Resistência aos compostos de amônio quaternário (QACs) de uso doméstico e hospitalar em patógenos prioritários multirresistentes / Resistance to quaternary ammonium compounds (QACs) for domestic and hospital use in multiresistant priority pathogens

Compostos de amônio quaternário (QACs) têm sido amplamente utilizados como desinfetantes e antissépticos, sendo essenciais na prevenção e controle de infecções bacterianas na medicina humana e veterinária. Embora patógenos prioritários multirresistentes têm sido muito bem caracterizados quanto ao perfil de suscetibilidade e contexto genético da resistência aos antibióticos, dados de resistência aos QACs são limitados. Assim, o objetivo do presente estudo foi avaliar a atividade in vitro dos QACs de uso doméstico e hospitalar [cloreto de benzalcônio (BAC), cloreto de cetilpiridinio (CPC) e brometo de cetiltrimetilamônio (CTAB)], contra patógenos prioritários multirresistentes, identificando os principais genes de resistência associados. Foram estudadas 100 cepas multirresistentes previamente sequenciados usando as plataformas Illumina MiSeq e NextSeq representativas de diferentes hospedeiros (humanos e animais) e fontes (ambientes e alimentos). As cepas foram identificadas como Klebsiella pneumoniae (n= 24), Escherichia coli (n= 30); Pseudomonas aeruginosa (n= 10), Enterobacter spp, (n= 8), Acinetobacter baumannii (n= 11) e Salmonella spp. (n= 17). Genes de resistência aos QACs foram identificados in silico através do alinhamento dos contigs obtidos de cada cepa sequenciada com genes de referência obtidos do GenBank, utilizando o programa Geneious versão 8 (Biomatters Ltd). A identidade de cada gene foi analisada utilizando o programa BLASTx, no qual um critério baseado em ≥90% identidade resultou na identificação dos genes mdfA (77%), qacE (44%), qacEΔ1 (43%), sugE(c) (29%), emrE (21%), qacA (19%), sugE(p) (5%), qacF (7%), qacH (7%) e qacL (7%) em 85 cepas; enquanto que 15 cepas não possuíam nenhum gene de resistência aos QACs. A concentração inibitória mínima (CIM) dos QACs para as 100 cepas foi determinada pelo método de microdiluição em caldo. Os resultados sugeriram que a resistência em patógenos prioritários circulando na interface humano-ambiente-animal não é restrita aos antibióticos, uma vez que a elevada ocorrência de genes qacE, qacEΔ1 e mdfA poderia estar associada com uma redução da suscetibilidade para QACs. Consequentemente, a resistência aos QACs poderia também contribuir para a persistência e adaptação destes patógenos nos seres humanos e outros animais, assim como em ambientes impactados antropogenicamente

Quaternary ammonium compounds (QACs) have been widely used as disinfectants and antiseptics, being applied as essential compounds in the prevention and control of bacterial infections in human-and veterinary hospital medicine. Although multiresistant priority pathogens have been well characterized with respect to their susceptibility profile and their genetic context of resistance for antibiotics, studies of resistance to QACs are limited. Thus, the objective of the present study was to evaluate the in vitro activity of QACs [(benzalkonium chloride (BAC), cetylpyridinium chloride (CPC) and cetyltrimethylammonium bromide (CTAB)] for household and hospital use against multiresistant priority pathogens, identifying the main resistance genes associated. A hundred multiresistant isolates (previously sequenced using the Illumina MiSeq and NextSeq platforms), representative of different hosts (humans and animals) and sources (environment and food) were studied. Isolates were identified as Klebsiella pneumoniae (n=24), Escherichia coli (n=30), Pseudomonas aeruginosa (n=10), Enterobacter spp. (n=8), Acinetobacter baumannii (n=11) and Salmonella spp. (n=17). In silico analysis for identification of genes conferring resistance to QACs were performed by aligning the contigs obtained from the strains with reference genes deposited in GenBank, using the Geneious version program (Biomatters Ltd). Similarities were analyzed using the BLASTx online program, considering the alignment criteria based on ≥ 90% identity. The result of these analysis revealed the presence of the following QAC genes: mdfA (77%), qacE (44%), qacEΔ1 (43%), sugE(c) (29%), emrE (21%), qacA (19%), sugE (p) (5%), qacF (7%), qacH (7%) e qacL (7%); while 15 strains showed no resistance genes for QACs. Determination of QACs minimum inhibitory concentration (MIC) for the 100 isolates, by the broth microdilution method. These results suggest that resistance to QACs in priority pathogens, circulating at the human-environment-animal interface, is not restricted to antibiotics, since the high occurrence of genes qacE, qacEΔ1 and mdfA were associated with a reduced susceptibility to QACs. Consequently, resistance to QACs could also contribute to the persistence and adaptation of these pathogens in humans and othes animals, as well as in anthropogenically impacted environments

