ABSTRACT: Sporotrichosis is a subcutaneous mycosis caused by fungus of the Sporothrix complex, and in Brazil the main species reported is Sporothrix brasiliensis, of which the diseased cat is the transmitter. Although, its occurrence has increased in the state of Paraíba, Brazil, since 2016, data on the disease in this state are limited. Therefore, this research aimed to identify molecularly isolates of Sporothrix spp. from domestic cats from cities in Paraíba, and in this way to expand the understanding of the disease in the state. Thirty-nine samples were analyzed, obtained from skin lesions of domestic felines, from the following cities in Paraíba: João Pessoa, Pilões, Patos, Areia, Bananeiras and Guarabira. Cytological analysis was performed to screen the samples, followed by fungal culture, and the molecular characterization of the isolates was performed, using the species-specific Polymerase Chain Reaction (PCR) or partial sequencing of the calmodulin gene. All isolates were identified as S. brasiliensis. The sequencing showed 100% similarity to the S. brasiliensis CBS 120339 strain. In view of this, it is concluded that in the study areas the species involved in cases of feline sporotrichosis is S. brasiliensis, its presence in Paraíba demonstrated the spread of the agent in regions distant from the epicenters in Brazil, alerting to the possible occurrence of zoonotic outbreaks similar to those found in the South and Southeast regions of the country. In addition, it highlights the emerging role of felines in the transmission of sporotrichosis in new endemic areas of Brazil.

RESUMO: A esporotricose é uma micose subcutânea causada por fungos do complexo Sporothrix, e no Brasil a principal espécie relatada é Sporothrix brasiliensis da qual o transmissor é o gato doente. Embora sua ocorrência tenha aumentado no estado da Paraíba, Brasil, desde 2016, os dados sobre a doença neste estado são limitados. Diante disso, esta pesquisa teve por objetivo identificar, molecularmente, isolados de Sporothrix spp. procedentes de felinos domésticos de cidades da Paraíba, e dessa maneira expandir a compreensão da enfermidade no estado. Foram analisadas 39 amostras, obtidas de lesões cutâneas de felinos domésticos, oriundos das seguintes cidades paraibanas: João Pessoa, Pilões, Patos, Areia, Bananeiras e Guarabira. Realizou-se análise citológica, para triagem das amostras, a seguir cultura fúngica, e posteriormente a caracterização molecular dos isolados, utilizando a Reação em Cadeia da Polimerase (PCR), espécie-específica ou sequenciamento parcial do gene calmodulina. Todos os isolados foram identificados molecularmente como S. brasiliensis. O sequenciamento demonstrou 100% de similaridade com a cepa S. brasiliensis CBS 120339. Contudo, conclui-se que nas áreas do estudo a espécie envolvida em casos de esporotricose felina é S. brasiliensis, sua presença na Paraíba demonstra a disseminação do agente em regiões distantes dos epicentros no Brasil, alertando para a possível ocorrência de surtos zoonóticos, semelhantes aos encontrados nas regiões Sul e Sudeste do país. Além disso, destaca o papel emergente dos felinos na transmissão da esporotricose em novas áreas endêmicas do Brasil.

Abstract We aimed with the present study to fill the gap on the performance and safety of stroke management and the costs related to hospitalizations, the relevant comorbidities associated with stroke patients, and the stroke patient outcomes health-related quality of life (HRQOL) progress. Our study investigated the clinical, neurological, and social impact of stroke in 220 patients in a tertiary hospital located in the countryside of the state of Santa Catarina, Brazil. Description of clinical and neurological characteristics of stroke patients between 2015 to 2020 was analyzed using electronic medical records. The most affected age group was 61-80 years, being female the most affected sex. Almost 89.5% of the patients had some risk factor, with a higher prevalence of ischemic stroke. This type of stroke was the expensive, in terms of hospitalization, with an average cost of $74.10. Considering the stroke-specific quality of life scale (SSQOL) score, 88.3% of patients who demonstrated some comorbidity and 47.6% of women had lower quality of life levels post-stroke. Our data could be useful to substantiate a data-base with epidemiology statistics characterization of stroke hospitalizations, indicating the severity of stroke for the patient.

