Epidemiological and Clinical Aspects of Cutaneous and Mucosal Leishmaniases in Portugal: Retrospective Analysis of Cases Diagnosed in Public Hospitals and Reported in the Literature between 2010 and 2020.

Leishmania infantum, a zoonotic vector-born parasite, is endemic in the Mediterranean region, presenting mostly as visceral (VL), but also as cutaneous (CL) and mucosal leishmaniasis (ML). This study aimed to describe the epidemiological and clinical aspects of the CL and ML cases diagnosed in mainland Portugal between 2010 and 2020. Collaboration was requested from every hospital of the Portuguese National Health System. Cases were screened through a search of diagnostic discharge codes or positive laboratory results for Leishmania infection. Simultaneously, a comprehensive literature search was performed. Descriptive statistics and hypothesis testing were performed using IBM® SPSS® Statistics. A total of 43 CL and 7 ML cases were identified, with a predominance of autochthonous cases (86%). In CL, immunosuppressed individuals constituted a significant proportion of patients (48%), and in this group, disseminated CL (22%) and simultaneous VL (54%) were common. In autochthonous cases, lesions, mostly papules/nodules (62%), were frequently observed on the head (48%). The approach to treatment was very heterogeneous. ML cases were all autochthonous, were diagnosed primarily in older immunosuppressed individuals, and were generally treated with liposomal amphotericin B. The findings suggest a need for enhanced surveillance and reporting, clinical awareness, and diagnostic capacity of these forms of leishmaniasis to mitigate underdiagnosis and improve patient outcomes. A holistic One Health approach is advocated to address the multifaceted challenges posed by leishmaniases in Portugal and beyond.

When the Lightning Strikes Twice: Navigating the Complex Terrain of Cerebral Tuberculosis.

Tuberculosis (TB), a Mycobacterium tuberculosis (Mtb) infection, remains a significant global health concern despite a declining incidence. This report highlights a complex case involving a 24-year-old patient from Angola who presented with a constellation of symptoms, including fever, weight loss, and neurological deficits. The patient had been on chronic corticosteroid therapy, a known risk factor for the reactivation of latent TB infection (LTBI). Her clinical course was marked by diagnostic challenges, such as a previous diagnosis of Kikuchi's disease and paradoxical progression despite appropriate tuberculostatic chemotherapy. Miliary TB, characterized by widespread dissemination of Mtb from the primary site of infection, can manifest in various extrapulmonary locations. Central nervous system (CNS) involvement, particularly TB meningitis, is the most severe form of TB, associated with significant morbidity and mortality. The diagnosis of miliary and CNS TB can be elusive due to nonspecific clinical presentations and imaging findings. This case underscores the importance of a high index of suspicion, especially in immunocompromised individuals, and the need for comprehensive microbiological analysis, including cerebrospinal fluid (CSF) examination, to confirm CNS involvement. Furthermore, this case illustrates the challenges associated with TB treatment, including the risk of drug toxicity, medication adherence, and the potential for drug resistance. Treatment duration for miliary TB is extended, typically lasting nine months to a year, and may require adaptation based on the patient's clinical response and drug penetration into the CNS. Corticosteroids play a critical role as adjuvant therapy, particularly in cases with perilesional edema or paradoxical reactions during treatment. This case underscores the complexity of diagnosing and managing miliary and CNS TB, emphasizing the importance of considering TB as a diagnostic possibility in patients with nonspecific symptoms and risk factors. Early identification, multidisciplinary collaboration, and tailored therapeutic strategies are essential for achieving optimal outcomes in such challenging cases. Additionally, screening for latent TB infection should be a priority for patients requiring immunosuppressive therapy to mitigate the risk of reactivation.

Supernumerary B Chromosomes of Tetragonisca fiebrigi Share Repeat Content with Standard Chromosome Set of both T. fiebrigi and Tetragonisca angustula (Apidae: Meliponini).

