Ensiling a dry bakery by-product: effect of hydration using acid whey or water associated or not at urea

SUMMARY The objective of this study was to investigate the characteristics of silages prepared with a dry bakery by-product hydrated with acid whey or water associated or not at urea. The trial was a 3 (hydration at three moisture levels: 250, 300, and 350 mL kg1 of by-product) × 3 (three application rates of urea: 0, 5, and 10 g kg1 of by-product) × 2 (liquid used at hydration: acid whey or water) factorial arrangement. Mini-silos were used as experimental silos and remained closed for 30 days. Overall, silages hydrated with acid whey had higher (P 0.01) production of lactic (19.6 vs. 18.5 g kg1 of dry matter [DM]), acetic (2.5 vs. 1.8 g kg1 of DM) and propionic acid (8.0 vs. 5.4 g kg1 of DM), and lower (P 0.05) fungi counts (5.35 and 5.01 cfu g1 of fresh silage) compared to silages hydrated with water. Increased hydration led to the higher production of total acids (P 0.01), decreasing silage pH; but increased hydration reduced DM recovery (P 0.01). Silages hydrated with water and acid whey at 250 mL kg1 and combined at 5 and 10 g kg1 of urea exhibited higher DM recovery (P 0.05). Silages of a dry bakery by-product hydrated with acid whey had a better fermentation pattern and could represent a practical strategy to avoid discarding both by-products into environment.

RESUMO O objetivo com este estudo foi investigar as características de silagens preparadas com um resíduo seco de padaria hidratado com soro ácido de leite ou água associado ou não à ureia. O experimento foi conduzido em um arranjo fatorial 3 (três níveis de hidratação: 250, 300 e 350 mL kg1 de subproduto) × 3 (três níveis de aplicação de ureia: 0, 5 e 10 g kg1 de subproduto) × 2 (dois líquidos usados na hidratação: soro ácido de leite ou água). Mini silos foram utilizados como silos experimentais e permaneceram fechados por 30 dias. Em geral, silagens hidratadas com soro ácido de leite apresentaram maior (P 0,01) produção de ácido lático (19,6 vs. 18,5 g kg1 de matéria seca [MS]), ácido acético (2,5 vs. 1,8 g kg1 de MS) e ácido propiônico (8,0 vs. 5,4 g kg1 de MS), e menores (P 0,05) contagens de fungos (5,35 e 5,01 ufg g1 de silagem fresca) comparadas às silagens hidratadas com água. A maior hidratação das silagens aumentou a produção total de ácidos (P 0,01) reduzindo os valores de pH; mas o aumento na hidratação reduziu (P 0,01) a recuperação de MS das silagens. Silagens hidratadas com água e soro ácido de leite a 250 mL kg1 e combinado com 5 e 10 g kg1 de ureia exibiram maior (P 0,05) recuperação de MS. Silagens de resíduo seco de padaria hidratadas com soro ácido de leite apresentaram melhor padrão fermentativo, podendo representar uma estratégia prática para evitar o descarte de ambos os subprodutos no ambiente.

Ensiling a dry bakery by-product: effect of hydration using acid whey or water associated or not at urea

SUMMARY The objective of this study was to investigate the characteristics of silages prepared with a dry bakery by-product hydrated with acid whey or water associated or not at urea. The trial was a 3 (hydration at three moisture levels: 250, 300, and 350 mL kg1 of by-product) × 3 (three application rates of urea: 0, 5, and 10 g kg1 of by-product) × 2 (liquid used at hydration: acid whey or water) factorial arrangement. Mini-silos were used as experimental silos and remained closed for 30 days. Overall, silages hydrated with acid whey had higher (P 0.01) production of lactic (19.6 vs. 18.5 g kg1 of dry matter [DM]), acetic (2.5 vs. 1.8 g kg1 of DM) and propionic acid (8.0 vs. 5.4 g kg1 of DM), and lower (P 0.05) fungi counts (5.35 and 5.01 cfu g1 of fresh silage) compared to silages hydrated with water. Increased hydration led to the higher production of total acids (P 0.01), decreasing silage pH; but increased hydration reduced DM recovery (P 0.01). Silages hydrated with water and acid whey at 250 mL kg1 and combined at 5 and 10 g kg1 of urea exhibited higher DM recovery (P 0.05). Silages of a dry bakery by-product hydrated with acid whey had a better fermentation pattern and could represent a practical strategy to avoid discarding both by-products into environment.

