Using bifidobacterium and propionibacterium strains in probiotic consortia to normalize the gastrointestinal tract / O uso de cepas de bifidobacterium e propionibacterium em consórcios probióticos para normalizar o trato gastrointestinal

The gastrointestinal microflora regulates the body's functions and plays an important role in its health. Dysbiosis leads to a number of chronic diseases such as diabetes, obesity, inflammation, atherosclerosis, etc. However, these diseases can be prevented by using probiotics ­ living microorganisms that benefit the microflora and, therefore, improve the host organism's health. The most common probiotics include lactic acid bacteria of the Bifidobacterium and Propionibacterium genera. We studied the probiotic properties of the following strains: Bifidobacterium adolescentis АС-1909, Bifidobacterium longum infantis АС-1912, Propionibacterium jensenii В-6085, Propionibacterium freudenreichii В-11921, Propionibacterium thoenii В-6082, and Propionibacterium acidipropionici В-5723. Antimicrobial activity was determined by the 'agar blocks' method against the following test cultures: Escherichia coli ATCC 25922, Salmonella enterica ATCC 14028, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa B6643, Proteus vulgaris ATCC 63, and Listeria monocytogenes ATCC 7644. Moderate antimicrobial activity against all the test cultures was registered in Bifidobacterium adolescentis АС-1909, Propionibacterium jensenii В-6085, and Propionibacterium thoenii В-6082. Antioxidant activity was determined by the DPPH inhibition method in all the lactic acid strains. Our study indicated that some Propionibacterium and Bifidobacterium strains or, theoretically, their consortia could be used as probiotic cultures in dietary supplements or functional foods to prevent a number of chronic diseases.

A microbiota gastrointestinal regula as funções do corpo e desempenha um papel importante na sua saúde. A disbiose leva a uma série de doenças crônicas, como diabetes, obesidade, inflamação, aterosclerose, etc. No entanto, essas doenças podem ser prevenidas pelo uso de probióticos − microrganismos vivos que beneficiam a microflora e, portanto, melhoram a saúde do organismo hospedeiro. Os probióticos mais comuns incluem bactérias do ácido láctico dos gêneros Bifidobacterium e Propionibacterium. Nós estudamos as propriedades probióticas das seguintes cepas: Bifidobacterium adolescentis АС-1909, Bifidobacterium longum infantis АС-1912, Propionibacterium jensenii В-6085, Propionibacterium freudenreichii В-11921, Propionibacterium thoenii В-6082 В-6082 acid e Propionibacterium thoenii В-6082 В-6082 acidibion. A atividade antimicrobiana foi determinada pelo método de 'blocos de ágar' contra as seguintes culturas de teste: Escherichia coli ATCC 25922, Salmonella enterica ATCC 14028, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa B6643, Proteus vulgaris ATCC 63 e Listeria monocytogenes moderada atividade ATCC 7644. Uma atividade antimicrobiana moderada contra todas as culturas de teste foi registrado em Bifidobacterium adolescentis АС-1909, Propionibacterium jensenii В-6085 e Propionibacterium thoenii В-6082. A atividade antioxidante foi determinada pelo método de inibição do DPPH em todas as cepas de ácido lático. Nosso estudo indicou que algumas cepas de Propionibacterium e Bifidobacterium − ou, teoricamente, seus consórcios − poderiam ser usadas ​​como culturas probióticas em suplementos dietéticos ou alimentos funcionais para prevenir uma série de doenças crônicas.

Organic acids for broilers: Effects on intestinal morphology and growth performance / Ácidos orgánicos para pollos de engorde: Efectos sobre la morfología intestinal y el rendimiento del crecimiento / Ácidos orgânicos para frangos de corte: Efeitos na morfologia intestinal e no desempenho de crescimento

Abstract Background: Organic acids and ammonium salts added to drinking water can optimize productivity of broiler chickens. Objective: To evaluate the effect of acidifying drinking water on productive performance, blood and intestinal pH, and intestinal morphology of broilers. Methods: 1,400 one-day-old broiler chicks were used to evaluate two pH levels (4 and 6) of drinking water during three periods (1-21, 1-28, and 1-42 days of age). The treatments consisted of water added with a blend of formic acid (31%), propionic acid (19%), and their salts ammonium formate (26%), and ammonium propionate (6%) compared to a control group (pH 8). Results: Compared to the control, the groups consuming water at pH 6 (0.038 moles) continuously for 42 days improved (p≤0.01) live weight (2.785 vs 2.691 kg), feed conversion ratio (1.430 vs 1.463 kg/kg), and increased the number of intestinal villi (59.0 vs 55.7). Additionally, blood and intestinal pH was reduced vs the control group (7.75 vs 7.89; 6.32 vs 6.41, respectively). Conclusion: The blend of formic and propionic acids and their ammonium salts in drinking water at pH 6 during the complete production cycle of broilers improves performance, increases the number of intestinal villi, and reduces the pH of blood, duodenum, and ileum.

