Fungal conversion of chicken-feather waste into biofortified compost / Conversão fungosa de resíduos de pena de frango em composto biofortificado

Poultry industry is amongst highly developed industries of Pakistan, fulfilling the protein demand of rapidly increasing population. On the other hand, the untreated poultry waste is causing several health and environmental problems. The current study was designed to check the potential of keratinolytic fungal species for the conversion of chicken-feather waste into biofortified compost. For the purpose, three fungal species were isolated from soil samples. These strains were pure cultured and then characterized phenotypically and genotypically. BLAST searches of 18S rDNA nucleotide sequence of the fungal isolates revealed that the two fungal isolates belonged to genus Aspergillus and one belonged to genus Chrysosporium. Optimum temperature for Aspergillus flavus, Aspergillus niger and Chrysosporium queenslandicum was 29, 26 and 25 oC, respectively. A. flavus showed maximum (53%) feather degradation, A. niger degraded feather waste up to 37%, while C. queenslandicum showed 21% keratinolytic activity on chicken feathers at their respective temperature optima. The degradation potential of these fungal species showed their ability to form compost that has agro-industrial importance.

A indústria avícola está entre as indústrias altamente desenvolvidas do Paquistão, atendendo a demanda de proteína da população em rápido crescimento. Por outro lado, os resíduos de aves não tratados estão causando diversos problemas de saúde e ambientais. O presente estudo foi desenhado para verificar o potencial de espécies de fungos queratinolíticos para a conversão de resíduos de penas de frango em composto biofortificado. Para tanto, três espécies de fungos foram isoladas de amostras de solo. Essas cepas foram cultivadas puramente e, em seguida, caracterizadas fenotipicamente e genotipicamente. As pesquisas do BLAST da sequência de nucleotídeos do rDNA 18S dos isolados de fungos revelaram que os dois isolados de fungos pertenciam ao gênero Aspergillus e um pertencia ao gênero Chrysosporium. A temperatura ótima para Aspergillus flavus, Aspergillus niger e Chrysosporium queenslandicum foi de 29, 26 e 25 oC, respectivamente. A. flavus apresentou degradação máxima de penas (53%), A. niger degradou resíduos de penas em até 37%, enquanto C. queenslandicum apresentou 21% de atividade queratinolítica em penas de frango em suas respectivas temperaturas ótimas. O potencial de degradação dessas espécies de fungos mostrou sua capacidade de formar composto de importância agroindustrial.

Aspergilose em calopsita (Nymphicus hollandicus) / Aspergillosis in a cockatiel (Nymphicus hollandicus)

Background: Aspergillosis is a defiance in avian medicine, however between of Aspergillus species, 3 are the most frequently observed (A. flavus, A. fumigatus and A. niger). Psittaciformes present medical conditions related to the 3 species, however A. fumigatus is the most observed. The defiance of the diagnosis is related to fungus characteristics (they are saprophytic in healthy birds), the clinical signs and lesions, which are common to various respiratory diseases, and the difficulty to access specific diagnostic methods. Thus, we report a case of aspergillosis in a cockatiel, whose clinical diagnosis was confirmed by fungal culture of cloacal swab, and treatment with a fungistatic drug. Case: A female cockatiel, 2-month-old, not dewormed, presented diarrhea, sneezing and difficulty to gain weight, although eating well (extruded feed and supplementation with fruits and vegetables). At a clinical examination revealed body score (BCS) of 3 (scale 1 at 5), cloacal temperature (CT) of 40 ºC, dilated nostrils and edema in the left eye region associated to evident hyperemia. Due to it has lived with another animal, was inferred that was hypovitaminosis A, by food dispute and/ or dominance, or endoparasitosis, being prescribed vitamin A supplementation and deworming with ivermectin. The bird returned 15 days later, showing maintenance of weight loss, BCS 2, ruffled feathers and areas suggestive of apteriolysis in both forelimbs, respiratory difficulty and pulmonary and cranial air sacs rales, in addition presented wound in the pygostyle region and ventus, occasioned by attempted attack of a dog, and aggravated by self-mutilation. Due the worsening of the clinical condition, an oral cavity and cloacal swabs were collected to bacteriological and fungal culture. As treatment was prescribed: a natural tranquilizer, seeking to control the self-mutilation; doxycycline oral suspension, to treat a probable bacterial respiratory infection; and oral itraconazole suspension, manipulated and with a dose calculated by allometric scaling, being placed an elizabethan collar in order to prevent biting the pygostyle and ventus. With a positive result of the cloacal swab fungal culture for A. fumigatus, the doxycycline was discontinued. After the first 30 days of treatment with itraconazole, the patient returns to alert, weighing 95 g, BCS 3, with absence of respiratory signs and with signs of recent molting process. New swabs (oral and cloacal) were collected for repetition of bacterial and fungal cultures, being maintained the treatment with itraconazole. The results of both bacterial and fungal cultures were negative. At the end of the treatment with itraconazole, the patient returns demonstrating weight gain, EEC 3, CT of 42ºC, uniform feather coverage and absence of any respiratory sign, being released by the clinician. Discussion: A preponderant factor in the treatment of various respiratory diseases in birds is the diagnosis, since many of diseases present the same clinical signs. Thus, the use of fungal culture was essential to determine the origin of the signs. Still, even with an accurate diagnosis, the clinician often comes up against the difficulty of treatment, especially in cases of mycotic infections. The knowledge about the best drug, considering the pharmacology, as well as the availability of concentrations and dosages marketed, in addition to the long period of treatment, can difficult or make impossible the case resolution. In fact, this is confirmed by the literature, where the most of aspergillosis diagnosis are post mortem, resulting from the delay in seeking veterinary care or in the administration of ineffective drugs. Thus, with this report, we sought to present a diagnostic tool, sometimes neglected, and a compilation with the main drugs and their effectiveness in the treatment of aspergillosis in Psittaciformes.

