Resumo

Abstract In the current context of emerging drug-resistant fungal pathogens such as Candida albicans and Candida parapsilosis, discovery of new antifungal agents is an urgent matter. This research aimed to evaluate the antifungal potential of 2-chloro-N-phenylacetamide against fluconazole-resistant clinical strains of C. albicans and C. parapsilosis. The antifungal activity of 2-chloro-N-phenylacetamide was evaluated in vitro by the determination of the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), inhibition of biofilm formation and its rupture, sorbitol and ergosterol assays, and association between this molecule and common antifungal drugs, amphotericin B and fluconazole. The test product inhibited all strains of C. albicans and C. parapsilosis, with a MIC ranging from 128 to 256 µg.mL-1, and a MFC of 512-1,024 µg.mL-1. It also inhibited up to 92% of biofilm formation and rupture of up to 87% of preformed biofilm. 2-chloro-N-phenylacetamide did not promote antifungal activity through binding to cellular membrane ergosterol nor it damages the fungal cell wall. Antagonism was observed when combining this substance with amphotericin B and fluconazole. The substance exhibited significant antifungal activity by inhibiting both planktonic cells and biofilm of fluconazole-resistant strains. Its combination with other antifungals should be avoided and its mechanism of action remains to be established.

Resumo No atual contexto de patógenos fúngicos resistentes emergentes tais como Candida albicans e Candida parapsilosis, a descoberta de novos agentes antifúngicos é uma questão urgente. Esta pesquisa teve como objetivo avaliar o potencial antifúngico da 2-cloro-N-fenilacetamida contra cepas clínicas de C. albicans e C. parapsilosis resistentes a fluconazol. A atividade antifúngica da substância foi avaliada in vitro através da determinação da concentração inibitória mínima (CIM), concentração fungicida mínima (CFM), ruptura e inibição da formação de biofilme, ensaios de sorbitol e ergosterol, e associação entre esta molécula e antifúngicos comuns, anfotericina B e fluconazol. O produto teste inibiu todas as cepas de C. albicans e C. parapsilosis, com uma CIM variando de 128 a 256 µg.mL-1, e uma CFM de 512-1,024 µg.mL-1. Também inibiu até 92% da formação de biofilme e causou a ruptura de até 87% de biofilme pré-formado. A 2-cloro-N-fenilacetamida não promoveu atividade antifúngica pela ligação ao ergosterol da membrana celular fúngica, tampouco danificou a parede celular. Antagonismo foi observado ao combinar esta substância com anfotericina B e fluconazol. A substância exibiu atividade antifúngica significativa ao inibir tanto as células planctônicas quanto o biofilme das cepas resistentes ao fluconazol. Sua combinação com outros antifúngicos deve ser evitada e seu mecanismo de ação deve ser estabelecido.

Resumo

In the current context of emerging drug-resistant fungal pathogens such as Candida albicans and Candida parapsilosis, discovery of new antifungal agents is an urgent matter. This research aimed to evaluate the antifungal potential of 2-chloro-N-phenylacetamide against fluconazole-resistant clinical strains of C. albicans and C. parapsilosis. The antifungal activity of 2-chloro-N-phenylacetamide was evaluated in vitro by the determination of the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), inhibition of biofilm formation and its rupture, sorbitol and ergosterol assays, and association between this molecule and common antifungal drugs, amphotericin B and fluconazole. The test product inhibited all strains of C. albicans and C. parapsilosis, with a MIC ranging from 128 to 256 µg.mL-1, and a MFC of 512-1,024 µg.mL-1. It also inhibited up to 92% of biofilm formation and rupture of up to 87% of preformed biofilm. 2-chloro-N-phenylacetamide did not promote antifungal activity through binding to cellular membrane ergosterol nor it damages the fungal cell wall. Antagonism was observed when combining this substance with amphotericin B and fluconazole. The substance exhibited significant antifungal activity by inhibiting both planktonic cells and biofilm of fluconazole-resistant strains. Its combination with other antifungals should be avoided and its mechanism of action remains to be established.

No atual contexto de patógenos fúngicos resistentes emergentes tais como Candida albicans e Candida parapsilosis, a descoberta de novos agentes antifúngicos é uma questão urgente. Esta pesquisa teve como objetivo avaliar o potencial antifúngico da 2-cloro-N-fenilacetamida contra cepas clínicas de C. albicans e C. parapsilosis resistentes a fluconazol. A atividade antifúngica da substância foi avaliada in vitro através da determinação da concentração inibitória mínima (CIM), concentração fungicida mínima (CFM), ruptura e inibição da formação de biofilme, ensaios de sorbitol e ergosterol, e associação entre esta molécula e antifúngicos comuns, anfotericina B e fluconazol. O produto teste inibiu todas as cepas de C. albicans e C. parapsilosis, com uma CIM variando de 128 a 256 µg.mL-1, e uma CFM de 512-1,024 µg.mL-1. Também inibiu até 92% da formação de biofilme e causou a ruptura de até 87% de biofilme pré-formado. A 2-cloro-N-fenilacetamida não promoveu atividade antifúngica pela ligação ao ergosterol da membrana celular fúngica, tampouco danificou a parede celular. Antagonismo foi observado ao combinar esta substância com anfotericina B e fluconazol. A substância exibiu atividade antifúngica significativa ao inibir tanto as células planctônicas quanto o biofilme das cepas resistentes ao fluconazol. Sua combinação com outros antifúngicos deve ser evitada e seu mecanismo de ação deve ser estabelecido.

Resumo

Candida albicans is the main fungal species involved in oral candidiasis, and its increasing resistance to pharmacological treatment encourages the search for improved antifungal agents. Lavandula dentata L. essential oil (LD-EO) has been recognized for its antimicrobial activity, but little is known about its role against oral C. albicans. This study evaluated the antifungal and antibiofilm activities, mechanisms of action, and toxicity of LD-EO from Brazil against oral strains of C. albicans. Antifungal activity was assessed based on Minimum Inhibitory Concentration (MIC), Minimum Fungicidal Concentration (MFC), association study with miconazole (Checkerboard method), and sorbitol and ergosterol assays. Inhibition of biofilm formation and disruption of preformed biofilm were considered when studying the effects of the product. Additionally, the toxicity of LD-EO was evaluated by a hemolysis assay on human erythrocytes. Phytochemical analysis by gas chromatography-mass spectrometry identified eucalyptol (33.1%), camphor (18.3%), and fenchone (15.6%) as major constituents. The test substance showed mainly fungicidal activity (MIC100 = 8 µg/mL; MFC = 16 µg/mL), including against two miconazole-resistant isolates of C. albicans. The effects of LD-EO were synergistic with those of miconazole and appeared not to involve damage to the fungal cell wall or plasma membrane. Its effectiveness in inhibiting biofilm formation was higher than the effect of disrupting preformed biofilm. Finally, the product exhibited low hemolytic activity at MIC. Based on the favorable and novel results described here, LD-EO could constitute a promising therapeutic alternative for oral candidiasis, including miconazole-resistant cases.