The dynamic structure of Spitzenkörpers of Trichosporon asahii examined by the fluorescent probe FM4-64

Abstract The Spitzenkörper is a dynamic and specialized multicomponent cell complex present in the tips of hyphal cells. The amphiphilic styryl dye FM4-64 was found to be ideal for imaging the dynamic changes of the apical vesicle cluster within growing hyphal tips. It is widely used as a marker of endocytosis and to visualize vacuolar membranes. Here we performed uptake experiments using FM4-64 to study the dynamic of the Spitzenkörper in Trichosporon asahii. We observed that Spitzenkörpers were present at the tip of the budding site of the spore, blastospore, and the germ tube of T. asahii. We also found that Spitzenkörpers were present at the tip of the hyphae as well as the subapical regions. Cytochalasin D, an inhibitor of actin polymerization, leads to abnormal Spitzenkörper formation and loss of cell polarity.

The influence of castanhão reservoir on nutrient and suspended matter transport during rainy season in the ephemeral Jaguaribe river (CE, Brazil) / Influência do açude Castanhão no transporte de nutrientes e materiais em suspensão durante a estação chuvosa no intermitente rio Jaguaribe (CE, Brasil)

Measurements of nutrient and suspended matter concentrations and loads entering and leaving the Castanhão reservoir during the rainy season were carried out to assess the influence of this large reservoir on land-sea fluvial transport in the ephemeral Jaguaribe river basin. Spatial variation indicated statistically significant attenuation of concentrations only for total phosphorous and suspended matter across the reservoir. Strong retention of nutrients and suspended matter loads by the reservoir was observed with average trapping efficiency of 89% for dissolved silicon, 98% of soluble reactive phosphorus, 71% for ammonium, 87% for total nitrogen, 98% for total phosphorus and 97% for suspended matter compared to the reservoir inflow. The dam operational procedure defined by the ephemeral conditions of the river reduced water releases compared to reservoir inflow and induced strong retention of nutrient and suspended matter loads within the reservoir when fluvial transfer occurs in this semiarid watershed.

Medidas de concentrações e fluxos de nutrientes e material particulado em suspensão de entrada e saída do Açude Castanhão foram efetuadas para determinar a influência de um grande reservatório no transporte fluvial entre o continente e o oceano na bacia hidrográfica intermitente do Rio Jaguaribe. A variação espacial indicou uma atenuação estatisticamente significativa ao longo do reservatório somente para as concentrações de fósforo total e material particulado em suspensão. Os resultados indicam uma forte retenção de nutrientes e do material particulado em suspensão com eficiências de retenção de 89% para sílica dissolvida, 98% para fósforo solúvel reativo, 71% para amônia, 87% para nitrogênio total, 98% para fósforo total e 97% para o material particulado em suspensão em relação aos fluxos de entrada no reservatório. A rotina de operação da barragem definida pela condição intermitente do Rio Jaguaribe reduz a descarga de água comparada com a entrada no reservatório e induz forte retenção do fluxo fluvial de nutrientes e material particulado em suspensão pelo reservatório.