Resumen Antecedentes: Agregar ácidos orgánicos y sus sales amoniacales en el agua de bebida de pollos de engorde puede optimizar su productividad. Objetivo: Evaluar el efecto de la acidificación del agua de bebida sobre el rendimiento productivo, pH sanguíneo e intestinal, y morfología intestinal del pollo de engorde. Métodos: Se utilizaron 1.400 pollos de engorde de un día de edad para evaluar dos niveles de pH (4 y 6) en agua de bebida durante tres períodos (1-21, 1-28 y 1-42 días de edad). Los tratamientos consistieron en agua con una mezcla de ácido fórmico (31%), ácido propiónico (19%) y sus sales formiato de amonio (26%) y propionato de amonio (6%) en comparación con un grupo control (pH 8). Resultados: En comparación con el control, los grupos que recibieron agua a pH 6 (0,038 moles) continuamente durante 42 días tuvieron mejor (p≤0,01) rendimiento en términos de peso vivo (2,785 vs 2,691 kg) y conversión alimenticia (1,430 vs 1,463 kg/kg) así como un mayor número de vellosidades intestinales (59,0 vs 55,7). Adicionalmente, el pH sanguíneo e intestinal disminuyó vs el grupo control (7,75 vs 7,89; 6,32 vs 6,41, respectivamente). Conclusión: La mezcla de ácidos fórmico y propiónico y sus sales de amonio en el agua de bebida a pH 6 durante todo el ciclo productivo mejora los parámetros productivos en pollos de engorde, aumenta el número de vellosidades intestinales y reduce el pH de la sangre, duodeno e íleon.

Resumo Antecedentes: Recomendações para a aplicação de mistura de ácidos orgânicos e sais de amônio na água potável são necessárias para otimizar a produtividade em frangos de corte. Objetivo: Avaliar o efeito da acidificação da água de bebida no desempenho produtivo, pH sanguíneo e intestinal e morfologia intestinal. Métodos: 1.400 pintos de corte entre 1 e 42 dias de idade foram utilizados para avaliar dois níveis de pH (4 e 6) na água de beber entre três períodos (1-21, 1-28 e 1-42 dias de idade) de tratamentos em água com uma mistura de ácido fórmico 31%, ácido propiônico 19%, e seus sais formato de amônio 26% e propionato de amônio 6% em relação ao controle (pH 8). Resultados: Os grupos que receberam água continuamente pH 6 (0,038 mols), por 42 dias tiveram melhor desempenho em relação ao controle (p≤0,01) em termos de peso vivo (2,785 vs 2,691 kg) e conversão alimentar (1,430 vs 1,463 kg/kg), bem como aumento do número de vilosidades intestinais (59,0 vs 55,7), sangue e pH intestinal reduzidos vs controle (7,75 vs 7,89; 6,32 vs 6,41). Conclusão: A mistura dos ácidos fórmico e propiônico e seus sais de amônio na água de bebida em pH 6 durante o ciclo completo de produção melhorou os parâmetros de produção em frangos de 42 dias de idade e aumentou o número de vilosidades intestinais e uma redução no sangue, duodeno, e pH do íleo.

Disruption of mitochondrial functions involving mitochondrial permeability transition pore opening caused by maleic acid in rat kidney.

Propionic acid (PA) predominantly accumulates in tissues and biological fluids of patients affected by propionic acidemia that may manifest chronic renal failure along development. High urinary excretion of maleic acid (MA) has also been described. Considering that the underlying mechanisms of renal dysfunction in this disorder are poorly known, the present work investigated the effects of PA and MA (1-5 mM) on mitochondrial functions and cellular viability in rat kidney and cultured human embryonic kidney (HEK-293) cells. Mitochondrial membrane potential (∆ψm), NAD(P)H content, swelling and ATP production were measured in rat kidney mitochondrial preparations supported by glutamate or glutamate plus malate, in the presence or absence of Ca2+. MTT reduction and propidium iodide (PI) incorporation were also determined in intact renal cells pre-incubated with MA or PA for 24 h. MA decreased Δψm and NAD(P)H content and induced swelling in Ca2+-loaded mitochondria either respiring with glutamate or glutamate plus malate. Noteworthy, these alterations were fully prevented by cyclosporin A plus ADP, suggesting the involvement of mitochondrial permeability transition (mPT). MA also markedly inhibited ATP synthesis in kidney mitochondria using the same substrates, implying a strong bioenergetics impairment. In contrast, PA only caused milder changes in these parameters. Finally, MA decreased MTT reduction and increased PI incorporation in intact HEK-293 cells, indicating a possible association between mitochondrial dysfunction and cell death in an intact cell system. It is therefore presumed that the MA-induced disruption of mitochondrial functions involving mPT pore opening may be involved in the chronic renal failure occurring in propionic acidemia.

Efficacy of weak acid preservatives on spoilage fungi of bakery products.