The stingless bees Tetragonisca angustula and Tetragonisca fiebrigi are widely distributed in Brazil, and both are commonly known as "jataí." Our goal was to investigate the possible origin of the B chromosomes in T. fiebrigi, a cytotaxonomic trait that differentiates T. fiebrigi from T. angustula. We analyzed diploid chromosome number (2n), B chromosome incidence, patterns of constitutive heterochromatin, and in situ localization of different repetitive DNA probes in T. angustula and T. fiebrigi. Both species displayed 2n = 34, with similar karyotype structures. One to three B chromosomes were observed in T. fiebrigi only. Constitutive heterochromatin was distributed on one arm of all chromosomes in both species, and T. fiebrigi B chromosomes were mainly heterochromatic with one euchromatic extremity. The (GA)15 and (CAA)10 microsatellite probes marked the euchromatic arms of all chromosomes in both species without marking the B chromosomes. The 18S ribosomal DNA (rDNA) probe marked 10 chromosomes in T. angustula and 6 A chromosomes in T. fiebrigi with an additional marking on 1B in individuals with 3B. The Tan-Bsp68I repetitive DNA probe marked the heterochromatic portion of all T. fiebrigi A and B chromosomes. This probe also marked the heterochromatic portion of all T. angustula chromosomes; therefore, both alternative hypotheses to the B chromosome origin are possible: (i) from the A chromosome complement of T. fiebrigi (intraspecific origin); or (ii) a by-product of genome reshuffling following the hybridization between T. fiebrigi and T. angustula (interspecific origin).

Validation of the Acute Physiology and Chronic Health Evaluation (APACHE) II Score in COVID-19 Patients Admitted to the Intensive Care Unit in Times of Resource Scarcity.

Introduction During the coronavirus disease 2019 (COVID-19) pandemic, a high number of patients needed to be admitted to the intensive care units (ICUs). Such a high demand led to periods where resources were insufficient and the triage of patients was needed. This study aims to evaluate the performance of the Acute Physiology and Chronic Health Evaluation (APACHE) II as a predictor of mortality in periods where triage protocols were implemented. Methods A single-center, longitudinal, retrospective cohort study was performed on patients admitted to the ICU between January 2020 and December 2021. Patients were divided into two periods: Period 1 (where patients needing ICU admission outnumbered the available resources) and Period 2 (where resources were adequate). The discriminative power of the APACHE II was checked using the receiver operating characteristic (ROC) curves. Calibration was accessed, and survival analysis was performed. Results Data from 428 patients were analyzed (229 in Period 1 and 199 in Period 2). The area under the ROC curve (AUROC) was 0.763 for Period 1 and 0.761 for Period 2, reflecting a good discriminative power. Logistic regression showed the APACHE II to be a significant predictor of mortality. The Hosmer-Lemeshow test demonstrated good calibration. The Youden index was determined, and a log-rank test showed a significantly lower survival for patients with higher APACHE II scores in both periods. Conclusions The APACHE II score is an effective tool in predicting mortality in patients with COVID-19 admitted to the ICU in a period where resource allocation and triage of patients are needed, paving a way for the future development of better and improved triage systems.

Shewanella algae, an Emerging Human Pathogen: A Series of Four Cases From a Portuguese Hospital.

Shewanella algae is a rod-shaped, Gram-negative bacterium that is considered an emerging human pathogen. Traditionally associated with warmer climates, S. algae has now been isolated from patients worldwide, and reports of infection are increasing. In a regional hospital on the outskirts of Lisbon, Portugal, four cases have been detected in the past 10 years. Two of the patients were migrants from African countries with daily contact with water; the other two patients were Portuguese, and no epidemiological risk factors were found among them. These are the first cases reported in Portugal. Risk factors associated with S. algae infection in patients discussed in this paper include the following: human immunodeficiency virus (HIV) infection, chronic venous insufficiency, lower limb ulcers, chronic kidney disease, diabetes, arterial hypertension, dilated cardiomyopathy, atrial fibrillation, chronic hepatic disease, and chronic pancreatitis. One patient died in the intensive care unit with septic shock and disseminated intravascular coagulation from a fulminant infection secondary to S. algae bacteraemia. The four clinical cases presented in this case series highlight the clinical features of this infection so that other physicians can successfully identify and treat S. algae infections.

