Effect of plasma and heat treatments on orange juice quality / Efeito de plasma e tratamento térmico na qualidade do suco de laranja

Heat treatment is used in the orange juice industry to neutralize the action of pathogenic microorganisms. However, it can reduce the nutritional value of the juice. Thus, our study assessed the cold plasma treatment as an alternative method against Escherichia coli and Candida albicans in 'Lima' orange juice. Both, plasma and heat treatments, reduced the amount of E. coli in the juice, inactivating 16.72 and 100%, respectively. Plasma did not inactivate C. albicans, but heat treatment inactivated 100%. Plasma and heat treatment increased Hue angle and luminosity (more yellowish juice). Plasma reduced vitamin C, carotenoids, and polyphenols content, while increased flavonoids. Heat treatment reduced the carotenoid content. However, neither heat nor plasma treatment altered the antioxidant activity. The plasma treatment reduced the intensity of color (chroma), the soluble solids content and the acidity ratio, total sugars, and the vitamin C content of juice compared to the heat-treated and control juices. Plasma-treated juice showed increased levels of yellow flavonoids, total phenolics and antioxidant activity until the 12th day of storage.

O tratamento térmico é utilizado na indústria de suco de laranja para neutralizar a ação de microrganismos patogênicos. No entanto, pode reduzir o valor nutricional do suco. Assim, o estudo avaliou o tratamento com plasma frio como método alternativo contra Escherichia coli e Candida albicans em suco de laranja 'Lima'. Ambos os tratamentos, plasma e calor, reduziram a quantidade de E. coli no suco, inativando 16.72 e 100%, respectivamente. O plasma não inativou C. albicans, mas o tratamento térmico inativou 100%. Plasma e tratamento térmico aumentaram o ângulo Hue e a luminosidade do suco (tom mais amarelo). O plasma reduziu o conteúdo de vitamina C, carotenóides e polifenóis, enquanto aumentou os flavonóides. O tratamento térmico reduziu o teor de carotenóides. No entanto, nem o calor nem o tratamento com plasma alteraram a atividade antioxidante. O tratamento com plasma reduziu a intensidade da cor (croma), o teor de sólidos solúveis e a razão de acidez, açúcares totais e o teor de vitamina C do suco em comparação com os sucos tratados termicamente e controle. O suco tratado com plasma apresentou maiores teores de flavonóides amarelos, fenólicos totais e atividade antioxidante até o 12º dia de armazenamento.

Candidose sistêmica em cães associada à cinomose / Systemic candidosis in dogs associated with canine distemper virus

Background: Candidosis is an opportunistic fungal infection caused by yeasts of the genus Candida, which normally reside on the surfaces of the mucous membranes and in the skin of several animal species and healthy humans; however the fungi can convert into pathogenic microorganisms and result in invasive infections with systemic involvement due to the impairment of the immune system. Systemic candidosis is rare in dogs and few reports of this mycosis are available in literature. Therefore, the objective of this study was to describe three cases of systemic candidosis associated to canine distemper virus in dogs, highlighting the main epidemiological, clinical and pathological aspects. Cases: Three cases of systemic infection by Candida sp. were diagnosed in dogs. The animals predominantly presented neurological clinical manifestations, followed by unspecific alterations with an evolution of 5 to 30 days. Macroscopically, the lesions were characterized by white-yellowish multifocal to coalescent areas surrounded by reddish borders in the kidneys and heart (cases 1, 2 and 3); liver and submandibular lymph node (case 1); and lung (case 2). In the brains of the three dogs were observed multifocal, blackened and/or reddish and friable areas. In case 1, was observed an increased volume of the right carpometacarpal joint, that when cut, released a reddish and turbid content. In case 2, there was deposition of a whitish lumpy material on the epicardial surface, aorta artery, pericardial sac and spleen; infarction on the right testicle, hydroureter, pyelonephritis and hydronephrosis. Microscopically, the lesions consisted of necrossupurative inflammation associated to hemorrhage, vasculitis, congestion, thrombosis, infarction and fungal structures with distinct morphological patterns which included blastoconidia, pseudohyphae and hyphae. The fungal structures were...

Candidose sistêmica em cães associada à cinomose / Systemic candidosis in dogs associated with canine distemper virus

Background: Candidosis is an opportunistic fungal infection caused by yeasts of the genus Candida, which normally reside on the surfaces of the mucous membranes and in the skin of several animal species and healthy humans; however the fungi can convert into pathogenic microorganisms and result in invasive infections with systemic involvement due to the impairment of the immune system. Systemic candidosis is rare in dogs and few reports of this mycosis are available in literature. Therefore, the objective of this study was to describe three cases of systemic candidosis associated to canine distemper virus in dogs, highlighting the main epidemiological, clinical and pathological aspects. Cases: Three cases of systemic infection by Candida sp. were diagnosed in dogs. The animals predominantly presented neurological clinical manifestations, followed by unspecific alterations with an evolution of 5 to 30 days. Macroscopically, the lesions were characterized by white-yellowish multifocal to coalescent areas surrounded by reddish borders in the kidneys and heart (cases 1, 2 and 3); liver and submandibular lymph node (case 1); and lung (case 2). In the brains of the three dogs were observed multifocal, blackened and/or reddish and friable areas. In case 1, was observed an increased volume of the right carpometacarpal joint, that when cut, released a reddish and turbid content. In case 2, there was deposition of a whitish lumpy material on the epicardial surface, aorta artery, pericardial sac and spleen; infarction on the right testicle, hydroureter, pyelonephritis and hydronephrosis. Microscopically, the lesions consisted of necrossupurative inflammation associated to hemorrhage, vasculitis, congestion, thrombosis, infarction and fungal structures with distinct morphological patterns which included blastoconidia, pseudohyphae and hyphae. The fungal structures were...(AU)