Optimization of oxalic acid production by fungi for biotechnological solubilization of rock phosphate

This study describes a biotechnological strategy for producing and applying oxalic acid to solubilize phosphorus (P) from rock phosphate (RP). We evaluated six fungal species (Aspergillus niger FS1, Penicillium islandicum FS41, Pleurotus ostreatus PO1, Rhizoctonia solani Rhiz555, Sclerotium rolfsii Sr25, and Sclerotinia sclerotiorum Scl134) and three culture media (potato dextrose broth, Tsao and Strasser media) to maximize oxalic acid production. Among the fungal isolates tested and culture media, S. rolfsii Sr25 and Tsao medium showed efficient oxalic acid production. Tsao medium was optimized following a response surface methodology after initial screening of factors affecting RP solubilization. The optimized concentrations were 1 g L-1 NaNO3, 100 g L-1 glucose, 2 g L-1 KH2PO4, 4.5 g L-1 yeast extract, and 25 mg L-1 MgSO4·7H2O used for 20 days of incubation. Under these conditions, 71 mmol L-1 oxalic acid was obtained, representing a three-fold increase over production under non-optimized conditions (20 mmol L-1). Under optimized conditions, oxalic acid produced by S. rolfsii Sr25 reacted with low-solubility RP and solubilized 100 % of the P contained in ore. Thus, using S. rolfsii Sr25 to produce oxalic acid seems a promising biotechnological alternative for P solubilization from RP.(AU)

Biomass of unconventional plants from Brazilian semiarid as substrate for hydrolytic enzymes production by Aspergillus niger under solid and submerged fermentation

Aspergillus niger KIJH was grown in solid and submerged fermentation using leaves and roots (with and without bark) of plants typically from Brazilian semiarid as substrate to produce a multienzymatic extract, which was characterised for its potential biotechnological applications. Solid-state fermentation (SSF) was applied to select the most promising plants biomass as induction substrates for the production of hydrolytic enzymes by fungus. The best biomasses were used as substrate in submerged fermentation (SmF) assays at two scales. Samples of up scale fermented culture were partially purified by ultrafiltration and activity and pH and temperature stability of CMCase and xylanase were evaluated. A. niger KIJH produced hydrolytic enzymes under SSF containing unconventional plants biomass from Brazilian semiarid. In SmF conditions, maximum CMCase (0.264 U mL-1) and xylanase (1.163 U mL-1) activities were induced by Jacaratia corumbensis. Scaling up the SmF to 500 mL of medium was able to maintain constant the production of CMCase (0.346 U mL-1) and xylanase (1.273 U mL-1) on the fermented culture. Ultrafiltered and concentrated extract presented CMCase activities practically constant in all temperature ranges (30-80°C) and pH (3.0-9.0), while xylanase optimum activity temperature was 50°C and pH in the range of 3.0 to 5.0. CMCase activity remained stable for 24 hours at 50°C

Biomass of unconventional plants from Brazilian semiarid as substrate for hydrolytic enzymes production by Aspergillus niger under solid and submerged fermentation

Aspergillus niger KIJH was grown in solid and submerged fermentation using leaves and roots (with and without bark) of plants typically from Brazilian semiarid as substrate to produce a multienzymatic extract, which was characterised for its potential biotechnological applications. Solid-state fermentation (SSF) was applied to select the most promising plants biomass as induction substrates for the production of hydrolytic enzymes by fungus. The best biomasses were used as substrate in submerged fermentation (SmF) assays at two scales. Samples of up scale fermented culture were partially purified by ultrafiltration and activity and pH and temperature stability of CMCase and xylanase were evaluated. A. niger KIJH produced hydrolytic enzymes under SSF containing unconventional plants biomass from Brazilian semiarid. In SmF conditions, maximum CMCase (0.264 U mL-1) and xylanase (1.163 U mL-1) activities were induced by Jacaratia corumbensis. Scaling up the SmF to 500 mL of medium was able to maintain constant the production of CMCase (0.346 U mL-1) and xylanase (1.273 U mL-1) on the fermented culture. Ultrafiltered and concentrated extract presented CMCase activities practically constant in all temperature ranges (30-80°C) and pH (3.0-9.0), while xylanase optimum activity temperature was 50°C and pH in the range of 3.0 to 5.0. CMCase activity remained stable for 24 hours at 50°C(AU)

Biomass of unconventional plants from Brazilian semiarid as substrate for hydrolytic enzymes production by Aspergillus niger under solid and submerged fermentation

Aspergillus niger KIJH was grown in solid and submerged fermentation using leaves and roots (with and without bark) of plants typically from Brazilian semiarid as substrate to produce a multienzymatic extract, which was characterised for its potential biotechnological applications. Solid-state fermentation (SSF) was applied to select the most promising plants biomass as induction substrates for the production of hydrolytic enzymes by fungus. The best biomasses were used as substrate in submerged fermentation (SmF) assays at two scales. Samples of up scale fermented culture were partially purified by ultrafiltration and activity and pH and temperature stability of CMCase and xylanase were evaluated. A. niger KIJH produced hydrolytic enzymes under SSF containing unconventional plants biomass from Brazilian semiarid. In SmF conditions, maximum CMCase (0.264 U mL-1) and xylanase (1.163 U mL-1) activities were induced by Jacaratia corumbensis. Scaling up the SmF to 500 mL of medium was able to maintain constant the production of CMCase (0.346 U mL-1) and xylanase (1.273 U mL-1) on the fermented culture. Ultrafiltered and concentrated extract presented CMCase activities practically constant in all temperature ranges (30-80°C) and pH (3.0-9.0), while xylanase optimum activity temperature was 50°C and pH in the range of 3.0 to 5.0. CMCase activity remained stable for 24 hours at 50°C a

Phytochemical profile and antifungal action of Anadenanthera colubrina extract on the quality of maize seeds

Maize (Zea mays L.) is among the most cultivated crops in the world and can be affected by several diseases, especially those transmitted by seeds. The study of alternatives to fungicides used for seed treatment has a promising field in essential oils. Thus, this study determined the phytochemical profile of the ethanolic extract from Anadenanthera colubrina (Vell.) Brenan and to evaluate its antifungal activity on the sanitary and physiological quality of maize seeds. The seeds used were of the Jaboatão cultivar, which were submitted to the following treatments: control (untreated seeds), commercial fungicide (dicarboximide) and A. colubrina extract at 200, 400, 600, 800, and 1,000 ppm. The seeds were subjected to sanitary and germination tests in a completely randomized experimental design. Phytochemical prospecting of A. colubrina extract indicated the presence of alkaloids, tannins, flavonoids and saponins, as well as the major compounds lupeol, gallic acid, ferulic acid, catechin and quercetin. The A. colubrina extract reduced the incidence of Aspergillus spp., including Aspergillus niger, Alternaria spp., Curvularia spp. and Fusarium spp. at all concentrations. The highest concentrations (800 and 1,000 ppm) of the A. colubrina extract reduced the incidence of Penicillium spp. and yielded an effective control of Rhizoctonia spp. The extract of A. colubrina did not present phytotoxic effect, guaranteeing the viability and vigor of maize seeds.