Vasculitis and Breast Cancer: Mind the Hint.

Diffuse alveolar haemorrhage related to an anti-neutrophil cytoplasmic antibody (ANCA)-associated small vessel vasculitis is an extremely rare form of presentation of breast cancer. Here we report the case of a 77-year-old woman with a histological diagnosis of a papillary ductal carcinoma of the breast presenting with a diffuse alveolar haemorrhage secondary to a perinuclear ANCA-associated vasculitis. To our knowledge, this is the first case ever reported of a diffuse alveolar haemorrhage related to an ANCA-associated small vessel vasculitis as a form of presentation of breast cancer. The therapeutic approach of this paraneoplastic vasculitis included the use of corticosteroids and plasmapheresis, a very useful technique to remove endothelial aggressors (circulating antibodies) as a strategy to earn time for a proper therapeutic decision specifically directed for disease modification, but that can also be associated with several severe adverse effects, which are illustrated in our case.

Microsatellite Marker Discovery in the Stingless Bee Uruçu-Amarela (Melipona rufiventris Group, Hymenoptera, Meliponini) for Population Genetic Analysis.

The species Melipona rufiventris Lepeletier, 1836 is a Brazilian native stingless bee that is part of a species complex known as the 'rufiventris group', making it difficult to distinguish between the different species. Populations in this group are facing a severe decline, leading to the risk of local extinction, and therefore, their conservation should be treated as a major concern. This study describes the first set of tri- and tetranucleotide microsatellite markers, using next-generation sequencing technology for use in the identification of genetic diversity and population structure in the 'rufiventris group'. A total of 16 microsatellite loci displayed polymorphism. Analysis of the whole data set (n = 50) detected 63 alleles in all loci, ranging from 2 to 7 with a mean of 3.9 alleles/locus. A genetic diversity analysis revealed high values for population differentiation estimates (FST = 0.252, RST = 0.317, and DEST = 0.284) between the Atlantic Forest, Cerrado, and Caatinga biomes. An additional evidence for genetic divergence among populations was also found in the 'rufiventris group'; these should be treated as separate conservation units or even as separate species. These microsatellite markers have demonstrated a strong potential for assessing population discrimination in this threatened stingless bee group.

The evolution of haploid chromosome numbers in Meliponini.

It is thought that two evolutionary mechanisms gave rise to chromosomal variation in bees: the first one points to polyploidy as the main cause of chromosomal evolution, while the second, Minimum Interaction Theory (MIT), is more frequently used to explain chromosomal changes in Meliponini and suggests that centric fission is responsible for variations in karyotype. However, differences in chromosome number between Meliponini and its sister taxa and in the karyotype patterns of the Melipona genus cannot be explained by MIT, suggesting that other events were involved in chromosomal evolution. Thus, we assembled cytogenetical and molecular information to reconstruct an ancestral chromosome number for Meliponini and its sister group, Bombini, and propose a hypothesis to explain the evolutionary pathways underpinning chromosomal changes in Meliponini. We hypothesize that the common ancestor shared by the Meliponini and Bombini tribes possessed a chromosome number of n = 18. The karyotype with n = 17 chromosomes was maintained in Meliponini, and variations of haploid numbers possibly originated through additional Robertsonian fissions and fusions. Thus, the low chromosome number would not be an ancestral condition, as predicted by MIT. We then conclude that Robertsonian fission and fusions are unlikely to be the cause of chromosomal rearrangements that originated the current karyotypes in Meliponini.

Cytogenetic Analysis and Chromosomal Mapping of Repetitive DNA in Melipona Species (Hymenoptera, Meliponini).