Atividade in vitro do extrato etanólico de própolis e do digluconato de clorexidina sobre as espécies de Candida isoladas da mucosa bucal de pacientes internados em Unidade de Terapia Intensiva (UTI) / In vitro activity of ethanolic extract of propolis and chlorhexidine digluconate on Candida species isolated from the oral mucosa of patients hospitalized in the Intensive Care Unit (ICU)

Avaliou-se a atividade dos extratos de própolis e digluconato de clorexidina em Candida sp isoladas da mucosa bucal de pacientes em UTI. Foram determinadas as concentrações fungicidas mínimas (CFM) e comparadas, nas doses sub-inibitórias, à produção de exoenzimas proteinase e fosfolipase e formação de franjas. Em 72 isolados foram avaliadas a atividade antifúngica pela técnica de microdiluição em série, na “base 2”, a produção das exoenzimas proteinase e fosfolipase, e a formação de franjas, antes e após a exposição às própolis e clorexidina. Dos 72 isolados, 53 eram C. albicans, 11 C. tropicalis, quatro C. guilhermondii e quatro sugestivas de C. dubliniensis. CFM 90% do extrato de própolis foi de 5% para C. albicans, 20% C. tropicalis, 0,625% C. guilhermondii e 0,312% sugestivas de C. dubliniensis. CFM 90% da clorexidina foi de 0,0018% para C. albicans, 0,012% C. tropicalis, de 0,0018% C. guilhermondii e de 0,00375% sugestivas de C. dubliniensis. Ocorreu inibição das exoenzimas e franjas, em ambos os produtos. Apesar da inibição da clorexidina ser menor que a da própolis, seu uso diário não causa efeitos colaterais indesejáveis como manchas nos dentes e na língua, perda do paladar e sensação de queimação na mucosa bucal.

The activity of propolis extract and chlorhexidine digluconate on Candida sp isolated from oral mucosa of patients in ICU was evaluated. The minimum fungicidal concentrations (MFC) were determined, and also the production of proteinase and phospholipase exoenzymes and the fringe formation. Seventy-two isolates were used and identified by the API 20C AUX® System. The antifungal activity was evaluated by “at base 2” serial microdilution technique. Also the exoenzymes production (proteinase and phospholipase), the fringes formation, before and after being exposed to propolis and chlorhexidine, were analysed. Of 72 isolates, 53 were C. albicans, 11 C. tropicalis, four C. guilhermondii and four suggestive C. dubliniensis. The MFC 90% of propolis extract was 5% C. albicans, 20% C. tropicalis, 0.625% C. guilhermondii; and 0.312% suggestive of C. dubliniensis. MFC 90% of chlorhexidine was 0.0018% C. albicans, 0.012% C. tropicalis, 0.0018% C. guilhermondii and 0.00375% suggestive of C. dubliniensis. The inhibition of exoenzymes and fringes occurred in the both products. Although the inhibition of chlorhexidine is lower than that showed by propolis, its daily use neither cause undesirable side effects as blemishes on the teeth and tongue, nor the loss of the taste and the burning sensation in the oral mucosa.

Atividade in vitro do extrato etanólico de própolis e do digluconato de clorexidina sobre as espécies de Candida isoladas da mucosa bucal de pacientes internados em Unidade de Terapia Intensiva (UTI) / In vitro activity of ethanolic extract of propolis and chlorhexidine digluconate on Candida species isolated from the oral mucosa of patients hospitalized in the Intensive Care Unit (ICU)

Avaliou-se a atividade dos extratos de própolis e digluconato de clorexidina em Candida sp isoladas da mucosa bucal de pacientes em UTI. Foram determinadas as concentrações fungicidas mínimas (CFM) e comparadas, nas doses sub-inibitórias, à produção de exoenzimas proteinase e fosfolipase e formação de franjas. Em 72 isolados foram avaliadas a atividade antifúngica pela técnica de microdiluição em série, na “base 2”, a produção das exoenzimas proteinase e fosfolipase, e a formação de franjas, antes e após a exposição às própolis e clorexidina. Dos 72 isolados, 53 eram C. albicans, 11 C. tropicalis, quatro C. guilhermondii e quatro sugestivas de C. dubliniensis. CFM 90% do extrato de própolis foi de 5% para C. albicans, 20% C. tropicalis, 0,625% C. guilhermondii e 0,312% sugestivas de C. dubliniensis. CFM 90% da clorexidina foi de 0,0018% para C. albicans, 0,012% C. tropicalis, de 0,0018% C. guilhermondii e de 0,00375% sugestivas de C. dubliniensis. Ocorreu inibição das exoenzimas e franjas, em ambos os produtos. Apesar da inibição da clorexidina ser menor que a da própolis, seu uso diário não causa efeitos colaterais indesejáveis como manchas nos dentes e na língua, perda do paladar e sensação de queimação na mucosa bucal.(AU)