Buffaloes (Bubalus bubalis) mycotic eye infection: reported herd cases in Thi-Qar province, Iraq / Infecção ocular micótica em búfalos (Bubalus bubalis): relato de casos em rebanho na província de Thi-Qar, Iraque

Buffaloes are one of the important farm animals in the south of Iraq and play an essential economical role mainly acting as dairy, meat, and draft animals. This study intended to diagnose buffalo mycotic eye infections in Thi-Qar province/Iraq. Some 250 buffaloes in the herd of 3,700 animals suffered from eye infections from December 2017 to November 2018. Eye swabs were collected from each infected eye of the affected buffaloes of both sexes before treatment. The animals were in different age groups. All samples were transferred to the laboratory in transfer media, and cultured on Sabouraud dextrose (SDA) agar with and without 0.05 g/mL and 0.4 g/mL chloramphenicol and cycloheximide, respectively. Later, the agars were incubated at 25oC and 37oC. The total percentage of eye infection was (6.75%), constituting (49.2%) mycotic infections. The predominant clinical manifestations that appeared on the infected buffaloes were eye inflammation represented by congestion, lacrimation, the opacity of cornea and edema, and reduced productivity of the infected animals. Different fungal isolates were identified from the samples including Aspergillus fumigates, Aspergillus flavus, Aspergillus niger, Penicillium spp., Alternaria spp., Fusarium spp., Candida spp., Cladosporium spp., Rhodotorula spp., Mucor spp. and Rhizopus spp. Calves buffaloes below one-year-old were more prone to mycotic infection than one-year-old or more. Additionally, male buffaloes were more susceptible to infection than females. In conclusion, this study isolated various types of fungus from the inflamed eyes of buffaloes. Fungal eye infection and the potential risk factors for fungal keratitis in buffaloes were also investigated. The study also approved the rapid diagnosis of fungi by direct microscopic detection and culture. The author recommends future studies including large numbers of the buffalo herd in Iraq to determine the epidemiology of this condition in the country.(AU)

Os búfalos são um dos animais de fazenda mais importantes no sul do Iraque e desempenham um papel econômico essencial, atuando principalmente na produção de leite, carne e como animal de tração. Este estudo objetivou diagnosticar infecções oculares micóticas em búfalos na província de Thi-Qar, Iraque. 250 búfalos no rebanho de um total de 3700 animais apresentaram infecção ocular durante o período compreendido entre dezembro de 2017 e novembro de 2018. Os esfregaços oculares foram colhidos dos olhos infectados dos búfalos afetados de ambos os sexos antes do tratamento. Os animais estavam em diferentes faixas etárias. Todas as amostras foram transferidas para o laboratório por meio de transferência e cultivadas em Ágar Sabouraud e Dextrose (SDA) com e sem 0,05 g/mL e 0,4 g/mL de cloranfenicol e cicloheximida, respectivamente. Posteriormente, os ágares foram incubados a 25ºC e 37ºC. A porcentagem total de infecção ocular foi de 6,75%, representando 49,2% de infecção micótica. As manifestações clínicas predominantes nos búfalos infectados foram inflamação ocular com congestão, lacrimejamento, opacidade da córnea e edema. Os animais acometidos também apresentaram redução de produtividade. Diferentes isolados de fungos foram identificados a partir das amostras, incluindo Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Penicillium spp., Alternaria spp., Fusarium spp., Candida spp., Cladosporium spp., Rhodotorula spp., Mucour spp. e Rizopus spp. Búfalos com menos de um ano de idade foram mais propensos a infecções micóticas do que com um ano ou mais. Além disso, os búfalos machos foram mais suscetíveis a infecção do que as fêmeas. Em conclusão, este estudo registrou o isolamento de vários tipos de fungos em olhos inflamados de búfalos. Além disso, a infecção ocular por fungos e os fatores de risco potenciais para ceratite fúngica em búfalos também foram observados. O estudo também aprovou o diagnóstico rápido de fungos por detecção microscópica direta e cultura. O autor recomenda outro estudo futuro, incluindo um grande número de rebanhos de búfalos no Iraque para determinar a epidemiologia desta condição no país.(AU)

Buffaloes (Bubalus bubalis) mycotic eye infection: reported herd cases in Thi-Qar province, Iraq / Infecção ocular micótica em búfalos (Bubalus bubalis): relato de casos em rebanho na província de Thi-Qar, Iraque

Buffaloes are one of the important farm animals in the south of Iraq and play an essential economical role mainly acting as dairy, meat, and draft animals. This study intended to diagnose buffalo mycotic eye infections in Thi-Qar province/Iraq. Some 250 buffaloes in the herd of 3,700 animals suffered from eye infections from December 2017 to November 2018. Eye swabs were collected from each infected eye of the affected buffaloes of both sexes before treatment. The animals were in different age groups. All samples were transferred to the laboratory in transfer media, and cultured on Sabouraud dextrose (SDA) agar with and without 0.05 g/mL and 0.4 g/mL chloramphenicol and cycloheximide, respectively. Later, the agars were incubated at 25oC and 37oC. The total percentage of eye infection was (6.75%), constituting (49.2%) mycotic infections. The predominant clinical manifestations that appeared on the infected buffaloes were eye inflammation represented by congestion, lacrimation, the opacity of cornea and edema, and reduced productivity of the infected animals. Different fungal isolates were identified from the samples including Aspergillus fumigates, Aspergillus flavus, Aspergillus niger, Penicillium spp., Alternaria spp., Fusarium spp., Candida spp., Cladosporium spp., Rhodotorula spp., Mucor spp. and Rhizopus spp. Calves buffaloes below one-year-old were more prone to mycotic infection than one-year-old or more. Additionally, male buffaloes were more susceptible to infection than females. In conclusion, this study isolated various types of fungus from the inflamed eyes of buffaloes. Fungal eye infection and the potential risk factors for fungal keratitis in buffaloes were also investigated. The study also approved the rapid diagnosis of fungi by direct microscopic detection and culture. The author recommends future studies including large numbers of the buffalo herd in Iraq to determine the epidemiology of this condition in the country.(AU)