Abstract Although a protocol for immediate postoperative thirst management has been applied clinically in a discrete way, most surgical and nursing teams remain neglected, unlike other complications in the postoperative period. This pilot study aimed to demonstrate if the ice popsicles promote thirst satiety and subsequently investigate the mechanism involved in this process. Twenty-five volunteers in the immediate postoperative period were recruited to participate in this study conducted in a tertiary medical center in southern Brazil. In this protocol, ice popsicles decreased the intensity and discomfort of thirst. Relevantly, all patients (100%) demonstrated satiety after ice popsicles administrated in the immediate postoperative period. Altogether, ice popsicles constitute a promising, innovative, and simple strategy for the management of thirst, mainly after surgery, adding to patient comfort and the security of the health team that manages it.

ABSTRACT Many infants are nurtured with milk supplied by human banks, whose bacteriological and physical-chemical profiles are a major issue. We investigated the bacteriological and physical-chemical characteristics, as well as genotypic and phenotypic and profiles of Staphylococcus species isolated from 240 samples of breast milk from a bank in a teaching hospital. Dornic acidity of milk revealed that 95.4% (229/240) had acceptable limits (< 8.0 oD). Caloric intake showed a wide variation in cream content (4%), fat (4%) and energy values (559.81 Kcal/L). Staphylococcus (105/186 or 56.5%) and Enterobacter (25/186 or 13.4%) were the most prevalent genera, although other microorganisms were identified, including Klebsiella pneumoniae and Pseudomonas aeruginosa. Amoxicillin/clavulanic acid (125/157 or 79.6%), vancomycin (115/157 or 73.2%), and cephalexin (112/157 or 71.3%) were the most effective antimicrobials. High resistance rates of isolates were found to penicillin G (141/157 or 89.8%), ampicillin (135/157 or 86%), and oxacillin (118/157 or 75.2%). Multidrug resistance to ≥ 3 antimicrobials occurred in 66.2% (123/186) of the isolates. Residues of microbial multiplication inhibitory substances were found in 85% (204/240) of samples. Among the coagulase-positive-CPS and negative-CoNS staphylococci, the mecA gene was detected in 53.3% (8/15) and 75% (30/40), respectively. Genes sea, seb and sec were detected in 20% (3/15) of CPS, while tsst-1 was detected in 13.34% (2/15). In addition, 13.3% (2/15) of S. aureus were toxin-producers. Genes sea, seb and sec were detected in 90% (36/40), 5% (2/40) and 15% (6/40) CoNS, respectively. Enterotoxin production was identified in 5% (2/40) of CoNS. The identification of multidrug-resistant bacteria, staphylococci species toxin-producers harboring methicillin-resistance genes, and residues of microbial multiplication inhibitory substances reinforce the need for a continuous vigilance of milk quality offered to infant consumption by human banks.

BACKGROUND Sporotrichosis is a subcutaneous mycosis caused by dimorphic pathogenic fungi belonging to the Sporothrix genus. Pathogenic Sporothrix species typically produce melanin, which is known to be a virulence factor. OBJECTIVES The aim of this study was to perform phenotypic, genotypic, and virulence analyses of two distinct Sporothrix brasiliensis strains isolated from the same lesion on a patient from Rio de Janeiro. METHODS AND FINDINGS Genotypic analyses by partial sequencing of the calmodulin, β-tubulin, and chitin synthase genes, as well as polymerase chain reaction (PCR)-fingerprinting by T3B, M13, and GACA, showed that the isolates were very similar but not identical. Both isolates had similar phenotypic characteristics and effectively produced melanin in their yeast forms, accounting for their ability of causing disease in a murine sporotrichosis model. Remarkably, isolate B was albino in its environmental form but caused more severe disease than the pigmented A isolate. CONCLUSIONS These findings indicate that the patient was infected by two genetically and biologically distinct S. brasiliensis that vary in their production of melanin in their environmental forms. The results underscore the importance of characterizing phenotypically different isolates found in the same clinical specimen or patient.