The activity of propolis extract and chlorhexidine digluconate on Candida sp isolated from oral mucosa of patients in ICU was evaluated. The minimum fungicidal concentrations (MFC) were determined, and also the production of proteinase and phospholipase exoenzymes and the fringe formation. Seventy-two isolates were used and identified by the API 20C AUX® System. The antifungal activity was evaluated by “at base 2” serial microdilution technique. Also the exoenzymes production (proteinase and phospholipase), the fringes formation, before and after being exposed to propolis and chlorhexidine, were analysed. Of 72 isolates, 53 were C. albicans, 11 C. tropicalis, four C. guilhermondii and four suggestive C. dubliniensis. The MFC 90% of propolis extract was 5% C. albicans, 20% C. tropicalis, 0.625% C. guilhermondii; and 0.312% suggestive of C. dubliniensis. MFC 90% of chlorhexidine was 0.0018% C. albicans, 0.012% C. tropicalis, 0.0018% C. guilhermondii and 0.00375% suggestive of C. dubliniensis. The inhibition of exoenzymes and fringes occurred in the both products. Although the inhibition of chlorhexidine is lower than that showed by propolis, its daily use neither cause undesirable side effects as blemishes on the teeth and tongue, nor the loss of the taste and the burning sensation in the oral mucosa.(AU)

Isolation of Candida spp. from denture-related stomatitis in Pará, Brazil

The aim of this study was to isolate and identify Candida species from the oral cavity of denture wearers with denture-related stomatitis who were attended at the University Federal of Pará (Belém City, Pará State, Brazil). A total of 36 denture wearers with denture-related stomatitis were included, and type I (50%), type II (33%) and type III (17%) stomatitis were observed. Candida spp. were isolated from 89% of the cases and included five different Candida species. C. albicans was the most frequently recovered species (78% of the cases), followed by C. famata and C. tropicalis. We observed a significant association between Candida species isolation and unsatisfactory denture condition (p = 0.0017). Our results demonstrated the highly frequency of Candida species isolation in denture wearers with denture-related stomatitis and showed the relationship between these species and poor denture maintenance.(AU)

Extracellular phospholipase production of oral Candida albicans isolates from smokers, diabetics, asthmatics, denture wearers and healthy individuals following brief exposure to polyene, echinocandin and azole antimycotics

Objective Candida albicans is the primary causative agent of oral candidosis, and one of its key virulent attributes is considered to be its ability to produce extracellular phospholipases that facilitate cellular invasion. Oral candidosis can be treated with polyenes, and azoles, and the more recently introduced echinocandins. However, once administered, the intraoral concentration of these drugs tend to be sub-therapeutic and rather transient due to factors such as the diluent effect of saliva and cleansing effect of the oral musculature. Hence, intra-orally, the pathogenic yeasts may undergo a brief exposure to antifungal drugs. We, therefore, evaluated the phospholipase production of oral C. albicans isolates following brief exposure to sub-therapeutic concentrations of the foregoing antifungals. Materials and methods Fifty C. albicans oral isolates obtained from smokers, diabetics, asthmatics using steroid inhalers, partial denture wearers and healthy individuals were exposed to sub-therapeutic concentrations of nystatin, amphotericin B, caspofungin, ketoconazole and fluconazole for one hour. Thereafter the drugs were removed and the phospholipase production was determined by a plate assay using an egg yolk-agar medium. Results The phospholipase production of these isolates was significantly suppressed with a percentage reduction of 10.65, 12.14, 11.45 and 6.40% following exposure to nystatin, amphotericin B, caspofungin and ketoconazole, respectively. This suppression was not significant following exposure to fluconazole. Conclusions Despite the sub-therapeutic, intra oral, bioavailability of polyenes, echinocandins and ketoconazole, they are likely to produce a persistent antifungal effect by suppressing phospholipase production, which is a key virulent attribute of this common pathogenic yeast.(AU)

Associação de megabacteriose, aspergilose e candidíase em periquitos australianos (Melopsittacus undulatus) em cativeiro, Marília, SP - Relato de caso / Association megabacteriosis, aspergillosis and candidiasis in australianparakeet (Melopsittacus undulatus) in captivity, Marilia, SP: case report

Infecções fúngicas causadas por Macrorhabdus ornithogaster, Aspergillus flavus, Candida albicans tem sido descritas em várias espécies aviárias sendo responsáveis por promover alterações gastrointestinais, respiratórias, neurológicas e dermatológicas entre outras. O presente artigo descreve aspectos clínicos, anatomopatológicos, procedimentos diagnósticos, medidas preventivas e terapêuticas de um caso crônico de três infecções fúngicas concomitantes em periquitos australianos (Melopsittacus undulatus) provenientes de um criatório comercial encaminhado ao laboratório de Ornitopatologia do Hospital Veterinário da Faculdade de Medicina Veterinária e Zootecnia da Universidade Estadual Paulista (FMVZ UNESP), campus Botucatu São Paulo, Brasil.(AU)