RESUMO O objetivo com este estudo foi investigar as características de silagens preparadas com um resíduo seco de padaria hidratado com soro ácido de leite ou água associado ou não à ureia. O experimento foi conduzido em um arranjo fatorial 3 (três níveis de hidratação: 250, 300 e 350 mL kg1 de subproduto) × 3 (três níveis de aplicação de ureia: 0, 5 e 10 g kg1 de subproduto) × 2 (dois líquidos usados na hidratação: soro ácido de leite ou água). Mini silos foram utilizados como silos experimentais e permaneceram fechados por 30 dias. Em geral, silagens hidratadas com soro ácido de leite apresentaram maior (P 0,01) produção de ácido lático (19,6 vs. 18,5 g kg1 de matéria seca [MS]), ácido acético (2,5 vs. 1,8 g kg1 de MS) e ácido propiônico (8,0 vs. 5,4 g kg1 de MS), e menores (P 0,05) contagens de fungos (5,35 e 5,01 ufg g1 de silagem fresca) comparadas às silagens hidratadas com água. A maior hidratação das silagens aumentou a produção total de ácidos (P 0,01) reduzindo os valores de pH; mas o aumento na hidratação reduziu (P 0,01) a recuperação de MS das silagens. Silagens hidratadas com água e soro ácido de leite a 250 mL kg1 e combinado com 5 e 10 g kg1 de ureia exibiram maior (P 0,05) recuperação de MS. Silagens de resíduo seco de padaria hidratadas com soro ácido de leite apresentaram melhor padrão fermentativo, podendo representar uma estratégia prática para evitar o descarte de ambos os subprodutos no ambiente.

Symbiotic nitrogen-fixing bacterial populations trapped from soils under agroforestry systems in the Western Amazon

Cowpea (Vigna unguiculata) is an important grain-producing legume that can forego nitrogen fertilization by establishing an efficient symbiosis with nitrogen-fixing bacteria. Although inoculating strains have already been selected for this species, little is known about the genotypic and symbiotic diversity of native rhizobia. Recently, Bradyrhizobium has been shown to be the genus most frequently trapped by cowpea in agricultural soils of the Amazon region. We investigated the genetic and symbiotic diversity of 148 bacterial strains with different phenotypic and cultural properties isolated from the nodules of the trap species cowpea, which was inoculated with samples from soils under agroforestry systems from the western Amazon. Sixty non-nodulating strains indicated a high frequency of endophytic strains in the nodules. The 88 authenticated strains had varying symbiotic efficiency. The SPAD (Soil Plant Analysis Development) index (indirect measurement of chlorophyll content) was more efficient at evaluating the contribution of symbiotic N2-fixation than shoot dry matter under axenic conditions. Cowpea-nodulating bacteria exhibited a high level of genetic diversity, with 68 genotypes identified by BOX-PCR. Sequencing of the 16S rRNA gene showed a predominance of the genus Bradyrhizobium, which accounted for 70 % of all strains sequenced. Other genera identified were Rhizobium, Ochrobactrum, Paenibacillus, Bosea, Bacillus, Enterobacter, and Stenotrophomonas. These results support the promiscuity of cowpea and demonstrate the high genetic and symbiotic diversity of rhizobia in soils under agroforestry systems, with some strains exhibiting potential for use as inoculants. The predominance of Bradyrhizobium in land uses with different plant communities and soil characteristics reflects the adaptation of this genus to the Amazon region.