Candida albicans é a principal espécie fúngica envolvida na candidíase bucal, e sua crescente resistência ao tratamento farmacológico encoraja a busca por melhores agentes antifúngicos. O óleo essencial da Lavandula dentata L. (LD-EO) tem sido reconhecido por sua atividade antimicrobiana, porém pouco se conhece o seu papel contra C. albicans bucal. Este trabalho avaliou as atividades antifúngica e antibiofilme, mecanismos de ação e toxicidade do LD-EO do Brasil contra cepas de C. albicans bucal. A atividade antifúngica foi avaliada baseando-se em: Concentração Inibitória Mínima (CIM), Concentração Fungicida Mínima (CFM), estudo de associação com o miconazol (método Checkerboard) e ensaios com sorbitol e ergosterol. A inibição da formação do biofilme e a ruptura do biofilme pré-formado foram considerados no estudo dos efeitos do produto. Adicionalmente, a toxicidade do LD-EO foi avaliada pelo ensaio de hemólise em eritrócitos humanos. A análise fitoquímica por cromatografia gasosa-espectrometria de massa identificou eucaliptol (33.1%), cânfora (18.3%) e fenchona (15.6%) como constituintes majoritários. A substância teste revelou atividade principalmente fungicida (CIM100 = 8 µg/mL; CFM = 16 µg/mL), inclusive contra dois isolados de C. albicans resistentes ao miconazol. Os efeitos do LD-EO foram sinérgicos aos do miconazol e parecem não envolver danos à parede celular ou à membrana plasmática fúngica. Sua efetividade em inibir a formação do biofilme foi maior que o efeito de eliminação do biofilme pré-formado. Finalmente, o produto exerceu baixa atividade hemolítica na CIM. Considerando os resultados favoráveis e inéditos aqui descritos, o LD-EO poderia constituir uma alternativa terapêutica promissora para a candidíase bucal, incluindo casos resistentes ao miconazol.

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Cats are susceptible to S. aureus, which mainly colonizes the nose and ears of these feline species. Otitis externa in cat ears is one of the illnesses produced by S. aureus in animals. Antibiotic therapy for affected animals is the conventional treatment for infections by S. aureus. Antibiotic use during prolonged treatment and given at the wrong doses can cause germs to become resistant. Given this context, research on S. aureus isolated from cat ears and tests for antibiotic resistance and the mecA gene is required. Samples of cat ears were obtained from the Amies media using a sterile cotton swab. Bacterial isolation was done on MSA media, and then the catalase and coagulase assays were used to identify the bacteria. S. aureus isolates were evaluated for sensitivity using disks of the antibiotics cefoxitin, tetracycline, erythromycin, gentamicin, and chloramphenicol connected to MHA media. All positive isolates of S. aureus underwent MRSA testing, and then the mecA gene was detected. The sample investigation revealed that 91% (91/100) were positive for S. aureus, and 3.30% (3/91) were confirmed to be multidrug-resistant (MDR) because they are resistant to 3­4 antibiotic classes. Out of the 12 MRSA isolates analyzed, the mecA gene was detected in one isolate. Inappropriate antibiotic use causes bacterial resistance in pets. Additionally, excessive antibiotic use in a population might develop acquired bacterial resistance to an antibiotic. Antibiotic use in animals must be assessed to administer medication and prevent the development of antibiotic resistance appropriately.(AU)

Gatos são suscetíveis a adquirir S.aureus que colonizam principalmente as narinas e os ouvidos de espécies de felinos. A otite externa no ouvido dos gatos é uma das doenças produzidas pelo S.aureus nos animais. A terapia com antibióticos é o tratamento convencional para as infecções produzidas pelo S.aureus. Os antibióticos utilizados durante o prolongado tratamento e o emprego de sub doses podem selecionar microorganismos resistentes. Com base em tais argumentos torna-se necessária a pesquisa de S.aureus isolados do ouvido dos gatos, bem como, a realização de testes para a resistência a antibióticos e do gene mecA. Empregando swabs estéreis de algodão foram obtidas amostras dos ouvidos dos gatos em meio de Amies. O isolamento bacteriano foi efetuado em meio MAS e os testes catalase e coagulase foram realizados para a identificação das bactérias. A sensibilidade dos isolados de S.aureus foi avaliada com o emprego de discos dos antibióticos cefoxitin, tetraxiclina, eritromicina, gentamicina e cloranfenicol, incorporados no meio MHA. Todos os isolados positivos de S.aureus foram submetidos ao test MRSA para a detecção do gene mecA. A amostra investigada revelou 91% (91/100) de positivos para S.aureus, dos quais, 3,30% (3/91) foram resistentes a múltiplas drogas (MDR) pois foram resistentes a 3-4 classes de antibióticos. De 12 MRSA isolados analisados o gene mecA foi detectado em um isolado. O uso inapropriado de antibióticos é a causa da resistência bacteriana em pets. Adicionalmente o emprego excessivo de antibióticos em uma população pode resultar no desenvolvimento de resistência bacteriana adquirida a antibióticos. O uso de antibióticos em animais deve ser ordenado por uma administração de medicamentos apropriada para prevenir o desenvolvimento da resistência.(AU)

Resumo

Infectious diseases are the leading cause of death worldwide. Thus, nanotechnology provides an excellent opportunity to treat drug-resistant microbial infections. Numerous antibiotics have been used to inhibit the growth and kill of microbes, but the development of resistance and the emergence of side effects have severely limited the use of these agents. Due to the development of the nanotechnology, nanoparticles are widely used as antimicrobials. Silver and chitosan nanoparticles have antifungal, antiviral and antibacterial properties, and many studies confirm the antifungal properties of silver nanoparticles. Nowadays, the use of nanoparticles in the diagnosis and treatment of infectious diseases has developed due to less side effects and also the help of these particles in effective drug delivery to the target tissue. Liposomes are also used as carriers of drug delivery, genes, and modeling of cell membranes in both animals and humans. The ability of these liposomes to encapsulate large amounts of drugs, minimize unwanted side effects, high effectiveness and low toxicity has attracted the interest of researchers. This review article examines recent efforts by researchers to identify and treat infectious diseases using antimicrobial nanoparticles and drug nano-carriers.

As doenças infecciosas são a principal causa de morte no mundo. Assim, a nanotecnologia oferece uma excelente oportunidade para tratar infecções microbianas resistentes a medicamentos. Numerosos antibióticos têm sido usados para inibir o crescimento e a morte de micróbios, mas o desenvolvimento de resistência e o surgimento de efeitos colaterais limitaram severamente o uso desses agentes. Devido ao desenvolvimento da nanotecnologia, as nanopartículas são amplamente utilizadas como antimicrobianos. As nanopartículas de prata e quitosana têm propriedades antifúngicas, antivirais e antibacterianas, e muitos estudos confirmam as propriedades antifúngicas das nanopartículas de prata. Atualmente, o uso de nanopartículas no diagnóstico e tratamento de doenças infecciosas tem se desenvolvido em razão do menor número de efeitos colaterais e também à ajuda dessas partículas na efetiva entrega de fármacos ao tecido-alvo. Os lipossomas também são usados como transportadores de entrega de drogas, genes e modelagem de membranas celulares em animais e humanos. Os lipossomas também são usados como transportadores de entrega de drogas, genes e modelagem de membranas celulares em animais e humanos. A capacidade desses lipossomas de encapsular grandes quantidades de fármacos, minimizar efeitos colaterais indesejados, alta eficácia e baixa toxicidade tem despertado o interesse de pesquisadores. Este artigo de revisão examina esforços recentes de pesquisadores para identificar e tratar doenças infecciosas usando nanopartículas antimicrobianas e nanotransportadores de drogas.