Fungal infections caused by Macrorhabdus ornithogaster, Aspergillus flavus, Candida albicans has been described as responsiblefor promoting gastrointestinal, respiratory, neurological and dermatological disorders in several avian species. The present paperdescribes the clinical signs, histopathological exams, treatment and preventions and of a chronic case of three fungal infectionsconcomitant in australian parakeet (Melopsittacus undulatus). The animals came from a commercial breeding facility and sent toOrnitopathology Laboratory of the Veterinary Hospital, Veterinary Medicine and Animal Science Faculty, Paulista State University(FMVZ - UNESP), Botucatu campus - São Paulo, Brazil.(AU)

Associação de megabacteriose, aspergilose e candidíase em periquitos australianos (Melopsittacus undulatus) em cativeiro, Marília, SP - Relato de caso / Association megabacteriosis, aspergillosis and candidiasis in australianparakeet (Melopsittacus undulatus) in captivity, Marilia, SP: case report

Infecções fúngicas causadas por Macrorhabdus ornithogaster, Aspergillus flavus, Candida albicans tem sido descritas em várias espécies aviárias sendo responsáveis por promover alterações gastrointestinais, respiratórias, neurológicas e dermatológicas entre outras. O presente artigo descreve aspectos clínicos, anatomopatológicos, procedimentos diagnósticos, medidas preventivas e terapêuticas de um caso crônico de três infecções fúngicas concomitantes em periquitos australianos (Melopsittacus undulatus) provenientes de um criatório comercial encaminhado ao laboratório de Ornitopatologia do Hospital Veterinário da Faculdade de Medicina Veterinária e Zootecnia da Universidade Estadual Paulista (FMVZ UNESP), campus Botucatu São Paulo, Brasil.

Fungal infections caused by Macrorhabdus ornithogaster, Aspergillus flavus, Candida albicans has been described as responsiblefor promoting gastrointestinal, respiratory, neurological and dermatological disorders in several avian species. The present paperdescribes the clinical signs, histopathological exams, treatment and preventions and of a chronic case of three fungal infectionsconcomitant in australian parakeet (Melopsittacus undulatus). The animals came from a commercial breeding facility and sent toOrnitopathology Laboratory of the Veterinary Hospital, Veterinary Medicine and Animal Science Faculty, Paulista State University(FMVZ - UNESP), Botucatu campus - São Paulo, Brazil.

MUCORMICOSE E CANDIDÍASE EM CÃES

Ultimamente, as doenças fúngicas tem sido responsáveis por elevada mortalidade na espécie canina. A zigomicose e a candidíase estão inseridas entre as principais micoses que eventualmente podem provocar infecções sistêmicas e fatais em indivíduos imunossuprmidos. Esta Dissertação, foi elaborada em dois capítulos, composta por dois artigos originais. O primeiro submetido à Journal of Comparative Pathology relata dois casos de infecções por fungos da ordem Mucorales com envolvimento sistêmico e hepatogástrico em cães, descrevendo os aspectos epidemiológicos, clínicos e patológicos. A doença ocorreu em cães Poodle, machos com idade de um e dez anos. As alterações clínicas incluíam sinais respiratórios, neurológicos e gastrointestinais que progrediram evoluindo para morte em três a dezenove dias. Macroscopicamente, no caso 1, havia áreas irregulares amareladas ou avermelhadas e elevadas que aprofundavam-se ao corte nos pulmões, coração e encéfalo. No caso 2, as alterações consistiam em ruptura de estômago com bordos e mucosa gástrica recobertos por espesso material brancacento e friável, além de múltiplas áreas extensas nodulares e cavitações profundas amarelo-brancacentas na superfície capsular do fígado. Histologicamente, foram observados piogranulomas em todos os órgãos afetados, associados a hifas fúngicas, vasculite e trombose. O diagnóstico de mucormicose foi realizado através das lesões microscópicas associadas às características morfotintoriais das hifas. As hifas foram fortemente imunomarcadas pelo anticorpo monoclonal anti-Rhizopus arrhizus. O segundo artigo, foi submetido à revista Arquivo Brasileiro de Medicina Veterinária e Zootecnia, descreve os aspectos epidemiológicos, clínicos e patológicos de três casos de candidíase sistêmica associada a cinomose em cães. Os cães acometidos tinham idade de quatro meses a cinco anos. Desses, dois eram cães sem raça definida e um Poodle. As manifestações clínicas eram predominantemente neurológicas, seguidas de alterações inespecíficas com evolução de cinco a trinta dias. Na necropsia havia áreas branco-amareladas circundadas por bordas avermelhadas nos rins, coração, fígado, linfonodo submandibular e pulmão. Adicionalmente, no caso 2, havia material grumoso brancacento aderido à superfície epicárdica, artéria aorta, saco pericárdico e baço. Além de infarto no testículo direito, hidroureter, pielonefrite e hidronefrose. No caso 1, havia aumento da articulação carpo-metacárpica direita com conteúdo avermelhado e turvo. No encéfalo, as lesões caracterizavam-se por áreas enegrecidas e/ou avermelhadas e friáveis. Microscopicamente, as lesões consistiam em áreas de necrose e hemorragia, circundadas por neutrófilos associada a vasculite, congestão, trombose, infarto e fungos com vários padrões morfológicos que incluíam leveduras, pseudo-hifas e hifas. Os três cães estavam imunossuprimidos decorrente da infecção concomitante pelo vírus da cinomose canina. Os casos 2 e 3 foram diagnosticados ainda com dermatofitose generalizada e linfoma, respectivamente. O diagnóstico de candidíase sistêmica foi realizado através das características morfotintoriais do agente e imunomarcação pelo anticorpo policlonal anti-Candida albicans. Com a realização desses trabalhos foi possível constatar que a mucormicose e a candidíase são doenças esporadicamente diagnosticadas na rotina do Laboratório de Patologia Animal do Hospital Veterinário Universitário Prof. Dr. Ivon Macêdo Tabosa da UFCG. Embora pouco frequentes, essas micoses constituem importantes causas de morte em cães.