Symbiotic nitrogen-fixing bacterial populations trapped from soils under agroforestry systems in the Western Amazon

Cowpea (Vigna unguiculata) is an important grain-producing legume that can forego nitrogen fertilization by establishing an efficient symbiosis with nitrogen-fixing bacteria. Although inoculating strains have already been selected for this species, little is known about the genotypic and symbiotic diversity of native rhizobia. Recently, Bradyrhizobium has been shown to be the genus most frequently trapped by cowpea in agricultural soils of the Amazon region. We investigated the genetic and symbiotic diversity of 148 bacterial strains with different phenotypic and cultural properties isolated from the nodules of the trap species cowpea, which was inoculated with samples from soils under agroforestry systems from the western Amazon. Sixty non-nodulating strains indicated a high frequency of endophytic strains in the nodules. The 88 authenticated strains had varying symbiotic efficiency. The SPAD (Soil Plant Analysis Development) index (indirect measurement of chlorophyll content) was more efficient at evaluating the contribution of symbiotic N2-fixation than shoot dry matter under axenic conditions. Cowpea-nodulating bacteria exhibited a high level of genetic diversity, with 68 genotypes identified by BOX-PCR. Sequencing of the 16S rRNA gene showed a predominance of the genus Bradyrhizobium, which accounted for 70 % of all strains sequenced. Other genera identified were Rhizobium, Ochrobactrum, Paenibacillus, Bosea, Bacillus, Enterobacter, and Stenotrophomonas. These results support the promiscuity of cowpea and demonstrate the high genetic and symbiotic diversity of rhizobia in soils under agroforestry systems, with some strains exhibiting potential for use as inoculants. The predominance of Bradyrhizobium in land uses with different plant communities and soil characteristics reflects the adaptation of this genus to the Amazon region.

Physiological and symbiotic diversity of Cupriavidus necator strains isolated from nodules of Leguminosae species

Biological nitrogen fixation, performed by diazotrophic bacteria, plays an important role in the maintenance of agricultural systems, as it contributes with significant amounts of the nitrogen (N) needed for plant growth, totally or partially exempting the use of industrial N fertilisers. Twenty-five bacterial strains isolated from nodules of Leucaena leucocephala and Phaseolus vulgaris trap plants were studied. These nodules were formed after inoculation with suspensions of soil samples collected close to the root system of Sesbania virgata. In previous studies, these bacteria were identified as Cupriavidus necator. This study aimed to evaluate the ability of these strains to fix N2 in the free-living state and to use carbon (C) sources; their resistance to antibiotics; growth in media with different pH values and salt concentrations and symbiotic efficiency with L. leucocephala and P. vulgaris. In each test, these strains were compared to C. taiwanensis LMG 19424T. Although a high variability regarding antibiotic resistance, salt tolerance and use of C sources were observed among the 25 C. necator strains, a large group behaved similar regarding salt tolerance (20 strains) and antibiotic resistance (22 strains). C. necator strains behaved in a different way of LMG 19424T. Only one of the 25 strains studied, UFLA02-69, was not able to establish symbiosis with its trap species, P. vulgaris. Only the strains LMG19424T and UFLA01-672 were efficient in symbiosis with L. leucocephala. The ability to use C sources, grow in different pHs and salt concentrations and resistance to several antibiotics, may grant high saprophytic competence and greater competitivity to these strains in relation to the native Leguminosae-nodulating bacterial populations, suggesting potential use in inoculant strain selection studies for legumes cultivated in soils with a wide range of pH and salt concentrations.

Physiological and symbiotic diversity of Cupriavidus necator strains isolated from nodules of Leguminosae species

Biological nitrogen fixation, performed by diazotrophic bacteria, plays an important role in the maintenance of agricultural systems, as it contributes with significant amounts of the nitrogen (N) needed for plant growth, totally or partially exempting the use of industrial N fertilisers. Twenty-five bacterial strains isolated from nodules of Leucaena leucocephala and Phaseolus vulgaris trap plants were studied. These nodules were formed after inoculation with suspensions of soil samples collected close to the root system of Sesbania virgata. In previous studies, these bacteria were identified as Cupriavidus necator. This study aimed to evaluate the ability of these strains to fix N2 in the free-living state and to use carbon (C) sources; their resistance to antibiotics; growth in media with different pH values and salt concentrations and symbiotic efficiency with L. leucocephala and P. vulgaris. In each test, these strains were compared to C. taiwanensis LMG 19424T. Although a high variability regarding antibiotic resistance, salt tolerance and use of C sources were observed among the 25 C. necator strains, a large group behaved similar regarding salt tolerance (20 strains) and antibiotic resistance (22 strains). C. necator strains behaved in a different way of LMG 19424T. Only one of the 25 strains studied, UFLA02-69, was not able to establish symbiosis with its trap species, P. vulgaris. Only the strains LMG19424T and UFLA01-672 were efficient in symbiosis with L. leucocephala. The ability to use C sources, grow in different pHs and salt concentrations and resistance to several antibiotics, may grant high saprophytic competence and greater competitivity to these strains in relation to the native Leguminosae-nodulating bacterial populations, suggesting potential use in inoculant strain selection studies for legumes cultivated in soils with a wide range of pH and salt concentrations.

