Synergistic effects of tacrolimus and azole antifungal compounds in fluconazole-susceptible and fluconazole-resistant Candida glabrata isolates

In vitro interaction between tacrolimus (FK506) and four azoles (fluconazole, ketoconazole, itraconazole and voriconazole) against thirty clinical isolates of both fluconazole susceptible and -resistant Candida glabrata were evaluated by the checkerboard microdilution method. Synergistic, indifferent or antagonism interactions were found for combinations of the antifungal agents and FK506. A larger synergistic effect was observed for the combinations of FK506 with itraconazole and voriconazole (43%), followed by that of the combination with ketoconazole (37%), against fluconazole-susceptible isolates. For fluconazole-resistant C. glabrata, a higher synergistic effect was obtained from FK506 combined with ketoconazole (77%), itraconazole (73%), voriconazole (63%) and fluconazole (60%). The synergisms that we observed in vitro, notably against fluconazole-resistant C. glabrata isolates, are promising and warrant further analysis of their applications in experimental in vivo studies.(AU)

Susceptibility variation of Malassezia pachydermatis to antifungal agents according to isolate source

Malassezia pachydermatisis associated with dermatomycoses and otomycosis in dogs and cats. This study compared the susceptibility of M. pachydermatis isolates from sick (G1) and healthy (G2) animals to azole and polyene antifungals using the M27-A3 protocol. Isolates from G1 animals were less sensitive to amphotericin B, nystatin, fluconazole, clotrimazole and miconazole.(AU)

Antibacterial activity of essential oil of cinnamon and trans-cinnamaldehyde against Staphylococcus spp. isolated from clinical mastitis of cattle and goats

Background: Milk production plays a role as a source of protein and constitutes an important socioeconomic factor for small farms. The lack of planning of the creators, together with the lack of specialized technical assistance, and the precarious hygienic and sanitary management, are barriers that hinder the development of the activity. Bovine and caprine mastitis is an inflammation process of the mammary gland caused by microorganisms. Staphylococcus spp. is the main causative agent of mastitis, with higher prevalence in cases of clinical and subclinical manifestations. The most common treatment is based on intramammary infusion of antibacterial agents. However, cure rates obtained with such drugs are not always effectives, because it may determine the emergence of resistant bacteria as well increase amounts of antibiotic residues in milk. Nevertheless, the treatment of bovine subclinical mastitis caused by Staphylococcus spp. in the lactation can be economically unviable. Alternative treatments of bovine mastitis with plant derived compounds have been described. The investigation of its antimicrobial activity against bacterial agents of mastitis is justifiable. Materials, Methods & Results: This study evaluated the antimicrobial activity of essential oil (EO) of Cinnamomum zeylanicum (cinnamon) and the fraction trans-cinnamaldehyde (TC) against 65 isolates of Staphylococcus spp. from mastitis of cows (n = 32) and goats (n = 33). The cinnamon EO was extracted from the leaves and was composed majority by isoeugenol (96%), while the TC was acquired commercially. The antimicrobial susceptibility tests were performed according to CLSI M31-A3 agar diffusion method. Based on these tests, the isolates were divided into subgroups according to the resistance profiles. Subsequently, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by broth microdilution technique CLSI M31-A3. The MICs (geometric mean, GM) for cinnamon EO were 2032 mg/L and 2102 mg/L, respectively, for cattle and goats, while the majority fraction TC was most active with MICs (GM) of 598 mg/L and 875 mg/L, respectively, for cattle and goats. Discussion: Some differences between these results and those reported by other authors can be attributed to the composition of OEs or techniques employed. In this study, we used the broth microdilution technique based on the document M31-A3, from CLSI, for the determination of MICs and MBCs. The lack of an internationally standardized technique for evaluating OEs and plant extracts that allow different protocols undermines the comparisons of results. The results of this study confirmed the antimicrobial activity of cinnamon EO and TC against clinical Staphylococcus spp. independent of the antibacterial susceptibility profile. The best antimicrobial activity was observed with TC than in the cinnamon EO, which deserves further research investigating the use of these compound in the treatment of mastitis. The TC can be found in high concentration in cinnamon bark. However, the essential oil used in this work was extracted from the leaves, and the major component was isoeugenol. The cinnamon EO composition can be varied depending on the part of the plant where it is extracted.

