Significant production of vanillin and in vitro amplification of ech gene in local bacterial isolates / Produção significativa de vanilina e amplificação in vitro do gene ech em isolados bacterianos locais

Abstract Vanillin is the major component which is responsible for flavor and aroma of vanilla extract and is produced by 3 ways: natural extraction from vanilla plant, chemical synthesis and from microbial transformation. Current research was aimed to study bacterial production of vanillin from native natural sources including sewage and soil from industrial areas. The main objective was vanillin bio-production by isolating bacteria from these native sources. Also to adapt methodologies to improve vanillin production by optimized fermentation media and growth conditions. 47 soil and 13 sewage samples were collected from different industrial regions of Lahore, Gujranwala, Faisalabad and Kasur. 67.7% bacterial isolates produced vanillin and 32.3% were non-producers. From these 279 producers, 4 bacterial isolates selected as significant producers were; A3, A4, A7 and A10. These isolates were identified by ribotyping as A3 Pseudomonas fluorescence (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) and A10 Bacillus subtilis (KT962919). Vanillin producers were further tested for improved production of vanillin and were grown in different fermentation media under optimized growth conditions for enhanced production of vanillin. The fermentation media (FM) were; clove oil based, rice bran waste (residues oil) based, wheat bran based and modified isoeugenol based. In FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36, and FM37, the selected 4 bacterial strains produced significant amounts of vanillin. A10 B. subtilis produced maximum amount of vanillin. This strain produced 17.3 g/L vanillin in FM36. Cost of this fermentation medium 36 was 131.5 rupees/L. This fermentation medium was modified isoeugenol based medium with 1% of isoeugenol and 2.5 g/L soybean meal. ech gene was amplified in A3 P. fluorescence using ech specific primers. As vanillin use as flavor has increased tremendously, the bioproduction of vanillin must be focused.

Resumo A vanilina é o principal componente responsável pelo sabor e aroma do extrato de baunilha e é produzida de três formas: extração natural da planta da baunilha, síntese química e transformação microbiana. A pesquisa atual teve como objetivo estudar a produção bacteriana de vanilina a partir de fontes naturais nativas, incluindo esgoto e solo de áreas industriais. O objetivo principal era a bioprodução de vanilina por meio do isolamento de bactérias dessas fontes nativas. Também para adaptar metodologias para melhorar a produção de vanilina por meio de fermentação otimizada e condições de crescimento. Foram coletadas 47 amostras de solo e 13 de esgoto de diferentes regiões industriais de Lahore, Gujranwala, Faisalabad e Kasur; 67,7% dos isolados bacterianos produziram vanilina e 32,3% eram não produtores. Desses 279 produtores, 4 isolados bacterianos selecionados como produtores significativos foram: A3, A4, A7 e A10. Esses isolados foram identificados por ribotipagem como fluorescência A3 Pseudomonas (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) e A10 Bacillus subtilis (KT962919). Os produtores de vanilina foram posteriormente testados para produção aprimorada de vanilina e foram cultivados em diferentes meios de fermentação sob condições de crescimento otimizadas para produção aprimorada de vanilina. Os meios de fermentação (FM) foram: à base de óleo de cravo, à base de resíduos de farelo de arroz (resíduos de óleo), à base de farelo de trigo e à base de isoeugenol modificado. Em FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36 e FM37, as 4 cepas bacterianas selecionadas produziram quantidades significativas de vanilina. A10 B. subtilis produziu quantidade máxima de vanilina. Essa cepa produziu 17,3 g / L de vanilina em FM36. O custo desse meio de fermentação 36 foi de 131,5 rúpias / L. Esse meio de fermentação foi um meio à base de isoeugenol modificado com 1% de isoeugenol e 2,5 g / L de farelo de soja. O gene ech foi amplificado em A3 P. fluorescence usando primers específicos para ech. Como o uso da vanilina como sabor aumentou tremendamente, a bioprodução da vanilina deve ser focada.

Fungal decomposition of chicken-feather waste in submerged and solid-state fermentation / Decomposição fúngica de resíduos de pena de frango em fermentação submersa e de estado sólido

Abstract Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.

Resumo A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.

Fungal decomposition of chicken-feather waste in submerged and solid-state fermentation

Abstract Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.

Resumo A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.

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.

Fungal decomposition of chicken-feather waste in submerged and solid-state fermentation / Decomposição fúngica de resíduos de pena de frango em fermentação submersa e de estado sólido

Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.

A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.

