Preservation of human gut microbiota inoculums for in vitro fermentations studies

The use of fecal inoculums for in vitro fermentation models requires a viable gut microbiota, capable of fermenting the unabsorbed nutrients. Fresh samples from human donors are used; however, the availability of fresh fecal inoculum and its inherent variability is often a problem. This study aimed to optimize a method of preserving pooled human fecal samples for in vitro fermentation studies. Different conditions and times of storage at -20 degrees C were tested. In vitro fermentation experiments were carried out for both fresh and frozen inoculums, and the metabolic profile compared. In comparison with the fresh, the inoculum frozen in a PBS and 30% glycerol solution, had a significantly lower (p < 0.05) bacterial count (<1 log CFU/mL). However, no significant differences (p < 0.05) were found between the metabolic profiles after 48 h. Hence, a PBS and 30% glycerol solution can be used to maintain the gut microbiota viability during storage at -20 degrees C for at least 3 months, without interfering with the normal course of colonic fermentation.info:eu-repo/semantics/publishedVersio

Suplementação dietética de glutamina ou ácido glutâmico para leitões recém-desmamados

The objective of this work was to evaluate the effects of dietary glutamine and glutamic acid supplementation on carbon turnover (δ13C) in the muscles of piglets, on animal performance, and on feed economic feasibility. The diets consisted of: control (C), 1% glutamine (G), and 1% glutamic acid (GA). In experiment I, 111 piglets were used to assess carbon turnover in the Masseter, Psoas major, and Abdominal internal oblique muscles in terms of their isotopic composition and 13C substitution over time. In experiment II, 72 piglets were assigned randomly to blocks (eight replicates per treatment and three animals per experimental unit), in order to evaluate animal performance and feed economic feasibility. The GA diet promoted the best 13C acceleration in the studied muscles. No diet effects were observed on performance variables. The G and GA diets presented higher costs than the C diet. Although supplementation with 1% glutamine or glutamic acid accelerates carbon turnover in the studied muscles, which suggests a faster recovery during post-weaning and proves the anabolic effect of these additives, its use is not economically viable for weanling piglets.O objetivo deste trabalho foi avaliar os efeitos da suplementação dietética de glutamina e ácido glutâmico no turnover do carbono (δ13C) em músculos de leitões, no desempenho dos animais e na viabilidade econômica das rações. As dietas consistiram em: controle (C), 1% de glutamina (G) e 1% de ácido glutâmico (AG). No experimento I, 111 leitões foram usados para avaliar o turnover do carbono nos músculos Masseter, Psoas major e Abdominal internal oblique, quanto à composição isotópica e à substituição do 13C no tempo. No experimento II, 72 leitões foram alocados em blocos ao acaso (oito repetições por tratamento e três animais por unidade experimental), para avaliar o desempenho dos animais e a viabilidade econômica das rações. A dieta AG promoveu a melhor aceleração de 13C nos músculos estudados. Não se constatou efeito das dietas nas variáveis de desempenho. As dietas G e AG apresentaram maiores custos que a dieta C. Embora a suplementação com 1% de glutamina ou ácido glutâmico acelere o turnover do carbono nos músculos estudados, o que sugere rápida recuperação no pós-desmame e comprova o efeito anabólico desses aditivos, seu uso não é economicamente viável para leitões recém-desmamados

Effect of Glutamine, Glutamic Acid and Nucleotides on the Turnover of Carbon (δC) in Organs of Weaned Piglets