Lately, fungal diseases have been responsible for high mortality in the canine species. The zygomycosis, and candidiasis are inserted between the main mycoses which eventually may cause systemic and fatal infections in immunosuppressed individuals. This Dissertation, was elaborated in two chapters, composed by two original articles. The first one submitted to the Journal of Comparative Pathology, describe two cases of fungal infection of the order Mucorales with systemic and hepatogastric involvement in dogs. The disease occurred in Poodle dogs, males aged one and ten years. The clinical signs included respiratory, neurological and gastrointestinal symptoms that progressed to death in three and nineteen days. Macroscopically, in case 1, there were irregular yellowish or reddish raised areas that deepened the cut in the lungs, heart and brain. In case 2, the alterations consisted of rupture of the stomach with borders and gastric mucosa covered by a thick and whitish, but also friable material, in addition to multiple extensive nodular areas and deep white-yellowish cavitations on the capsular surface of the liver. Histologically, piogranulomas were observed in all affected organs, associated with fungal hyphae, vasculitis and thrombosis. The diagnosis of mucormycosis was based on the microscopic lesions associated with the morphotinctorial features of the hyphae. The hyphae were strongly immunomarked by the anti-Rhizopus arrhizus monoclonal antibody. The second article, was submitted to the journal Arquivo Brasileiro de Medicina Veterinária e Zootecnia, describes the epidemiological, clinical and pathological aspects of three cases of systemic candidiasis associated with distemper in dogs. The affected dogs were between four months and five years. Of these, two were without defined breed dogs and one Poodle. The clinical manifestations were predominantly neurological, followed by nonspecific changes with evolution from five to thirty days. At necropsy there were yellowish-white areas surrounded by reddish edges in the kidneys, heart, liver, submandibular lymph node and lung. In addition, in case 2, there were lumpy materials whitish adhered to the epicardial surface, aortic artery, pericardial sac and spleen. In addition to infarction in the right testicle, hydroureter, pyelonephritis and hydronephrosis. In case 1, there was an increase in the right carpal-metacarpal joint, that when cutting reddish and cloudy content flowed. In the encephalon, the lesions were characterized by blackened and/or reddish and friable areas. Microscopically, the lesions consisted of areas of necrosis and hemorrhage, surrounded by neutrophils associated with vasculitis, congestion, thrombosis, infarction, and fungi with various morphological patterns including yeasts, pseudo-hyphae and hyphae. The three dogs were immunosuppressed due to the concomitant infection with the distemper virus. Cases 2 and 3 were diagnosed still with generalized dermatophytosis and lymphoma, respectively. The diagnosis of systemic candidiasis was made through the morphotinctorial characteristics of the agent and immunostaining by the polyclonal anti-Candida albicans antibody. With the accomplishment of these works it was possible to verify that mucormycosis and candidiasis are diseases sporadically diagnosed in the routine of the Laboratório de Patologia Animal of the Hospital Veterinário Universitário Prof. Dr. Ivon Macêdo Tabosa of the UFCG. Although uncommon, these mycoses are important causes of death in dogs.

Antimicrobial effect of para-alkoxyphenylcarbamic acid esters containing substituted N-phenylpiperazine moiety

In current research, nine basic esters of para-alkoxyphenylcarbamic acid with incorporated 4-(4fluoro-/3-trifluoromethylphenyl)piperazin-1-yl fragment, 6i-6m and 8f-8i, were screened for their in vitro antimicrobial activity against Candida albicans, Staphylococcus aureus and Escherichia coli, respectively. Taking into account the minimum inhibitory concentration assay (MIC), as the most active against given yeast was evaluated 8i (MIC = 0.20 mg/mL), the most lipophilic structure containing para-butoxy and trifluoromethyl substituents. Investigating the efficiency of the compounds bearing only a single atom of fluorine and appropriate para-alkoxy side chain against Candida albicans, the cut-off effect was observed. From evaluated homological series, the maximum of the effectiveness was noticed for the stucture 6 k (MIC = 0.39 mg/mL), containing para-propoxy group attached to phenylcarbamoyloxy fragment, beyond which the compounds ceased to be active. On the contrary, all the tested molecules were against Staphylococcus aureus and Escherichia coli (MICs > 1.00 mg/mL) practically inactive.(AU)