Fungal decomposition of chicken-feather waste in submerged and solid-state fermentation / Decomposição fúngica de resíduos de pena de frango em fermentação submersa e de estado sólido

Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.(AU)

A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.(AU)

Evaluation of Lactobacillus Plantarum Additive on Growth Performance, Excreta Microbiota, Nutrient Digestibility, Gas Emission, and Meat Quality in Ross308-Broilers

The intention of this study was to analyze the effects of Lactobacillus plantarum (L. plantarum) additive with different nutrient density diets on growth performance, excreta microbiota, nutrient digestibility, gas emission, and meat quality in Ross308-broilers. A total of 576 mixed-sex, 1-d old Ross-308 chicks were randomly allocated to one of four treatment groups with 8 replication and 18 chicks/cage. For a period of 35 days, HD and LD group chicks were fed with commercial corn and soybean meal-based basal diet which contains high and low nutrient density diet, respectively. The other treatment groups LP1 and LP2 chicks were fed with LD+ 0.05% and 0.01 % of L. plantarum, respectively. During day 21 and the overall experimental period, the body weight gain of broilers significantly increased (p<0.05) in HD and L. plantarum groups compared to the LD group. On day 35, broilers fed L. plantarum additive had significantly increased (p<0. 05) the nutrient digestibility of dry matter and nitrogen compared to those fed HD and LD diets. Moreover, dietary inclusion of L. plantarum additive had significantly increased (p<0.05) lactobacillus population and decreased (p>0.05) E. coli and ammonium emission. However, the meat quality traits were not affected by experimental diets. In conclusion, we infer that a low-density diet with 0.1% of L. plantarum additive could serve as an excellent alternative feed additive to enhance the performance of broilers.(AU)

Inclusion of Organic Acids in the Drinking Water and Feed for the Control of Salmonella Heidelberg in Broilers

Poultry products may be a source of foodborne human salmonellosis. The use of alternatives to antimicrobials that are not harmful to humans may reduce the presence of Salmonella spp. in poultry production. Among the products used, organic acids stand out. In the present study, three different organic acid (OA) blends were evaluated for the control of Salmonella Heidelberg (SH) in commercial broilers. Day-old chicks (n = 114) were randomly assigned to four treatments, with three replicates of 12 birds each. Birds in treatments A and B received SCFA (0.2mL/L) and SCFA + MCFA (0.2mL/L), respectively, in the drinking water, while birds in treatment C received SCFA + MCFA in the feed (2g/Kg of feed). Birds from treatment D did not receive OAs (control group). At 8 days of age, each bird was orally inoculated with SH at 108 CFU/mL, and cloacal swabs and SH enumeration of the cecal content were performed 24-, 48-, and 72-hours post-inoculation (hpi). The results show a reduction of both SH shedding and counts in the birds fed OAs at all pi times relative to the control birds. Fecal shedding was significantly lower in the OA-treated groups compared with the control group. As for SH presence in the cecum, significant differences were detected between groups C and D at 24 and 72 hpi, and between groups B and D at 72 hpi. The results of this study indicate that the use of feeding OAs to broilers may contribute to reduce the incidence of SH in the poultry production chain, allowing better flock health management, provided an efficient biosecurity program is employed.(AU)

Ingestive behavior of lambs fed relocated and inoculated whole-plant corn silage

We aimed to determine the effect of relocation of whole-plant corn silage (WPCS) with different fermentation profiles on ingestive behavior of lambs. Twenty-four male Santa Inês lambs, were used in a completely randomized block design based on initial bodyweight (17.5 ± 1.8 kg) for the following treatments: CS: WPCS (not inoculated and not relocated); R-12h: WPCS relocated for 12h; IR-12h: WPCS inoculated with Lactobacillus plantarumand relocated for 12h; and R-24h: WPCS not inoculated and relocated for 24h. Ingestive behavior was observed as continuous (feeding, ruminating, and idling) and specific (urination, defecation, and water and salt access frequency) activities for 48h (3h intervals for a total of eight periods in 24h). The behavioral activities were affected only by periods (p = 0.0001). Feeding peaks was high at 8:00–11:00am(54.2%) and 5:00-8:00pm(54.9%) in compare to ruminating and idling at the same period. Rumination time was intense at night, at 63.7% (2:00–5:00am) and 69.7% (5:00–8:00am). Idling time (49.7%) was long after silage supply. The specific activities were intense during the morning periods. Supply of WPCS relocated up to 24h did not alter the ingestive behavior.(AU)