BACKGROUND Sporotrichosis is caused by species of the genus Sporothrix. From 1998 to 2015, 4,703 cats were diagnosed at the Fundação Oswaldo Cruz (Fiocruz), Rio de Janeiro, Brazil. Even after the description of the Sporothrix species, the characterisation of feline isolates is not performed routinely. OBJECTIVES To characterise the clinical isolates from cats at the species level and correlate them with the clinical and epidemiological characteristics of the cats. METHODS Forty seven Sporothrix spp. isolates from cats assisted at Fiocruz from 2010 to 2011 were included. Medical records were consulted to obtain the clinical and epidemiological data. The isolates were identified through their morphological and physiological characteristics. T3B polymerase chain reaction (PCR) fingerprinting was used for molecular identification of the species. FINDINGS In phenotypic tests, 34 isolates were characterised as S. brasiliensis, one as S. schenckii and 12 as Sporothrix spp. PCR identified all isolates as S. brasiliensis. MAIN CONCLUSIONS S. brasiliensis is the only etiological agent of feline sporotrichosis in Rio de Janeiro to date. None association was found between the isolates and the clinical and epidemiological data. In addition, we strongly recommend the use of molecular techniques for the identification of isolates of Sporothrix spp.

There is a paucity of studies on the cell biology of Sporothrix luriei, the less common of the pathogenic Sporothrix species worldwide. The production of DHN-melanin, eumelanin, and pyomelanin were evaluated on the mycelial and yeast forms of the S. luriei ATCC 18616 strain. The mycelial form of this species produced only pyomelanin, which protected the fungus against environmental stressors such as ultraviolet light, heat, and cold. The yeast form was unable to produce any of the tested melanin types. The lack of melanin in the parasitic form of S. luriei may be an explanation for its low frequency in human infections.

BACKGROUND Sporothrix brasiliensis is the most virulent sporotrichosis agent. This species usually responds to antifungal drugs, but therapeutic failure can occur in some patients. Antifungal susceptibility tests have been performed on this species, but no clinical breakpoints (CBPs) are available. In this situation, minimal inhibitory concentration (MIC) distributions and epidemiological cutoff values (ECVs) support the detection of identification of resistant strains. OBJECTIVES To study the MIC distributions of five antifungal drugs against S. brasiliensis and to propose tentative ECVs. METHODS MICs of amphotericin B (AMB), itraconazole (ITR), ketoconazole (KET), posaconazole (POS), and terbinafine (TRB) against 335 S. brasiliensis strains were determined by the Clinical and Laboratory Standards Institute broth microdilution method. FINDINGS The proposed ECV, in µg/mL, for AMB, ITR, KET, POS, and TRB were 4.0, 2.0, 1.0, 2.0, and 0.25, respectively. Percentages of wild-type strains in our population for the above antifungal drugs were 98.48, 95.22, 95.33, 100, and 97.67%, respectively. MAIN CONCLUSIONS These ECVs will be useful to detect strains with resistance, to define CBPs, and to elaborate specific therapeutic guidelines for S. brasiliensis. Rational use of antifungals is strongly recommended to avoid the emergence of resistant strains and ensure the therapeutic effectiveness of sporotrichosis.

RESUMO: Para acessar a prevalência de MRSP na pele e nas narinas de cães com piodermite superficial, determinar a suscetibilidade in vitro, e correlacionar estes dados com a presença do gene mecA, foram selecionados 43 cães. Amostras de lesões de pele e narinas foram coletadas, cultivadas, e analisadas fenotipica e genotipicamente. Em 62 amostras (91%), os microrganismos foram classificados como Staphylococcus pseudintermedius. A taxa de resistência a antibióticos variou entre 7% (amicacina; 4/62) e 77% (sulfamethoxazol + trimetoprim; 48/62). Resistência a oxacilina foi observada em 34% das amostras (21/62). Vinte e cinco amostras (37%), eram cepas portadoras do gene mecA. Correlação significativa (P<0,01) foi observada entre a presença do gene mecA e a resistência à oxacilina. Considerando os cães, 17 eram portadores de cepas com gene mecA e 8(47%) delas carreavam este gene nas amostras de lesão de pele e nas narinas. Correlação significativa (P>0,01) foi observada entre a presença do gene mecA nas lesões de pele e nas narinas. Sendo assim, resistência à oxacilina in vitro pode ser aferida com segurança para indicar a presença do gene mecA em amostras de MRSP, e as narinas podem constituir em um reservatório dos microorganismos em cães.

