Análise microbiológica e físico-química do gelo utilizado na conservação de pescado em supermercados de Rio Branco - AC / Microbiological and physicochemical analysis of ice used in fish conservation in supermarkets in Rio Branco - AC

O gelo é fundamental para a conservação adequada do pescado, pois retarda o crescimento microbiano, preservando a qualidade físico-química, microbiológicas e características organolépticas. No entanto, o gelo pode ser fonte de risco para a saúde dos consumidores quando produzido sob más condições sanitárias. Pontos críticos devem ser controlados durante a etapa de produção e armazenagem do gelo, como, uso de água potável e condições higiênicas-sanitárias. A presença de altas populações de microrganismos heterotróficos e coliformes e a má qualidade físico-química do gelo utilizado para conservação de pescado pode representar um risco potencial ao consumidor, podendo ser veiculador de doenças como Salmonelose, infecção por Escherichia coli e intoxicação por Staphylococcus aureus, além de reduzir a vida útil de prateleira do produto. Neste contexto, a realização deste estudo teve como objetivo avaliar a qualidade microbiológica e físico-química do gelo utilizado na conservação de pescados comercializados em Rio Branco - AC. Foram analisadas 15 amostras de gelo coletadas em três diferentes supermercados. Foi preconizado a coleta de gelo fabricado com maquinário próprio e que fossem utilizados na comercialização do pescado fresco, que seriam expostos e resfriados em bancas de gelo no local. Foram colhidas 300g de gelo manualmente com uso de luvas e acondicionadas em frascos de vidros esterilizados, em caixa isotérmica. A qualidade microbiológica destas amostras foi analisada pela contagem de microrganismos heterotróficos mesófilos e psicrotróficos. Além disso, foi realizada a determinação do número mais provável de coliformes totais á 30°C, termotolerantes e a quantificação do pH e cloro residual livre das amostras. Com relação a determinação de microrganismos mesófilos, foram registrados valores entre 1,0x10¹ e 9,15x103 UFC/mL com amostras fora do padrão estabelecido pela Portaria MG/MS n°888/2021. A quantidade de psicrotróficos variou entre 1,0x10¹ e 89,8x10³ UFC/mL. Foi registrada a presença de coliformes totais á 30°C em 6 amostra e de termotolerantes em 4 amostras. Portanto, foi registrado variação nos padrões de qualidade dos gelos fabricados nos supermercados para conservação do pescado, constituindo risco potencial para a saúde dos consumidores.

The ice is essential for the proper conservation of fish, as it delays microbial growth, preserving the physical-chemical, microbiological and organoleptic characteristics. However, ice can be a source of health risk for consumers when produced under poor sanitary conditions. Critical points must be controlled during the ice production and storage stage, such as the of potable water and hygienic-sanitary conditions. The presence of high populations of heterotrophic and coliform microorganisms and the poor physicochemical quality of the ice used for fish conservation can represent a potential risk to the consumer, which can be a carrier of diseases such as Salmonellosis, Escherichia coli infection and Staphylococcus aureus poisoning, in addition to reducing the shelf life of the product. In this context, this study aimed to evaluate the microbiological and physical-chemical quality of the ice used in the conservation of fish marketed in Rio Branco - AC. 15 ice samples collected in three different supermarkets were analyzed. It was recommended the collection of ice manufactured with its own machinery and that it be used in the commercialization of fresh fish, which would be exposed and cooled in ice stalls on site 300g ice manually with the use of gloves and packed in sterilized glass bottles, in an isothermal box. The microbiological quality of these samples was analyzed by counting mesophilic and psychrotrophic heterotrophic microorganisms. In addition, the determination of the most probable number of total coliforms at 30°C, thermotolerant and the quantification of pH and free residual chlorine of the samples were performed. Regarding the determination of mesophilic microorganisms, values between 1,00x10¹ and 9,15x10³ UFC/mL were recorded with samples (60%) out of the standard established by Ordinance MG/MS No. 888/2021. The amount of psychrotrophs varied between 1,00x10¹ and 8,98x103 CFU/mL. The presence of total coliforms at 30°C was recorded in 6 samples and of thermotolerants in 4 samples. Therefore, there was a variation in the quality standards of ices manufactured in supermarkets for fish conservation, constituting a potential risk to the health of consumers.

