that the white-yellowish coloration is common for this local population.