ABSTRACT: In order to assess the prevalence of Methicillin-resistant Staphylococcus pseudintermedius from skin and nostrils of dogs with pyoderma, to determine its in vitro susceptibility, and to correlate these data with the presence of the mecA gene, 43 dogs were selected. Samples were collected from secretion of their skin lesions and right nostril, cultured, and analyzed for phenotypic and genotypic characteristics of the bacteria studied. In 62 samples (91%) the microorganism was classified as S. pseudintermedius. The rate of resistance against antibiotics ranged from 7% (amikacin; 4/62) to 77% (sulfamethoxazole + trimethoprim; 48/62). Resistance against oxacillin was found in 34% of the samples (21/62). Twenty-five samples (37%) were strains that carried the mecA gene. A significant correlation (P<0.01) was found between presence of the mecA gene and oxacillin resistance. Seventeen dogs were mecA gene carriers, and 8 (47%) of them had the gene in the skin lesions and nostril. A significant correlation (P<0.01) was also observed between the presence of mecA gene in the skin lesions and nostrils. Oxacillin resistance in vitro can be safely used to indicate the presence of mecA gene in MRSP samples. The nostrils can be a reservoir of MRSP in dogs.

The Piper species chemistry has been widely investigated and the phytochemical analyses have led to the isolation of a number of active compounds like alkaloids, terpenes and flavones among others. The aim of this study was to evaluate the leishmanicidal activity of 2-[1-hydroxy-3-phenyl-(Z,2E)-2-propenylidene]-4-methyl-4-cyclopentene-1,3-dione (DCPC), a cyclopentenedione derivative isolated from the roots of Piper carniconnectivum C. DC., Piperaceae. Leishmanicidal activity against Leishmania amazonensis promastigotes was assessed, and the risk to host cell was assessed by measuring the cytotoxicity to peritoneal macrophages from BALB/c mice in vitro. L. amazonensis promastigotes and host macrophages were cultured in the presence of 100, 50, 25, 12.5 and 6 µg/mL of the cyclopentenedione derivative for up to 96 h. At the end of this period, the inhibitory concentrations (IC50) were compared with those from untreated cultures. The IC50 for promastigotes was 4.4 µg/mL after 96 h of treatment with the derivative. The 50% cytotoxic concentration (CC50) against murine peritoneal macrophages was 129 µg/mL. These results indicate that DCPC is a promising molecule for the development of leishmanicidal drugs.

A cavidade oral apresenta vários genótipos de Streptococcus mutans, que podem possuir diferentes capacidades de virulência. Entretanto, pouco se sabe sobre a diversidade e virulência de genótipos de S. mutans isolados in vivo sob uma condição controlada de alto desafio cariogênico. Este estudo avaliou a diversidade genotípica de S. mutans identificados no biofilme dental formado in vivo na presença de sacarose, assim como a acidogenicidade e aciduricidade desses genótipos. Para possibilitar formação de biofilme, voluntários bochecharam com água destilada ou solução de sacarose 8x/dia durante 3 dias. S. mutans isolados da saliva e do biofilme dental foram genotipados por PCR com primers-arbitrários. Genótipos isolados do biofilme foram avaliados em relação à habilidade de reduzir o pH da suspensão devido à glicólise, em relação à susceptibilidade a ácidos e também atividade F-ATPase. A maioria dos voluntários apresentou apenas 1 genótipo na saliva, que foram detectados em quase todas as amostras de biofilme em altas proporções. Genótipos isolados somente na presença de sacarose apresentaram maior acidogenicidade do que aqueles genótipos isolados apenas na presença de água. Genótipos de biofilmes formados na presença de sacarose foram mais acidúricos após 30 e 60 min de incubação em pH 2,8 e 5,0, respectivamente. Os resultados do presente estudo sugerem que biofilmes formados sob condição de alto desafio cariogênico podem apresentar genótipos de S. mutans mais acidúricos e mais acidogênicos.