Candida albicans morphologies revealed by scanning electron microscopy analysis

Scanning electron microscope (SEM) observations were used to analyze particular morphologies of Candida albicans clinical isolate (strain 82) and mutants defective in hyphae-promoting genes EFG1 (strain HLC52) and/ or CPH1 (strains HLC54 and Can16). Transcription factors Efg1 and Cph1 play role in regulating filamentation and adhesion of C. albicans' morphologies. Comparative analysis of such mutants and clinical isolate showed that Efg1 is required for human serum-induced cell growth and morphological switching. In the study, distinct differences between ultrastructural patterns of clinical strain's and null mutants' morphologies were observed (spherical vs tube-like blastoconidia, or solid and fragile constricted septa vs only the latter observed in strains with EFG1 deleted). In addition, wild type strain displayed smooth colonies of cells in comparison to mutants which exhibited wrinkled phenotype. It was observed that blastoconidia of clinical strain exhibited either polarly or randomly located budding. Contrariwise, morphotypes of mutants showed either multiple polar budding or a centrally located single bud scar (mother-daughter cell junction) distinguishing tube-like yeast/ pseudohyphal growth (the length-to-width ratios larger than 1.5). In their planktonic form of growth, blastoconidia of clinical bloodstream isolate formed constitutively true hyphae under undiluted human serum inducing conditions. It was found that true hyphae are essential elements for developing structural integrity of conglomerate, as mutants displaying defects in their flocculation and conglomerate-forming abilities in serum. While filamentation is an important virulence trait in C. albicans the true hyphae are the morphologies which may be expected to play a role in bloodstream infections.(AU)

Cancer drugs inhibit morphogenesis in the human fungal pathogen, Candida albicans

Candida infections are very common in cancer patients and it is a common practice to prescribe antifungal antibiotics along with anticancer drugs. Yeast to hyphal form switching is considered to be important in invasive candidiasis. Targeting morphogenetic switching may be useful against invasive candidiasis. In this study, we report the antimorphogenetic properties of thirty cancer drugs.(AU)

Effect of alcohols on filamentation, growth, viability and biofilm development in Candida albicans

In this study we report the potential of alcohols as morphogenetic regulators in Candida albicans. All the alcohols tested influenced various modes of growth like planktonic as well as biofilm forms. Viability was affected at high concentrations. Among the alcohols, the response of C. albicans to amyl alcohol (pentanol) was noteworthy. Amyl alcohol at a concentration 0.5% which was not inhibitory to growth and viability specifically inhibited morphogenetic switching from yeast to hyphal forms. It also inhibited normal biofilm development favoring yeast dominated biofilms. Based on this study we hypothesize that alcohols produced under anaerobic conditions may not favor biofilm development and support dissemination of yeast cells. Since anaerobic conditions are not found to favor production of quorum sensing molecules like farnesol, the alcohols may play a role in morphogenetic regulation.

Effect of alcohols on filamentation, growth, viability and biofilm development in Candida albicans

In this study we report the potential of alcohols as morphogenetic regulators in Candida albicans. All the alcohols tested influenced various modes of growth like planktonic as well as biofilm forms. Viability was affected at high concentrations. Among the alcohols, the response of C. albicans to amyl alcohol (pentanol) was noteworthy. Amyl alcohol at a concentration 0.5% which was not inhibitory to growth and viability specifically inhibited morphogenetic switching from yeast to hyphal forms. It also inhibited normal biofilm development favoring yeast dominated biofilms. Based on this study we hypothesize that alcohols produced under anaerobic conditions may not favor biofilm development and support dissemination of yeast cells. Since anaerobic conditions are not found to favor production of quorum sensing molecules like farnesol, the alcohols may play a role in morphogenetic regulation.

COMBINAÇÃO DE TÉCNICAS FENOTÍPICAS PARA DISTINGUIR Candida albicans DE Candida dubliniensis E UMA INVESTIGAÇÃO DOS MECANISMOS DE RESISTÊNCIA A DERIVADOS AZÓLICOS EM ISOLADOS DE Candida albicans DE ORIGEM ANIMAL