Os búfalos são um dos animais de fazenda mais importantes no sul do Iraque e desempenham um papel econômico essencial, atuando principalmente na produção de leite, carne e como animal de tração. Este estudo objetivou diagnosticar infecções oculares micóticas em búfalos na província de Thi-Qar, Iraque. 250 búfalos no rebanho de um total de 3700 animais apresentaram infecção ocular durante o período compreendido entre dezembro de 2017 e novembro de 2018. Os esfregaços oculares foram colhidos dos olhos infectados dos búfalos afetados de ambos os sexos antes do tratamento. Os animais estavam em diferentes faixas etárias. Todas as amostras foram transferidas para o laboratório por meio de transferência e cultivadas em Ágar Sabouraud e Dextrose (SDA) com e sem 0,05 g/mL e 0,4 g/mL de cloranfenicol e cicloheximida, respectivamente. Posteriormente, os ágares foram incubados a 25ºC e 37ºC. A porcentagem total de infecção ocular foi de 6,75%, representando 49,2% de infecção micótica. As manifestações clínicas predominantes nos búfalos infectados foram inflamação ocular com congestão, lacrimejamento, opacidade da córnea e edema. Os animais acometidos também apresentaram redução de produtividade. Diferentes isolados de fungos foram identificados a partir das amostras, incluindo Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Penicillium spp., Alternaria spp., Fusarium spp., Candida spp., Cladosporium spp., Rhodotorula spp., Mucour spp. e Rizopus spp. Búfalos com menos de um ano de idade foram mais propensos a infecções micóticas do que com um ano ou mais. Além disso, os búfalos machos foram mais suscetíveis a infecção do que as fêmeas. Em conclusão, este estudo registrou o isolamento de vários tipos de fungos em olhos inflamados de búfalos. Além disso, a infecção ocular por fungos e os fatores de risco potenciais para ceratite fúngica em búfalos também foram observados. O estudo também aprovou o diagnóstico rápido de fungos por detecção microscópica direta e cultura. O autor recomenda outro estudo futuro, incluindo um grande número de rebanhos de búfalos no Iraque para determinar a epidemiologia desta condição no país.(AU)

Phytochemical profile and antifungal action of Anadenanthera colubrina extract on the quality of maize seeds

ABSTRACT Maize (Zea mays L.) is among the most cultivated crops in the world and can be affected by several diseases, especially those transmitted by seeds. The study of alternatives to fungicides used for seed treatment has a promising field in essential oils. Thus, this study determined the phytochemical profile of the ethanolic extract from Anadenanthera colubrina (Vell.) Brenan and to evaluate its antifungal activity on the sanitary and physiological quality of maize seeds. The seeds used were of the Jaboatão cultivar, which were submitted to the following treatments: control (untreated seeds), commercial fungicide (dicarboximide) and A. colubrina extract at 200, 400, 600, 800, and 1,000 ppm. The seeds were subjected to sanitary and germination tests in a completely randomized experimental design. Phytochemical prospecting of A. colubrina extract indicated the presence of alkaloids, tannins, flavonoids and saponins, as well as the major compounds lupeol, gallic acid, ferulic acid, catechin and quercetin. The A. colubrina extract reduced the incidence of Aspergillus spp., including Aspergillus niger, Alternaria spp., Curvularia spp. and Fusarium spp. at all concentrations. The highest concentrations (800 and 1,000 ppm) of the A. colubrina extract reduced the incidence of Penicillium spp. and yielded an effective control of Rhizoctonia spp. The extract of A. colubrina did not present phytotoxic effect, guaranteeing the viability and vigor of maize seeds.

Preparation and process optimization of microbial organic copper as a feed additive / Preparação e otimização do processo de cobre orgânico microbiano como aditivo alimentar

As an essential trace element for animals, copper significantly contributes to the growth and health of animals. Compared to inorganic trace elements, organic trace elements are better supplements; notably, they are acquired through microbial transformation. Therefore, we screened for copper-enriched microorganisms from high copper content soil to obtain organic copper. Sodium diethyldithio carbamate trihydrate was applied as a chromogenic agent for determining micro amounts of intracellular copper through spectrophotometry. In total, 50 fungi were isolated after the successful application of the screening platform for copper-rich microbes. Following morphological and molecular biology analyses, the N-2 strain, identified as Aspergillus niger sp. demonstrated showed better copper enrichment potential than others. Notably, the strain tolerance to copper was nearly thrice that of Saccharomyces cerevisiae, up to 1600mg/L. The content of the organic bound copper was 22.84mg Cu/g dry cell. Using the Central Composite Design (CCD) response surface method, we optimized the fermentation condition (inoculation amount, 13%; temperature, 28(C; pH, 5.0). Compared to the original strain results under the single factor fermentation condition, we reported an increase by 24.18% under the optimized conditions. Collectively, these findings provide a reference for uncovering new and low-cost organic copper additives.(AU)