Organic acids and their salts are usually the first choice in the bread industry to restrict fungal spoilage, but their efficacy is pH-dependent and spoilage by fungi remains as a common threat. Therefore, this study aimed to evaluate the susceptibility of spoilage fungi of bakery products to acetic, sorbic, and propionic acids at different pH. Penicillium roqueforti, Penicilium paneum, Aspergillus pseudoglaucus, Aspergillus montevidensis and Hyphopichia burtonii strains isolated from spoiled products had their minimum inhibitory concentration (MIC) defined by macrodilution. The concentrations tested were: (i) sorbic acid up to 32 mM; (ii) propionic acid up to 1024 mM and (iii) acetic acid up to 800 mM with pH adjusted in 4.5, 5.0, 5.0 and 6.0 after setting the agent concentration. The lowest MICs for all agents were obtained at pH 4.5, usually doubling with every 0.5 pH increase. P. roqueforti strains isolated from spoiled products were the most resistant to all tested preservatives; while strains of the related species P. paneum, showed similar tolerance to acetic and propionic acids but was double more susceptible to sorbic acid. Strains of A. pseudoglaucus and A. montevidensis were indistinctly susceptible to the preservatives and were the most susceptible species to propionic and acetic acids. H. burtonii strains demonstrated the most variable behaviour in comparison to the other strains being the most susceptible to sorbic acid, were like Aspergillus strains regarding propionic acid, but tolerate well acetic acid. Propionic acid concentrations usually allowed in baked goods are lower than the concentrations required to inhibit the most tolerant isolates tested in this study. The same is true for sorbic acid at higher pH levels. Spoilage species of bakery ware presents a distinct susceptibility profile to the preservatives commonly used in this sector, but the high tolerance observed is a cause of concern.

A novel peptide-based electrochemical biosensor for breast cancer characterization over a poly 3-(3-aminophenyl) propionic acid matrix.

In this work, a new electrochemical biosensor was developed using peptides selected by Phage Display as biorecognition phase to Breast Cancer (BC) characterization. Phage clones were selected against MCF-7 (ER-positive BC) proteins, in order to characterize patients with aggressive luminal BC. Biotin-C3 and biotin-H2 peptides were chemically synthesized and validated by flow cytometry, immunofluorescence assays, and ELISA assays, being more reactive to the MCF-7 lineage. Furthermore, a new matrix for the coupling of biomolecules on the surface of graphite electrodes was generated, through electrochemical modification with a new material derived from 3-(3-aminophenyl)propionic acid (3-3-APPA). Electrochemical and morphological characterizations were carried out, and the mechanism of electropolymerization of poly(3-3-APPA) was proposed, in which the carboxylate groups are kept in the structure of the formed polymer. Then, a biosensor was developed by immobilizing the biotin-C3 and biotin-H2 peptides in the SPE/poly(3-3-APPA)/avidin system for the detection of BC tumor markers in serological samples. Finally, peptides were validated using samples from patients with BC and Benign Breast Disease. Biotin-C3 peptide characterized luminal BC according to p53 status and to HER2 expression, being the biosensor a better strategy when compared to ELISA test. This new biosensor will open a new perspective for a rapid and electrochemical platform for the characterization of BC and its molecular subtypes.

New Insights into Controlling Homoacetogenesis in the Co-digestion of Coffee Waste: Effect of Operational Conditions and Characterization of Microbial Communities.

In this research batch reactors were operated with coffee processing waste and autochthonous microbial consortium, and a taxonomic and functional analysis was performed for phase I of stabilization of maximum H2 production and for phase II of maximum H2 consumption. During phase I, the reactor's operating conditions were pH 4.84 to 8.18, headspace 33.18% to 66.82%, and pulp and husk from 6.95 to 17.05 g/L. These assays continued for phase II, with initial pH conditions of 5.8-8.1, headspace of 33.18-66.82%, and pulp and husk remaining from phase I. The highest homoacetogenesis was observed in assay 5 with pH 7.7, 40% headspace, and 15 g/L of pulp and husk (initial concentrations of phase I). A relative abundance of Clostridium 41%, Lactobacillus 20% and Acetobacter 14% was observed in phase I. In phase II, there was a change in relative abundance of 21%, 63%, and 1%, respectively, and functional genes involved with autotrophic (formyltetrahydrofolate synthase) and heterotrophic (enolase) homoacetogenesis, butanol (3-hydroxybutyryl-CoA dehydrogenase), and propionic acid (propionate CoA-transferase) were identified. This study provides a new and amplified insight into the physicochemical and microbiological factors, which can be used to propose adequate operational conditions to maximize the bioenergy production and reduce homoacetogenesis in biological reactors.

Prenatal exposure to propionic acid induces altered locomotion and reactive astrogliosis in male rats.

Autism spectrum disorder (ASD) is a range of neurodevelopmental disorders characterized by movement and social deficits with rapidly increasing incidence worldwide. Propionic acid (PPA) is a histone deacetylase inhibitor that regulates neuronal plasticity in the brain. Evaluation of the behavioral and cellular consequences of PPA exposure during a critical neurodevelopmental window is required. Therefore, in the present study we aimed to evaluate the effects of prenatal PPA exposure on locomotor behavior and astrocyte number, as well as on levels of nitric oxide (NO), synaptophysin (SYP; a marker of synaptic plasticity), and metallothionein 3 (MT-III; a marker of reactive oxygen species and zinc metabolism), in the prefrontal cortex (PFC) of male rats. All parameters were evaluated at three critical ages of development: postnatal days (PD) 21 (weaning age), PD35 (pre-pubertal age) and PD70 (post-pubertal age). Prenatal PPA exposure induced hypolocomotion and decreased rearing events at weaning age. Moreover, astrogliosis in the PFC was observed in PPA-treated rats at pre- and post-pubertal age. SYP levels were dramatically decreased in PPA-treated rats with simultaneous astrogliosis, suggesting reduced synaptic plasticity. MT-III expression was deregulated in PPA-treated rats. Finally, the expression of NO in the PFC remained unaltered in PPA-treated rats. These results mimic behavioral, neuronal and astrocytic characteristics observed in ASD patients.