Leveduras do gênero Candida constituem os principais patógenos fúngicos para homens e animais. Embora Candida albicans ainda ocupe uma posição de destaque entre os casos de infeções, espécies não-albicans vem tendo importância aumentada nas últimas décadas. Além disso, isolados de C. albicans resistentes a derivados azólicos tem sido descritos na prática clínica humana e em cepas de origem veterinária. O presente estudo teve como objetivo reabordar a identificação laboratorial de isolados veterinários de C. albicans resistentes a derivados azólicos, propondo combinação de métodos fenotípicos para distinguir as espécies crípticas C. albicans e Candida dubliniensis. Além disso, almejou-se investigar os mecanismos de resistência das cepas estudadas. Para tanto, trinta e sete cepas veterinárias de C. albicans resistentes a derivados azólicos e três cepas humanas de C. dubliniensis foram submetidas à identificação fenotípica por prova do tubo germinativo, microcultivo em ágar cornmeal, semeadura em ágar cromogênico, semeadura em ágar semente de girassol, prova de opacificação em ágar Tween 80 e reação em cadeia da polimerase (PCR) espécie-específica. Para estudo dos mecanismos de resistência, cepas de C. albicans foram estudadas com testes de efluxo de rodamina 6G (n = 11), determinação do conteúdo de ergosterol (n = 6) e testes de expressão dos genes CDR1, CDR2, MDR1 e ERG11 (n = 30). As cepas estudadas apresentaram prova do tubo germinativo positiva e produção de clamidoconídios em ágar cornmeal. Trinta e cinco cepas (35/37) apresentaram coloração verde em ágar cromogênico. Em ágar semente de girassol, 36 cepas (36/37) apresentaram padrão de colônia lisa, e no ágar Tween 80, 34 (34/37) apresentaram opacificação em menos de 5 dias. Todas as cepas testadas apresentaram produto específico na PCR, sendo confirmadas como C. albicans. As cepas de C. dubliniensis apresentaram colônias de coloração verde no ChromoAgar Candida, aspecto rugoso no ágar girassol, não opacificaram o meio em ágar Tween 80 em até 12 dias e não apresentaram produto específico na PCR. Nos estudos de resistência, as cepas resistentes aos azólicos apresentaram valores de diferença de unidade de fluorescência relativa maiores que os isolados sensíveis nos estudos com rodamina 6G, demonstrando assim maior atividade de bombas de efluxo ATP-dependentes. Não houve diferença significativa no conteúdo de ergosterol entre as cepas estudadas. Foi verificado que 73,3% (22/30) dos isolados apresentaram superexpressão de um ou mais genes, sendo 40,9%, 18,2%, 59,1% e 54,5% superexpressão de CDR1, CDR2, MDR1 e ERG11, respectivamente. Nenhum isolado superexpressou os quatro genes simultaneamente. Por fim, conclui-se que a combinação de técnicas fenotípicas leva à distinção confiável entre C. albicans e C. dubliniensis, reservando as técnicas moleculares para contextos mais específicos. O padrão de resistência a derivados azólicos apresentado pelas cepas estudadas é multifatorial, incluindo, no mínimo, atividade elevada de bombas de efluxo e a superexpressão gênica.

Yeasts of genus Candida are the main fungal pathogens for humans and animals. Although Candida albicans still occupies a prominent position among cases of infections, non-albicans species have increased in the last decades. In addition, azole-resistant C. albicans have been described in human clinical practice and in strains of veterinary origin. The objective of this study was to reevaluate laboratory identification of azole-resistant C. albicans from veterinary isolates, proposing a combination of phenotypic methods for the reliable identification of the species. In addition, this study aimed to investigate the resistance mechanisms of the strains. Thirty-seven veterinary strains of azole-resistant C. albicans and three human strains of C. dubliniensis were submitted to phenotypic identification by germ tube test, micromorphological analysis on cornmeal agar, growth on chromogenic agar, growth on sunflower seed agar, opacification test on Tween 80 agar and species-specific PCR. For the study of resistance mechanisms, C. albicans strains were evaluated for efflux of 6G rhodamine (n=11), determination of ergosterol content (n=6) and expression of CDR1, CDR2, MDR1 and ERG11 genes (n=30). All studied strains presented positive germ tube test and production of chlamydoconidia on cornmeal agar. Thirty five strains (35/37) showed green colonies on chromogenic agar. On sunflower seed agar, 36 strains (36/37) presented a smooth colony pattern, and on Tween 80 agar, 34 (34/37) had opacification in less than 5 days. All strains tested showed specific product on PCR, which confirmed their identity as C. albicans. C. dubliniensis strains showed green colonies on ChromoAgar Candida, a rough appearance on sunflower agar, did not form opacification zone on Tween 80 agar, after up to 12 days, and did not present specific products in PCR. In resistance studies, azole resistant strains showed higher relative fluorescence unit difference values than sensitive isolates in efflux of 6G rhodamine tests, thus demonstrating increased activity of ATP-dependent efflux pumps. There was no significant difference in ergosterol content among the studied strains. It was verified that 73.3% (22/30) of the isolates presented overexpression of one or more genes, of which 40.9%, 18.2%, 59.1% and 54.5% strains overexpressed CDR1, CDR2, MDR1 and ERG11, respectively. No isolate overexpressed simultaneously the four genes. Finally, we conclude that the combination of phenotypic techniques leads to reliable differentiation of C. albicans and C. dubliniensis, reserving molecular techniques for more specific contexts. Resistance mechanisms to azole derivatives of the studied strains is multifactorial, including, at a minimum, the enhanced activity of efflux pump activity and the gene overexpression.

