RESUMO
This study aimed to evaluate the microbiological and physicochemical characteristics of the ruminal fluid and histological characteristics of the gastrointestinal tract (GIT) of sheep on a high-grain diet containing the probiotic fungi Aspergillus terreus and Rhizomucor spp. The study included four treatment groups (without probiotic, with Rhizomucor spp., with Aspergillus terreus, and with a mixture of both fungi), and two types of corn (ground/whole), in a Completely Randomized Design (CRD) arranged in 4 x 2 factorial design. Santa Inês x Dorper lambs were housed in eight pens with five lambs each for 75 days. Rumen fluid was collected to study the rumen microbiological profile, macroscopic characteristics, ammonia nitrogen concentration, and microbiological activity. In addition, GIT samples were taken for histological analysis. Fluid analyses showed that the animals presented a low acidosis index. The samples presented a predominantly aromatic odor and blackish-brown color, indicating a neutral pH and high microbial activity. The rumen pH differed (P < 0.05) according to the level of processed corn consumed, being higher for ground grain corn (GGC). There was no difference for any of the microbiological communities analyzed (P > 0.05) (Lac+ and Lac- bacteria, fungi, yeasts, and protozoa). Six genera of facultative anaerobic fungi were identified in 15 observations. Cladosporium spp. was the most prevalent genus (46.66%), followed by Aspergillus spp. (26,66%). The width of the base of rumen papillae showed significant correlation being greater for GCG (P < 0.05) with Rhizomucor and for the control (P < 0.05). The rumen fluid of sheep on a high-grain diet with added Aspergillus terreus and Rhizomucor spp. showed no microbiological and physicochemical changes.
Objetivou-se avaliar as características microbiológicas e físico-químicas do fluido ruminal e histológicas do trato gástrico intestinal (TGI) de ovinos sob dieta de alto grão com probiótico fungos Aspergillus terreus e/ou Rhizomucor spp. Analisou-se quatro probióticos (sem inóculos, com Rhizomucor spp., com Aspergillus terreus e com mistura dos dois fungos) e dois processamentos de milho (moído/inteiro), em fatorial 4x2 em em Delineamento Inteiramente Casualizados (DIC). Borregos Santa Inês/Dorper foram alojados em oito baias com cinco borregos em cada, durante 75 dias. Coletou-se fluido ruminal para o estudo do perfil microbiológico do rúmen, da característica macroscópica, da concentração de nitrogênio amoniacal e da atividade microbiológica, além dos fragmentos do TGI para análises histológicas. Pelas análises dos fluidos, os animais apresentaram baixo índice de acidose. O odor aromático e a cor castanho-enegrecido predominaram, o que caracteriza ambiente com pH neutro. As amostras do fluido apresentaram alta atividade microbiana. O pH ruminal diferenciou-se (P<0,05) quanto ao tipo de processamento, sendo maior para milho grão moído (MGM). Não houve diferença para nenhuma das comunidades microbiológicas analisadas (P>0,05) (bactérias Lac+ e Lac-, fungos, leveduras e protozoários). Seis gêneros de fungos anaeróbicos facultativos foram identificados num total de 15 observações. O Cladosporium spp. foi o gênero mais prevalente (46,66%), seguido do Aspergillus spp. (26,66%). A largura da base das papilas ruminais apresentou interação significativa, sendo maior para MGM (P<0,05) com Rhizomucor e o controle (P<0,05). O fluido ruminal de ovinos sob dieta de alto concentrado de grão com adição dos fungos Aspergillus terreus e Rhizomucor spp. não tiveram afetadas as características microbiológicas e físico-químicas.