Lower methane emissions were associated with higher abundance of ruminal Prevotella in a cohort of Colombian buffalos.

BACKGROUND: Ruminants burp massive amounts of methane into the atmosphere and significantly contribute to the deposition of greenhouse gases and the consequent global warming. It is therefore urgent to devise strategies to mitigate ruminant's methane emissions to alleviate climate change. Ruminal methanogenesis is accomplished by a series of methanogen archaea in the phylum Euryarchaeota, which piggyback into carbohydrate fermentation by utilizing residual hydrogen to produce methane. Abundance of methanogens, therefore, is expected to affect methane production. Furthermore, availability of hydrogen produced by cellulolytic bacteria acting upstream of methanogens is a rate-limiting factor for methane production. The aim of our study was to identify microbes associated with the production of methane which would constitute the basis for the design of mitigation strategies. RESULTS: Moderate differences in the abundance of methanogens were observed between groups. In addition, we present three lines of evidence suggesting an apparent higher abundance of a consortium of Prevotella species in animals with lower methane emissions. First, taxonomic classification revealed increased abundance of at least 29 species of Prevotella. Second, metagenome assembly identified increased abundance of Prevotella ruminicola and another species of Prevotella. Third, metabolic profiling of predicted proteins uncovered 25 enzymes with homology to Prevotella proteins more abundant in the low methane emissions group. CONCLUSIONS: We propose that higher abundance of ruminal Prevotella increases the production of propionic acid and, in doing so, reduces the amount of hydrogen available for methanogenesis. However, further experimentation is required to ascertain the role of Prevotella on methane production and its potential to act as a methane production mitigator.

Short-chain fatty acids production by Bifidobacterium species in the presence of salep

BACKGROUND: Salep is obtained by grinding dried orchid tubers and used as a valuable ingredient in the food industry. Because of the glucomannan content of salep, it is thought to have prebiotic potential. However, there is little information in studies concerning the fermentation characteristics and potential prebiotic properties of salep. The objective of this study was to investigate the effect of salep on bifidobacterial growth by measuring the highest optical density (OD), calculating the specific growth rates, and determining the production of lactic acid and short-chain fatty acids (acetic, propionic, and butyric acid) as a result of bacterial fermentation. RESULT: The OD and pH values obtained in this study showed that salep was utilized as a source of assimilable carbon and energy by the Bifidobacterium species (BS). All Bifidobacterium strains produced lactic, acetic, propionic, and butyric acid, indicating that salep is readily fermented by these bacteria. Salep at 1% (w/v) showed a similar effect on bifidobacterial growth as that promoted by 1% (w/v) glucose used as a traditional carbon source. CONCLUSIONS: Bifidobacterium species can develop in media containing salep as well as in glucose and exhibit the potential to be used as new sources of prebiotics.

Disturbance of bioenergetics and calcium homeostasis provoked by metabolites accumulating in propionic acidemia in heart mitochondria of developing rats.

Propionic acidemia is caused by lack of propionyl-CoA carboxylase activity. It is biochemically characterized by accumulation of propionic (PA) and 3-hydroxypropionic (3OHPA) acids and clinically by severe encephalopathy and cardiomyopathy. High urinary excretion of maleic acid (MA) and 2-methylcitric acid (2MCA) is also found in the affected patients. Considering that the underlying mechanisms of cardiac disease in propionic acidemia are practically unknown, we investigated the effects of PA, 3OHPA, MA and 2MCA (0.05-5 mM) on important mitochondrial functions in isolated rat heart mitochondria, as well as in crude heart homogenates and cultured cardiomyocytes. MA markedly inhibited state 3 (ADP-stimulated), state 4 (non-phosphorylating) and uncoupled (CCCP-stimulated) respiration in mitochondria supported by pyruvate plus malate or α-ketoglutarate associated with reduced ATP production, whereas PA and 3OHPA provoked less intense inhibitory effects and 2MCA no alterations at all. MA-induced impaired respiration was attenuated by coenzyme A supplementation. In addition, MA significantly inhibited α-ketoglutarate dehydrogenase activity. Similar data were obtained in heart crude homogenates and permeabilized cardiomyocytes. MA, and PA to a lesser degree, also decreased mitochondrial membrane potential (ΔΨm), NAD(P)H content and Ca2+ retention capacity, and caused swelling in Ca2+-loaded mitochondria. Noteworthy, ΔΨm collapse and mitochondrial swelling were fully prevented or attenuated by cyclosporin A and ADP, indicating the involvement of mitochondrial permeability transition. It is therefore proposed that disturbance of mitochondrial energy and calcium homeostasis caused by MA, as well as by PA and 3OHPA to a lesser extent, may be involved in the cardiomyopathy commonly affecting propionic acidemic patients.