Candidemia in a brazilian tertiary hospital: microbiological and clinical features over a six-year period

Yeasts are becoming a common cause of nosocomial fungal infections in immunocompromised patients. Such infections often develop into sepsis with high mortality rates. The aim of this study was to evaluate some of the numerous factors associated with the development of candidemia. Medical records were retrospectively analyzed of 98 Candida spp. patients. Results showed that the most prevalent risk factors for developing candidemia were: antibiotics and antifungal agents (93.9% and 79.6%, respectively); the use of central venous catheter (93.9%); mechanical ventilation (73.5%); and parenteral nutrition (60.2%). The main species of Candida found were: C. parapsilosis (37.76%), C. albicans (33.67%); and others (28.57%). C. glabrata showed the highest mortality rate (75%), followed by C. tropicalis (57.1%) and C. albicans (54.5%). The elevated mortality rate found in this study indicates that preventive measures against candidemia must be emphasized in hospitals.(AU)

Candidemia in a brazilian tertiary hospital: microbiological and clinical features over a six-year period

Yeasts are becoming a common cause of nosocomial fungal infections in immunocompromised patients. Such infections often develop into sepsis with high mortality rates. The aim of this study was to evaluate some of the numerous factors associated with the development of candidemia. Medical records were retrospectively analyzed of 98 Candida spp. patients. Results showed that the most prevalent risk factors for developing candidemia were: antibiotics and antifungal agents (93.9% and 79.6%, respectively); the use of central venous catheter (93.9%); mechanical ventilation (73.5%); and parenteral nutrition (60.2%). The main species of Candida found were: C. parapsilosis (37.76%), C. albicans (33.67%); and others (28.57%). C. glabrata showed the highest mortality rate (75%), followed by C. tropicalis (57.1%) and C. albicans (54.5%). The elevated mortality rate found in this study indicates that preventive measures against candidemia must be emphasized in hospitals.(AU)

Antimicrobial activity of meta-alkoxyphenylcarbamates containing substituted N-phenylpiperazine fragment

In the present investigation, the basic esters of meta-alkoxyphenylcarbamic acid bearing variously substituted N-phenylpiperazine fragment were screened for their in vitro antimicrobial activity against Staphylococcus aureus, Escherichia coli and Candida albicans, respectively. The most effective against Escherichia coli was found the compound 6d (MIC=195,3 g/mL) bearing simultaneously para-fluoro substituent at the 4phenylpiperazin-1-yl core and meta-methoxy side chain in the lipophilic part of the molecule. From whole analyzed set of the molecules the substance 8e with propoxy side chain forming meta-alkoxyphenylcarbamoyl fragment and lipophilic, sterically bulky meta-trifluoromethyl group attached at N-phenylpiperazine moiety was evaluated as the most active against Candida albicans (MIC=97,7 g/mL). On the contrary, all investigated structures were practically inactive against Staphylococcus aureus (MIC>1000 g/mL)

In vitro study on the antimicrobial effect of hydroalcoholic extracts from Mentha arvensis L. (Lamiaceae) against oral pathogens / Estudo in vitro do efeito antimicrobiano dos extratos hidroalcólicos de Mentha arvensis L. (Lamiaceae) contra patógenos orais

In vitro tests could be a valuable tool for the evaluation of medicinal plants' antimicrobial activity. Mentha arvensis of the Lamiaceae family is one of the most frequently traditional plants used in Brazil. Hydroalcoholic extracts of M. arvensis were analyzed for antimicrobial activity on Streptococcus mutans, Streptococcus sobrinus and Candida albicans. Three different assays (agar diffusion, broth macro- and micro-dilution methods) were used to evaluate antimicrobial activity. Although hydroalcoholic extracts of M. arvensis did not show any antibacterial effect, its antifungal activity against C. albicans was revealed. According to the micro-dilution broth assay, MIC of the hydroalcoholic extract from leaves of M. arvensis on Candida albicans strains ranged between 625 and 2500 µg mL-1. Results suggest that M. arvensis hydroalcoholic extract may be considered a potentially antifungal agent against C. albicans, and a possible item for human antibiotic therapy. However, further biological tests on the plant's efficacy and side-effects are necessary before its use on humans.

Testes in vitro podem ser uma ferramenta valiosa para a avaliação da atividade antimicrobiana de plantas medicinais. Mentha arvensis é uma das plantas medicinais brasileiras mais frequentemente utilizadas e pertence à família Lamiaceae. No presente estudo, extratos hidroalcólicos de M. arvensis foram analisados quanto à sua atividade antimicrobiana sobre Streptococcus mutans, Streptococcus sobrinus e Candida albicans. Três diferentes ensaios (métodos de difusão em ágar, macro e microdiluição em caldo) foram utilizados para avaliação da atividade antimicrobiana. Embora os extratos hidroalcólicos de M. arvensis não demonstraram qualquer efeito antibacteriano, eles apresentaram atividade antifúngica contra C. albicans. Baseado no ensaio de microdiluição em caldo, a CIM do extrato hidroalcólico das folhas de M. arvensis sobre cepas de C. albicans variaram de 625 a 2500 µg mL-1. Estes achados sugerem que o extrato hidroalcólico de M. arvensis pode ser considerado um agente antifúngico em potencial contra C. albicans, e um possível candidato para antibioticoterapia humana. Contudo, mais testes biológicos sobre a eficácia e efeitos adversos desta planta são necessários antes do seu uso em humanos.