Sugarcane total mixed ration silage ensiling with chitosan and homolactic microbial inoculant: characteristics of silage and animal digestion / Silagem de dieta total de cana de açúcar ensilada com quitosana e inoculante microbiano homoláctico: características da silagem e digestão animal

This study aimed to evaluate total mixed ration silages with sugarcane and the additives microbial inoculant and chitosan. Thirty mini-silos were used in a completely randomized design, with three treatments and ten replications. Silages were composed of sugarcane mixed with corn bran, whole soybean, urea, and mineral mixtures at a 50:50 roughage to concentrate ratio. Treatments consisted of control silage, microbial additive (Lactobacillus plantarum + Pediococcus acidilactici, 4 g/t of KeraSil, Kera Nutrição Animal), and chitosan (10 g/kg of natural matter). Silages were evaluated for fermentation and microbiological profile, fermentation losses, aerobic stability, chemical-bromatological composition, intake, and digestibility. Fermentation profile showed no significant difference between treatments for pH values, with a mean value of 4.79. Production of acetic and propionic acids showed no difference between treatments, with mean values of 7.34 and 0.053 mmol/kg DM, respectively. Dry matter, organic matter, and crude protein intake of the total mixed ration silage differed statistically from the other treatments (P<0.05), but fresh sugarcane and sugarcane silage intake did not differ from each other (P>0.05). Digestibility values of DM, OM, and NDF were higher in the total mixed ration silage (P<0.05), while sugarcane silage and fresh sugarcane showed no difference from each other (P>0.05). Total mixed ration silage increased nutrient intake and digestibility, with a better fermentation pattern when added with the microbial inoculant.

O objetivo foi avaliar as silagens de ração total com cana de açúcar e aditivos inoculante microbiano e quitosana. Foram utilizados trinta mini-silos utilizando o delineamento inteiramente casualizado, com três tratamentos e dez repetições. As silagens foram compostas por cana-de-açúcar misturada com farelo de milho, soja integral, uréia e misturas minerais na proporção de 50:50. Os tratamentos foram: silagem controle, aditivo microbiano (Lactobacillus plantarum + Pediococcus acidilactici, 4 g/t de KeraSil, Kera Nutrição Animal) e quitosana (10 g/kg da matéria natural). As silagens foram avaliadas quanto à fermentação e perfil microbiológico, perdas de fermentação, estabilidade aeróbica, composição químico-bromatológica, consumo e digestibilidade. O perfil de fermentação não mostrou diferença significativa entre tratamentos para os valores de pH, com um valor médio de 4,79. A produção de ácidos acéticos e propiônicos não mostrou diferença entre os tratamentos, com valores médios de 7,34 e 0,053 mmol/kg DM, respectivamente. A matéria seca, a matéria orgânica e a ingestão de proteína bruta da mistura total de ração silagem diferiram estatisticamente dos outros tratamentos (P<0,05), mas a ingestão de cana-de-açúcar fresca e de silagem de cana-de-açúcar não diferiram uma da outra (P>0,05). Os valores de digestibilidade de DM, OM e NDF foram maiores na silagem de ração mista total (P<0,05), enquanto a silagem de cana de açúcar e a cana de açúcar fresca não mostraram diferença entre si (P>0,05). A silagem de ração mista total aumentou a ingestão de nutrientes e a digestibilidade, com um melhor padrão de fermentação quando adicionado o inoculante microbiano.

Silage quality of two energy-cane cultivars produced with different additives / Qualidade da silagem de duas cultivares de cana-energia produzidas com diferentes aditivos

This study examined the quality of silage from two cultivars of energy cane (Saccharum spontaneum) produced with different additives. The experiment was laid out in a completely randomized design with a 2×5 factorial arrangement of treatments consisting of two cultivars (VG3, VG1126) and four additives (1% urea; 1% NaOH; 1% CaO; and 1.0 × 1011 CFU g-1 of Lactobacillus plantarum- LP) + control, with eight replicates. Experimental mini-silos were used, which were opened 72 days after ensiling. The dry matter (DM), crude protein (CP), organic matter (OM), mineral matter (MM), ether extract (EE), neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents, and silage degradability were. We identified significant interactions effect between cultivar and type of additive for DM, CP, EE, NDF. ADF. Organic and mineral matter percentage were similar between the two cultivars. LP was the least beneficial additive for DM degradation.After 72 h of fermentation, in vitro NDF degradation of cultivar VG3 increased due to urea, NaOH, and CaO, whereas degradation cultivar VG1126 decreased when inoculated with LP or CaO. The NaOH and CaO improved silage degradability thus improving its quality. The use of LP in the present study did not benefit silage for energy-cane cultivars. The use of NaOH and CaO improves the quality of the silage, two varieties of sugarcane energy VG3 and VG1126, enabling its use with better efficiency for animal feed.