Airways exposure of bacterial superantigen SEB enhances bone marrow eosinophil population and facilitates its egress to blood and lung tissue.

BACKGROUND: Differentiation of bone marrow eosinophils (BM-EO) and its trafficking to peripheral blood and respiratory mucosa are a hallmark of inflammatory diseases. Staphylococcal enterotoxin B (SEB) has been shown to aggravate airways eosinophilic inflammation. This study aimed to investigate the effects of mouse airways SEB exposure on BM-EO population, as well as its adhesive properties and release of cytokines/chemokines that orchestrate BM-EO trafficking to lungs. METHODS: Male BALB/c mice were intranasally exposed to SEB (1 µg), and at 4, 16, 24 and 48 h thereafter, bone marrow (BM), circulating blood and bronchoalveolar lavage (BAL) fluid were collected. Levels of cytokines/chemokines and expressions of VLA-4 and CCR3 in BM were evaluated. Adhesion of BM to ICAM-1 and VCAM-1 were also evaluated. RESULTS: SEB exposure promoted a marked eosinophil influx to BAL at 16 and 24 h after exposure, which was accompanied by significant increases in counts of immature (16 h) and mature (4 to 48 h) forms of eosinophil in BM, along with blood eosinophilia (16 h). In BM, higher levels of eotaxin, IL-5, IL-4, IL-3 and IL-7 were detected at 16 to 48 h. SEB also significantly increased CCR3 expression and calcium levels in BM-EO, and enhanced the cell adhesion to ICAM-1 (24 h) and ICAM-1 (48 h). CONCLUSION: Airways SEB exposure increases the number of eosinophils in BM by mechanisms involving a network of cytokine and chemokine release, facilitating the BM-EO adhesion to ICAM-1 and VCAM-1 to gain access to the peripheral blood and lung tissues.

Airway exposure to Staphylococcal enterotoxin type B (SEB) enhances the number and activity of bone marrow neutrophils via the release of multiple cytokines.

BACKGROUND: The lung infections by Staphylococcus aureus are strongly associated with its ability to produce enterotoxins. However, little is known about the mechanisms underlying trafficking of bone marrow (BM) neutrophils during airway inflammation induced by Staphylococcal enterotoxin B (SEB). We therefore aimed to investigate the effects of mouse airways SEB exposure on BM neutrophil counts and its adhesive properties as well as on the release of cytokines/chemokines that orchestrate BM neutrophils trafficking to lung tissue. METHODS: Male BALB/c mice were intranasally exposed to SEB (1 µg), and at 4, 16 and 24 h thereafter, BM, circulating blood, bronchoalveolar lavage (BAL) fluid and lung tissue were collected. BM neutrophils adhesion, MAC-1 and LFA1-α expressions (by flow cytometry) as well as measurement of cytokine and/or chemokines levels were assayed after SEB-airway exposure. RESULTS: Prior exposure to SEB promoted a marked influx of neutrophils to BAL and lung tissue, which was accompanied by increased counts of BM immature neutrophils and blood neutrophilia. BM neutrophil expressions of LFA1-α and MAC-1 were unchanged by SEB exposure whereas a significant enhancement of adhesion properties to VCAM-1 was observed. The early phase of airway SEB exposure was accompanied by high levels of GM-CSF, G-CSF, IFN-γ, TNF-α and KC/CXCL1, while the latter phase by the equilibrated actions of SDF1-α and MIP-2. CONCLUSION: Mouse airways exposure to SEB induces BM cytokines/chemokines release and their integrated actions enhance the adhesion of BM neutrophils leading to acute lung injury.

Prevalence of the genus Cladosporium on the integument of leaf-cutting ants characterized by 454 pyrosequencing.