Sesbania virgata stimulates the occurrence of its microsymbiont in soils but does not inhibit microsymbionts of other species

The legume species Sesbania virgata establishes a specific and efficient symbiosis with Azorhizobium doebereinerae. Previous studies have shown that A. doebereinerae occurrence correlates to the presence of S. virgata. This work aimed to evaluate the occurrence of A. doebereinerae and of other nitrogen-fixing Leguminosae-nodulating bacteria (NFLNB) in soil samples collected adjacent to and 10 m away from the stems of five S. virgata plants in pasture areas. Symbiotic characteristics of isolates from these NFLNB populations were also studied. S. virgata and the four promiscuous legume species Leucaena leucocephala, Macroptilium atropurpureum, Phaseolus vulgaris and Vigna unguiculata were inoculated with soil samples to trap A. doebereinerae and other NFLNB. NFLNB capable of inducing nodulation in at least one of these legumes were found in all samples. M. atropurpureum was the most promiscuous species, as it trapped the highest number of NFLNB cultural types from soil suspensions. The other species were less promiscuous in the following order: V. unguiculata, P. vulgaris, and L. leucocephala. Isolates of the promiscuous legumes were classified into seven cultural groups. One of these groups, isolated from all promiscuous species, showed fast-growth alkali-reaction in culture medium (like Azorhizobium); it was identified as Cupriavidus. This is the first report of symbiosis of Cupriavidus with Papilionoideae species. The symbiotic efficiency of promiscuous hosts with NFLNB varied, but it was always less than that of controls with mineral nitrogen or an inoculant strain. S. virgata was efficiently nodulated only by A. doebereinerae, which occurred mainly in samples collected close to the plant stem, corroborating a high stimulus by its host species. A high diversity of NFLNB occurs as saprophytes close to the S. virgata root system.

A espécie de leguminosa Sesbania virgata estabelece uma simbiose especifica e eficiente com Azorhizobium doebereinerae. Estudos prévios indicam que a ocorrência de A. doebereinerae esta relacionada à presença de S. virgata. Avaliou-se a ocorrência de A. doebereinerae e de outras Bactérias Fixadoras de Nitrogênio Nodulíferas em Leguminosas (BFNNL) em amostras de solos coletadas próximo e a 10 metros do caule de cinco plantas de S. virgata em áreas de pastagem. As características simbióticas de isolados das populações dessas BFNNL foram também estudadas. Para captura de A. doebereinerae e de outras BFNNL, essas amostras de solos foram inoculadas em S. virgata e nas leguminosas promíscuas Leucaena leucocephala, Macroptilium atropurpureum, Phaseolus vulgaris e Vigna unguiculata. Todas as amostras de solos apresentaram BFNNL capazes de nodular, pelo menos, uma espécie. M. atropurpureum foi a espécie mais promíscua capturando um grande número de tipos culturais de BFNNL das suspensões de solo. As outras espécies foram menos promíscuas na seguinte ordem: V. unguiculata, P. vulgaris, e L. leucocephala. Os isolados das espécies hospedeiras consideradas promíscuas foram agrupados em sete tipos culturais. Um desses grupos, isolado de todas as espécies promiscuas, apresentou crescimento rápido e reação alcalina em meio de cultura (como Azorhizobium) e foi identificado como Cupriavidus. Este é o primeiro relato da simbiose de Cupriavidus com espécies de Papilionoideae. A simbiose de BFNNL com as demais espécies teve eficiência variável, mas foi sempre menor que a da adubação nitrogenada e da estirpe recomendada como inoculante. S. virgata formou simbiose somente com A. doebereinerae, a qual foi mais freqüente nas amostras de solos coletadas próximo ao caule da planta, corroborando alto estímulo deste microsimbionte por sua espécie hospedeira. Uma alta diversidade de BFNNL ocorre saprofiticamente próximo ao sistema radicular de S. virgata.