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The increasing use of antimicrobial drugs has been linked to the rise of drug-resistant fungus in recent years. Antimicrobial resistance is being studied from a variety of perspectives due to the important clinical implication of resistance. The processes underlying this resistance, enhanced methods for identifying resistance when it emerges, alternate treatment options for infections caused by resistant organisms, and so on are reviewed, along with strategies to prevent and regulate the formation and spread of resistance. This overview will focus on the action mechanism of antifungals and the resistance mechanisms against them. The link between antibacterial and antifungal resistance is also briefly discussed. Based on their mechanism action, antifungals are divided into three distinct categories: azoles, which target the ergosterol synthesis; 5-fluorocytosine, which targets macromolecular synthesis and polyenes, which interact physiochemically with fungal membrane sterols. Antifungal resistance can arise through a wide variety of ways. Overexpression of the target of the antifungal drug, changes to the drug target, changes to sterol biosynthesis, decreased intercellular concentration of the target enzyme, and other processes. A correlation exists between the mechanisms of resistance to antibacterial and antifungals, despite the fact that the comparison between the two is inevitably constrained by various parameters mentioned in the review. Drug extrusion via membrane pumps has been thoroughly documented in both prokaryotic and eukaryotic cells, and development of new antifungal compounds and strategies has also been well characterized.

O uso crescente de medicamentos antimicrobianos tem sido associado ao aumento de fungos resistentes aos medicamentos nos últimos anos. A resistência antimicrobiana está sendo estudada sob diversas perspectivas devido à importante implicação clínica da resistência. Os processos subjacentes a esta resistência, os métodos melhorados para identificar a resistência quando esta surge, as opções alternativas de tratamento para infecções causadas por organismos resistentes, etc., são revistos, juntamente com estratégias para prevenir e regular a formação e propagação da resistência. Esta visão geral focará no mecanismo de ação dos antifúngicos e nos mecanismos de resistência contra eles. A ligação entre resistência antibacteriana e antifúngica também é brevemente discutida. Com base no seu mecanismo de ação, os antifúngicos são divididos em três categorias distintas: azóis, que têm como alvo a síntese do ergosterol; 5-fluorocitosina, que tem como alvo a síntese macromolecular, e polienos, que interagem físicoquimicamente com esteróis da membrana fúngica. A resistência antifúngica pode surgir de várias maneiras. Superexpressão do alvo do medicamento antifúngico, alterações no alvo do medicamento, alterações na biossíntese de esterol, diminuição da concentração intercelular da enzima-alvo e outros processos. Existe uma correlação entre os mecanismos de resistência aos antibacterianos e aos antifúngicos, apesar de a comparação entre os dois ser inevitavelmente limitada por vários parâmetros mencionados na revisão. A extrusão de fármacos através de bombas de membrana foi exaustivamente documentada em células procarióticas e eucarióticas, e o desenvolvimento de novos compostos e estratégias antifúngicas também foi bem caracterizado.

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ABSTRACT: The constituents of the hydroethanolic extract ofAzadiractaindicaroot were investigated using ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-QTOOF-MSE). Acute toxicity was evaluated in an experimental animal model. We investigated the antibacterial activities ofA. indicaroots againstSalmonella typhimuriumandStaphylococcus aureusand the antifungal activities against strains ofTrichophyton rubrum, Candida albicansandCandida tropicalis. We identified nine secondary metabolites in the hydroethanolic extract by UPLC-QTOOF-MSE. The extract was highly effective in inhibiting the growth of T. rubrum strains, so it can be effective in combating the dermatophyte tested,but it had no inhibition potential on any bacterial strains orCandidaspecies evaluated. It was possible to infer that the extract had no acute toxicity in relation to the animal model Danio rerio. Therefore, since neem has a high bioactive potential and adapts well to the climate of semiarid regions, growing this species could become a source of income for farmers by its use to produce naturals fungicide and drug, as alternatives to conventional products, which can cause microbiological resistance and/or are toxic to the environment, besides being expensive.

RESUMO: Os constituintes do extrato hidroetanólico da raiz deA. indicaforam investigados por cromatografia líquida de ultra-alta performance acoplada à espectrometria de massas do tipo quadrupolo-tempo de voo (UPLC-QTOOF-MSE). A toxicidade aguda foi avaliada em modelo animal. Investigamos as atividades antibacterianas contra Salmonella typhimuriumeStaphylococcus aureuse as atividades antifúngicas contra cepas deTrichophyton rubrum, Candida albicanseCandida tropicalis. Identificamos nove metabólitos secundários no extrato etanólico por UPLC-QTOOF-MSE. O extrato foi altamente eficaz na inibição do crescimento de cepas de T. rubrum, podendo ser eficaz no combate ao dermatofito avaliado, mas não apresentou potencial de inibição em nenhuma cepa bacteriana ou espécies deCandidaavaliadas. Também foi possível inferir que o extrato não apresentou toxicidade aguda em relação ao modelo animalDanio rerio. Portanto, como o Neem tem alto potencial bioativo e se adapta bem ao clima das regiões semiáridas, o cultivo dessa espécie pode se tornar uma fonte de renda para os agricultores a partir da utilização da planta para produção de fungicida e/ou fármaco naturais como alternativa aos produtos convencionais, que podem causar resistência microbiológica e/ou são tóxicas ao meio ambiente, além de serem caros.

Resumo

The constituents of the hydroethanolic extract ofAzadiractaindicaroot were investigated using ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-QTOOF-MSE). Acute toxicity was evaluated in an experimental animal model. We investigated the antibacterial activities ofA. indicaroots againstSalmonella typhimuriumandStaphylococcus aureusand the antifungal activities against strains ofTrichophyton rubrum, Candida albicansandCandida tropicalis. We identified nine secondary metabolites in the hydroethanolic extract by UPLC-QTOOF-MSE. The extract was highly effective in inhibiting the growth of T. rubrum strains, so it can be effective in combating the dermatophyte tested,but it had no inhibition potential on any bacterial strains orCandidaspecies evaluated. It was possible to infer that the extract had no acute toxicity in relation to the animal model Danio rerio. Therefore, since neem has a high bioactive potential and adapts well to the climate of semiarid regions, growing this species could become a source of income for farmers by its use to produce naturals fungicide and drug, as alternatives to conventional products, which can cause microbiological resistance and/or are toxic to the environment, besides being expensive.