The relationship of attine ants with their mutualistic fungus and other microorganisms has been studied during the last two centuries. However, previous studies about the diversity of fungi in the ants' microenvironment are based mostly on culture-dependent approaches, lacking a broad characterization of the fungal ant-associated community. Here, we analysed the fungal diversity found on the integument of Atta capiguara and Atta laevigata alate ants using 454 pyrosequencing. We obtained 35,453 ITS reads grouped into 99 molecular operational taxonomic units (MOTUs). Data analysis revealed that A. capiguara drones had the highest diversity of MOTUs. Besides the occurrence of several uncultured fungi, the mycobiota analysis revealed that the most abundant taxa were the Cladosporium-complex, Cryptococcus laurentii and Epicoccum sp. Taxa in the genus Cladosporium were predominant in all samples, comprising 67.9 % of all reads. The remarkable presence of the genus Cladosporium on the integument of leaf-cutting ants alates from distinct ant species suggests that this fungus is favored in this microenvironment.

Staphylococcal enterotoxin A regulates bone marrow granulocyte trafficking during pulmonary inflammatory disease in mice.

Pulmonary neutrophil infiltration produced by Staphylococcal enterotoxin A (SEA) airway exposure is accompanied by marked granulocyte accumulation in bone marrow (BM). Therefore, the aim of this study was to investigate the mechanisms of BM cell accumulation, and trafficking to circulating blood and lung tissue after SEA airway exposure. Male BALB/C mice were intranasally exposed to SEA (1µg), and at 4, 12 and 24h thereafter, BM, circulating blood, bronchoalveolar lavage (BAL) fluid and lung tissue were collected. Adhesion of BM granulocytes and flow cytometry for MAC-1, LFA1-α and VLA-4 and cytokine and/or chemokine levels were assayed after SEA-airway exposure. Prior exposure to SEA promoted a marked PMN influx to BAL and lung tissue, which was accompanied by increased counts of immature and/or mature neutrophils and eosinophils in BM, along with blood neutrophilia. Airway exposure to SEA enhanced BM neutrophil MAC-1 expression, and adhesion to VCAM-1 and/or ICAM-1-coated plates. Elevated levels of GM-CSF, G-CSF, INF-γ, TNF-α, KC/CXCL-1 and SDF-1α were detected in BM after SEA exposure. SEA exposure increased production of eosinopoietic cytokines (eotaxin and IL-5) and BM eosinophil VLA-4 expression, but it failed to affect eosinophil adhesion to VCAM-1 and ICAM-1. In conclusion, BM neutrophil accumulation after SEA exposure takes place by integrated action of cytokines and/or chemokines, enhancing the adhesive responses of BM neutrophils and its trafficking to lung tissues, leading to acute lung injury. BM eosinophil accumulation in SEA-induced acute lung injury may occur via increased eosinopoietic cytokines and VLA-4 expression.

Leaf-cutting ants: an unexpected microenvironment holding human opportunistic black fungi.

Fungus-growing ants of the genus Atta are known for their leaf-cutting habit, a lifestyle they have maintained since their 50-million-year-old co-evolution with a mutualistic fungus, cultivated as food. Recent studies have highlighted that, in addition to the mutualistic fungus, nests of ants harbor a great diversity of microbial communities. Such microorganisms include the dematiaceous fungi, which are characterized by their melanized cell walls. In order to contribute to the knowledge of fungal ecology, as well as opportunistic strains that may be dispersed by these social insects, we isolated and identified fungi carried by gynes of Atta capiguara and Atta laevigata, collected from colonies located in Fazenda Santana, Botucatu (São Paulo, Brazil). The isolation was carried out using the oil flotation technique, which is suitable for the growth of black fungi. Inoculated plates were incubated at 25 and 35 °C until black cultures were visible (20-45 days). Isolates were identified based on microscopic and molecular characteristics. Some isolated genera were: Cladophialophora, Cladosporium, Exophiala, Ochroconis, Phaeococcomyces, Phialophora and Penidiella. Hyaline species were also found. The results obtained from this work showed that leaf-cutting gynes may contribute to the dispersal of opportunistic dematiaceous fungi. It is suggested that more attention should be paid to this still unexplored subject.

Effect of age at disease onset in the clinical profile of spondyloarthritis: a study of 1424 Brazilian patients.