Subprodutos da uva como promotores de crescimento em dietas de frangos de corte de 1 a 21 dias de idade / Grape by-products as growth promoters for broilers from 1 to 21 days of age

Foi realizado um experimento para se avaliar os efeitos da utilizacao de compostos fenolicos contidos em subprodutos da uva (Vitis vinifera) como promotores de crescimento em dietas de frango de corte. Foram utilizados 600 pintos de corte machos Ross, de um a 21 dias de idade, distribuidos em delineamento inteiramente casualizado, com seis tratamentos, inoculados ou nao, com cepas de Escherichia coli, constituindo-se fatorial 6 x 2 com cinco repeticoes de dez aves cada uma. Foi utilizada uma dieta isonutritiva composta por racao inicial basal, com subproduto de uva ou antibiotico, de acordo com os seguintes tratamentos: T1-controle negativo; T2-0,05% de flavomicina e sulfato de colistina; T3-0,04% extrato de semente de uva desengordurada; T4-0,1% de semente de uva integral (SUI); T5-0,47% de SUI e T6-2,35% de SUI. As variaveis analisadas foram: ganho de peso, consumo de racao, ganho de peso medio, peso final, conversao alimentar e indice de eficiencia produtiva. A inclusao de subprodutos da uva nao influenciou as variaveis zootecnicas nem apresentou atividade promotora de crescimento. A inoculacao com E. coli piorou o ganho de peso e o peso final. Os resultados nao recomendam a utilizacao de subprodutos de uva.(AU)

An experiment was conducted to determine the effects of phenolic compounds from grape by-products (Vitis vinifera) as growth promoters in the diets of broilers between 1 and 21 days of age. Six hundred Ross male chicks were raised from 1 to 21 days of age. The experimental design was completely randomized a 6 x 2 factorial scheme (six diets, inoculated or not inoculated with strains of Escherichia coli), and five repetitions consisting of 10 animals each. Each bird received an isonutritive basal diet consisting of grape by-products or antibiotics as follows: T1-negative control; T2 positive control-0.05% of flavomycin and colistin sulphate; T3 0.04% defatted grape seed extract; T4 0.1% grape seed (WGS); T5 0.47% WGS; and T6 2.35% WGS. Analyzed variables were: weight gain, feed intake, average weight gain, body weight, feed conversion and production efficiency index. Including grape by-products did not influence parameters of rate growth or show any growth promoter activity. Inoculating with E. coli decreased weight gain and body weight. The results did not recommend the use of grape by-product.(AU)

Subprodutos da uva como promotores de crescimento em dietas de frangos de corte de 1 a 21 dias de idade / Grape by-products as growth promoters for broilers from 1 to 21 days of age

Foi realizado um experimento para se avaliar os efeitos da utilizacao de compostos fenolicos contidos em subprodutos da uva (Vitis vinifera) como promotores de crescimento em dietas de frango de corte. Foram utilizados 600 pintos de corte machos Ross, de um a 21 dias de idade, distribuidos em delineamento inteiramente casualizado, com seis tratamentos, inoculados ou nao, com cepas de Escherichia coli, constituindo-se fatorial 6 x 2 com cinco repeticoes de dez aves cada uma. Foi utilizada uma dieta isonutritiva composta por racao inicial basal, com subproduto de uva ou antibiotico, de acordo com os seguintes tratamentos: T1-controle negativo; T2-0,05% de flavomicina e sulfato de colistina; T3-0,04% extrato de semente de uva desengordurada; T4-0,1% de semente de uva integral (SUI); T5-0,47% de SUI e T6-2,35% de SUI. As variaveis analisadas foram: ganho de peso, consumo de racao, ganho de peso medio, peso final, conversao alimentar e indice de eficiencia produtiva. A inclusao de subprodutos da uva nao influenciou as variaveis zootecnicas nem apresentou atividade promotora de crescimento. A inoculacao com E. coli piorou o ganho de peso e o peso final. Os resultados nao recomendam a utilizacao de subprodutos de uva.