Como elemento traço essencial para os animais, o cobre contribui significativamente para o crescimento e saúde dos animais. Comparado aos oligoelementos inorgânicos, os oligoelementos orgânicos são melhores suplementos; notavelmente, eles são adquiridos através de transformação microbiana. Portanto, nós selecionamos microorganismos enriquecidos com cobre de solos com alto teor de cobre para obter cobre orgânico. O carbamato de sódio diethyldithio trihidratado foi aplicado como agente cromogênico para a determinação de micro quantidades de cobre intracelular através da espectrofotometria. No total, 50 fungos foram isolados após a aplicação bem sucedida da plataforma de triagem para micróbios ricos em cobre. Após análises morfológicas e de biologia molecular, a cepa N-2, identificada como Aspergillus niger sp. demonstrou um melhor potencial de enriquecimento de cobre do que outras. Notavelmente, a tolerância da estirpe ao cobre foi quase três vezes maior que a da Saccharomyces cerevisiae, até 1600mg/L. O conteúdo de cobre ligado orgânico era de 22,84mg Cu/g de célula seca. Usando o método de superfície de resposta Central Composite Design (CCD), nós otimizamos a condição de fermentação (quantidade de inoculação, 13%; temperatura, 28C; pH, 5,0). Em comparação com os resultados da deformação original sob a condição de fermentação de fator único, relatamos um aumento de 24,18% sob as condições otimizadas. Coletivamente, estas descobertas fornecem uma referência para descobrir novos aditivos de cobre orgânico de baixo custo.(AU)

Biological control in the germination of seeds from two species native of the Cerrado region / Controle biológico na germinação de sementes de duas espécies nativas da região do Cerrado

Microorganisms have been efficiently used for the biological control of phytopathogens through the production of antimicrobial substances. However, the objectives of this work were: to study the germination of Butia purpurascens Glassman and Butia archeri Glassman seeds in different substrates, to select and identify the endophytic and rhizospheric bacterial isolates of B. purpurascens and B. archeri, and to perform antibiosis tests based on the isolated microorganisms of these tree species. No difference was found between the cultivation substrates for the percentages of germination, hard seeds, and fungal contamination in the B. purpurascens seeds. The Bacillus subtilis isolated showed the best capacity for suppressing the growth of the two deteriorative fungi tested in B. purpurascens seeds. No difference was found for inhibition of the growth of Aspergillus niger fungus (deteriorative fungus of B. archeri seeds) between the microorganisms with Bacillus sp. and Brevibacillus brevis compared to the control. In the microbiolization of B. purpurascens and B. archeri seeds performed with microbiological solutions produced from the endophytic and rhizospheric strains of Bacillus sp., no differences were observed in the percentages of germination and contamination by fungi. For B. archeri seeds, there was contamination by fungi and bacteria after one day of cultivation, primarily in the regions with lesions caused by the extraction and scarification process.(AU)

Microorganismos específicos tem sido usados eficientemente para o controle biológico de fitopatógenos através da produção de substâncias antimicrobianas. Portanto, os objetivos deste estudo foram: estudar a germinação de sementes de Butia purpurascens Glassman e Butia archeri Glassman em diferentes substratos; selecionar e identificar isolados bacterianos rizosféricos e endofíticos de B. purpurascens e B. archeri, e realizar testes de antibiose a partir de microorganismos isolados dessas espécies arbóreas. Não houve diferença entre os substratos de cultivo para as porcentagens de germinação, de sementes duras e de contaminação por fungos nas sementes de B. purpurascens. O isolado bacteriano Bacillus subtilis apresentou melhor capacidade de supressão do crescimento dos dois fungos deterioradores testados nas sementes de B. purpurascens. Não foi encontrada diferença para a inibição do crescimento do fungo Aspergillus niger (fungo deteriorador de sementes de B. archeri) entre os microorganismos com Bacillus sp. e Brevibacillus brevis em comparação ao controle. Na microbiolização das sementes de B. purpurascens e B. archeri realizada com soluções microbiológicas produzidas a partir de cepas endofíticas e rizosféricas de Bacillus sp. não foram observadas diferenças na porcentagem de germinação e contaminação por fungos. Para B. archeri houve contaminação por fungos e bactérias após um dia de cultivo, principalmente nas regiões de lesão proporcionadas pelo processo de extração e escarificação.(AU)

Iron Uptake by Fungi Isolated from Arcelor Mittal -Annaba- in the Northeast of Algeria

The Metal pollution is one of the major risks in the world nowadays. Iron is an essential metal for growth and proliferation of a vast majority of organisms, but it can be toxic to human health and other living beings in the environment at high concentrations due to its increased industrial activity. Fungi have a remarkable capacity to uptake and detoxify iron metal using different mechanisms such as bioaccumulation. Thus, the aim of this work is to study the ability of iron uptake by the fungal strains isolated from Arcelor Mittal -Annaba- in Algeria. Three strains were screened at high concentration of iron (1 g/l) and their capacity to uptake iron has been studied on Czapek Yeast Agar medium. The amounts of uptaken iron ions were estimated in the same liquid medium using Atomic Absorption Spectroscopy (AAS). The results of the iron uptake by these screened strains showed that Cladosporium cladosporioides uptakes the highest concentration of iron (347.7 ppm), Aspergillus niger was able to accumulate up to 170 ppm of this metal while, the lowest uptake of this metal was shown by Penicillium citrinum with 106.43 ppm. It was found that the spore germination of three fungal strains was low when the medium is supplemented with high concentrations of iron. This indicated the potential of these fungal strains as biological agents for removal of iron from the industrial effluents containing high concentrations of it.(AU)

Iron Uptake by Fungi Isolated from Arcelor Mittal -Annaba- in the Northeast of Algeria

The Metal pollution is one of the major risks in the world nowadays. Iron is an essential metal for growth and proliferation of a vast majority of organisms, but it can be toxic to human health and other living beings in the environment at high concentrations due to its increased industrial activity. Fungi have a remarkable capacity to uptake and detoxify iron metal using different mechanisms such as bioaccumulation. Thus, the aim of this work is to study the ability of iron uptake by the fungal strains isolated from Arcelor Mittal -Annaba- in Algeria. Three strains were screened at high concentration of iron (1 g/l) and their capacity to uptake iron has been studied on Czapek Yeast Agar medium. The amounts of uptaken iron ions were estimated in the same liquid medium using Atomic Absorption Spectroscopy (AAS). The results of the iron uptake by these screened strains showed that Cladosporium cladosporioides uptakes the highest concentration of iron (347.7 ppm), Aspergillus niger was able to accumulate up to 170 ppm of this metal while, the lowest uptake of this metal was shown by Penicillium citrinum with 106.43 ppm. It was found that the spore germination of three fungal strains was low when the medium is supplemented with high concentrations of iron. This indicated the potential of these fungal strains as biological agents for removal of iron from the industrial effluents containing high concentrations of it.