Os constituintes do extrato hidroetanólico da raiz deA. indicaforam investigados por cromatografia líquida de ultra-alta performance acoplada à espectrometria de massas do tipo quadrupolo-tempo de voo (UPLC-QTOOF-MSE). A toxicidade aguda foi avaliada em modelo animal. Investigamos as atividades antibacterianas contra Salmonella typhimuriumeStaphylococcus aureuse as atividades antifúngicas contra cepas deTrichophyton rubrum, Candida albicanseCandida tropicalis. Identificamos nove metabólitos secundários no extrato etanólico por UPLC-QTOOF-MSE. O extrato foi altamente eficaz na inibição do crescimento de cepas de T. rubrum, podendo ser eficaz no combate ao dermatofito avaliado, mas não apresentou potencial de inibição em nenhuma cepa bacteriana ou espécies deCandidaavaliadas. Também foi possível inferir que o extrato não apresentou toxicidade aguda em relação ao modelo animalDanio rerio. Portanto, como o Neem tem alto potencial bioativo e se adapta bem ao clima das regiões semiáridas, o cultivo dessa espécie pode se tornar uma fonte de renda para os agricultores a partir da utilização da planta para produção de fungicida e/ou fármaco naturais como alternativa aos produtos convencionais, que podem causar resistência microbiológica e/ou são tóxicas ao meio ambiente, além de serem caros.

Resumo

The aim of the current report is to describe a cutaneous candidiasis case affecting a canine individual treated at the University Veterinary Hospital of State University of Maranhão (UEMA), in São Luís City. The patient had three-month history of skin diseases; it had been previously subjected to several treatments based on antibiotics, corticosteroids and antifungal drugs that have failed to show clinical improvements. Dermatological assessment has indicated generalized moist dermatitis, intense skin desquamation, alopecia, pruritus and meliceric crusts along the animal's body, mainly in its dorsal region. Complementary tests, such as skin cytology and microscopy, trichogram, qualitative PCR and serology for canine visceral leishmaniasis, as well as fungal culture and antifungigram were requested based on this scenario. Serology recorded inconclusive results for leishmaniasis, whereas PCR recorded negative results in the presence of the agent's DNA. Cytology, microscopy and trichogram results have evidenced fungal infection in the assessed samples. Moreover, mycological culture and antifungigram resulted in the growth of Candida sp. specimens capable of resisting antifungal agents such as amphotericin B, fluconazole, itraconazole and nystatin. The therapy adopted after candidiasis diagnosis confirmation comprised oral doses of manipulated ketoconazole, in combination to topical therapy with shampoo based on moisturizing formulas associated with Miconazole and Chlorhexidine (at 2%), for four weeks. After 30 days, when the adopted therapy was over, the aforementioned animal presented remission of the previously observed lesions and fully improved condition.

The aim of the current report is to describe a cutaneous candidiasis case affecting a canine individual treated at the University Veterinary Hospital of State University of Maranhão (UEMA), in São Luís City. The patient had three-month history of skin diseases; it had been previously subjected to several treatments based on antibiotics, corticosteroids and antifungal drugs that have failed to show clinical improvements. Dermatological assessment has indicated generalized moist dermatitis, intense skin desquamation, alopecia, pruritus and meliceric crusts along the animal's body, mainly in its dorsal region. Complementary tests, such as skin cytology and microscopy, trichogram, qualitative PCR and serology for canine visceral leishmaniasis, as well as fungal culture and antifungigram were requested based on this scenario. Serology recorded inconclusive results for leishmaniasis, whereas PCR recorded negative results in the presence of the agent's DNA. Cytology, microscopy and trichogram results have evidenced fungal infection in the assessed samples. Moreover, mycological culture and antifungigram resulted in the growth of Candida sp. specimens capable of resisting antifungal agents such as amphotericin B, fluconazole, itraconazole and nystatin. The therapy adopted after candidiasis diagnosis confirmation comprised oral doses of manipulated ketoconazole, in combination to topical therapy with shampoo based on moisturizing formulas associated with Miconazole and Chlorhexidine (at 2%), for four weeks. After 30 days, when the adopted therapy was over, the aforementioned animal presented remission of the previously observed lesions and fully improved condition.

Resumo

Sporotrichosis is a chronic fungal infection caused by Sporothrix species. The occurrence of cases that are resistant to long-term treatment, especially in the nasal planum of cats, emphasizes the importance of studying its pathogenesis. The purpose of this study was to analyze and compare the inflammatory process of cutaneous lesions of feline refractory sporotrichosis to clinical aspects through cytopathological and histopathological examination. Moreover, the study included 13 cats with cutaneous lesions that had been resistant to itraconazole treatment for more than a year. Cutaneous lesions samples were collected for cytopathological, histopathological, and fungal culture analyses. Tissue fragments were processed and stained with hematoxylin-eosin (HE) and Grocott methenamine silver (GMS). Further, two clinical presentations had the highest occurrence: the localized cutaneous form in animals with good general condition and stable disease (n=9, 69.2%) and the disseminated cutaneous form in cats with poor general condition (n=4, 30.8%). In cats with refractory sporotrichosis, the nasal planum (84.6%) was the most common location of lesions. In the cytopathological study, cats with fewer than two lesions and in good general condition (n=9, 69.2%) showed absence or mild yeast intensity (up to 5 yeasts per field), lower intensity of macrophages and neutrophils, and higher intensity of epithelioid cells, lymphocytes, plasma cells, and eosinophils. On the other hand, (n=4, 30.8%) of the cats with disseminated sporotrichosis and a poor general condition had a marked intensity of yeasts, which were mostly phagocytosed by an increased number of macrophages and neutrophils. Of those animals with good general condition, the majority (n=6, 66.7%) had higher eosinophil intensity. In histopathology, malformed suppurative granuloma was the predominant type (n=9, 69.2%) in feline sporotrichosis lesions, followed by well-formed granulomas (n=4, 30.8%). Malformed granulomas showed mild to moderate fungal intensity (55.6%) in animals with good general condition and localized lesions while marked fungal intensity (44.4%) in cats with the disseminated form of the disease and poor general condition. Well-formed granulomas (n=4, 30.7%) had mild to moderate intensity of fungal load, and 75% of the animals with this type of granuloma had only one lesion and were in good general condition. Long-term itraconazole treatment in these cats with refractory sporotrichosis can keep the infection under control and localized lesions stable; however, fungus reactivation can occur, resulting in an exuberant and inefficient immune response.