Sesbania virgata stimulates the occurrence of its microsymbiont in soils but does not inhibit microsymbionts of other species

The legume species Sesbania virgata establishes a specific and efficient symbiosis with Azorhizobium doebereinerae. Previous studies have shown that A. doebereinerae occurrence correlates to the presence of S. virgata. This work aimed to evaluate the occurrence of A. doebereinerae and of other nitrogen-fixing Leguminosae-nodulating bacteria (NFLNB) in soil samples collected adjacent to and 10 m away from the stems of five S. virgata plants in pasture areas. Symbiotic characteristics of isolates from these NFLNB populations were also studied. S. virgata and the four promiscuous legume species Leucaena leucocephala, Macroptilium atropurpureum, Phaseolus vulgaris and Vigna unguiculata were inoculated with soil samples to trap A. doebereinerae and other NFLNB. NFLNB capable of inducing nodulation in at least one of these legumes were found in all samples. M. atropurpureum was the most promiscuous species, as it trapped the highest number of NFLNB cultural types from soil suspensions. The other species were less promiscuous in the following order: V. unguiculata, P. vulgaris, and L. leucocephala. Isolates of the promiscuous legumes were classified into seven cultural groups. One of these groups, isolated from all promiscuous species, showed fast-growth alkali-reaction in culture medium (like Azorhizobium); it was identified as Cupriavidus. This is the first report of symbiosis of Cupriavidus with Papilionoideae species. The symbiotic efficiency of promiscuous hosts with NFLNB varied, but it was always less than that of controls with mineral nitrogen or an inoculant strain. S. virgata was efficiently nodulated only by A. doebereinerae, which occurred mainly in samples collected close to the plant stem, corroborating a high stimulus by its host species. A high diversity of NFLNB occurs as saprophytes close to the S. virgata root system.

A espécie de leguminosa Sesbania virgata estabelece uma simbiose especifica e eficiente com Azorhizobium doebereinerae. Estudos prévios indicam que a ocorrência de A. doebereinerae esta relacionada à presença de S. virgata. Avaliou-se a ocorrência de A. doebereinerae e de outras Bactérias Fixadoras de Nitrogênio Nodulíferas em Leguminosas (BFNNL) em amostras de solos coletadas próximo e a 10 metros do caule de cinco plantas de S. virgata em áreas de pastagem. As características simbióticas de isolados das populações dessas BFNNL foram também estudadas. Para captura de A. doebereinerae e de outras BFNNL, essas amostras de solos foram inoculadas em S. virgata e nas leguminosas promíscuas Leucaena leucocephala, Macroptilium atropurpureum, Phaseolus vulgaris e Vigna unguiculata. Todas as amostras de solos apresentaram BFNNL capazes de nodular, pelo menos, uma espécie. M. atropurpureum foi a espécie mais promíscua capturando um grande número de tipos culturais de BFNNL das suspensões de solo. As outras espécies foram menos promíscuas na seguinte ordem: V. unguiculata, P. vulgaris, e L. leucocephala. Os isolados das espécies hospedeiras consideradas promíscuas foram agrupados em sete tipos culturais. Um desses grupos, isolado de todas as espécies promiscuas, apresentou crescimento rápido e reação alcalina em meio de cultura (como Azorhizobium) e foi identificado como Cupriavidus. Este é o primeiro relato da simbiose de Cupriavidus com espécies de Papilionoideae. A simbiose de BFNNL com as demais espécies teve eficiência variável, mas foi sempre menor que a da adubação nitrogenada e da estirpe recomendada como inoculante. S. virgata formou simbiose somente com A. doebereinerae, a qual foi mais freqüente nas amostras de solos coletadas próximo ao caule da planta, corroborando alto estímulo deste microsimbionte por sua espécie hospedeira. Uma alta diversidade de BFNNL ocorre saprofiticamente próximo ao sistema radicular de S. virgata.