Este estudo avaliou a qualidade da silagem de duas cultivares de cana-de-energia (Saccharumspontaneum) produzidas com diferentes aditivos. O experimento foi instalado em delineamento inteiramente casualizado, com arranjo fatorial 2 × 5 de tratamentos composto por duas cultivares (VG3, VG1126) e quatro aditivos (1% uréia; 1% NaOH; 1% CaO; e 1,0 × 1011 UFC g -1 de Lactobacillus plantarum- LP) + controle, com oito repetições. Foram utilizados minisilos experimentais, os quais foram abertos 72 dias após a ensilagem. Os teores de matéria seca (MS), proteína bruta (PB), matéria orgânica (MO), matéria mineral (MM), extrato etéreo (EE), fibra em detergente neutro (FDN) e fibra em detergente ácido (FDA) e degradabilidade da silagem foram avaliados. Identificou-se efeito de interação significativo entre cultivar e tipo de aditivo para MS, PB, EE, FDN. ADF. As porcentagens de matéria orgânica e mineral foram semelhantes entre as duas cultivares. LP foi o aditivo menos benéfico para a degradação da MS. Após 72 h de fermentação, a degradação in vitro da FDN da cultivar VG3 aumentou devido à uréia, NaOH e CaO, enquanto a degradação da cultivar VG1126 diminuiu quando inoculada com LP ou CaO. O NaOH e o CaO melhoraram a degradabilidade da silagem, melhorando assim sua qualidade. O uso da LP no presente estudo não beneficiou a silagem para cultivares de cana-de-energia. O uso de NaOH e CaO melhora a qualidade da silagem, duas variedades de energia da cana-de-açúcar VG3 e VG1126, possibilitando seu uso com melhor eficiência para ração animal.

Whole-plant soybean ensiling with chitosan and homolactic microbial inoculant: fermentative profile, aerobic stability, and sheep intake and digestibility / Ensilagem de planta inteira de soja com quitosana e inoculante microbiano homolático: perfil fermentativo, estabilidade aeróbia e consumo e digestibilidade em ovinos

This study aimed to evaluate the effects of chitosan and homolactic microbial inoculant on fermentative losses, chemical composition, fermentative profile, and aerobic stability of whole-plant soybean silage (WPSS). Additionally, it was evaluated nutrients intake and digestibility of sheep fed increasing levels of WPSS. Thirty experimental silos were randomly allocated to one of the following treatments: 1) CON: control, WPSS without additives; 2) LPPA: WPSS with Lactobacillus plantarum and Pediococcus acidilactici; and 3) CHI: chitosan, WPSS with 5 g/kg of chitosan. Ten male sheep were used to evaluate increasing dietary levels of WPSS: 0, 200, 400, 600, and 800 g/kg of diet dry matter (DM). Additives increased silage lactic acid bacteria and decreased the count of mold and yeast, gas, and total losses. Silages treated with additives had lower pH, NH3-N, and ethanol concentrations and higher lactic and propionic acids relative to CON. LPPA-treated silos showed higher organic matter and non-fiber carbohydrates content than CHI-ones. Additives increased the aerobic stability of WPSS. The addition of WPSS in sheep diets linearly increased nutrients intake and digestibility. Chitosan and LPPA improve WPSS fermentation, aerobic stability, and nutritional value. The WPSS in substitution to Cynodon hay increases sheep feed intake and nutrients digestibility.