A esporotricose é uma infecção fúngica crônica causada por espécies do gênero Sporothrix. A ocorrência de casos refratários ao tratamento de longo prazo, especialmente na região nasal de gatos, alerta para a importância do estudo de sua patogênese. Este estudo teve como objetivo analisar o processo inflamatório das lesões cutâneas de esporotricose felina refratária ao tratamento, por meio de exames citopatológico e histopalógico, e comparar com aspectos clínicos. Treze gatos com lesões cutâneas refratárias ao tratamento com Itraconazol por mais de um ano foram incluídos no estudo. Amostras de lesões cutâneas foram coletadas para análises citopatológicas, histopatológicas e cultura fúngica. Fragmentos de tecidos foram processados e corados pela hematoxilina-eosina (HE) e Prata Metenamina de Grocott (GMS). Duas apresentações clínicas tiveram maior ocorrência: a forma localizada cutânea em animais com bom estado geral e doença estável (n=9, 69,2%); e a forma disseminada cutânea em gatos com estado geral ruim (n=4, 30,8%). A região nasal (84,6%) foi a localização mais frequente das lesões nos gatos com esporotricose refratária ao tratamento. No estudo citopatológico, felinos com menos de duas lesões, e em bom estado geral, (n=9, 69,2%) revelaram ausência ou leve intensidade de leveduras (até 5 leveduras por campo), menor intensidade de macrófagos e neutrófilos, e maior intensidade de células epitelióides, linfócitos, plasmócitos e eosinófilos. Enquanto, (n=4, 30,8%) dos felinos que apresentavam a forma disseminada da esporotricose associada a um estado geral ruim, revelaram acentuada intensidade de leveduras em sua maioria fagocitadas por acentuado número de macrófagos e neutrófilos. Dos animais com bom estado geral, a maioria (n=6, 66,7%) apresentava maior intensidade de eosinófilos. Na histopatologia, o granuloma supurativo mal formado foi o tipo predominante (n=9, 69,2%) nas lesões de esporotricose felina, e (n=4, 30,8%) foram de granulomas bem formados. Granulomas mal formados apresentaram leve a moderada intensidade fúngica (55,6%) nos animais com bom estado geral e lesões localizadas, e acentuada intensidade fúngica (44,4%) nos gatos com a forma disseminada da doença e estado geral ruim. Granulomas bem formados (n=4, 30,7%) apresentaram leve a moderada intensidade de carga fúngica, sendo 75% dos animais com esse tipo de granuloma com apenas uma lesão e bom estadogeral. O tratamento de longo prazo com itraconazol, nesses gatos com esporotricose refratária, pode manter a infecção controlada e lesões localizadas estáveis de forma temporária, contudo a reativação do fungo pode ocorrer levando a uma resposta imunológica exuberante e ineficiente.

Resumo

Sporotrichosis is a chronic fungal infection caused by Sporothrix species. The occurrence of cases that are resistant to long-term treatment, especially in the nasal planum of cats, emphasizes the importance of studying its pathogenesis. The purpose of this study was to analyze and compare the inflammatory process of cutaneous lesions of feline refractory sporotrichosis to clinical aspects through cytopathological and histopathological examination. Moreover, the study included 13 cats with cutaneous lesions that had been resistant to itraconazole treatment for more than a year. Cutaneous lesions samples were collected for cytopathological, histopathological, and fungal culture analyses. Tissue fragments were processed and stained with hematoxylin-eosin (HE) and Grocott methenamine silver (GMS). Further, two clinical presentations had the highest occurrence: the localized cutaneous form in animals with good general condition and stable disease (n=9, 69.2%) and the disseminated cutaneous form in cats with poor general condition (n=4, 30.8%). In cats with refractory sporotrichosis, the nasal planum (84.6%) was the most common location of lesions. In the cytopathological study, cats with fewer than two lesions and in good general condition (n=9, 69.2%) showed absence or mild yeast intensity (up to 5 yeasts per field), lower intensity of macrophages and neutrophils, and higher intensity of epithelioid cells, lymphocytes, plasma cells, and eosinophils. On the other hand, (n=4, 30.8%) of the cats with disseminated sporotrichosis and a poor general condition had a marked intensity of yeasts, which were mostly phagocytosed by an increased number of macrophages and neutrophils. Of those animals with good general condition, the majority (n=6, 66.7%) had higher eosinophil intensity. In histopathology, malformed suppurative granuloma was the predominant type (n=9, 69.2%) in feline sporotrichosis lesions, followed by well-formed granulomas (n=4, 30.8%). Malformed granulomas showed mild to moderate fungal intensity (55.6%) in animals with good general condition and localized lesions while marked fungal intensity (44.4%) in cats with the disseminated form of the disease and poor general condition. Well-formed granulomas (n=4, 30.7%) had mild to moderate intensity of fungal load, and 75% of the animals with this type of granuloma had only one lesion and were in good general condition. Long-term itraconazole treatment in these cats with refractory sporotrichosis can keep the infection under control and localized lesions stable; however, fungus reactivation can occur, resulting in an exuberant and inefficient immune response.(AU)

A esporotricose é uma infecção fúngica crônica causada por espécies do gênero Sporothrix. A ocorrência de casos refratários ao tratamento de longo prazo, especialmente na região nasal de gatos, alerta para a importância do estudo de sua patogênese. Este estudo teve como objetivo analisar o processo inflamatório das lesões cutâneas de esporotricose felina refratária ao tratamento, por meio de exames citopatológico e histopalógico, e comparar com aspectos clínicos. Treze gatos com lesões cutâneas refratárias ao tratamento com Itraconazol por mais de um ano foram incluídos no estudo. Amostras de lesões cutâneas foram coletadas para análises citopatológicas, histopatológicas e cultura fúngica. Fragmentos de tecidos foram processados e corados pela hematoxilina-eosina (HE) e Prata Metenamina de Grocott (GMS). Duas apresentações clínicas tiveram maior ocorrência: a forma localizada cutânea em animais com bom estado geral e doença estável (n=9, 69,2%); e a forma disseminada cutânea em gatos com estado geral ruim (n=4, 30,8%). A região nasal (84,6%) foi a localização mais frequente das lesões nos gatos com esporotricose refratária ao tratamento. No estudo citopatológico, felinos com menos de duas lesões, e em bom estado geral, (n=9, 69,2%) revelaram ausência ou leve intensidade de leveduras (até 5 leveduras por campo), menor intensidade de macrófagos e neutrófilos, e maior intensidade de células epitelióides, linfócitos, plasmócitos e eosinófilos. Enquanto, (n=4, 30,8%) dos felinos que apresentavam a forma disseminada da esporotricose associada a um estado geral ruim, revelaram acentuada intensidade de leveduras em sua maioria fagocitadas por acentuado número de macrófagos e neutrófilos. Dos animais com bom estado geral, a maioria (n=6, 66,7%) apresentava maior intensidade de eosinófilos. Na histopatologia, o granuloma supurativo mal formado foi o tipo predominante (n=9, 69,2%) nas lesões de esporotricose felina, e (n=4, 30,8%) foram de granulomas bem formados. Granulomas mal formados apresentaram leve a moderada intensidade fúngica (55,6%) nos animais com bom estado geral e lesões localizadas, e acentuada intensidade fúngica (44,4%) nos gatos com a forma disseminada da doença e estado geral ruim. Granulomas bem formados (n=4, 30,7%) apresentaram leve a moderada intensidade de carga fúngica, sendo 75% dos animais com esse tipo de granuloma com apenas uma lesão e bom estadogeral. O tratamento de longo prazo com itraconazol, nesses gatos com esporotricose refratária, pode manter a infecção controlada e lesões localizadas estáveis de forma temporária, contudo a reativação do fungo pode ocorrer levando a uma resposta imunológica exuberante e ineficiente.(AU)