Morphological and physiological alterations occur in the digestive system of weanling piglets, compromising the performance in subsequent phases. This experiment aimed at verifying the influence of glutamine, glutamate and nucleotides on the carbon turnover in the pancreas and liver of piglets weaned at 21 days of age. Four diets were evaluated: glutamine, glutamic acid or nucleotides-free diet (CD); containing 1% glutamine (GD); containing 1% glutamic acid (GAD) and containing 1% nucleotides (ND). One hundred and twenty-three piglets were utilized with three pigs slaughtered at day zero (weaning day) and three at each one of the experimental days (1, 2, 4, 5, 7, 9, 13, 20, 27, and 49 post-weaning), in order to collect organ samples, which were analyzed for the δ13C isotopic composition and compared by means of time. No differences were found (p>0.05) among treatments for the turnover of the 13C in the pancreas (T50% = 13.91, 14.37, 11.07, and 9.34 days; T95% = 46.22, 47.73, 36.79, and 31.04 days for CD, GD, GAD, and ND, respectively). In the liver, the ND presented accelerated values of carbon turnover (T50% = 7.36 and T95% = 24.47 days) in relation to the values obtained for the GD (T50% = 10.15 and T95% = 33.74 days). However, the values obtained for the CD (T50% = 9.12 and T95% = 30.31 days) and GAD (T50% = 7.83 and T95% = 26.03 days) had no differences (p>0.05) among other diets. The technique of 13C isotopic dilution demonstrated trophic action of nucleotides in the liver

Metodologia analítica para extração ultra-sônica de minerais em rações de suínos

No presente trabalho é proposto um método para determinação de cálcio, magnésio, sódio e potássio em amostras de rações de leitões utilizando ultra-som no processo de extração dos analitos e posterior quantificação, respectivamente, por espectrometria de absorção atômica por chama (FAAS) e espectrometria de emissão atômica (AES). As condições de extração otimizadas foram: massa de amostra de 100 mg; granulometria da amostra < 60 mm; solução extratora HCl 0,10 mol L-1; tempo de sonificação de cinco ciclos de 10 s e potência de sonificação de 102 W. O método proposto foi aplicado em estudos de digestibilidade desses nutrientes em amostras de rações utilizadas na criação comercial de leitões e os resultados obtidos mostraram-se concordantes com os resultados provenientes da mineralização ácidaThe aim of the present work brings a developed and an optimized method for determination of calcium, magnesium, sodium and potassium in piglets feed and feces by using ultrasound in the extraction process of nutrients and subsequent quantification by flame atomic absorption spectrometry (FAAS) and atomic emission spectrometry (AES). The optimum established conditions of extraction were: mass of sample: 100 mg, sample particle size: < 60 mm, extractor solution: HCl 0.10 mol L-1, sonication time: five cycles of 10 s and ultrasound power: 102 W. The proposed method was applied in studies of digestibility of those nutrients in different feed used in piglets diets and their results proved to be compatible with those obtained from mineralized samples by acid digestionCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Ação de beta-glucanos em leitões recém-desmamados alimentados com dietas com diferentes densidades nutricionais