Effect of rehydration with whey and inoculation with Lactobacillus plantarum and Propionibacterium acidipropionici on the chemical composition, microbiological dynamics, and fermentative losses of corn grain silage / Efeito da reidratação com soro de leite e inoculação com Lactobacillus plantarum e Propionibacterium acidipropionici sobre a composição química, dinâmica microbiológica e perdas fermentativas da silagem de grão de milho

The objective of this study was to compare rehydrated corn grain silages using water or whey and inoculated (Lactobacillus plantarum and Propionibacterium acidipropionici) or not. We also verified whether rehydration with whey associated with the bacterial inoculant improves material conservation. The treatments were as follows: silages rehydrated with water without inoculant (SWa); silages rehydrated with water and inoculated (SWaI); silages rehydrated with whey without inoculant (SWe); silages rehydrated with whey and inoculated (SWeI). A completely randomized design was used, with three replications, treatments in a 2 × 2 factorial scheme (RE: rehydration with water or whey, and IN: inoculation or addition of water without chlorine), during the following storage periods (T): 0, 4, 8, 16, 32, and 64 days. There was an RE x IN x T interaction (P < 0.001) for dry matter (DM), with lower values over time for SWe and higher values in SWeI. An interaction between RE x IN (P < 0.001) and IN x T (P < 0.001) was observed for pH, with higher values for SWe at 64 days of storage and a reduction from the first days of ensiling for SWaI and SWeI. The microbiological variables showed an RE x IN x T interaction (P < 0.001), with the highest counts of lactic acid bacteria for SWaI and SWeI up to 8 days of storage and subsequent higher counts in SWa and SWe. Higher counts were obtained in SWe. For the variables of fermentative losses, there was an RE x IN x T interaction (P < 0.001), with SWeI showing lower losses. The inoculation associated with whey for rehydration of corn grain improved the fermentation profile of the silage, with lower pH values and reduced losses.(AU)

O objetivo desse estudo foi comparar as silagens de grão de milho reidratado utilizando água ou soro de leite e inoculados (Lactobacillus plantarum e Propionibacterium acidipropionici) ou não, e verificar se a reidratação com soro associada ao inoculante bacteriano melhora a conservação do material. Os tratamentos foram: silagens reidratadas com água sem inoculante (SA); silagens reidratadas com água com inoculante (SAI); silagens reidratadas com soro de leite inoculante (SS); silagens reidratadas com soro de leite com inoculante (SSI). Foi utilizado um delineamento inteiramente casualizado, com três repetições, tratamentos em esquema fatorial 2 × 2 (RE: reidratação com água ou soro de leite, e IN: inoculação ou adição de água sem cloro), durante os períodos de estocagem (T): 0, 4, 8, 16, 32 e 64dias. Houve interação RE x IN x T (P < 0,001) para variável matéria seca (MS) com menores valores ao longo do tempo para SS e valores mais elevados em SSI. Foi observada interação RE x IN (P < 0,001)e IN x T (P < 0,001) para pH, com maiores valores para SS aos 64 dias de estocagem e redução desde os primeiros dias de ensilagem para SAI e SSI. As variáveis microbiológicas apresentaram interação RE x IN x T (P < 0,001), sendo a maior contagem de bactérias ácido láticas para SAI e SSI até os 8dias de estocagem e após com maiores contagens em SA e SS; maiores contagens foram obtidas em SS. Para as variáveis de perdas fermentativas houve interação RE x IN x T (P < 0,001), em geral, com SSI apresentando menores perdas. A inoculação associada ao uso de soro para reidratação de grão de milho melhorou o perfil fermentativo da silagem, com menores valores de pH e redução das perdas.(AU)

Effect of rehydration with whey and inoculation with Lactobacillus plantarum and Propionibacterium acidipropionici on the chemical composition, microbiological dynamics, and fermentative losses of corn grain silage / Efeito da reidratação com soro de leite e inoculação com Lactobacillus plantarum e Propionibacterium acidipropionici sobre a composição química, dinâmica microbiológica e perdas fermentativas da silagem de grão de milho

The objective of this study was to compare rehydrated corn grain silages using water or whey and inoculated (Lactobacillus plantarum and Propionibacterium acidipropionici) or not. We also verified whether rehydration with whey associated with the bacterial inoculant improves material conservation. The treatments were as follows: silages rehydrated with water without inoculant (SWa); silages rehydrated with water and inoculated (SWaI); silages rehydrated with whey without inoculant (SWe); silages rehydrated with whey and inoculated (SWeI). A completely randomized design was used, with three replications, treatments in a 2 × 2 factorial scheme (RE: rehydration with water or whey, and IN: inoculation or addition of water without chlorine), during the following storage periods (T): 0, 4, 8, 16, 32, and 64 days. There was an RE x IN x T interaction (P < 0.001) for dry matter (DM), with lower values over time for SWe and higher values in SWeI. An interaction between RE x IN (P < 0.001) and IN x T (P < 0.001) was observed for pH, with higher values for SWe at 64 days of storage and a reduction from the first days of ensiling for SWaI and SWeI. The microbiological variables showed an RE x IN x T interaction (P < 0.001), with the highest counts of lactic acid bacteria for SWaI and SWeI up to 8 days of storage and subsequent higher counts in SWa and SWe. Higher counts were obtained in SWe. For the variables of fermentative losses, there was an RE x IN x T interaction (P < 0.001), with SWeI showing lower losses. The inoculation associated with whey for rehydration of corn grain improved the fermentation profile of the silage, with lower pH values and reduced losses.