Resumo

ABSTRACT Dermatophytes are hyaline fungi that parasitize the keratinized tissue of humans and animals causing mycotic infections. Natural products are promising molecules for the development of new antifungal drugs, due to the phenomenon of resistance and toxicity. This study reports the isolation and identification of lachnophyllum ester and evaluates its antioxidant, antifungal and modulatory activities against dermatophytes fungi. Lachnophyllum ester was obtained using a silica gel column chromatography of the essential oil from the aerial parts of Baccharis trinervis and analyzed by gas chromatography/mass spectrometry. Antimicrobial activity was determined by the broth microdilution method. The modulatory activity assays were performed by the checkerboard technique using lachnophyllum ester and ketoconazole as standard. The lachnophyllum ester exhibited good antioxidant activity as measured by a -carotene/linoleic acid bleaching system, with 71.43% ± 0.01% inhibition rate. In addition, it showed antifungal activity against Trichophyton rubrum and Microsporum canis strains. In the modulatory assay, interaction between lachnophyllum ester and ketoconazole was synergistic, reducing the minimum inhibitory concentration (MIC) values of the antifungal drug and modulating its antifungal action against dermatophyte strains. In conclusion, lachnophyllum ester has been shown to act as a natural antioxidant compound, as well as an antimicrobial alternative against dermatophyte fungi of the genus Trichophyton and Microsporum.

RESUMO Dermatófitos são fungos hialinos que parasitam o tecido queratinizado de humanos e animais causando infecções fúngicas. Os produtos naturais são moléculas promissoras para o desenvolvimento de novos fármacos antifúngicos, devido ao fenômeno de resistência e a toxicidade. Este estudo relata o isolamento e a identificação do éster lacnofilum e avalia sua atividade antioxidante, antifúngica e modulatória contra fungos dermatófitos. O éster lacnofilum foi obtido por cromatografia em coluna de sílica gel do óleo essencial das partes aéreas da Baccharis trinervis e analisado por cromatografia gasosa/espectrometria de massa. A atividade antimicrobiana foi determinada pelo método de microdiluição em caldo. Os ensaios de atividade modulatória foram realizados pela técnica de checkerboard, utilizando éster lacnofilum e cetoconazol como padrão. O éster lacnofilum exibiu boa atividade antioxidante, medida pelo sistema -caroteno/ácido linoléico, com taxa de inibição de 71,43% ± 0,01%. Além disso, mostrou atividade antifúngica contra as cepas de Trichophyton rubrum e Microsporum canis. No ensaio modulatório, a interação entre éster lacnofilum e cetoconazol foi sinérgica, reduzindo os valores concentração inibitória mínima (CIM) do antifúngico e modulando sua ação antifúngica contra cepas de dermatófitos. Em conclusão, o éster lacnofilum demonstrou atuar como um composto antioxidante natural, bem como uma alternativa antimicrobiana contra fungos dermatófitos do gênero Trichophyton e Microsporum.

Resumo

The aim of study to investigate inhibition effects of cinnamic acid (volatile compound of cinnamon), limonene (essential oil of lemon) and eugenol (essential oil of clove, cinnamon) on Metschnikowia fructicola, Candida oleophila, Schisosaccharomyces pombe, Saccharomyces uvarum and Kloeckera apiculata. In this study, tube dilution method was used. Among eugenol, limonene and cinnamic acid, it was found that cinnamic acid has the most inhibition effect with low concentrations (%2.8, %3.84, %4.36, %5,4) at tube dilution methods. Also it was found that test yeast have different resistance against test materials.(AU)

O objetivo do estudo foi investigar os efeitos de inibição do ácido cinâmico (composto volátil da canela), limoneno (óleo essencial de limão) e eugenol (óleo essencial de cravo, canela) sobre Metschnikowia fructicola, Candida oleophila, Schisosaccharomyces pombe, Saccharomyces uvarum e Kloeckera apiculata. Neste estudo, foi utilizado o método de diluição em tubo. Entre eugenol, limoneno e ácido cinâmico, verificou-se que o ácido cinâmico tem o maior efeito de inibição com baixas concentrações (% 2,8,% 3,84,% 4,36,% 5,4) nos métodos de diluição em tubo. Também foi descoberto que a levedura testada tem diferentes resistências contra os materiais de teste.(AU)

Resumo

Dermatophytes are hyaline fungi that parasitize the keratinized tissue of humans and animals causing mycotic infections. Natural products are promising molecules for the development of new antifungal drugs, due to the phenomenon of resistance and toxicity. This study reports the isolation and identification of lachnophyllum ester and evaluates its antioxidant, antifungal and modulatory activities against dermatophytes fungi. Lachnophyllum ester was obtained using a silica gel column chromatography of the essential oil from the aerial parts of Baccharis trinervis and analyzed by gas chromatography/mass spectrometry. Antimicrobial activity was determined by the broth microdilution method. The modulatory activity assays were performed by the checkerboard technique using lachnophyllum ester and ketoconazole as standard. The lachnophyllum ester exhibited good antioxidant activity as measured by a β-carotene/linoleic acid bleaching system, with 71.43% ± 0.01% inhibition rate. In addition, it showed antifungal activity against Trichophyton rubrum and Microsporum canis strains. In the modulatory assay, interaction between lachnophyllum ester and ketoconazole was synergistic, reducing the minimum inhibitory concentration (MIC) values of the antifungal drug and modulating its antifungal action against dermatophyte strains. In conclusion, lachnophyllum ester has been shown to act as a natural antioxidant compound, as well as an antimicrobial alternative against dermatophyte fungi of the genus Trichophyton and Microsporum.(AU)

Dermatófitos são fungos hialinos que parasitam o tecido queratinizado de humanos e animais causando infecções fúngicas. Os produtos naturais são moléculas promissoras para o desenvolvimento de novos fármacos antifúngicos, devido ao fenômeno de resistência e a toxicidade. Este estudo relata o isolamento e a identificação do éster lacnofilum e avalia sua atividade antioxidante, antifúngica e modulatória contra fungos dermatófitos. O éster lacnofilum foi obtido por cromatografia em coluna de sílica gel do óleo essencial das partes aéreas da Baccharis trinervis e analisado por cromatografia gasosa/espectrometria de massa. A atividade antimicrobiana foi determinada pelo método de microdiluição em caldo. Os ensaios de atividade modulatória foram realizados pela técnica de checkerboard, utilizando éster lacnofilum e cetoconazol como padrão. O éster lacnofilum exibiu boa atividade antioxidante, medida pelo sistema β-caroteno/ácido linoléico, com taxa de inibição de 71,43% ± 0,01%. Além disso, mostrou atividade antifúngica contra as cepas de Trichophyton rubrum e Microsporum canis. No ensaio modulatório, a interação entre éster lacnofilum e cetoconazol foi sinérgica, reduzindo os valores concentração inibitória mínima (CIM) do antifúngico e modulando sua ação antifúngica contra cepas de dermatófitos. Em conclusão, o éster lacnofilum demonstrou atuar como um composto antioxidante natural, bem como uma alternativa antimicrobiana contra fungos dermatófitos do gênero Trichophyton e Microsporum.(AU)