Stingless bees of the genus Melipona are subdivided into 4 subgenera called Eomelipona, Melikerria, Melipona sensu stricto, and Michmelia according to species morphology. Cytogenetically, the species of the genus Melipona show variation in the amount and distribution of heterochromatin along their chromosomes and can be separated into 2 groups: the first with low content of heterochromatin and the second with high content of heterochromatin. These heterochromatin patterns and the number of chromosomes are characteristics exclusive to Melipona karyotypes that distinguish them from the other genera of the Meliponini. To better understand the karyotype organization in Melipona and the relationship among the subgenera, we mapped repetitive sequences and analyzed previously reported cytogenetic data with the aim to identify cytogenetic markers to be used for investigating the phylogenetic relationships and chromosome evolution in the genus. In general, Melipona species have 2n = 18 chromosomes, and the species of each subgenus share the same characteristics in relation to heterochromatin regions, DAPI/CMA3 fluorophores, and the number and distribution of 18S rDNA sites. Microsatellites were observed only in euchromatin regions, whereas the (TTAGG)6 repeats were found at telomeric sites in both groups. Our data indicate that in addition to the chromosome number, the karyotypes in Melipona could be separated into 2 groups that are characterized by conserved cytogenetic features and patterns that generally are shared by species within each subgenus, which may reflect evolutionary constraints. Our results agree with the morphological separation of the Melipona into 4 subgenera, suggesting that they must be independent evolutionary lineages.

Looking into Enteric Virome in Patients with IBD: Defining Guilty or Innocence?

BACKGROUND: Although there is some evidence suggesting that certain viruses may be involved in the onset of inflammatory bowel disease (IBD), data regarding viral prevalence and viral load in blood and mucosa of patients with IBD are scarce. The main aim of this study is to evaluate the prevalence and viral load of common Epstein-Barr virus (EBV), cytomegalovirus (CMV), and human herpes virus 6 in blood and mucosa of adult patients with endoscopic active IBD. METHODS: From January to December 2014, ulcerative colitis and Crohn's disease patients with active endoscopic disease were consecutively enrolled. Subjects undergoing colonoscopy for colorectal cancer screening served as healthy controls (HCs). Paired blood and mucosal samples from each patient and HC were collected for EBV, CMV, and human herpes virus 6 quantitative real time polymerase chain reaction assessment of the viral load. RESULTS: One hundred forty-five subjects were included; 95 IBD patients with active endoscopic disease (43 ulcerative colitis and 52 Crohn's disease) and 50 healthy subjects. CMV and EBV DNA were detected more frequently in the mucosa of patients with IBD compared with HCs (CMV P = 0.017; EBV P < 0.001), irrespective of IBD type. The frequency of human herpes virus 6 DNA detection both in the blood and in the mucosa did not differ between patients with IBD and HCs. EBV median viral load was similar in the inflamed and noninflamed mucosa was not affected by the use of immunomodulators and/or anti-tumor necrosis factor alpha agents, and did not correlate with endoscopic disease activity. CONCLUSIONS: EBV, and to a lesser extent CMV, were more prevalent in patients with IBD than in HCs. Mucosal viral load was not influenced by the therapeutic regimen, did not differ between inflamed and noninflamed mucosa, and did not seem to be influenced by the endoscopic activity of the disease, suggesting that EBV may be more involved in the onset of IBD than in its severity and clinical evolution.

Integrating Paleodistribution Models and Phylogeography in the Grass-Cutting Ant Acromyrmex striatus (Hymenoptera: Formicidae) in Southern Lowlands of South America.