Assuntos
Animais , Aspergillus/patogenicidade , Rúmen/microbiologia , Ruminantes/crescimento & desenvolvimento , Cladosporium/patogenicidade , Probióticos/efeitos adversos , Rhizomucor/patogenicidade , Trato Gastrointestinal/anatomia & histologiaRESUMO
Thermophilic fungi constitute an ecologically well-defined group, commonly found in environments wherever decomposition of organic matter takes place, making them self-heating. The importance of thermophilic fungus in ecosystems contrasts with the incompleteness of our understanding of the group's biogeography patterns, phylogenies and coevolution relationships. Actually, the lack of data about thermophilic fungi from the Brazil is a limiting factor that also contributes for this scenario. In order to reduce this gap of knowledge, we aimed to characterize thermophilic filamentous fungi in Araucaria Forest, Atlantic Forest biome. Species identification was achieved by using internal transcribed spacers (ITS) as molecular ribosomal markers. In total, 240 heat-tolerant fungal strains were isolated and identified as Thermothielavioides terrestris, Thielavia sp., Thermoascus crustaceus, Aspergillus fumigatus, Rhizomucor miehei, Rhizomucor pusillus, and Rhizopus microsporus. All thermophilic strains exhibited optimal growth at 45 °C. T. crustaceus, T. miehei e R. pusillus were the dominant species, with the frequencies of occurrence of 35.00%, 28.33% and 23.33%, respectively. Our data reveals the apparent diversity of the Neotropical realm and may serve as reference to future studies that will try to elucidate important aspects of group.
Assuntos
Araucaria , Ecossistema , Brasil , Eurotiales , Florestas , Fungos/genética , Rhizomucor , Rhizopus , SordarialesRESUMO
This work aimed the application of a new biocatalyst for biodiesel production from residual agro-industrial fatty acids. A recombinant Pichia pastoris displaying lipase from Rhizomucor miehei (RML) on the cell surface, using the PIR-1 anchor system, were prepared using glycerol as the carbon source. The biocatalyst, named RML-PIR1 showed optimum temperature of 45 °C (74.0 U/L). The stability tests resulted in t1/2 of 3.49 and 2.15 h at 40 and 45 °C, respectively. RML-PIR1 was applied in esterification reactions using industrial co-products as substrates, palm fatty acid distillate (PFAD) and soybean fatty acid distillate (SFAD). The highest productivity was observed for SFAD after 48 h presenting 79.1% of conversion using only 10% of biocatalyst and free-solvent system. This is about ca. eight times higher than commercial free RML in the same conditions. The stabilizing agents study revealed that the treatment using glutaraldehyde (GA) and poly(ethylene glycol) (PEG) enabled increased stability and reuse of biocatalyst. It was observed by SEM analysis that the treatment modified the cell morphology. RML-PIR1-GA presented 87.9% of the initial activity after 6 reuses, whilst the activity of unmodified RML-PIR decreased by 40% after the first use. These results were superior to those obtained in the literature, making this new biocatalyst promising for biotechnological applications, such as the production of biofuels on a large scale.
Assuntos
Agricultura , Biocombustíveis/microbiologia , Resíduos Industriais , Lipase/metabolismo , Rhizomucor/enzimologia , Saccharomycetales/metabolismo , Biocatálise , Esterificação , Especificidade por Substrato , TemperaturaRESUMO
El objetivo fue evaluar los fungal ruminales como probióticos potenciales en asociación con dietas ricas en granos en el actuación y rendimiento de la canal y la acidosis en corderos Santa Inês/Dorper. Adoptó un diseño completamente aleatorizado en factorial 4x2, con dos procesos (granos de maíz molidos o integral) y cuatro inoculantes (sin inóculo, con Rhizomucor spp., con Aspergillus terreus y con una mezcla de los dos hongos). Durante 75 días se utilizaron ocho establos con cinco corderos/puesto. Las dietas con 85% de maíz asociado con 15% de núcleo granos se proporcionaron ad libitum dos veces al día, con inoculantes rociados sobre el alimento. Durante la prueba se realizaron cinco recoleccion es de muestras de los ingredientes de las dietas y sobras para realizar los análisis centesimales. Los animales se pesaron cada das semanas para evaluar los índices de rendimiento. Al final de la prueba, en el momento del sacrificio, se calcularon los rendimientos en canal de cinco animales por tratamiento. Los datos se sometieron a análisis de variación y las medias de los datos se compararon mediante la prueba de Tukey al 5%. No hubo interacción entre los factores de procesamiento y probióticos para las variables de desempeño (P > 0,05), así como tampoco hubo efecto de agregar los probióticos sobre el desempeño de las ovejas alimentadas con los diferentes tratamientos...(AU)