Resumo

Endemic systemic mycoses remain a health challenge, since these opportunistic diseases are increasingly infecting immunosuppressed patients. The simultaneous use of antifungal compounds and other drugs to treat infectious or non-infectious diseases has led to several interactions and undesirable effects. Thus, new antifungal compounds should be investigated. The present study aimed to evaluate the activity of liriodenine extracted from Annona macroprophyllata on agents of systemic mycoses, with emphasis on the genus Paracoccidioides. Methods: The minimum inhibitory concentration (MIC) and minimum fungicide concentration (MFC) were determined by the microdilution method. The cellular alterations caused by liriodenine on a standard P. brasiliensis (Pb18) strain were evaluated by transmission and scanning electron microscopy. Results: Liriodenine was effective only in 3 of the 8 strains of the genus Paracoccidioides and in the Histoplasma capsulatum strain, in a very low concentration (MIC of 1.95 µg.mL-1); on yeasts of Candida spp. (MIC of 125 to 250 µg.mL-1), including C. krusei (250 µg.mL-1), which has intrinsic resistance to fluconazole; and in Cryptococcus neoformans and Cryptococcus gattii (MIC of 62.5 µg.mL-1). However, liriodenine was not effective against Aspergillus fumigatus at the studied concentrations. Liriodenine exhibited fungicidal activity against all standard strains and clinical isolates that showed to be susceptible by in vitro tests. Electron microscopy revealed cytoplasmic alterations and damage to the cell wall of P. brasiliensis (Pb18). Conclusion: Our results indicate that liriodenine is a promising fungicidal compound that should undergo further investigation with some chemical modifications.(AU)

Resumo

Susceptibility testing is essential to inform the correct management of Aspergillus infections. In this study we present antifungal susceptibility profile of A. fumigatus isolates recovered from lungs of birds with and without aspergillosis. Fifty three isolates were tested for their antifungal susceptibility to voriconazole (VRC), itraconazole (ITZ), amphotericin (AMB) and caspofungin (CSP) using the M38-A2 broth microdilution reference method. Five isolates were resistant to more than one antifungal drug (CSP + AMB, VRC + ITZ and AMB + ITZ). Fifteen (28%) isolates with susceptible increased exposure (I) to ITZ were sensible to VRC. Resistance to AMB (>2µg/mL) was observed in only four isolates. Eleven (21%) A. fumigatus present resistance to ITZ (13%) and VRC (8%). Fungal isolation from respiratory samples has been regarded as being of limited usefulness in the ante mortem diagnosis of aspergillosis in birds. However, the results suggest that the detection and antifungal susceptibility profile may be helpful for monitoring of therapy for avian species and where antifungal resistance might be emerging and what conditions are associated to the event.(AU)

Os testes de suscetibilidade são essenciais para informar o correto manejo das infecções por Aspergillus. Neste estudo apresentamos o perfil antifúngico de isolados de A. fumigatus provenientes de pulmões de aves com e sem aspergilose. Cinqüenta e três isolados foram testados quanto à susceptibilidade antifúngica ao voriconazol (VRC), itraconazol (ITZ), anfotericina B (AMB) e caspofungina (CSP) pelo método de referência de microdiluição do caldo M38-A2. Cinco isolados foram resistentes a mais de um antifúngico (CSP + AMB, VRC + ITZ e AMB + ITZ). Quinze (28%) isolados suscetíveis - com exposição aumentada (I) ao ITZ foram sensíveis ao VRC. A resistência ao AMB (>2µg/mL) foi observada em apenas quatro isolados. Onze (21%) A. fumigatus apresentaram resistência a ITZ (13%) e VRC (8%). O isolamento de fungos de amostras respiratórias tem sido considerado de utilidade limitada no diagnóstico ante mortem de aspergilose em aves. No entanto, os resultados sugerem que a detecção e o perfil de suscetibilidade a antifúngicos podem ser úteis para o monitoramento da terapia de espécies aviárias, assim como a emergência da resistência antifúngica e quais condições podem estar associadas ao evento.(AU)

Resumo

The early blight (caused by the fungus Alternaria solani) results in significant damage to the tomato crop, directly affecting productivity. An alternative to the frequent use of pesticides is the use of essential oils, which can act in defense against phytopathogens. The objective of this work was to evaluate the toxic activity in vitro of the bergamot orange (Citrus aurantium ssp. bergamia) essential oil against A. solani, the control of the early blight, and the activity of defense enzymes in tomatoes treated with this oil and inoculated with A. solani. Mycelial discs of A. solani were added to dishes with V8 culture mediato which essential oil at concentrations of 0, 500, 1000, 1500, 2000, and 2500 µL L-¹, in addition to a standard fungicide treatment (azoxystrobin + difenoconazole, 200 + 125 g L-¹, respectively) was added. The Petri dishes were incubated at 25 °C in the dark. Mycelial growth was evaluated daily for 19 days, when all treatments reached maximum growth. Sporulation analysis was performed there after. Tomato plants were treated with bergamot essential oil, 30 days after transplanting, in the concentrations and fungicide mentioned, in the second pair of leaves. After 72 hours, the pathogen was inoculated using a spray bottle, on treated leaves (second pair of leaves) and untreated leaves (third pair of leaves). The area under the disease progress curve (AUDPC) was calculated based on five severity assessments.The activity of peroxidase (POX), polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL) was evaluated in leaves treated with the concentration of 2500 µL L-¹ of essential oil. The concentration of 2500 μL L-¹ reduced mycelial growth and sporulation of the pathogen by 68.15% and 29.48%, respectively. In treatments with application of essential oil, lower AUDPC was observed for a concentration of 2500 μL L-¹, which was statistically similar to that [...].

A pinta preta (causada pelo fungo Alternaria solani) causa prejuízos significativos na cultura do tomateiro, afetando diretamente a produtividade. Uma alternativa ao uso frequente de agrotóxico é a utilização de óleos essenciais, que podem atuar na defesa contra fitopatógenos. O objetivo deste trabalho foi avaliar a atividade tóxica in vitro do óleo essencial de bergamota (Citrus aurantium subsp. bergamia) contra A. solani, o controle da pinta preta e a atividade de enzimas de defesa em tomateiros tratados com esse óleo e inoculados com A. solani. Discos miceliais de A. solani foram adicionados em placas com meio de cultura V8 adicionado de óleo essencial nas concentrações de 0, 500, 1000, 1500, 2000 e2500 µL L-¹, além de um tratamento padrão com fungicida (azoxistrobina + difenoconazol, 200 + 125g L-¹ respectivamente). As placas de Petri foram incubadas a 25 °C no escuro. O crescimento micelial foi avaliado diariamente, por 19 dias, quando todos os tratamentos atingiram o máximo crescimento, sendo então feita a análise da esporulação. Aos 30 dias após o transplantio, o segundo par de folhas de plantas de tomate foram tratadas com óleo essencial de bergamota e de fungicida nas concentrações de óleo citados. Após 72 horas o patógeno foi inoculado com borrifador, nas folhas tratadas (segundo par de folhas) e nas não tratadas (terceiro par de folhas). A área abaixo da curva de progresso da doença(AACPD) foi calculada com base em cinco avaliações de severidade. A atividade de peroxidase (POX), polifenoloxidase (PPO) e fenilalaninaamônia-liase (PAL) foi avaliada em folhas tratadas coma concentração de 2500 µL L-¹ de óleo essencial. A concentração de 2500 μL L-¹ reduziu o crescimento micelial e a esporulação do patógeno, em 68,15% e 29,48%, respectivamente. Nos tratamentos com aplicação de óleo essencial, a menor AACPD foi observada para a concentração de 2500 μL L-¹, sendo semelhante estatisticamente ao observado para a aplicação de [...] .