Este estudo objetivou avaliar os efeitos da adição de quitosana e inoculante homolático sobre as perdas fermentativas, composição química, perfil fermentativo e estabilidade aeróbia da silagem de planta inteira de soja (SPIS). Em adição, foi avaliado o consumo e a digestibilidade de nutrientes em ovinos alimentados com dietas contendo níveis crescentes de SPIS. Trinta silos experimentais foram aleatoriamente alocados a um dos seguintes tratamentos: 1) CON: controle, SPIS sem aditivos; 2) LPPA: SPIS com Lactobacillus plantarum e Pediococcus acidilactici; e 3) QUI: quitosana, SPIS com 5 g/kg de quitosana. Dez ovinos machos foram usados para avaliar os níveis dietéticos de SPIS: 0, 200, 400, 600 e 800 g/kg da matéria seca (MS). Os aditivos aumentaram a contagem de bactérias láticas e reduziram a contagem de fungos e leveduras e as perdas fermentativas totais da SPIS. Silagens tratadas com aditivos tiveram menores pH, N-NH3 e etanol e maiores concentrações de ácido lático e propiônico, quando comparadas ao tratamento controle. Silos tratados com LPPA apresentaram maiores teores de matéria orgânica e carboidratos não fibrosos do que aqueles do tratamento QUI. Os aditivos aumentaram a estabilidade aeróbia da SPIS. A adição de SPIS na dieta de ovinos aumentou linearmente o consumo e a digestibilidade dos nutrientes. Quitosana e inoculante microbiano homolático melhoram a fermentação, estabilidade aeróbia e o valor nutricional da SPIS. A substituição de feno de Cynodon por SIPS aumenta o consumo e a digestibilidade dos nutrientes em ovinos.

Clinical safety of oral licomycin treatment in Nile tilapia (Oreochromis niloticus) / Segurança clínica do tratamento via oral com lincomicina em tilapias (Oreochromis niloticus)

Lincomycin is a broad-spectrum antimicrobial acting against Gram-positive bacteria, widely used in veterinary medicine. In fish, there are only limited in vitro data, thus requiring the design of effective therapeutic protocols for their use in aquatic organisms. In this context, the objective was to evaluate the clinical safety of lincomycin treatment, administered orally in tilapia, through hematological, biochemical and somatic index evaluation. A total of 136 tilapia (+100g) were randomly distributed in 17 tanks (100L of water, n=8) to constitute the following treatments: TO (control group, not treated with lincomycin); T1, T2, T3 (treated with 10, 20 and 40mg/kg¹ of lincomycin b.w., respectively) and T4 physiological standard (reference values). Eight animals were sampled per treatment in 4 periods: 2, 4 and 8 days post-treatment (DPT), and a group that was treated for 8 days with the drug and then treated only with commercial feed until the 12th day (recovery period). Tilapia treated with lincomycin had no difference in the hematological and leukocyte evaluation, in the hepatic, renal and splenic somatic index. However, they presented a transient increase in the values of ALT, AST, cholesterol, triglycerides and creatinine, which returned to normal levels after the period of recovery (12DPT). Furthermore, an increase in total protein, albumin and globulin levels was observed in treated animals. It is concluded that although there were some transient changes during the experiment, lincomycin has a good clinical safety margin at doses of 10, 20 and 40mg/Kg¹ b.w. for Nile tilapia.

A lincomicina é um antimicrobiano de amplo espectro atuando contra bactérias gram-positivas, amplamente utilizada na medicina veterinária. Em peixes existem apenas dados limitados in-vitro, necessitando assim, de delineamento de protocolos terapêuticos eficazes para seu uso em organismos aquáticos. Desse modo, objetivou-se avaliar a segurança clínica do tratamento com lincomicina, administrada por via oral em tilápias, por meio da avaliação hematológica, bioquímica e indice somático hepático, renal e esplênico. Foram utilizadas 136 tilápias (±100g), distribuídas aleatoriamente em 17 tanques (100L de água, n=8) para constituir as repetições dos diferentes tratamentos: TO (grupo controle, não tratado com lincomicina); T1, T2, T3 (tratados com 10, 20 e 40mg/kg de p.v. de lincomicina, respectivamente) e T4 padrão fisiológico (valores de referência). Oito animais foram amostrados por tratamento em 4 períodos: 2, 4 e 8 dias pós-tratamento (DPT) com lincomicina, e um grupo que foi tratado por 8 dias com o fármaco e após isso tratado apenas com ração comercial até o 12º dia (período de recuperação). Tilápias tratadas com lincomicina não tiveram diferença na avaliação hematológica e leucocitária, no índice somático hepático, renal e esplênico, entretanto, apresentaram um aumento transitório nos valores de ALT, AST, colesterol, triglicérides e creatinina, que retornaram aos níveis normais após o período de recuperação (12DPT). Ademais, foram observados um incremento nos níveis de proteína total, albumina e globulina nos animais tratados. Conclui-se que embora tenha ocorrido algumas alterações transitórias ao decorrer do experimento, a lincomicina apresenta boa margem de segurança clinica nas doses de 10, 20 e 40mg/Kg¹ p.v para tilápia do Nilo.