The objective of this study was to evaluate ruminal fungi as potential probiotics in association with high-grain diets in carcass performance and yield, and acidosis in Santa Inês/Dorper lambs. A 4x2 factorial was adopted in CRD, with two processes (ground or whole grain) and four inoculants (without inoculants, with Rhizomucor spp., With Aspergillus terreus and with a mixture of fungi). Eight stalls with five lambs/stall were used for 75 days. Diets with 85 % corn plus 15 % nucleus are provided twice a day, with inoculants sprayed at the time of food handling. Five collections of ingredients of the ingredients of the diets, diets and leftovers were carried out during the experiment. The animals were weighed every two weeks. At the end of the experiment, on slaughter, the carcass index of five animals/treatment was calculated. Data were submitted to ANOVA and the data averages compared by the Tukey test at 5 %. There was no interaction between the processing and probiotic factors for the performance variables (P > 0.05), just as there was no effect of the addition of probiotics on the performance of the sheep fed with the different treatments (P > 0.05). Higher consumption of DM, MM, EE, NDF and NFC was observed for animals fed with whole grain corn (MGI) (P < 0.05). There was no significant difference (P > 0.05) for the consumption of OM, CP, MDG...(AU)
Objetivou-se avaliar fungos ruminais como potenciais probióticos em associação a dietas de alto grão no desempenho e rendimento de carcaça, e quadro de acidose em borregos Santa Inês/Dorper. Adotou-se fatorial 4x2 em DIC, sendo dois processamentos (grão moído ou inteiro) e quatro inoculantes (sem inóculos, com Rhizomucor spp., com Aspergillus terreus e com mistura dos dois fungos). Utilizou-se oito baias com cinco borregos/baia, durante 75 dias. Dietas com 85% de milho associadas à 15% de núcleo foram fornecidas ad libitum duas vezes ao dia, com inóculos aspergidos no arraçoamento. Realizou-se cinco coletas de amostras dos ingredientes das dietas e das sobras durante o ensaio para realização das análises centesimais. Pesou-se os animais quinzenalmente para avaliação dos índices de desempenho. Ao final do ensaio, na ocasião do abate, foram calculados os rendimentos de carcaça de cinco animais por tratamento. Os dados foram submetidos à ANOVA e as médias dos dados comparadas pelo teste de Tukey a 5 %. Não houve interação entre os fatores processamento e probióticos para as variáveis de desempenho (P > 0.05), assim como, não houve efeito da adição dos probióticos sobre o desempenho dos ovinos alimentados com os diferentes tratamentos (P > 0.05). Foi observado maior consumo de MS, MM, EE, FDN e CNF para os animais alimentados com milho grão inteiro (MGI)...(AU)
Assuntos
Animais , Probióticos/administração & dosagem , Fungos , Ovinos , Rhizomucor , Aspergillus , Grão Comestível , Zea mays , AcidoseRESUMO
El objetivo fue evaluar los fungal ruminales como probióticos potenciales en asociación con dietas ricas en granos en el actuación y rendimiento de la canal y la acidosis en corderos Santa Inês/Dorper. Adoptó un diseño completamente aleatorizado en factorial 4x2, con dos procesos (granos de maíz molidos o integral) y cuatro inoculantes (sin inóculo, con Rhizomucor spp., con Aspergillus terreus y con una mezcla de los dos hongos). Durante 75 días se utilizaron ocho establos con cinco corderos/puesto. Las dietas con 85% de maíz asociado con 15% de núcleo granos se proporcionaron ad libitum dos veces al día, con inoculantes rociados sobre el alimento. Durante la prueba se realizaron cinco recoleccion es de muestras de los ingredientes de las dietas y sobras para realizar los análisis centesimales. Los animales se pesaron cada das semanas para evaluar los índices de rendimiento. Al final de la prueba, en el momento del sacrificio, se calcularon los rendimientos en canal de cinco animales por tratamiento. Los datos se sometieron a análisis de variación y las medias de los datos se compararon mediante la prueba de Tukey al 5%. No hubo interacción entre los factores de procesamiento y probióticos para las variables de desempeño (P > 0,05), así como tampoco hubo efecto de agregar los probióticos sobre el desempeño de las ovejas alimentadas con los diferentes tratamientos...