Resumo

In this study, we described the antifungal activity of three Brazilian propolis extracts: brown, green and from jataí bees against Sporothrix brasiliensis. The extracts were obtained from ethanolic extraction and their chemical composition was determined by high-performance liquid chromatography coupled to mass spectrometry. The cellular toxicity was measured in MDBK (Madin-Darby Bovine Kidney) cells and quantified by the MTT assay (3- (4,5 dimethylthiazol-2yl -2,5-diphenyl-2H bromine tetrazolato). For antifungal activity, the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were determined by broth microdilution. The results showed that cell toxicity was not observed at lower concentrations (0.097 to 0.39μg/ml) for all extracts in comparison to cell control. Among the chemical compounds identified, caffeic acid, p-coumaric acid, chlorogenic acid, ferulic acid and rutin were quantified. In antifungal activity, green and jataí did not exhibit activity against the isolates (MIC and MFC greater than 0.78mg/ml). However, all isolates of S. brasiliensis were sensitive to brown propolis (MIC of 0.09 to 0.78mg/ml), including the standard strain (P<0.001). Among the Brazilian propolis studied, the brown propolis showed activity against the S. brasiliensis isolates and more studies should be undertaken in order to evaluate its promising use in the treatment of sporotrichosis.(AU)

Neste estudo, descreveu-se a atividade antifúngica de três extratos de própolis brasileiras: marrom, verde e de abelhas jataí (Tetragonisca angustula), contra Sporothrix brasiliensis. Os extratos foram obtidos de extração etanólica, e a sua composição química foi determinada por cromatografia líquida de alta eficiência, acoplada à espectrometria de massa. A toxicidade celular foi medida em células MDBK (Madin-Darby Bovine Kidney), avaliada por observação microscópica e quantificada pelo ensaio MTT (3- (4,5-dimetiltiazol-2-ilo -2,5-difenil-2H bromo tetrazolato). Para a atividade antifúngica, determinou-se a concentração inibitória mínima (CIM) e a concentração fungicida mínima (CFM) por meio de microdiluição em caldo. Os resultados mostraram que a toxicidade celular não foi observada em concentrações menores (0,097 a 0,39μg/ml). Entre os compostos químicos identificados, foram quantificados o ácido cafeico, ácido p-cumárico, ácido clorogênico, ácido ferúlico e a rutina. Na atividade antifúngica, as própolis verde e jataí não apresentaram atividade contra os isolados (CIM e CFM maior que 0,78mg/ml), porém todos os isolados de S. brasiliensis foram sensíveis à própolis marrom (CIM de 0,09 a 0,78mg/ml), incluindo a cepa padrão (P<0,001). Entre as própolis brasileiras estudadas, a marrom mostrou atividade contra S. brasiliensis, e mais estudos devem ser realizados para avaliar seu uso promissor no tratamento da esporotricose.(AU)

Resumo

Mycotoxin levels in Fusarium head blight (FHB) infections can be difficult to quantify. The relationship between mycotoxin and disease is not consistent and it is not clear if wheat-pathogen interaction is of significance in regions where more than one Fusarium species with dis-tinct trichothecene production ability co-exists. This study aimed to investigate whether a set of eight Brazilian wheat genotypes, varying in resistance according to classification by the breeder, exhibit a common or differential resistance to Fusarium graminearum (Fgra) (deoxynivalenol-producing) and Fusarium meridionale (nivalenol-producing) (Fmer) using full-spike and central spikelet inoculation (type II resistance). Fgra was generally more aggressive than Fmer based on the percentage of diseased spikes (99 and 84 %, respectively) and number of diseased spikelets (mean 2.8 and 2.0, respectively) below the central spikelet. The genotype-pathogen species interaction was not significant, but there were differences between the genotypes, with BRS 194 and BRS 327 being the least and most resistant, respectively, based on severity ratings. The incidence of Fusarium-damaged kernel (FDK, %) was not affected by species, but two genotypes (BRS Parrudo and BRS 327) showed a lower incidence of FDK. There were substantial variations in the accumulation of deoxynivalenol and nivalenol among the genotypes, reaching maxima of 691.2 μg g–1 and 355.2 μg g–1, respectively, suggesting that Fgra is a more potent producer of trichothecene. Our data confirm prior resistance classifications by the breeders and suggest that the use of a single highly aggressive Fgra isolate may be sufficient for effective screening for FHB resistance. However, further studies are needed to elucidate the accumulation of resistance to trichothecene.

Resumo

Background: Sporotrichosis is a zoonosis caused by fungi of the Sporothrix schenckii complex. Cats have important zoonotic potential due to the high parasite load found in the cutaneous lesions. Refractory cases to antifungal therapy are increasing, becoming a public health problem. Miltefosine is a drug with high in vitro activity against Sporothrix brasiliensis. However, this efficacy has not been confirmed in vivo yet and administration has not been studied in the feline species. This study aimed to evaluate the effectiveness and safety of miltefosine administration in cats with resistant sporotrichosis.Materials, Methods & Results: Ten cats with refractory sporotrichosis were included in this study. Inclusion criteria were previous diagnosis for sporotrichosis associated with therapy with oral antifungals (itraconazole and/or potassium iodide) for more than one year, followed by one of these conditions: incomplete clinical remission, worsening of disease during treatment or recurrence after conventional treatment. Cats underwent clinical and dermatological examinations. Data on lifestyle and previous treatments were provided by the owners. Fungal culture and histochemical techniques were performed in all cats before miltefosine treatment. Blood samples were collected before and during the study for laboratory tests, such as blood count, blood urea nitrogen (BUN), serum creatinine, alanine aminotransferase (ALT) and alkaline phosphatase, as well as serology for Feline Immunodeficiency (FIV) and Feline Leukemia (FeLV). Miltefosine was prescribed at a dosage of 2 mg/kg orally every 24 h. The animals were monitored by phone during the whole process and reassessed on days 0, 15, 30 and 45 of the treatment. Out of 10 cats, lesions were present on the nose in nine, ears in four, periocular areas in one and limbs in four. One cat received treatment for 45 days, six for 30 days, one for 21 days, one for 15 days and one for 3 days.[...]