Biosorption potential of lead tolerant fungi isolated from refuse dumpsite soil in Nigeria

Metals are non-biodegradable and recurrent in the environs. Heavy metals tolerant fungiwere isolated from refuse dumpsite soil using pour plate method. These fungiwere identified as Aspergillus niger, Penicillium chrysogenumandRhizomucor sp. The fungal isolates were screened for cadmium (Cd), lead (Pb) and zinc (Zn) with concentration of 200ppm, 400ppm and 600ppm. Aspergillus nigerand Penicillium chrysogenumshowed high tolerance for the metals in contrast to the control. The fungiwith high tolerance were used for biosorption study. However, Penicillium chrysogenumshowed higher lead removal or biosorption potential of 1.07ppm, 3.35ppm and 4.19ppm as compared with Aspergillus nigerwith lead removal of 0.67ppm, 3.11ppm and 3.79ppm at 5th, 10thand 15thday respectively. One-way Analysis of Variance was used to interpret the data generated from the biosorption study which revealed that there was no significant different (p>0.05)between the lead removal of Aspergillus nigerandPenicillium chrysogenumon the 5thday but there was significant difference (p<0.05)in the lead removal of Aspergillus nigerand Penicillium chrysogenumon the 10thand 15thday. This study suggests the use of these fungal isolates for removal and biotreatment of heavy metal contaminated and polluted environment.

Biological control in the germination of seeds from two species native of the Cerrado region

Abstract Microorganisms have been efficiently used for the biological control of phytopathogens through the production of antimicrobial substances. However, the objectives of this work were: to study the germination of Butia purpurascens Glassman and Butia archeri Glassman seeds in different substrates, to select and identify the endophytic and rhizospheric bacterial isolates of B. purpurascens and B. archeri, and to perform antibiosis tests based on the isolated microorganisms of these tree species. No difference was found between the cultivation substrates for the percentages of germination, hard seeds, and fungal contamination in the B. purpurascens seeds. The Bacillus subtilis isolated showed the best capacity for suppressing the growth of the two deteriorative fungi tested in B. purpurascens seeds. No difference was found for inhibition of the growth of Aspergillus niger fungus (deteriorative fungus of B. archeri seeds) between the microorganisms with Bacillus sp. and Brevibacillus brevis compared to the control. In the microbiolization of B. purpurascens and B. archeri seeds performed with microbiological solutions produced from the endophytic and rhizospheric strains of Bacillus sp., no differences were observed in the percentages of germination and contamination by fungi. For B. archeri seeds, there was contamination by fungi and bacteria after one day of cultivation, primarily in the regions with lesions caused by the extraction and scarification process.

Resumo Microorganismos específicos tem sido usados eficientemente para o controle biológico de fitopatógenos através da produção de substâncias antimicrobianas. Portanto, os objetivos deste estudo foram: estudar a germinação de sementes de Butia purpurascens Glassman e Butia archeri Glassman em diferentes substratos; selecionar e identificar isolados bacterianos rizosféricos e endofíticos de B. purpurascens e B. archeri, e realizar testes de antibiose a partir de microorganismos isolados dessas espécies arbóreas. Não houve diferença entre os substratos de cultivo para as porcentagens de germinação, de sementes duras e de contaminação por fungos nas sementes de B. purpurascens. O isolado bacteriano Bacillus subtilis apresentou melhor capacidade de supressão do crescimento dos dois fungos deterioradores testados nas sementes de B. purpurascens. Não foi encontrada diferença para a inibição do crescimento do fungo Aspergillus niger (fungo deteriorador de sementes de B. archeri) entre os microorganismos com Bacillus sp. e Brevibacillus brevis em comparação ao controle. Na microbiolização das sementes de B. purpurascens e B. archeri realizada com soluções microbiológicas produzidas a partir de cepas endofíticas e rizosféricas de Bacillus sp. não foram observadas diferenças na porcentagem de germinação e contaminação por fungos. Para B. archeri houve contaminação por fungos e bactérias após um dia de cultivo, principalmente nas regiões de lesão proporcionadas pelo processo de extração e escarificação.

Etiology, multidrug resistance, and acute-phase proteins biomarkers as in equine septic arthritis / Etiologia, resistência antimicrobiana e proteínas de fase aguda como biomarcadores na artrite séptica equina

Etiology, multiple antibiotic resistance index, and acute-phase protein profile in the serum and synovial fluid of 44 horses presenting septic arthritis were investigated. Young animals up to 24 months of age not yet submitted to taming were sampled. Synovial samples from the horses were submitted to culture and bacterial isolates subjected to 18 antimicrobials to investigate the in vitro multidrug resistance pattern. Also, hematological aspects, fibrinogen, and serum levels of some acute-phase proteins (total protein, albumin, globulin, amyloid A, and C-reactive protein) were assessed. To statistical analysis, microbiological isolation were divided into four groups: G1 = animals with negative isolation, G2 = isolation of Gram-negative bacteria, G3 = isolation of Gram-positive bacteria), and G4 = isolation of fungi. Microbial isolation was obtained from 77.2% (32/44) of the synovial samples. Escherichia coli (7/44 = 15.9%), Staphylococcus aureus (5/44 = 11.4%), Streptococcus equi subsp. equi (3/44 = 6.8%), Aspergillus niger (3/44 = 6.8%), Pseudomonas aeruginosa (2/44 = 4.5%), Streptococcus equi subsp. zooepidemicus (1/44 = 2.3%), Trueperella pyogenes (1/44 = 2.3%), and Rhodococcus equi (1/44 = 2.3%) were prevalent bacterial agents. Aspergillus niger is reported by the first time as a primary agent of septic arthritis in horses. Gentamicin (84.9%), marbofloxacin (79.3%) and ceftriaxone (72.4%) were the most effective antimicrobials, whereas high resistance of the isolates (>50%) was found to erythromycin (75.9%), clarithromycin (75.9%), penicillin (69%), clindamycin (58.6%), and streptomycin (55.2%). Multiple antibiotic resistance index ( 0.3) was identified in 50% (16/32) of the isolates, with highest resistance in Gram-negative bacterial isolates. Serum levels of amyloid A, C-reactive protein, fibrinogen, globulin, and total protein, as well as the number of nucleated cells in the synovial fluid had values above the reference for horses, reinforcing acute-phase proteins as biomarkers in diagnosis of septic arthritis. We highlighted the diversity of microorganisms that may be involved in equine septic arthritis, the high resistance of bacterial isolates to conventional antimicrobials, the high lethality of young horses with septic arthritis, and importance of in vitro antimicrobial susceptibility pattern tests to therapeutic approaches in the disease.(AU)