OBJECTIVES: To analyse demographic and clinical variables in patients with disease onset before and after 40, 45 and 50 years in a large series of Brazilian SpA patients. METHODS: A common protocol of investigation was prospectively applied to 1424 SpA patients in 29 centres distributed through the main geographical regions in Brazil. The mean age at disease onset was 28.56 ± 12.34 years, with 259 patients (18.2%) referring disease onset after 40 years, 151 (10.6%) after 45 years and 81 (5.8%) after 50 years. Clinical and demographic variables and disease indices (BASDAI, BASFI, BASRI, MASES, ASQoL) were investigated. Ankylosing spondylitis was the most frequent disease (66.3%), followed by psoriatic arthritis (18%), undifferentiated SpA (6.7%), reactive arthritis (5.5%), and enteropathic arthritis (3.5%). RESULTS: Comparing the groups according to age of disease onset, those patients with later onset presented statistical association with female gender, peripheral arthritis, dactylitis, nail involvement and psoriasis, as well as negative statistical association with inflammatory low back pain, alternating buttock pain, radiographic sacroiliitis, hip involvement, positive familial history, HLA-B27 and uveitis. BASDAI, BASFI and quality of life, as well as physicians and patient's global assessment, were similar in all the groups. Radiographic indices showed worse results in the younger age groups. CONCLUSIONS: There are two different clinical patterns in SpA defined by age at disease onset: one with predominance of axial symptoms in the group with disease onset ≤ 40 years and another favouring the peripheral manifestations in those with later disease onset.

Interação entre transgênicos (Bt) e inseticidas no controle de pragas-chave em híbridos de milho-safrinha / Interaction between transgenics and insecticides in the control of key pests on off-season maize hybrids

A cultura do milho é atacada por diversos lepidópteros-praga. Recentemente, foi liberada no Brasil a comercialização de híbridos de milho Bt, integrando em seu genoma o gene Cry1Ab, proveniente de Bacillus thuringiensis subsp. kurstaki, que codifica a proteína com efeito tóxico sobre insetos da ordem Lepidooptera. Dessa forma, o objetivo deste trabalho foi avaliar, em condições de campo, a eficácia de híbridos de milho convencionais e transgênicos para o controle de pragas, submetidos ou não à pulverização com inseticidas. O ensaio foi realizado em área experimental do Polo Apta Centro Norte, em Pindorama, Estado de São Paulo. Foram avaliados cinco híbridos de milho convencionais em comparação com os respectivos híbridos isogênicos Bt. A avaliação dos danos visuais de Spodoptera frugiperda (Smith) foi realizada a cada 10 dias, em 20 plantas das duas linhas centrais da parcela, e para Helicoverpa zea (Bod.) e Diatraea saccharalis (Fab.) na pré-colheita. As interações entre os híbridos e a transgenia ocorreram em todas as avaliações a partir dos 40 dia após emergência (DAE). Os híbridos transgênicos (Bt) avaliados apresentaram as menores notas de danos visuais de S. frugiperda em todas as avaliações. A transgenia também proporcionou menor percentagem de espigas danificadas por H. zea e S. frugiperda e de colmos atacados por D. saccharalis. Conclui-se que a transgenia proporciona redução nos danos visuais ocasionados pelas pragas, embora apresente desempenho diferente entre os híbridos.

Diverse lepidopteran pests attack the maize crop. The commercialization of hybrids of transgenic Bt maize, integrating in its genome the gene Cry1Ab, from Bacillus thuringiensis subsp. kurstaki, which codifies the protein with toxic effect on insects of the Lepidoptera order, has been recently introduced in Brazil. The objective of this work was to evaluate, under field conditions, the effectiveness of conventional and transgenic maize hybrids for the control of pests, sprayed or not with insecticides. The assay was carried out in an experimental area of the Polo Apta Centro Norte, in Pindorama, São Paulo State, Brazil. Five conventional maize hybrids were evaluated in comparison with the respective isogenic Bt hybrids. The evaluation was carried out every 10 days, in 20 plants of the 2 central lines of the parcel for Spodoptera frugiperda (Smith) damage. For damages of Helicoverpa zea (Bod.) and Diatraea saccharalis (Fab.) the evaluations were carried out in the pre-harvest. The interactions between the hybrids and the transgeny occurred in all evaluations starting at the 40th day after emergence (DAE). The Bt hybrids evaluated presented the lowest scores for visual S. frugiperda damages in all the evaluations. The transgenic maize also provided the lowest percentage of cobs damaged by H. zea and S. frugiperda, and of stalks attacked by D. saccharalis. In conclusion, the transgenic maize provided reduction in visual damages caused by the pests, however, it presented different performance among the hybrids.