A nutrição exerce influência na modulação do sistema imune e na melhoria da resistência dos animais à infecção, visto que os nutrientes são necessários para a multiplicação celular durante a resposta imune e síntese de moléculas relacionadas à comunicação celular. Dois experimentos foram conduzidos com leitões desmamados aos 21 dias de idade e submetidos ao desafio com 150 μg kg-1 PV de lipopolissacarídeo (LPS) de Escherichia coli sorotipo 055:B5. Objetivou-se avaliar efeitos da adição de beta-glucanos originários da levedura Saccharomyces cerevisiae (0 e 300 ppm) em dietas com densidade nutricional baixa e elevada (3300 e 3450 kcal de EM kg-1) sobre o desempenho (Experimento I) e morfometria intestinal (altura de vilosidades: AV, profundidade de criptas: PC, relação AV:PC e área das placas de Peyer: PP) e peso de órgãos (fígado, timo e baço) dos leitões (Experimento II). No experimento I foram utilizados 96 leitões em delineamento de blocos ao acaso (oito repetições), em esquema fatorial 2 x 2 (2 níveis de beta-glucanos x 2 densidades nutricionais), enquanto no segundo experimento, 40 leitões foram distribuídos em delineamento de blocos ao acaso (cinco repetições) com arranjo fatorial dos tratamentos 2 x 2 x 2 (2 níveis de beta-glucanos x 2 densidades nutricionais x 2 épocas de abate: uma semana antes da primeira inoculação com LPS e uma semana após a segunda inoculação com LPS). Nos experimentos foram realizadas análises estatísticas dos dados utilizando o procedimento GLM (Experimento I) e análise multivariada (Experimento II) do SAS, sendo as médias comparadas pelo teste de Bonferroni P(0,05) dos beta-glucanos e da densidade nutricional da dieta no desempenho, PC do jejuno e na área...Nutrition influences the modulation of the immune system and improves the resistance to infection, because the nutrients are required for cell proliferation during the immune response. Two experiments were conducted with piglets weaned at 21 days old and immunologically challenged with 150 μg kg-1 BW lipopolysaccharide of Escherichia coli serotype 055:B5 to evaluate the effects of two levels of beta-glucans (0 and 300 ppm) of yeast Saccharomyces cerevisiae and two levels of nutrient density (3300 and 3450 kcal ME kg-1) on growth performance (Experiment I) and intestinal morphometry (villus height: VH, cript depth: CD, VH:CD, Peyer’s patches area: PP) and organs weight (liver, spleen and thymus) of weanling pigs (Experiment II). In Experiment I, 96 weanling pigs were randomly assigned in a complete block design in a 2 x 2 factorial arrangement (2 levels of beta-glucans: 0 and 300 ppm, and 2 levels of nutrient density: 3300 and 3450 kcal ME kg-1) with 8 replications per treatment and 3 pigs per pen. In Experiment II, 40 weanling pigs were randomly assigned in a complete block design in a 2 x 2 x 2 factorial arrangement with the same levels of beta-glucans and nutrient density of experiment I at two slaughter moments with 5 replications per treatment and one pig per pen. Data were analyzed in accordance with GLM procedure (Experiment I) and multivariate procedure (Experiment II) by SAS, and the treatment means were compared by Bonferroni test (P0.05) on growth performance, and CD in jejunum neither in PP ileal area. The addition of beta-glucans and high nutrient density diets are not viable for pigs from 21 to 59 days of age, once no effects were observed on growth... (Complete abstract click electronic access below

Avaliação do consumo de energia elétrica em duas granulometrias de moagem de grãos de milho de textura dentada e dura

O trabalho objetivou avaliar o consumo de energia elétrica por meio de medidas de amperagem, sobre dois graus de moagem de grãos de milho para alimentação animal. O delineamento experimental utilizado foi inteiramente ao acaso em Esquema Fatorial, com quatro tratamentos e dez repetições. Foram utilizados dois diâmetros geométricos médios (DGM) de 865 e 570mm, grosso e fino, respectivamente, com texturas dentada e dura. Constatou-se maior consumo de energia elétrica na granulometria fina, com aumento na demanda de energia de 14 % para a moagem dos grãos de textura dura em relação aos grãos de textura dentada. Portanto, em granulometria de menor DGM (570mm) deve-se dar preferência por grãos de milho de textura dentada pelo menor consumo de energia elétrica na moagem, enquanto que em granulometria de 865mm, o consumo de energia para ambos os híbridos foi o mesmo

Importance of gastrointestinal in vitro models for the poultry industry and feed formulations

The animal’s diet is a crucial factor, as poultry feed formulations influences greatly their development, well-being and final products quality, i.e., meat and eggs. Therefore, the search for feed additives that provide concomitantly better performances, low-cost usage, guarantying the animal well-being and products safety, became a priority to the poultry industry. Although in an early stage, research has been focused on developing the optimal cost-efficient feed formulations, taking into account the chicken’s physiology and function of the gastrointestinal tract and intestinal microbiota. This review discusses a number of concepts and novel approaches towards the optimization of poultry’s feed formulations, by critically encompassing the animal’s growth and performance. Additionally, it highlights the in vitro gastrointestinal models capabilities as a potential solution to test highly nutritive, well-balanced and efficient feed formulations within a circular economy framework.info:eu-repo/semantics/publishedVersio