An experiment was conducted to determine the effects of phenolic compounds from grape by-products (Vitis vinifera) as growth promoters in the diets of broilers between 1 and 21 days of age. Six hundred Ross male chicks were raised from 1 to 21 days of age. The experimental design was completely randomized a 6 x 2 factorial scheme (six diets, inoculated or not inoculated with strains of Escherichia coli), and five repetitions consisting of 10 animals each. Each bird received an isonutritive basal diet consisting of grape by-products or antibiotics as follows: T1-negative control; T2 – positive control-0.05% of flavomycin and colistin sulphate; T3 – 0.04% defatted grape seed extract; T4 – 0.1% grape seed (WGS); T5 – 0.47% WGS; and T6 – 2.35% WGS. Analyzed variables were: weight gain, feed intake, average weight gain, body weight, feed conversion and production efficiency index. Including grape by-products did not influence parameters of rate growth or show any growth promoter activity. Inoculating with E. coli decreased weight gain and body weight. The results did not recommend the use of grape by-product.

Zoosporogênese in vitro entre isolados do oomiceto Pythium insidiosum / In vitro zoosporogenesis among oomycetes Pythium insidiosum isolates

Pythium insidiosum é um oomiceto aquático, responsável pela etiologia da pitiose, uma enfermidade crônica, observada freqüentemente em eqüinos. A produção de zoósporos móveis por este microrganismo se constitui no fator determinante da ocorrência da enfermidade. Este estudo avaliou a zoosporogênese e quantificou a produção de zoósporos de 32 amostras de Pythium insidiosum isoladas de eqüinos naturalmente infectados. Pythium insidiosum foi cultivado em meio Corn Meal Agar acrescido de fragmentos de grama, durante 5 dias, a 37ºC. Posteriormente, os fragmentos de grama parasitados foram incubados em Meio de Indução a 37ºC, por 24 horas. Observou-se que 16 amostras (50 por cento) produziram 20.000 zoósporos mL-1, 12 isolados (37,5 por cento) produziram acima de 20.000 zoósporos mL-1, enquanto quatro amostras (12,5 por cento) produziram menos de 20.000 zoósporos mL-1. O período de maior produção de zoósporos foi entre 6 e 8 horas de incubação. O protocolo utilizado na indução da zoosporogênese mostrou-se eficiente e representa uma importante ferramenta, tanto para a identificação do Pythium insidiosum, como para a obtenção de zoósporos em quantidades suficientes para a inoculação em animais experimentais e aplicação no desenvolvimento de testes de suscetibilidade.(AU)

Pythium insidiosum is an aquatic oomycete and the etiology of a chronic disease called pythiosis, commonly found in the skin of horses. The production of mobile zoospores by this microorganism is the determinant factor of this disease. This study evaluated the zoosporogenesis and quantification of zoospores in 32 samples of Pythium insidiosum isolated from horses with pythiosis. The assay used culture of the Pythium insidiosum in Corn Meal Agar plus grass blades for 5 days at 37ºC. The grass blades were incubated in Induction Medium at 37ºC for 24 hours. The findings showed 16 samples (50 percent) yielded 20,000 zoospores mL-1, 12 (37.5 percent) samples yielded over 20,000 zoospores mL-1 and 4 samples (12.5 percent) yielded less than 20,000 zoospores mL-1. The zoospores production was higher between 6 and 8 hours of incubation. The protocol used in the zoosporogenesis induction was efficient and represents an important tool for Pythium insidiosum identification and the attainment of zoospores in adequate amounts for inoculation in experimental animals and application in the development of susceptibility tests.(AU)