The objective of this study was to evaluate ruminal fungi as potential probiotics in association with high-grain diets in carcass performance and yield, and acidosis in Santa Inês/Dorper lambs. A 4x2 factorial was adopted in CRD, with two processes (ground or whole grain) and four inoculants (without inoculants, with Rhizomucor spp., With Aspergillus terreus and with a mixture of fungi). Eight stalls with five lambs/stall were used for 75 days. Diets with 85 % corn plus 15 % nucleus are provided twice a day, with inoculants sprayed at the time of food handling. Five collections of ingredients of the ingredients of the diets, diets and leftovers were carried out during the experiment. The animals were weighed every two weeks. At the end of the experiment, on slaughter, the carcass index of five animals/treatment was calculated. Data were submitted to ANOVA and the data averages compared by the Tukey test at 5 %. There was no interaction between the processing and probiotic factors for the performance variables (P > 0.05), just as there was no effect of the addition of probiotics on the performance of the sheep fed with the different treatments (P > 0.05). Higher consumption of DM, MM, EE, NDF and NFC was observed for animals fed with whole grain corn (MGI) (P 0.05) for the consumption of OM, CP, MDG...
Objetivou-se avaliar fungos ruminais como potenciais probióticos em associação a dietas de alto grão no desempenho e rendimento de carcaça, e quadro de acidose em borregos Santa Inês/Dorper. Adotou-se fatorial 4x2 em DIC, sendo dois processamentos (grão moído ou inteiro) e quatro inoculantes (sem inóculos, com Rhizomucor spp., com Aspergillus terreus e com mistura dos dois fungos). Utilizou-se oito baias com cinco borregos/baia, durante 75 dias. Dietas com 85% de milho associadas à 15% de núcleo foram fornecidas ad libitum duas vezes ao dia, com inóculos aspergidos no arraçoamento. Realizou-se cinco coletas de amostras dos ingredientes das dietas e das sobras durante o ensaio para realização das análises centesimais. Pesou-se os animais quinzenalmente para avaliação dos índices de desempenho. Ao final do ensaio, na ocasião do abate, foram calculados os rendimentos de carcaça de cinco animais por tratamento. Os dados foram submetidos à ANOVA e as médias dos dados comparadas pelo teste de Tukey a 5 %. Não houve interação entre os fatores processamento e probióticos para as variáveis de desempenho (P > 0.05), assim como, não houve efeito da adição dos probióticos sobre o desempenho dos ovinos alimentados com os diferentes tratamentos (P > 0.05). Foi observado maior consumo de MS, MM, EE, FDN e CNF para os animais alimentados com milho grão inteiro (MGI)...
Assuntos
Animais , Aspergillus , Fungos , Grão Comestível , Ovinos , Probióticos/administração & dosagem , Rhizomucor , Acidose , Zea maysRESUMO
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.
Assuntos
Análise do Solo , Fungos/fisiologia , Intoxicação por Chumbo , Resíduos de Alimentos , Aspergillus niger , Penicillium chrysogenum , RhizomucorRESUMO
In this work, the synthesis of two fruit flavor esters, namely methyl and ethyl butyrate, by lipase from Rhizomucor miehei immobilized onto chitosan in the presence of the surfactant sodium dodecyl sulfate SDS was investigated. In the optimized conditions, maximum esterification yield for ethyl butyrate and methyl butyrate was (92 ± 1%) and (89 ± 1%), respectively. Esterification yields for both reactions were comparable or even superior to the ones achieved when the synthesis was catalyzed by a commercial enzyme, Lipozyme®, at the same reaction conditions. For ethyl butyrate, the developed biocatalyst was used for seven consecutive cycles of reaction with retention of its catalytic activity. For methyl butyrate synthesis the biocatalyst was used for four consecutive cycles without loss of its catalytic activity. The results show that chitosan may be employed in obtaining biocatalysts with high catalytic efficiency and can successfully replace the currently commercial available biocatalysts.