Past climate changes often have influenced the present distribution and intraspecific genetic diversity of organisms. The objective of this study was to investigate the phylogeography and historical demography of populations of Acromyrmex striatus (Roger, 1863), a leaf-cutting ant species restricted to the open plains of South America. Additionally, we modeled the distribution of this species to predict its contemporary and historic habitat. From the partial sequences of the mitochondrial gene cytochrome oxidase I of 128 A. striatus workers from 38 locations we estimated genetic diversity and inferred historical demography, divergence time, and population structure. The potential distribution areas of A. striatus for current and quaternary weather conditions were modeled using the maximum entropy algorithm. We identified a total of 58 haplotypes, divided into five main haplogroups. The analysis of molecular variance (AMOVA) revealed that the largest proportion of genetic variation is found among the groups of populations. Paleodistribution models suggest that the potential habitat of A. striatus may have decreased during the Last Interglacial Period (LIG) and expanded during the Last Maximum Glacial (LGM). Overall, the past potential distribution recovered by the model comprises the current potential distribution of the species. The general structuring pattern observed was consistent with isolation by distance, suggesting a balance between gene flow and drift. Analysis of historical demography showed that populations of A. striatus had remained constant throughout its evolutionary history. Although fluctuations in the area of their potential historic habitat occurred during quaternary climate changes, populations of A. striatus are strongly structured geographically. However, explicit barriers to gene flow have not been identified. These findings closely match those in Mycetophylax simplex, another ant species that in some areas occurs in sympatry with A. striatus. Ecophysiological traits of this species and isolation by distance may together have shaped the phylogeographic pattern.

Genetic diversity and structure of Atta robusta (Hymenoptera, Formicidae, Attini), an endangered species endemic to the restinga ecoregion.

The genetic diversity and structure of the ant Atta robusta were assessed by ISSR (inter-simple sequence repeats) in 72 colonies collected from 10 localities in the Brazilian states of Espírito Santo (48 colonies) and Rio de Janeiro (24 colonies). The ISSR pattern included 67 bands, 51 of them (76.1%) polymorphic. Analysis of molecular variance (AMOVA) revealed a high level (57.4%) of inter-population variation, which suggested a high degree of genetic structure that was confirmed by UPGMA (unweighted pair-group method using an arithmetic average) cluster analysis. The significant correlation between genetic and geographic distances (r = 0.64, p < 0.05) indicated isolation that reflected the distance between locations. Overall, the populations were found to be genetically divergent. This finding indicates the need for management plans to preserve and reduce the risk of extinction of A. robusta.

Could pseudogenes be widespread in ants? Evidence of numts in the leafcutter ant Acromyrmex striatus (Roger, 1863) (Formicidae: Attini).

The incorporation of fragments of mitochondrial DNA (mtDNA) in the nuclear genome, known as numts (nuclear mitochondrial pseudogenes), undermines general assumptions concerning the use of mtDNA in phylogenetic and phylogeographic studies. Accidental amplifications of these nuclear copies instead of the mitochondrial target can lead to crucial misinterpretations, thus the correct identification of numts and their differentiation from true mitochondrial sequences are important in preventing this kind of error. Our goal was to describe the existence of cytochrome b (cytb) numts in the leafcutter ant Acromyrmex striatus (Roger, 1863). PCR products were directly sequenced using a pair of universal primers designed to amplify the cytb gene of these insects. Other species of leafcutter ants were also sequenced. The sequences were analyzed and the numts were identified by the presence of double peaks, indels and premature stop codons. Only A. striatus clearly showed the presence of numts, while the other species displayed the expected amplification of the mtDNA cytb gene target using the same primer pair. We hope that our report will highlight the benefits and challenges of using mtDNA in the molecular phylogenetic reconstruction and phylogeographic studies of ants, while establishing the importance of numts reports for future studies.

Possible introgression of B chromosomes between bee species (Genus Partamona ).

The origin of supernumerary (B) chromosomes is still a debated topic, with intra- and interspecific origins being the most plausible options. In the bee Partamona helleri, a sequence-characterized amplified region (SCAR) marker being specific to B chromosomes suggested the possibility of interspecific origin. Here, we search for this marker in 3 close relative species and perform DNA sequence comparison between species. The SCAR sequence does not show homology with other sequences in the databases, but does contain an open reading frame with sequence homology with a reverse transcriptase. Dot-blot hybridization using the SCAR marker as a probe confirmed that it is present in B-carrying, but not B-lacking larvae of P. helleri, and indicated its presence in adult individuals of P. cupira and P. criptica. Additionally, PCR amplification of the SCAR marker was successful on genomic DNA obtained from P. helleri and P. rustica larvae carrying B chromosomes, and on genomic DNA obtained from adult individuals of P. cupira, P. criptica and P. rustica. Finally, a comparison of the DNA sequence of the SCAR markers amplified from these 4 species showed very few nucleotide differences between the species. The complete association between B chromosome and SCAR presence and the scarce divergence observed for this DNA sequence between the 4 species analyzed suggest the possibility that this B chromosome has recently been transferred between species through several episodes of interspecific hybridization.