Responses of Broiler Chickens Fed Low or High Non-Starch Polysaccharide Diets and the Addition of Humic Substances from a Worm Compost

The objective was to evaluate the productivity, microbiology and histopathology of the jejunum, ceca and liver in broilers fed a control or a high non-starch polysaccharide (HNSPs) diet added with an extract of humic substances (EHS). 240 broilers individually allocated, from 21-42 d of age were assigned to a factorial arrangement of 2 types of diets: 1) A corn/soybean meal diet (Control) and 2) As Control plus 7% distillers dried grain with solubles and 12% of wheat bran (HNSPs), and 3 growth promoters: 1) Antibiotic growth promoters (AGP); 2) without AGP (NAGP) and C) with 0.5% of EHS. At the end of the trial, the breast and carcass were weighed and samples of the intestine, ceca and liver were taken for microbial and histopathology analysis. Results were subjected to ANOVA. EHS-fed broilers had lower feed conversion ratio (FCR) and total aerobic bacterial (TAB) counts in the liver and higher ashes digestibility with the Control diet, but negative responses were seen with the HNSPs diet (Type of diet and growth promoter interaction, p≤0.05). The lesion scores in the jejunum were lower in EHS-fed broilers with the Control and HNSPs diet (p≤0.05). The TAB and E. coli were lower in the jejunum and ceca of AGP-fed broilers (p<0.01) compared to NAGP and EHS groups. EHS-fed broilers showed improved FCR and ileal ashes digestibility and lower TAB in the liver with the Control diet and had lower lesion score in the jejunum and similar weight gain compared to the AGP-fed broilers.(AU)

Effects of Medium Chain Fatty Acid Supplementation in Broiler Diet on Microbiological Quality of Litter

In our experiment, coconut and palm oil supplementation was added to the diet of broiler chickens to prove the hypothesis that medium chain fatty acids (MCFAs) can reduce the occurrence of potential pathogens (Clostridium perfingens, Coliform) in gut microflore and therefore improve bird welfare. Cobb 500 cockerels were divided in five groups. Control birds were fed commercial broiler diet, while birds in the four treatment group diets were supplemented with coconut, palm oil, or a combination of the two, respectively. As a positive control, sunflower oil supplementation was included in the diet of the fifth group. During the 28 days of the study (from 14 to 42 days of life), 5 samplings were scheduled, when excreta samples were taken from the litter to analyse total microbial count and the number of Clostridia, Coliforms, and Salmonella. According to the results of microbiological analysis, coconut oil supplementation led to the continuous decline of Clostridium perfringens numbers until they vanished by the end of the experimental period. A similar but faster decline was found as a result of palm oil and the combined (palm oil+coconut oil) treatment. However, the number of Coliforms and total microbial count changed only slightly by the end of the study. No Salmonella spp. was present in the samples throughout the experiment. Altogether, coconut and palm oil supplementation has a beneficial effect on the microbiological composition of poultry litter.(AU)

Effects of Medium Chain Fatty Acid Supplementation in Broiler Diet on Microbiological Quality of Litter

In our experiment, coconut and palm oil supplementation was added to the diet of broiler chickens to prove the hypothesis that medium chain fatty acids (MCFAs) can reduce the occurrence of potential pathogens (Clostridium perfingens, Coliform) in gut microflore and therefore improve bird welfare. Cobb 500 cockerels were divided in five groups. Control birds were fed commercial broiler diet, while birds in the four treatment group diets were supplemented with coconut, palm oil, or a combination of the two, respectively. As a positive control, sunflower oil supplementation was included in the diet of the fifth group. During the 28 days of the study (from 14 to 42 days of life), 5 samplings were scheduled, when excreta samples were taken from the litter to analyse total microbial count and the number of Clostridia, Coliforms, and Salmonella. According to the results of microbiological analysis, coconut oil supplementation led to the continuous decline of Clostridium perfringens numbers until they vanished by the end of the experimental period. A similar but faster decline was found as a result of palm oil and the combined (palm oil+coconut oil) treatment. However, the number of Coliforms and total microbial count changed only slightly by the end of the study. No Salmonella spp. was present in the samples throughout the experiment. Altogether, coconut and palm oil supplementation has a beneficial effect on the microbiological composition of poultry litter.