Teste de ELISA indireto para o diagnóstico sorológico de pitiose / Indirect ELISA for the serodiagnostic of pythiosis

A pitiose, doença granulomatosa de eqüinos causada pelo oomiceto Pythium insidiosum, tem como característica a evolução rápida seguida de morte dos animais. Estas mortes muitas vezes são causadas por diagnósticos errôneos ou demorados quando os doentes já não respondem ao tratamento. Este trabalho teve por objetivo a padronização do ensaio imunoenzimático indireto (ELISA) para diagnóstico sorológico de pitiose em eqüinos e coelhos, visando a diminuição de erros e de tempo necessário para o diagnóstico. Para o desenvolvimento e validação do teste foram utilizadas 72 amostras de soro de eqüinos saudáveis e 44 soros de eqüinos com pitiose confirmada. Os resultados da validação do ELISA para eqüinos foram: sensibilidade 97,72 por cento, especificidade 90,27 por cento, valor preditivo positivo 86 por cento, valor preditivo negativo 98,4 por cento e eficiência de 93,1 por cento. Para coelhos, o teste foi padronizado com 48 amostras de soro de animais saudáveis e 24 amostras de coelhos imunizados com antígenos de P. insidiosum. Os resultados foram: sensibilidade 91,66 por cento, especificidade 95,83 por cento, valor preditivo positivo 91,66 por cento, valor preditivo negativo 95,83 por cento e eficiência de 94,44 por cento. Os resultados deste trabalho demonstram que o ensaio imunoenzimático indireto é um método seguro e eficaz para o diagnóstico sorológico da pitiose.(AU)

Pythiosis is a granulomatous disease caused by the oomycete Pythium insidiosum that affects humans and animals, especially horses. Deaths are very often the consequence of incorrect or late diagnosis when animals no longer respond to treatment. This study aimed standardization of the ELISA assay for the serodiagnostic of pythiosis in horses and rabbits, in order to minimize errors and delays in the diagnosis of the disease. Sera of 72 healthy and 44 of by pythiosis affected horses were used for development and evaluation of the test. The ELISA for equine diagnostic showed 97.72% sensitivity, 90.27% specificity, 86% positive predictive value, 98.4% negative predictive value, and 93.1% efficiency. The rabbit test was standardized with 48 sera of healthy rabbits and 24 sera of rabbits immunized with P. insidiosum antigens. The results were 91.66 % sensitivity, 95.83% specificity, 91.66% positive predictive value, 95.83% negative predictive value, and 94.44% efficiency. It can be concluded that ELISA is a reliable test for diagnostic and serological monitoring of pythiosis.(AU)

Fungal microbiota in drinking water and domestic sewage / Microbiota fúngica em amostras de água potável e esgoto doméstico

The aim of this study was to evaluate the fungal microflora present in drinking water and domestic sewage from different districts of Santa Maria, Rio Grande do Sul, Brazil. Water and sewage samples were collected during the four seasons and analyzed by the technique of Colony Forming Units (CFU). Yeasts and fungi of the genera Penicillium and Aspergillus were observed in the water samples. The genus Geotrichum was also found in the sewage. Therefore, it is concluded that treatment of water held in the municipality is unable to remove these agents.

O objetivo deste estudo foi investigar a microbiota fúngica presente na água potável e no esgoto doméstico de distintos bairros de Santa Maria, Rio Grande do Sul, Brasil. Amostras de água e esgoto foram colhidas durante as quatro estações do ano e analisadas pela técnica de Unidades Formadoras de Colônias (UFC). Foi observada a presença dos gêneros Penicillium, Aspergillus e leveduras nas amostras de água. No esgoto, além dos agentes fúngicos reportados na água foi identificado o gênero Geotrichum. Dessa forma, conclui-se que o tratamento da água realizado no município não é capaz de eliminar estes agentes.