Morphological and morphometric studies of the antennal sensilla from two populations of Atta robusta (Borgmeier 1939) (Hymenoptera: Formicidae).

The ant Atta robusta is endemic to the "restinga" ecosystems where it has an important role in the dynamics of seed dispersal. Despite its importance, A. robusta is considered a threatened species. In this study we analyzed the antennal sensory organs of two different populations of A. robusta (from the cities of São Mateus and Maricá in in Espírito Santo and Rio de Janeiro States, respectively) using a scanning electron microscope (SEM). SEM revealed different types of sensilla in the A. robusta antennae, i.e., curved and straight trichoid, basiconic, ampullacea and coeloconic, which were highly abundant found in the distal flagellomeres (F) compared with other antenna regions. There were differences in samples collected from two locations in terms of the sensilla number and length. The average numbers of straight and curved trichoid sensillae numbers were different in F9 and F8, respectively, while the average length of the curved trichoid sensilla was only different in F9. These variations in sensory organs between two populations of A. robusta may indicate an adaptation of this species to different environmental conditions. The number of straight trichoid sensilla was only significantly different in F9.

Morphological and morphometric studies of the antennal sensilla from two populations of Atta robusta (Borgmeier 1939) (Hymenoptera: Formicidae) / Estudo morfológico e morfométrico das sensilas antenais de Atta robusta (Borgmeier 1939) (Hymenoptera: Formicidae)

The ant Atta robusta is endemic to the “restinga” ecosystems where it has an important role in the dynamics of seed dispersal. Despite its importance, A. robusta is considered a threatened species. In this study we analyzed the antennal sensory organs of two different populations of A. robusta (from the cities of São Mateus and Maricá in in Espírito Santo and Rio de Janeiro States, respectively) using a scanning electron microscope (SEM). SEM revealed different types of sensilla in the A. robusta antennae, i.e., curved and straight trichoid, basiconic, ampullacea and coeloconic, which were highly abundant found in the distal flagellomeres (F) compared with other antenna regions. There were differences in samples collected from two locations in terms of the sensilla number and length. The average numbers of straight and curved trichoid sensillae numbers were different in F9 and F8, respectively, while the average length of the curved trichoid sensilla was only different in F9. These variations in sensory organs between two populations of A. robusta may indicate an adaptation of this species to different environmental conditions. The number of straight trichoid sensilla was only significantly different in F9.

Atta robusta é uma espécie de formiga endêmica, com papel importante na dinâmica dos ecossistemas de restinga. Apesar de sua importância como dispersora de sementes nesse tipo de ecossistema, essa espécie encontra-se ameçada de extinção. O presente trabalho lançou mão da microscopia eletrônica de varredura (MEV) para descrever as sensilas antenais de A. robusta e investigar diferenças entre sensilas antenais dessa espécie em indivíduos coletados em localidades diferentes, nos municípios de São Mateus (Estado do Espírito Santo) e Maricá (Estado do Rio de Janeiro). Diferentes tipos de sensilas como tricoideas retas e curvadas, basicônicas, ampuláceas e celocônicas foram encontradas nas antenas, sendo que todas são mais numerosas no flagelomero nove (F9) ou mais distal. Houve diferença em número e em comprimento das sensilas tricoideas retas e curvatas em F8 e F9 entre os indivíduos das duas localidades. Os números médios das sensilas tricoides retas e curvadas foram diferentes para os F9 e F8, respectivamente, enquanto que o comprimento médio das sensilas tricoides curvadas foi diferente para o F9. Essa variação na quantidade de sensilas antenais entre as populações é, possivelmente, uma plasticidade fenotípica, característica importante para a adaptação de espécies a variações ambientais. Esse trabalho representa um estudo preliminar na tentativa de se estudar a variação nos órgãos sensoriais entre diferentes populações de A. robusta.