Assuntos
Butiratos/química , Rhizomucor/metabolismo , Quitosana , Enzimas Imobilizadas , Esterificação , Ésteres/síntese química , Aromatizantes/síntese química , Proteínas Fúngicas , Cinética , Lipase/metabolismo , Lipase/farmacologia , Dodecilsulfato de Sódio/química , TensoativosRESUMO
We present an efficient approach to the synthesis of a series of glycyrrhetinic acid derivatives. Six derivatives, five of them new compounds, were obtained through chemoenzymatic reactions in very good to excellent yield. In order to find the optimal reaction conditions, the influence of various parameters such as enzyme source, nucleophile:substrate ratio, enzyme:substrate ratio, solvent and temperature was studied. The excellent results obtained by lipase catalysis made the procedure very efficient considering their advantages such as mild reaction conditions and low environmental impact. Moreover, in order to explain the reactivity of glycyrrhetinic acid and the acetylated derivative to different nucleophiles in the enzymatic reactions, molecular docking studies were carried out. In addition, one of the synthesized compounds exhibited remarkable antiviral activity against TKâ¯+â¯and TK- strains of Herpes simplex virus type 1 (HSV-1), sensitive and resistant to acyclovir (ACV) treatment.
Assuntos
Antivirais/farmacologia , Ácido Glicirretínico/farmacologia , Herpesvirus Humano 1/efeitos dos fármacos , Lipase/metabolismo , Simulação de Acoplamento Molecular , Aciclovir/farmacologia , Antivirais/química , Antivirais/metabolismo , Biocatálise , Candida/enzimologia , Carica/enzimologia , Relação Dose-Resposta a Droga , Eurotiales/enzimologia , Ácido Glicirretínico/química , Ácido Glicirretínico/metabolismo , Testes de Sensibilidade Microbiana , Estrutura Molecular , Rhizomucor/enzimologia , Rhizopus/enzimologia , Relação Estrutura-AtividadeRESUMO
Lipase from Rhizomucor miehei (RML) was immobilized onto chitosan support in the presence of some surfactants added at low levels using two different strategies. In the first approach, the enzyme was immobilized in the presence of surfactants on chitosan supports previously functionalized with glutaraldehyde. In the second one, after prior enzyme adsorption on chitosan beads in the presence of surfactants, the complex chitosan beads-enzyme was then cross-linked with glutaraldehyde. The effects of surfactant concentrations on the activities of free and immobilized RML were evaluated. Hexadecyltrimethylammonium bromide (CTAB) promoted an inhibition of enzyme activity while the nonionic surfactant Triton X-100 caused a slight increase in the catalytic activity of the free enzyme and the derivatives produced in both methods of immobilization. The best derivatives were achieved when the lipase was firstly adsorbed on chitosan beads at 4 °C for 1 h, 220 rpm followed by cross-link the complex chitosan beads-enzyme with glutaraldehyde 0.6% v.v-1 at pH 7. The derivatives obtained under these conditions showed high catalytic activity and excellent thermal stability at 60° and 37 °C. The best derivative was also evaluated in the synthesis of two flavor esters namely methyl and ethyl butyrate. At non-optimized conditions, the maximum conversion yield for methyl butyrate was 89%, and for ethyl butyrate, the esterification yield was 92%. The results for both esterifications were similar to those obtained when the commercial enzyme Lipozyme® and free enzyme were used in the same reaction conditions and higher than the one achieved in the absence of the selected surfactant.
Assuntos
Quitosana/sangue , Enzimas Imobilizadas/química , Proteínas Fúngicas/química , Lipase/química , Rhizomucor/enzimologia , Tensoativos/química , Estabilidade Enzimática , Concentração de Íons de HidrogênioRESUMO
In this work, the synthesis of acylglycerides with high nutritional value was carried out by enzymatic esterification at sn-2 position of 1,3-dicaprin with palmitic acid. A comparative study of the performance of several biocatalysts according to the obtained products was carried out. The results obtained with several of the biocatalysts evaluated are very interesting, and it would be possible to use them to obtain a mixture of acylglycerides to act as a fat substitute. The final product was composed of about 90% of nutritionally attractive glycerides. These glycerides were medium-chain length triglycerides, medium-long chain triglycerides (mainly triglycerides with medium chain fatty acids at sn-1 and sn-3 positions and long chain fatty acid at sn-2 position), and 1,3-diglycerides. Pseudomonas fluorescens lipase and Burkholderia cepacia lipase immobilized on chitosan demonstrated unusual high activity in the sn-2 esterification of 1,3-dicaprin with palmitic acid at 45 °C and 12 h with 33% yield to 1,3-dicaproyl-2-palmitoyl glycerol. Burkholderia cepacia lipase has the advantage of being immobilized; however, BCL/chitosan has the advantages of being immobilized and therefore its easy recovery from the reaction media.