Nutrient and phytoplankton biomass in the Amazon River shelf waters

The Amazon River estuary is notable at the Amazon Continental Shelf, where the presence of the large amount of water originating from the Amazon during the river's falling discharge period was made evident by the low salinity values and high nutrient levels. Even so, the presence of oceanic waters in the shelf area was significant. Dissolved organic nitrogen was the predominant species of the nitrogen cycle phases, followed by total particulate nitrogen, nitrate, ammonium and nitrite. The chlorophyll a data in the eutrophic area indicated that there is sufficient nitrogen in the area to withstand productivity, though dissolved inorganic nitrogen removal processes are faster than regeneration or mineralization. The anomalous amounts of inorganic dissolved nitrogen showed more removal than addition. The simulations with the bidimensional MAAC-2D model confirmed that high nutrient waters are displaced northwest-ward (two cores at 2.5ºN-50ºW and 4ºN-51ºW) by the stronger NBC during falling river discharge. During high river flow period these nutrient-rich lenses are distributed around 0.5ºN-48.5ºW as well as along the shallow Amazonian shelf (20m-50m depth, 1ºN-3.5ºN), as a result of the spreading of Amazon freshwater outflow.

O estuário do rio Amazonas é notável na Plataforma Continental do Amazonas, onde a presença das águas fluviais foi detectada, mesmo durante o período da diminuição da descarga desse rio, pelos baixos valores de salinidade e altos valores de nutrientes. Contudo, a presença das águas oceânicas também foi marcante. Em relação às fases do ciclo do nitrogênio, o nitrogênio orgânico dissolvido foi a forma predominante, seguido do nitrogênio total particulado, nitrato, amônia e nitrito. Os dados de clorofila a indicaram uma área eutrófica onde há nitrogênio embora os valores da anomalia do nitrogênio inorgânico dissolvido tenham mostrado que ocorre maior remoção do que adição dessa forma nitrogenada na área em estudo. Os resultados das simulações com o modelo bidimensional MAAC-2D confirmaram que as águas ricas em nutrientes são deslocadas para noroeste (dois núcleos em 2,5°N-50°W e 4°N-51°W), pela ação de uma CNB mais forte durante o período de decaimento da vazão dos rios. No período de vazão máxima, estas lentes de águas ricas de nutrientes distribuem-se próximo de 0,5°N-48,5°W, bem como ao longo da plataforma Amazônica rasa (20m-50m profundidade, 1°N-3,5°N), como resultado do espalhamento da água doce de origem continental.

The importance of excretion by Chironomus larvae on the internal loads of nitrogen and phosphorus in a small eutrophic urban reservoir / Excreção por quironomídeos em reservatório urbano

Measurements of ammonium and phosphate excretion by the Chironomus larvae were conducted in order to evaluate the importance of these chironomids for the internal loads of a small eutrophic urban reservoir. Ammonium and phosphate excretion rates by Chironomus larvae of small size (6-10 mm total length) were significantly higher than those of the Chironomids having medium (9-11 mm) and large (11-16 mm) sizes. A dependence in relation to temperature was recorded for the ammonium and phosphate excretions that was significantly higher at 25 °C than at 20 and 15 °C. Through a linear relation between biomass (dry weight) and total length and, between excretion and biomass and, data on chironomids densities, after an intense sampling in 33 sites distributed all along the reservoir bottom, the mean phosphate and ammonium excretion rates corresponded to 2,014 ± 5,134 µg.m-2/day and 1,643 ± 3,974 µg.m-2/day, respectively. Considering the mean biomass (34 mg.m-2) of Chironomus, the lake area (88,156 m²) and the mean excretion rates, the contribution of benthic chironomids to the internal loads would be 181 KgP and 147 KgN. for the sampling months (October-November 1998). These values showed that the internal loads by excretion from Chironomus larvae correspond to approximately 33 percent of the external loads of phosphorus in the lake and, in the case of nitrogen, to only 5 percent.