Foram investigados a etiologia, o índice de resistência múltipla aos antibióticos e o perfil de proteínas de fase aguda, no soro ou líquido sinovial, de 44 equinos com sinais clínicos compatíveis com artrite séptica. Foram amostrados animais até 24 meses de idade não submetidos a doma. As amostras de líquido sinovial dos 44 animais foram submetidas à cultura bacteriológica e fúngica, e os isolados bacterianos submetidos in vitro diante de 18 antimicrobianos visando investigar a multirresistência. Também foram avaliados os aspectos hematológicos e o fibrinogênio sérico, bem como os níveis séricos de certas proteínas de fase aguda (proteína total, albumina, globulina, amilóide sérico A e proteína C reativa). O isolamento microbiano das amostras foi dividido em quatro grupos: G1 = animais com isolamento negativo; G2 = isolamento de bactérias Gram-negativas; G3 = isolamento de bactérias Gram-positivas e G4 = isolamento fúngico. O isolamento microbiano foi obtido em 77,2% (32/44) das amostras de líquido sinovial. Escherichia coli (7/44 = 15,9%), Staphylococcus aureus (5/44 = 11,4%), Streptococcus equi subsp. equi (3/44 = 6,8%), Aspergillus niger (3/44 = 6,8%), Pseudomonas aeruginosa (2/44 = 4,5%), Streptococcus equi subsp. zooepidemicus (1/44 = 2,3%), Trueperella pyogenes (1/44=2,3%) e Rhodococcus equi (1/44 = 2,3%) foram os patógenos mais frequentes. Aspergillus niger é relatado pela primeira vez como agente primário de artrite séptica em cavalos. Gentamicina (84,9%), marbofloxacino (79,3%) e ceftriaxona (72,4%) foram os antimicrobianos mais eficazes, enquanto elevada resistência dos isolados (>50%) foi encontrada para eritromicina (75,9%), claritromicina (75,9%), penicilina (69%), clindamicina (58,6%) e estreptomicina (55,2%). O índice de resistência múltipla aos antibióticos ( 0,3) foi identificado em 50% (16/32) dos isolados, com maior resistência obervada em bactérias Gram-negativas. Os níveis séricos de amilóide A, proteína C reativa, fibrinogênio, globulina e concentração total de proteínas, bem como o número de células nucleadas no líquido sinovial apresentaram valores acima da referência para equinos saudáveis, reforçando o uso destas proteínas de fase aguda como biomarcadores no diagnóstico da doença. O presente estudo enfatiza a diversidade de microrganismos que podem estar envolvidos na artrite séptica equina, a alta resistência dos isolados aos antimicrobianos convencionais, a alta mortalidade de equinos jovens com artrite séptica e a importância de realizar o tratamento da afecção com respaldo em testes de sensibilidade microbiana in vitro.(AU)

Mold species and fungi load of washed and unwashed table eggs / Espécies de bolores e contaminação fúngica de ovos de galinha lavados e não lavados

Egg quality has been widely studied, mainly because defects in quality can pose risks to public health, as well as economic losses. Nevertheless, studies about fungi in eggs are scarce. The objective was to compare the fungal microbiota from washed and unwashed eggs in the rainy season and dry season of the year. This exploratory research consisted in the analysis of large size white table eggs acquired from 48 different lots. Two manufacturers were sampled considering the main characteristic of washed or unwashed eggs. From each lot, a 30-egg pack were purchased and six of those eggs were used for mycological analyzes. The eggs were analyzed externally with 0.1% peptone salt solution wash of the eggshells and internally with aliquots being sampled from a pool made from the six eggs content. Samples were inoculated in Potato Dextrose Agar and isolated colonies were passed to test tubes. When sporulated, the isolates were subjected to decimal dilutions using 0.1% Tween 80 to dissociate the conidia. Microcultures were carried out for optical microscopy observation of the reproductive structures of fungi, stained with lactophenol. Aspergillus spp. was the most frequently isolated fungi isolated, with A. niger and A. flavus predominant in the dry season, while A. fumigatus and A. terreus in the rainy season. Low numbers of fungi were identified from egg shells, with a higher

A qualidade do ovo já foi largamente estudada, especialmente por causa de defeitos na qualidade que podem significar riscos para a saúde pública, além de perdas econômicas. Entretanto, estudos sobre fungos em ovos são escassos. O objetivo foi comparar a microbiota fúngica de ovos lavados e ovos não lavados nas estações chuvosa e seca do ano. As amostras consistiram de ovos comerciais, do tipo branco e tamanho grande, com caráter exploratório pela análise de 48 lotes. Dois fabricantes foram amostrados considerando a característica principal de ovos lavados ou não lavados. De cada lote, foram adquiridos 30 ovos, sendo seis ovos utilizados nas análises micológicas. Os ovos foram analisados externamente fazendo lavagem com solução salina peptonada e internamentecom alíquotas retiradas de pools de seis ovos. Amostras foram inoculadas em Ágar Dextrose Batata e os isolados foram passados para tubos de ensaio. Posteriormente as colônias foram replicadas para tubos de ensaio contendo meio Ágar Dextrose Batata. Quandojá esporulados, os isolados foram submetidos a diluições utilizando Tween 80 0,1% para desagregar os conídios. Foram realizados microcultivos para observação em microscópio óptico das estruturas de reprodução dos fungos, corados com lactofenol. Aspergillusspp. foi a espécie mais frequentemente isolada, com predominância de A. niger e A. flavus na estação seca e A. fumigatuseA. terreusna chuvosa. Baixas quantidades de fungos foram observadas nas cascas dos ovos, com quantidades maiores em ovos não lavados.