O objetivo desse estudo foi comparar as silagens de grão de milho reidratado utilizando água ou soro de leite e inoculados (Lactobacillus plantarum e Propionibacterium acidipropionici) ou não, e verificar se a reidratação com soro associada ao inoculante bacteriano melhora a conservação do material. Os tratamentos foram: silagens reidratadas com água sem inoculante (SA); silagens reidratadas com água com inoculante (SAI); silagens reidratadas com soro de leite inoculante (SS); silagens reidratadas com soro de leite com inoculante (SSI). Foi utilizado um delineamento inteiramente casualizado, com três repetições, tratamentos em esquema fatorial 2 × 2 (RE: reidratação com água ou soro de leite, e IN: inoculação ou adição de água sem cloro), durante os períodos de estocagem (T): 0, 4, 8, 16, 32 e 64dias. Houve interação RE x IN x T (P < 0,001) para variável matéria seca (MS) com menores valores ao longo do tempo para SS e valores mais elevados em SSI. Foi observada interação RE x IN (P < 0,001)e IN x T (P < 0,001) para pH, com maiores valores para SS aos 64 dias de estocagem e redução desde os primeiros dias de ensilagem para SAI e SSI. As variáveis microbiológicas apresentaram interação RE x IN x T (P < 0,001), sendo a maior contagem de bactérias ácido láticas para SAI e SSI até os 8dias de estocagem e após com maiores contagens em SA e SS; maiores contagens foram obtidas em SS. Para as variáveis de perdas fermentativas houve interação RE x IN x T (P < 0,001), em geral, com SSI apresentando menores perdas. A inoculação associada ao uso de soro para reidratação de grão de milho melhorou o perfil fermentativo da silagem, com menores valores de pH e redução das perdas.

The Role of Chitosan as a Possible Agent for Enteric Methane Mitigation in Ruminants.

Livestock production is a main source of anthropogenic greenhouse gases (GHG). The main gases are CH4 with a global warming potential (GWP) 25 times and nitrous oxide (N2O) with a GWP 298 times, that of carbon dioxide (CO2) arising from enteric fermentation or from manure management, respectively. In fact, CH4 is the second most important GHG emitted globally. This current scenario has increased the concerns about global warming and encouraged the development of intensive research on different natural compounds to be used as feed additives in ruminant rations and modify the rumen ecosystem, fermentation pattern, and mitigate enteric CH4. The compounds most studied are the secondary metabolites of plants, which include a vast array of chemical substances like polyphenols and saponins that are present in plant tissues of different species, but the results are not consistent, and the extraction cost has constrained their utilization in practical animal feeding. Other new compounds of interest include polysaccharide biopolymers such as chitosan, mainly obtained as a marine co-product. As with other compounds, the effect of chitosan on the rumen microbial population depends on the source, purity, dose, process of extraction, and storage. In addition, it is important to identify compounds without adverse effects on rumen fermentation. The present review is aimed at providing information about chitosan for dietary manipulation to be considered for future studies to mitigate enteric methane and reduce the environmental impact of GHGs arising from livestock production systems. Chitosan is a promising agent with methane mitigating effects, but further research is required with in vivo models to establish effective daily doses without any detrimental effect to the animal and consider its addition in practical rations as well as the economic cost of methane mitigation.

Experimental evidence that maleic acid markedly compromises glutamate oxidation through inhibition of glutamate dehydrogenase and α-ketoglutarate dehydrogenase activities in kidney of developing rats.

Maleic acid (MA), which has been reported to be highly excreted in propionic acidemia (PAcidemia), was demonstrated to cause nephropathy by bioenergetics impairment and oxidative stress, but the effects on kidney mitochondrial respiration has not yet been properly investigated. Therefore, the present study investigated the effects of MA (0.05-5 mM), as well as of propionic (PA) and 3-hydroxypropionic (3OHPA) acids (5 mM) that accumulate in PAcidemia, on mitochondrial respiration supported by glutamate, glutamate plus malate or succinate in mitochondrial fractions and homogenates from rat kidney, as well as in permeabilized kidney cells. MA markedly decreased oxygen consumption in state 3 (ADP-stimulated) and uncoupled (CCCP-stimulated) respiration in glutamate and glutamate plus malate-respiring mitochondria, with less prominent effects when using succinate. We also found that PA significantly decreased state 3 and uncoupled respiration in glutamate- and glutamate plus malate-supported mitochondria, whereas 3OHPA provoked milder or no changes. Furthermore, glutamate dehydrogenase and α-ketoglutarate dehydrogenase activities necessary for glutamate oxidation were significantly inhibited by MA in a dose-dependent and competitive fashion. The MA-induced decrease of state 3 and uncoupled respiration found in mitochondrial fractions were also observed in homogenates and permeabilized renal cells that better mimic the in vivo cellular milieu. Taken together, our data indicate that MA, and PA to a lesser extent, disturb mitochondrial-oxidative metabolism in the kidney with the involvement of critical enzymes for glutamate oxidation. It is postulated that our present findings may be possibly involved in the chronic renal failure observed in patients with PAcidemia.