Medidas de excreção de amônia e fosfato por larvas de Chironomus foram executadas com a finalidade de avaliar a importância desses quironomídeos nas cargas internas de um pequeno reservatório urbano eutrófico. Taxas de excreção de amônia e fosfato significativamente mais elevadas foram encontradas nas larvas de Chironomus de pequeno (6-10 mm) tamanho, em relação às de tamanho médio (9-11 mm) e grande (11-16 mm). Uma dependência em relação à temperatura foi registrada para a excreção de amônia e fosfato, significativamente mais alta a 25 °C do que a 20 e 15 °C. Depois da obtenção das relações lineares entre biomassa (peso seco) e comprimento total e excreção com biomassa e dos dados de densidade de quironomídeos, após amostragem em 33 pontos distribuídos por todo o reservatório, foi possível estimar uma taxa média de excreção de fosfato de 2014 ± 5134 µg.m-2/dia e, de amônia em 1643 ± 3974 µg.m-2/dia. Considerando a biomassa média (34 mg.m-2) de quironomídeos, a área do lago (88.156 m²) e as taxas médias de excreção, a contribuição dos quironomídeos bênticos para as cargas internas seria de 181 KgP e 147 KgN, para o mês da amostragem (outubro-novembro de 1998). Esses valores mostraram que as cargas internas por excreção pelas larvas de Chironomus correspondem aproximadamente a 33 por cento das cargas externas de fósforo para o lago e, somente 5 por cento no caso de nitrogênio.

A study on major inorganic ion composition of atmospheric aerosols.

Atmospheric aerosol samples were collected from Akola and Buldana region covering around 40 sqkm area during October-November 2002 and were analyzed for ten major inorganic ions namely F-, Cl-, NO3-, SO4(2-), PO4(2-), Na+, K+, Ca2+, Mg2+ and NH4+ using ion chromatographic technique. The average mass of aerosols was found to be 225.81 microg/m3 with standard deviation of 31.29 and average total water soluble load of total cations and anions was found to be 4.32 microg/m3. The concentration of ions in samples showed a general pattern as SO4(2-) > NO3- > Cl- > PO4(2-) > F- for anions and Na+ > Ca2+ > NH4+ > Mg2+ > K+ for cations. The overall composition of the aerosols was taken into account to identify the sources. The trend showed higher concentration of sodium followed by calcium, sulfate, nitrate, phosphate and ammoinum and found to be influenced by terrestrial sources. The presence of SO4(2-) and NO3- in aerosols may be due to re-suspension of soil particles. Ca2+, Mg2+ and Cl- are to be derived from soil materials. The presence of NH4+ may be attributed to the reaction of NH3 vapors with acidic gases may react or condense on an acidic particle surface of anthropogenic origin. The atmospheric aerosol is slightly acidic due to neutralization of basicity by SO2 and NO(x).

Studies on phyto-and-zooplankton composition and its relation to fish productivity in a west coast fish pond ecosystem.

A fish pond ecosystem on the west coast of Korea was studied for biotic and abiotic factors. The occurrence rate of phytoplankton at the embanked fish farm was 84 times higher to that of shore waters. Phytoplankton species which occurred during the study period were categorized into Cyanophyta, Dinophyta and Bacillariophyta with a total of 26 subgeneric taxa (22 species of Dinophyta, 2 species of Bacillariophyta and 2 species of Cyanophyta). The total cell density during the study period ranged from16 x 103 to 13.59 x 10(5) cells l(-1). As for the zooplankton, the monthly density ranged from 26 - 8.684 x 10(3) cells m(-3) with an average of 126.0 x 10(3) cells m(-3) for April. In the case of May, it ranged from 41 - 3.331 x 10(3) cells m(-3) and its average of 50.3 x 10(3) cells m(-3) which was comparatively less. The varying environmental factors in Seosan embankment A, B and Chonsu Bay waters could have affected the fish farm. According to the results, the concentration of inorganic nitrogen was 0.067-0.106 (average: 0.083) mg l(-1) and the concentration of organic nitrogen was 0.008-0.022 (average: 0.014) mgl(-1). Also due to the release of brackish water at Seosan embankment A and B, the inorganic nitrogen showed an increasing trend. The survival rate of Sabastes schlegeli, and Acanthopagrus schlegeli was high in stations 2H and I (20.87%, 33%) and relatively low in stations 2B to 2F.