Mold species and fungi load of washed and unwashed table eggs / Espécies de bolores e contaminação fúngica de ovos de galinha lavados e não lavados

Egg quality has been widely studied, mainly because defects in quality can pose risks to public health, as well as economic losses. Nevertheless, studies about fungi in eggs are scarce. The objective was to compare the fungal microbiota from washed and unwashed eggs in the rainy season and dry season of the year. This exploratory research consisted in the analysis of large size white table eggs acquired from 48 different lots. Two manufacturers were sampled considering the main characteristic of washed or unwashed eggs. From each lot, a 30-egg pack were purchased and six of those eggs were used for mycological analyzes. The eggs were analyzed externally with 0.1% peptone salt solution wash of the eggshells and internally with aliquots being sampled from a pool made from the six eggs content. Samples were inoculated in Potato Dextrose Agar and isolated colonies were passed to test tubes. When sporulated, the isolates were subjected to decimal dilutions using 0.1% Tween 80 to dissociate the conidia. Microcultures were carried out for optical microscopy observation of the reproductive structures of fungi, stained with lactophenol. Aspergillus spp. was the most frequently isolated fungi isolated, with A. niger and A. flavus predominant in the dry season, while A. fumigatus and A. terreus in the rainy season. Low numbers of fungi were identified from egg shells, with a higher(AU)

A qualidade do ovo já foi largamente estudada, especialmente por causa de defeitos na qualidade que podem significar riscos para a saúde pública, além de perdas econômicas. Entretanto, estudos sobre fungos em ovos são escassos. O objetivo foi comparar a microbiota fúngica de ovos lavados e ovos não lavados nas estações chuvosa e seca do ano. As amostras consistiram de ovos comerciais, do tipo branco e tamanho grande, com caráter exploratório pela análise de 48 lotes. Dois fabricantes foram amostrados considerando a característica principal de ovos lavados ou não lavados. De cada lote, foram adquiridos 30 ovos, sendo seis ovos utilizados nas análises micológicas. Os ovos foram analisados externamente fazendo lavagem com solução salina peptonada e internamentecom alíquotas retiradas de pools de seis ovos. Amostras foram inoculadas em Ágar Dextrose Batata e os isolados foram passados para tubos de ensaio. Posteriormente as colônias foram replicadas para tubos de ensaio contendo meio Ágar Dextrose Batata. Quandojá esporulados, os isolados foram submetidos a diluições utilizando Tween 80 0,1% para desagregar os conídios. Foram realizados microcultivos para observação em microscópio óptico das estruturas de reprodução dos fungos, corados com lactofenol. Aspergillusspp. foi a espécie mais frequentemente isolada, com predominância de A. niger e A. flavus na estação seca e A. fumigatuseA. terreusna chuvosa. Baixas quantidades de fungos foram observadas nas cascas dos ovos, com quantidades maiores em ovos não lavados.(AU)

Atividade in vitro do óleo essencial de Origanum vulgare L. em isolados clínicos de Aspergillus spp / In vitro activity of essential oil of Origanum vulgare L. in clinical isolates from Aspergillus spp

Aspergillus fumigatus, A. flavus e A. niger são os mais importantes agentes etiológicos da aspergilose, relevante micose aviária, com tratamento ineficaz e altas taxas de mortalidade. Em vista da importância da aspergilose, da necessidade de prospectar novos fármacos e do potencial terapêutico do óleo essencial de Origanum vulgare L. (OEO), o orégano, objetivou-se avaliar a sensibilidade in vitro de isolados clínicos de Aspergillus spp. em relação ao OEO. O óleo foi obtido por hidrodestilação em Clevenger, e a análise química realizada por cromatografia de massa (GC/MS). Observaram-se 15 diferentes compostos ativos, sendo 4-terpineol, hidrato de sabinene e timol os majoritários. Nos testes de microdiluição em caldo (Reference..., 2008), todos os isolados (n= 23) foram sensíveis ao OEO: A. fumigatus teve CIM entre 28,125mg/mL (0,1875%) e 450mg/mL (3,0%), A. flavus entre 112,5mg/mL (0,75%) e 450mg/mL, e A. niger 112,5mg/mL. CFM variou de 112,5mg/mL a 450mg/mL nos isolados de A. fumigatus, de 225mg/mL (1,5%) a 450mg/mL em A. flavus, e foi de 450mg/mL em A. niger. CIM e CFM foram idênticos em 6/14 isolados, o que demonstra que o óleo com a mesma concentração pode ter capacidade fungistática e fungicida. CIM 90 correspondeu à CIM máxima. Os resultados demonstram a atividade anti-Aspergillus do OEO, com CIM 90 de 450mg/mL (3%).(AU)

Aspergillus fumigatus, A. flavus and A. niger are the most important etiological agents of aspergillosis, a relevant avian mycosis, with innefective treatment and high mortality rates. Due the importance of aspergillosis, the necessity of prospection of new drugs and the therapeutic potential of the essential oil of Origanum vulgare L. (OEO), popularly known as oregano, aims to evaluate the in vitro sensitivity of Aspergillus spp. opposing to OEO. The oil was obtained by hydrodistillation in Clevenger, and the chemical analysis performed by mass chromatography (GC/MS). 15 different active compounds were observed, being 4-terpineol (18.4%), sabinene hydrate (15.6%) and thymol (13.6%), the majority components. In the in vitro susceptibility test, all strains showed sensitivity to OEO, MIC of Aspergillus fumigatus ranged from 28,125mg/mL (0,1875%) to 450mg/mL (3,0%), A. flavus 112,5mg/mL (0,75%) to 450mg/mL, and A. niger 112,5mg/mL. MFC ranged from 112,5mg/mL to 450mg/mL in the A. fumigatus isolates, 225mg/mL (1,5%) to 450mg/mL in A. flavus, and 450mg/mL for A. niger. The MIC and FMC values were identical in 6/14 of the isolated subjects, demonstrating that the oil with the same concentration can have fungistatic and fungicidal capacity. The results demonstrates anti-Aspergillus activities of OEO with CIM90 de 450mg/mL (3%).(AU)