Development of a Chicken Gastrointestinal Tract (GIT) Simulation Model: Impact of Cecal Inoculum Storage Preservation Conditions

A chicken gastrointestinal tract (GIT) simulation model was developed to help predict the potential effects of feed additives supplementation on chicken’ microbiota. The chemical and enzymatic conditions for oral, gastric, intestinal, and cecum fermentation phases were designed to closely resemble the chicken GIT conditions. For cecum fermentation, the inoculum was obtained from the cecal contents of 18 38-day broiler chickens. The impact of inoculum preservation on bacteria viability was assessed by comparing two methods of preservation with fresh inoculum: (1) 5% dimethyl sulfoxide (DMSO) at −80 °C and (2) 30% glycerol at −20 °C. The fermentation with fresh and frozen (DMSO method) inoculums was performed and compared using standard chicken feed (SCF) and SCF with 1% fructooligosaccharides (FOS), and inoculum control (IC) condition without feed matrix was used as a baseline. Inoculum’s viability was assessed throughout 90 days of storage by culture media platting, while bacterial growth and metabolites production during fermentation was evaluated by quantitative polymerase chain reaction (qPCR), high-performance liquid chromatography (HPLC), and total ammonia nitrogen quantification. The DMSO method was shown to be the most suitable for cecal inoculum storage. Higher growth of beneficial cecal bacteria for fresh inoculum was observed in SCF while for frozen inoculum, was the SCF + FOS condition. Also, frozen inoculum had lower activity of butyrate producers and proteolytic bacteria, showing different fermentation profiles. The GIT model developed showed to be useful to test the effect of feed additives supplementation

Effect of digestible threonine level of dietar on productivo performance, commercial ego grading and ego Quality of 55 to 61 week-old playing heno

The aim of this study was to establish the digestible threonine (dig. Thr) requirements and the ideal proportion between Thr and Lys for Isa Brown laying hens on productive traits and egg quality characteristics from 55- to 58- and 59- to 61 weeks of age. Increased levels of L-Thr were added to a basal diet that contained 3.7 g/kg of dig. Thr. Therefore, diets containing 3.7, 4.0, 4.3, 4.6, 4.9 and 5.2 g/kg of dig. Thr and a relation between Thr and Lys of 0.58, 0.63, 0.67, 0.72, 0.77 and 0.81, respectively were provided to laying hens. A total of 216 hens were distributed in a randomized design among the 6 experimental diets. There were 9 replicates per treatment, and 4 birds per replicate. Data were analyzed by repeated measures analysis using the MIXED procedure of SAS. The main effects (dig. Thr level and experimental period) and their interaction were studied. Neither the dig. Thr level nor the experimental period had effect on performance traits and egg quality. Also, commercial egg grading was not influenced by period. However, there was a quadratic effect (P<0.001) of dig. Thr level within each scale egg weight. In conclusion, diets containing more than 3.7 g/kg of dig. Thr do not improve laying hens productivity at 55- to 61-week-old

GFAAS Determination of Zinc in Fish Feed and Feces Using Slurry Sampling

This paper presents a simple, fast, and sensitive method to determine zinc in samples of feces and fish feed by electrothermal atomic absorption spectrometry through the direct introduction of slurries of the samples into the spectrometer's graphite tube. The procedure is based on the injection of 10 mu L of an acidified aqueous solution containing 0.50% w/v of feces or feed and 0.50% v/v HNO(3) into graphite tube. The limits of detection and quantification calculated for 20 readings of the blank of the standard slurries (0.50% w/v of feces or feed devoid of zinc) were 0.04 and 0.13 mu g L(-1) for the standard feces slurries and 0.05 and 0.17 mu g L(-1) for the standard feed slurries. The proposed method was applied in studies of digestibility of zinc in different fish feeds, and their results proved compatible with that obtained from samples mineralized by acid digestion using microwave oven