The oral cavity harbors several Streptococcus mutans genotypes, which could present distinct virulence properties. However, little is known about the diversity and virulence traits of S. mutans genotypes isolated in vivo under controlled conditions of high cariogenic challenge. This study evaluated the genotypic diversity of S. mutans isolated from dental biofilms formed in vivo under sucrose exposure, as well as their acidogenicity and aciduricity. To form biofilms, subjects rinsed their mouths with distilled water or sucrose solution 8 times/day for 3 days. S. mutans collected from saliva and biofilms were genotyped by arbitrarily-primed PCR. Genotypes identified in the biofilms were evaluated regarding their ability to lower the suspension pH through glycolysis and their acid susceptibility and F-ATPase activity. Most subjects harbored only one genotype in saliva, which was detected in almost all biofilm samples at high proportions. Genotypes isolated only in the presence of sucrose had higher acidogenicity than those isolated only in the presence of water. Genotypes from biofilms formed with sucrose were more aciduric after 30 and 60 min of incubation at pH 2.8 and 5.0, respectively. The present results suggest that biofilms formed under high cariogenic conditions may harbor more aciduric and acidogenic S. mutans genotypes.

An epidemic of sporotrichosis, a subcutaneous mycosis caused by the fungus Sporothrix schenckii, is ongoing in Rio de Janeiro, Brazil, in which cases of human infection are related to exposure to cats. In an attempt to demonstrate the zoonotic character of this epidemic using molecular methodology, we characterised by DNA-based typing methods 19 human and 25 animal S. schenckii isolates from the epidemic, as well as two control strains. To analyse the isolates, the random amplified polymorphic DNA (RAPD) technique was performed using three different primers, together with DNA fingerprinting using the minisatellite derived from the wild-type phage M13 core-sequence. The analyses generated amplicons with considerable polymorphism. Although isolates exhibited high levels of genetic relatedness, they could be clustered into 5-10 genotypes. The RAPD profiles of epidemic S. schenckii isolates could be distinguished from that of the United States isolate, displaying 20% similarity to each primer and 60% when amplified with the M13 primer. DNA fingerprinting of S. schenckii isolated from the nails (42.8%) and the oral cavities (66%) of cats were identical to related human samples, suggesting that there is a common infection source for animals and humans in this epidemic. It is clear that cats act as a vehicle for dissemination of S. schenckii.

INTRODUÇÃO: A mudança na marca da heparina rotineiramente utilizada nas cirurgias cardíacas no Brasil tem sido acompanhada por aumento do número de casos de discrasia sanguínea, aumento de reoperações e efeitos adversos em nossa Instituição e em outras. MÉTODOS: Foram avaliadas no Laboratório de Tecido Conjuntivo do HUCFF/UFRJ, quatro preparações disponíveis e comparadas à heparina retirada do mercado (Liquemine) e ao padrão de controle internacional. As preparações de heparina foram submetidas à ressonância nuclear magnética para avaliação da integridade estrutural, bem como avaliação de sua eficácia anticoagulante. RESULTADOS: Houve diferença significativa quanto à atividade anticoagulante entre as amostras. Também se observou a presença de contaminação com dermatam sulfato, amostras degradadas quimicamente e com significativa alteração do peso molecular. CONCLUSÃO: Das amostras estudadas, nenhuma atendeu aos requisitos de segurança para utilização em cirurgias cardíacas com circulação extracorpórea. Nenhuma delas apresentou a qualidade semelhante ao Liquemine, não mais disponível no mercado brasileiro.

INTRODUCTION: The change in the heparin solution trade mark in Brazil that had been commonly used in cardiac surgery has shown increased number in the coagulopathy, re-exploration and other side effects in our Institution and others. METHODS: All four different heparin solutions available in the Brazilian market were studied in the Connective Tissue Lab, HUCFF, UFRJ and compared to the Liquemine (out of the market) and the international control solution. All samples were evaluated by magnetic nuclear resonance as well as their anticoagulant effectiveness. RESULTS: There were significant differences among them regarding the anticoagulant activity. It was also observed contamination with other dermatan sulfate, samples chemically degraded and with significant change in the molecular weight. CONCLUSION: Among the studied samples, none of them can offer security in cardiac surgeries on pump. None of them has demonstrated similar quality to Liquemine, which is not available in the Brazilian market.