Cytogenetic and molecular analyses reveal a divergence between Acromyrmex striatus (Roger, 1863) and other congeneric species: taxonomic implications.

The leafcutter ants, which consist of Acromyrmex and Atta genera, are restricted to the New World and they are considered the main herbivores in the neotropics. Cytogenetic studies of leafcutter ants are available for five species of Atta and 14 species of Acromyrmex, both including subspecies. These two ant genera have a constant karyotype with a diploid number of 22 and 38 chromosomes, respectively. The most distinct Acromyrmex species from Brazil is A. striatus, which is restricted to the southern states of Santa Catarina and Rio Grande do Sul. Several cytogenetic and phylogenetic studies have been conducted with ants, but the karyotypic characterization and phylogenetic position of this species relative to leafcutter ants remains unknown. In this study, we report a diploid number of 22 chromosomes for A. striatus. The phylogenetic relationship between A. striatus and other leafcutter ants was estimated based on the four nuclear genes. A. striatus shared the same chromosome number as Atta species and the majority of metacentric chromosomes. Nuclear data generated a phylogenetic tree with a well-supported cluster, where A. striatus formed a different clade from other Acromyrmex spp. This combination of cytogenetic and molecular approaches provided interesting insights into the phylogenetic position of A. striatus among the leafcutter ants and the tribe Attini.

Honey physicochemical properties of three species of the Brazilian Melipona.

Physicochemical analyses were carried out to evaluate 27 samples of honeys from three species of the Brazilian genus Melipona (M. capixaba, M. rufiventris and M. mondury) from Espírito Santo and Minas Gerais States. The parameters water activity (Aw), percentage of soluble solids (Brix %), pH, acidity (meq/Kg) and moisture (%) were evaluated. The honey characteristics obtained from these samples were very similar to the ones from other Melipona species. However, regarding the honey from Apis (honey bee), only the pH values were similar. The low pH value and the high acidity detected in Melipona honey are potential factors for increasing the honey shelf life because they do not provide favorable conditions for the microbial development. On the other hand, the high level of water activity favors the growth of microorganisms, especially yeast, which demands a more careful handled and storage. The observed differences between Melipona and Apis honey reinforce the need for specific quality settings for stingless bee honey.

Microsatellite loci for the stingless bee Melipona rufiventris (Hymenoptera: Apidae).

Eight microsatellite primers were developed from ISSR (intersimple sequence repeats) markers for the stingless bee Melipona rufiventris. These primers were tested in 20 M. rufiventris workers, representing a single population from Minas Gerais state. The number of alleles per locus ranged from 2 to 5 (mean = 2.63) and the observed and expected heterozygosity values ranged from 0.00 to 0.44 (mean = 0.20) and from 0.05 to 0.68 (mean = 0.31), respectively. Several loci were also polymorphic in M. quadrifasciata, M. bicolor, M. mandacaia and Partamona helleri and should prove useful in population studies of other stingless bees.

[Visitation of orchid by Melipona capixaba Moure & Camargo (Hymenoptera: Apidae), bee threatened with extinction]. / Visitação de orquídea por Melipona capixaba Moure & Camargo (Hymenoptera: Apidae), abelha ameaçada de extinção.

The stingless bee Melipona capixaba Moure & Camargo is a species restricted to the Atlantic forest in the Domingos Martins, Conceição do Castelo, Venda Nova do Imigrante and Afonso Cláudio County, in the Espírito Santo State, Brazil. Despite its cological importance as pollinator few studies have examined the ecology and biology of this bee. This note relates a case of the M. capixaba workers carrying pollinarium attached to the scuttellum. The pollinaria were identified as belonging to the orchid subtribe Maxillariinae species possibly of the genus Maxillaria sensu lato or Xylobium.