Silage Quality of two energy-cane cultivars produced with different additives

ABSTRACT This study examined the quality of silage from two cultivars of energy cane (Saccharum spontaneum) produced with different additives. The experiment was laid out in a completely randomized design with a 2×5 factorial arrangement of treatments consisting of two cultivars (VG3, VG1126) and four additives (1% urea; 1% NaOH; 1% CaO; and 1.0 × 1011 CFU g-1 of Lactobacillus plantarum- LP) + control, with eight replicates. Experimental mini-silos were used, which were opened 72 days after ensiling. The dry matter (DM), crude protein (CP), organic matter (OM), mineral matter (MM), ether extract (EE), neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents, and silage degradability were. We identified significant interactions effect between cultivar and type of additive for DM, CP, EE, NDF. ADF. Organic and mineral matter percentage were similar between the two cultivars. LP was the least beneficial additive for DM degradation.After 72 h of fermentation, in vitro NDF degradation of cultivar VG3 increased due to urea, NaOH, and CaO, whereas degradation cultivar VG1126 decreased when inoculated with LP or CaO. The NaOH and CaO improved silage degradability thus improving its quality. The use of LP in the present study did not benefit silage for energy-cane cultivars. The use of NaOH and CaO improves the quality of the silage, two varieties of sugarcane energy VG3 and VG1126, enabling its use with better efficiency for animal feed.

RESUMO Este estudo avaliou a qualidade da silagem de duas cultivares de cana-de-energia (Saccharumspontaneum) produzidas com diferentes aditivos. O experimento foi instalado em delineamento inteiramente casualizado, com arranjo fatorial 2 × 5 de tratamentos composto por duas cultivares (VG3, VG1126) e quatro aditivos (1% uréia; 1% NaOH; 1% CaO; e 1,0 × 1011 UFC g -1 de Lactobacillus plantarum- LP) + controle, com oito repetições. Foram utilizados minisilos experimentais, os quais foram abertos 72 dias após a ensilagem. Os teores de matéria seca (MS), proteína bruta (PB), matéria orgânica (MO), matéria mineral (MM), extrato etéreo (EE), fibra em detergente neutro (FDN) e fibra em detergente ácido (FDA) e degradabilidade da silagem foram avaliados. Identificou-se efeito de interação significativo entre cultivar e tipo de aditivo para MS, PB, EE, FDN. ADF. As porcentagens de matéria orgânica e mineral foram semelhantes entre as duas cultivares. LP foi o aditivo menos benéfico para a degradação da MS. Após 72 h de fermentação, a degradação in vitro da FDN da cultivar VG3 aumentou devido à uréia, NaOH e CaO, enquanto a degradação da cultivar VG1126 diminuiu quando inoculada com LP ou CaO. O NaOH e o CaO melhoraram a degradabilidade da silagem, melhorando assim sua qualidade. O uso da LP no presente estudo não beneficiou a silagem para cultivares de cana-de-energia. O uso de NaOH e CaO melhora a qualidade da silagem, duas variedades de energia da cana-de-açúcar VG3 e VG1126, possibilitando seu uso com melhor eficiência para ração animal

Effect of phytase and protease combination on performance, metabolizable energy, and amino acid digestibility of broilers fed nutrient-restricted diets

Three experiments were carried out to evaluate the effect of dietary combination of different enzymes (phytase + protease) on performance, metabolizable energy, and amino acid digestibility of broiler chickens fed diets with nutritional reduction. A total of 1,400, 336, and 384 male chickens were distributed in a completely randomized design, in the experiments of performance, metabolism, and digestibility, respectively. Treatments were divided as follows: positive control (PC), negative control - NC1 (PC minus 0.16% Ca, 0.15% available P (aP), and 0.5% crude protein (CP)), NC2 (PC minus 0.16% Ca, 0.15% aP, and 1% CP), NC1 added with phytase deriving from citrobacter and protease deriving from Bacillus licheniformis (CBE), NC1 added with phytase deriving from E. coli and protease deriving from microbial fermentation (SE), NC2 added with CBE, and NC2 added with SE. A protein-free diet was included in the digestibility experiment. The nutritional restriction did not affect feed intake of birds in the first experiment; however, the restriction inhibited body weight gain and feed conversion ratio in all phases. In experiment 2, the nutritional restriction decreased AME and AMEn values, although the addition of phytase and protease in diets improved both parameters, mainly in NC2. Treatment NC2 impaired the standardized digestibility of total essential amino acids of animals subjected to experiment 3, although the addition of enzymes helped to recover the digestibility to levels similar to PC. Supplementation of phytase in association with protease is effective to improve performance, energy metabolism, and standardized amino acid digestibility of broilers fed diets with nutrient restriction.(AU)