Organic acid effects on Brassica napus L. var. oleifera seed germination and seedling growth / Efeitos de ácidos orgânicos na germinação e crescimento inicial de Brassica napus L. var. oleifera

Canola (Brassica napus L. var. oleifera) is cultivated in the Southern Brazil as an alternative winter crop in the rotation system. No-tillage and hydromorphic soils favor the formation of aliphatic organic acids with short chain and low molecular weight, as acetic, butyric, and propionic acids. These acids may cause negative effects on the canola production. Thus, this study aimed to evaluate the response of five canola hybrid genotypes under the effect of acetic, butyric and propionic acids on the germination and initial phase of seedling growth. The results showed the characters evaluated respond differently to treatments applied within the same genotype. Therefore, the selection of a genotype with a resilient performance for the characters evaluated under stress is complex. Nevertheless, a sensitive profile response was observed in Hyola 433 genotype under different organic acids. Furthermore, acetic acid showed no negative impact on germination and initial growth of the genotypes.(AU)

A Canola (Brassica napus L. var. oleifera) é cultivada no Sul do Brasil como uma alternativa para a estação fria no sistema de rotação de culturas. O sistema de plantio direto e a ocorrência de solos hidromórficos favorece a formação de ácidos orgânicos alifáticos com cadeia curta e baixo peso molecular, como os ácidos acético, butírico e propiônico. Estes ácidos podem causar efeitos negativos na produção de canola. Assim, este estudo teve por objetivo avaliar a resposta de cinco genótipos híbridos de canola sob o efeito dos ácidos acético, butírico e propiônico na germinação e na fase inicial do crescimento da plântula. Os resultados demonstraram que os caracteres avaliados respondem de forma diferente em relação aos tratamentos aplicados dentro do mesmo genótipo. Dessa forma, a seleção de um genótipo que manteve um comportamento padrão nos caracteres avaliados sob estresse é dificultado. Entretanto, em uma visão geral, o genótipo Hyola 433 manteve respostas características de sensibilidade nos caracteres avaliados em relação aos ácidos utilizados. Ainda, foi detectado que o ácido acético teve menor efeito na germinação e na fase inicial de crescimento dos genótipos estudados.(AU)

Organic acid effects on Brassica napus L. var. oleifera seed germination and seedling growth / Efeitos de ácidos orgânicos na germinação e crescimento inicial de Brassica napus L. var. oleifera

Canola (Brassica napus L. var. oleifera) is cultivated in the Southern Brazil as an alternative winter crop in the rotation system. No-tillage and hydromorphic soils favor the formation of aliphatic organic acids with short chain and low molecular weight, as acetic, butyric, and propionic acids. These acids may cause negative effects on the canola production. Thus, this study aimed to evaluate the response of five canola hybrid genotypes under the effect of acetic, butyric and propionic acids on the germination and initial phase of seedling growth. The results showed the characters evaluated respond differently to treatments applied within the same genotype. Therefore, the selection of a genotype with a resilient performance for the characters evaluated under stress is complex. Nevertheless, a sensitive profile response was observed in Hyola 433 genotype under different organic acids. Furthermore, acetic acid showed no negative impact on germination and initial growth of the genotypes.

A Canola (Brassica napus L. var. oleifera) é cultivada no Sul do Brasil como uma alternativa para a estação fria no sistema de rotação de culturas. O sistema de plantio direto e a ocorrência de solos hidromórficos favorece a formação de ácidos orgânicos alifáticos com cadeia curta e baixo peso molecular, como os ácidos acético, butírico e propiônico. Estes ácidos podem causar efeitos negativos na produção de canola. Assim, este estudo teve por objetivo avaliar a resposta de cinco genótipos híbridos de canola sob o efeito dos ácidos acético, butírico e propiônico na germinação e na fase inicial do crescimento da plântula. Os resultados demonstraram que os caracteres avaliados respondem de forma diferente em relação aos tratamentos aplicados dentro do mesmo genótipo. Dessa forma, a seleção de um genótipo que manteve um comportamento padrão nos caracteres avaliados sob estresse é dificultado. Entretanto, em uma visão geral, o genótipo Hyola 433 manteve respostas características de sensibilidade nos caracteres avaliados em relação aos ácidos utilizados. Ainda, foi detectado que o ácido acético teve menor efeito na germinação e na fase inicial de crescimento dos genótipos estudados.

Propionic acid metabolism and poly-3-hydroxybutyrate-co-3-hydroxyvalerate production by a prpC mutant of Herbaspirillum seropedicae Z69.