A composição do biofilme dental in situ exposto a açúcares é bem conhecida, mas o efeito dos açúcares na diversidade genotípica de S. mutans no biofilme dental ainda não foi explorada. Este estudo avaliou a diversidade genotípica de S. mutans no biofilme dental formado in situ sob frequente exposição à sacarose e seus monossacarídeos constituintes (glicose e frutose). Primeiramente, saliva de voluntários foi coletada para isolamento de S. mutans e os mesmos voluntários usaram um dispositivo intraoral palatino, contendo blocos de esmalte, que foram submetidos 8x/dia aos seguintes tratamentos: água destilada e deionizada (controle negativo), solução de glicose 10% + frutose 10% (carboidratos fermentáveis) e solução de sacarose 20% (fermentável e indutor de PEC). Após 3, 7 e 14 dias, os biofilmes foram coletados e colônias de S. mutans foram isoladas. A técnica de reação em cadeia de polimerase usando primers arbitrários (AP-PCR) demonstrou que o genótipo salivar foi detectado em quase todas as amostras de biofilme, independente do tratamento, e parece refletir aqueles genótipos presentes em maiores proporções no biofilme. Além do genótipo salivar, outros foram encontrados nos biofilmes, mas em uma menor proporção e foram distintos entre os tratamentos. Os dados sugerem que o modelo in situ é útil para a avaliação da diversidade genotípica de S. mutans. Porém, nas condições do presente estudo, não foi possível demonstrar que genótipos específicos foram detectados no biofilme devido ao estresse induzido pelo metabolismo da sacarose ou fermentação de seus monossacarídeos.

In situ dental biofilm composition under sugar exposure is well known, but sugar effect on the genotypic diversity of S. mutans in dental biofilm has not been explored. This study evaluated S. mutans genotypic diversity in dental biofilm formed in situ under frequent exposure to sucrose and its monosaccharide constituents (glucose and fructose). Saliva of 7 volunteers was collected for isolation of S. mutans and the same volunteers wore intraoral palatal appliances, containing enamel slabs, which were submitted to the following treatments: distilled and deionized water (negative control), 10% glucose + 10% fructose (fermentable carbohydrates) solution or 20% sucrose (fermentable and EPS inductor) solution, 8x/day. After 3, 7 and 14 days, the biofilms were colleted and S. mutans colonies were isolated. Arbitrarily primed polymerase chain reaction (AP-PCR) of S. mutans showed that salivary genotypes were also detected in almost all biofilm samples, independently of the treatment, and seemed to reflect those genotypes present at higher proportion in biofilms. In addition to the salivary genotypes, others were found in biofilms but in lower proportions and were distinct among treatment. The data suggest that the in situ model seems to be useful to evaluate genotypic diversity of S. mutans, but, under the tested conditions, it was not possible to clearly show that specific genotypes were selected in the biofilm due to the stress induced by sucrose metabolism or simple fermentation of its monosaccharides.

Reportamos um caso raro de histoplasmose crônica disseminada com diversas lesões ulceradas na cavidade oral de um paciente alcoólatra, sem o vírus da imunodeficiência humana, com nenhum outro sinal detectável ou sintomas de doença sistêmica ou manifestações extra-orais. Análise histopatológica revelou processo inflamatório crônico com granulomas contendo organismos fúngicos. O isolamento do Histoplasma capsulatum forneceu o diagnóstico definitivo. Tratamento com itraconazol resultou numa remissão completa das lesões orais. De acordo com nosso conhecimento, este é o segundo caso reportado de histoplasmose oral em um paciente HIV negativo descrito no Brasil.

We report a rare case of chronic disseminated histoplasmosis with several ulcerated lesions in the oral cavity in an alcoholic patient without human immunodeficiency virus infection, with no detectable signs and symptoms of systemic disease or extraoral manifestations. Histopathological analysis revealed chronic inflammatory process with granulomas containing Histoplasma-like organisms. The isolation of Histoplasma capsulatum provided the definitive diagnosis. Treatment with itraconazole resulted in complete remission of oral lesions. As far we aware, this is the second case report of oral histoplasmosis in an HIV negative patient described in Brazil.