Nutritional and metabolic parameters of ewe lambs fed yeast in the diet containing fibrolytic enzyme / Parâmetros nutricionais e metabólicos de borregas alimentadas com leveduras na ração contendo enzima fibrolítica

This study aimed to evaluate the dry matter digestibility, intake and metabolic profile of ewe lambs fed active or inactive yeasts in the diet containing fibrolytic enzyme. Twenty crossbred animals with an average weight of 33.4kg and an average age of six months were distributed in a completely randomized design. The treatments consisted of yeasts: Control (no yeast), Milk Sacc X® (active yeast), Active Flora®(active yeast + inactive yeast) and Rumen Yeast®(inactive yeast). Analysis of variance and SNK test were applied with a significance level of 5% for type I error. There was no difference for dry matter intake between treatments (P>0.05). There was a significant trend (P=0.0596) for dry matter digestibility, with higher values for the control treatment. There was a significant trend for blood metabolites (P=0.0705), in which the diet containing Milk Sacc® was superior to the other treatments for total protein concentration. In addition, there was a statistical difference (P<0.05) for urea concentration, in which control and Active Flora® treatments were superior to the others. The inclusion of active or inactive yeasts in the diet for ewe lambs containing fibrolytic enzyme reduces dietary digestibility, without affecting the dry matter intake, in addition, it increases urea and total protein levels without causing liver or kidney damage in ewe lambs.

Objetivou-se avaliar a digestibilidade da matéria seca, consumo e perfil metabólico de borregas alimentadas com leveduras vivas e inativadas na ração contendo enzima fibrolítica. Foram utilizados vinte animais mestiços com peso médio de 33,4kg e idade média de seis meses, distribuídas em delineamento inteiramente casualizado. Os tratamentos consistiram nas leveduras: Controle (sem levedura), Milk Sacc X® (levedura ativa), Active Flora®(levedura viva mais levedura inativada) e Rúmen Yeast® (levedura inativa). Foi realizada à análise variância e teste SNK com nível de significância de 5% para o erro tipo I. Não houve diferença entre tratamentos para consumo de matéria seca com uso ou não de levedura na dieta (P>0,05). Houve tendência significativa (P=0,0596) para a digestibilidade da matéria seca sendo superior para com o tratamento sem inclusão de leveduras. Para os metabólitos sanguíneos houve tendência significativa (P=0,0705) sendo o Milk Sacc® superior aos demais tratamentos para concentração de proteínas totais. Além disso, houve diferença estatística (P<0,05) para concentração de ureia cujos tratamentos controle e Active Flora® foram superiores aos demais. A inclusão de leveduras vivas ou inativadas na dieta para borregas contendo enzima fibrolítica diminui a digestibilidade da dieta, sem afetar o consumo de matéria seca, além disso, aumenta teores de ureia e proteínas totais sem causar prejuízos hepáticos ou renais as borregas.

Effect of essential oils and aqueous extracts of plants on in vitro rumen fermentation and methane production

The objective of this study was to evaluate in vitro rumen fermentation and methane production under the influence of two sources of phytochemicals: essential oils (EOs) and aqueous extracts (AEs). Treatments were set up in a completely randomized block design, with 4×2+1 factorial arrangement of four species, S (garlic, G; cinnamon, C; rosemary, R; eucalyptus; EU) × two types of presentation, P (essential oil, EO; aqueous extract, AE) and a basal diet, BD (50% concentrate, 20% alfalfa and 30% corn silage). Rumen fermentation was evaluated using the in vitro gas production technique. All experimental units were incubated with 500 mg of BD for 72 hours. Treatments were added at a single dose of 900 mg/L of rumen inoculum. Gas pressure was recorded at 0, 2, 4, 6, 8, 10, 14, 18, 24, 30, 36, 42, 48, 60 and 72 h postincubation. There was an interaction effect (P × S) between plant extract presentation (P) and plant species (S) for all variables. Treatments GEO, CEO, REO decreased volatile fatty acids (mmol/200 mg), microbial mass production (mg/g), CH4 production (mL/g), in vitro dry matter digestibility (P < 0.05), and total gas production at 24 and 72 h post-incubation (P < 0.05; mL/g DM, mL/g OM). No differences (P > 0.05) were observed between AEs and BD. In conclusion, the use of EOs negatively affected rumen fermentation parameters and the production of CH4. Garlic and cinnamon EOs effectively reduced methane emissions; however, they also reduced in vitro dry matter digestibility.