Polyhydroxyalkanoates (PHAs) are thermoplastic polyesters produced by a wide range of bacteria as carbon and energy reserves. PHA accumulation is typically increased under unbalanced growth conditions and with carbon source in excess. Although polyhydroxybutyrate (PHB) could be used for specific applications, it is brittle and not a useful alternative for plastics like polypropylene. Far more useful polypropylene-like PHAs, are copolymers composed of 3-hydroxybutyrate and 3-hydroxyvalerate, P(3HB-co-3HV). Propionic acid is one of the carbon sources that can be used to generate 3HV. A mutant derived from Herbaspirillum seropedicae Z69, a strain previously described as capable of producing P(3HB-co-3HV) from propionic acid, was constructed to increase 3HV biosynthetic efficiency. The strategy involved elimination of a catabolic route for propionyl-CoA by deficiency marker exchange of a selected gene. The mutant (Z69Prp) was constructed by elimination of the 2-methylcitrate synthase (PrpC) gene of the 2-methylcitrate cycle for propionate catabolism. Strain Z69Prp was unable to grow on sodium propionate, but in cultures with glucose-propionate accumulated 50% of its dry weight as copolymer. Z69Prp had 14.1 mol% 3HV; greater than that of strain Z69 (2.89 mol%). The 3HV yield from propionic acid (Y3HV/prop) was 0.80 g g-1, and below the maximum theoretical value (1.35 g g-1).

Synthesis, molecular docking and biological evaluation of novel phthaloyl derivatives of 3-amino-3-aryl propionic acids as inhibitors of Trypanosoma cruzi trans-sialidase.

In the last two decades, trans-sialidase of Trypanosoma cruzi (TcTS) has been an important pharmacological target for developing new anti-Chagas agents. In a continuous effort to discover new potential TcTS inhibitors, 3-amino-3-arylpropionic acid derivatives (series A) and novel phthaloyl derivatives (series B, C and D) were synthesized and molecular docking, TcTS enzyme inhibition and determination of trypanocidal activity were carried out. From four series obtained, compound D-11 had the highest binding affinity value (-11.1 kcal/mol) compared to reference DANA (-7.8 kcal/mol), a natural ligand for TS enzyme. Furthermore, the 3D and 2D interactions analysis of compound D-11 showed a hydrogen bond, π-π stacking, π-anion, hydrophobic and Van der Waals forces with all important amino acid residues (Arg35, Arg245, Arg314, Tyr119, Trp312, Tyr342, Glu230 and Asp59) on the active site of TcTS. Additionally, D-11 showed the highest TcTS enzyme inhibition (86.9% ±â€¯5) by high-performance ion exchange chromatography (HPAEC). Finally, D-11 showed better trypanocidal activity than the reference drugs nifurtimox and benznidazole with an equal % lysis (63 ±â€¯4 and 65 ±â€¯2 at 10 µg/mL) and LC50 value (52.70 ±â€¯2.70 µM and 46.19 ±â€¯2.36 µM) on NINOA and INC-5 strains, respectively. Therefore, D-11 is a small-molecule with potent TcTS inhibition and a strong trypanocidal effect that could help in the development of new anti-Chagas agents.

Fermented non-digestible fraction from combined nixtamalized corn (Zea mays L.)/cooked common bean (Phaseolus vulgaris L.) chips modulate anti-inflammatory markers on RAW 264.7 macrophages.

Chronic non-communicable diseases (NCDs) are low-level inflammation processes affected by several factors including diet. It has been reported that mixed whole grain and legume consumption, e.g. corn and common bean, might be a beneficial combination due to its content of bioactive compounds. A considerable amount would be retained in the non-digestible fraction (NDF), reaching the colon, where microbiota produce short-chain fatty acids (SCFAs) and phenolic compounds (PC) with known anti-inflammatory effect. The aim of this study was to estimate the anti-inflammatory potential of fermented-NDF of corn-bean chips (FNDFC) in RAW 264.7 macrophages. After 24 h, FNDFC produced SCFAs (0.156-0.222 mmol/l), inhibited nitric oxide production > 80% and H2O2 > 30%, up-regulated anti-inflammatory cytokines (I-TAC, TIMP-1) > 2-fold, and produced angiostatic and protective factors against vascular/tissue damage, and amelioration of tumor necrosis factor signalling and inflammatory bowel disease. These results confirm the anti-inflammatory potential derived from healthy corn-bean chips.

Valorization of the Crude Glycerol for Propionic Acid Production Using an Anaerobic Fluidized Bed Reactor with Grounded Tires as Support Material.

This study evaluated the propionic acid (HPr) production from crude glycerol (CG) (5000 mg L-1) in an anaerobic fluidized bed reactor (AFBR). Grounded tire particles (2.8-3.35 mm) were used as support material for microbial adhesion. The reactor was operated with hydraulic retention times (HRT) varying from 8 to 0.5 h under mesophilic (30 °C) conditions. The HPr was the main metabolite produced, increasing in composition from 66.5 to 99.6% by decreasing the HRT from 8 to 0.5 h. Other metabolic products were 1,3-propanediol, with a maximum of 29.4% with an HRT of 6 h, ethanol, acetic, and butyric acids. The decrease in HRT from 8 to 0.5 h decreased the HPr yield, with a maximum of 0.48 ± 0.06 g HPr g COD-1 and an HRT of 6 h, and favored HPr productivity, with a maximum of 4.09 ± 1.24 g L-1 h-1 and HRT of 0.5 h. In the biogas, the H2 content increased from 12.5 to 81.2% by decreasing the HRT from 8 to 0.5 h. These results indicate the potential application of the AFBR for HPr production using an immobilized mixed culture.