INTERAÇÃO ENTRE TRANSGÊNICOS (BT) E INSETICIDAS NO CONTROLE DE PRAGAS-CHAVE EM HÍBRIDOS DE MILHO-SAFRINHA

ABSTRACT Diverse lepidopteran pests attack the maize crop. The commercialization of hybrids of transgenic Bt maize, integrating in its genome the gene Cry1Ab, from Bacillus thuringiensis subsp. kurstaki, which codifies the protein with toxic effect on insects of the Lepidoptera order, has been recently introduced in Brazil. The objective of this work was to evaluate, under field conditions, the effectiveness of conventional and transgenic maize hybrids for the control of pests, sprayed or not with insecticides. The assay was carried out in an experimental area of the Polo Apta Centro Norte, in Pindorama, São Paulo State, Brazil. Five conventional maize hybrids were evaluated in comparison with the respective isogenic Bt hybrids. The evaluation was carried out every 10 days, in 20 plants of the 2 central lines of the parcel for Spodoptera frugiperda (Smith) damage. For damages of Helicoverpa zea (Bod.) and Diatraea saccharalis (Fab.) the evaluations were carried out in the pre-harvest. The interactions between the hybrids and the transgeny occurred in all evaluations starting at the 40th day after emergence (DAE). The Bt hybrids evaluated presented the lowest scores for visual S. frugiperda damages in all the evaluations. The transgenic maize also provided the lowest percentage of cobs damaged by H. zea and S. frugiperda, and of stalks attacked by D. saccharalis. In conclusion, the transgenic maize provided reduction in visual damages caused by the pests, however, it presented different performance among the hybrids.

RESUMO A cultura do milho é atacada por diversos lepidópteros-praga. Recentemente, foi liberada no Brasil a comercialização de híbridos de milho Bt, integrando em seu genoma o gene Cry1Ab, proveniente de Bacillus thuringiensis subsp. kurstaki, que codifica a proteína com efeito tóxico sobre insetos da ordem Lepidooptera. Dessa forma, o objetivo deste trabalho foi avaliar, em condições de campo, a eficácia de híbridos de milho convencionais e transgênicos para o controle de pragas, submetidos ou não à pulverização com inseticidas. O ensaio foi realizado em área experimental do Polo Apta Centro Norte, em Pindorama, Estado de São Paulo. Foram avaliados cinco híbridos de milho convencionais em comparação com os respectivos híbridos isogênicos Bt. A avaliação dos danos visuais de Spodoptera frugiperda (Smith) foi realizada a cada 10 dias, em 20 plantas das duas linhas centrais da parcela, e para Helicoverpa zea (Bod.) e Diatraea saccharalis (Fab.) na pré-colheita. As interações entre os híbridos e a transgenia ocorreram em todas as avaliações a partir dos 40 dia após emergência (DAE). Os híbridos transgênicos (Bt) avaliados apresentaram as menores notas de danos visuais de S. frugiperda em todas as avaliações. A transgenia também proporcionou menor percentagem de espigas danificadas por H. zea e S. frugiperda e de colmos atacados por D. saccharalis. Conclui-se que a transgenia proporciona redução nos danos visuais ocasionados pelas pragas, embora apresente desempenho diferente entre os híbridos.

[A new quaternary alkaloid from Aspidosperma gilbertii]. / Um novo alcalóide quaternário de Aspidosperma gilbertii.

The present work describes the structure determination of a new quaternary carbozol type alkaloid, isolated from Aspidosperma gilbertii: N,N-Dimethyltetrahydroellipticinium Hydroxyde.

Um novo alcaloide quaternario de Aspidosperma gilbertii. / A new quaternary alkaloid from Aspidosperma gilbertii

O presente trabalho descreve a determinacao da estrutura de um novo alcaloide quaternario, isolado de A, gilbertii: N,N-Dimetiltetraidroecelipticino, isolado da forma de hidroxido