Changes in rumen microbes in sheep in response to dietary forage quality and relationships with fermentation characteristics

An in vitro experiment was carried out using the Hohenheim gas production technique to evaluate 24-h gas production, apparently and truly degraded dry matter (DM), partitioning factor (PF), short chain fatty acids, crude protein (CP) and carbohydrate (CHO) fractionation of grass and multipurpose tree species (MPTS) foliage diets. Four grasses and three MPTS were used to formulate 12 diets of equal mixtures (0.5:0.5 on DM basis) of each grass with each MPTS. In vitro gas production was terminated after 24 h for each diet. True DM degradability was measured from incubated samples and combined with gas volume to estimate PF. Diets had greater (P<0.001) CP (102–183 g/kg DM) content than sole grasses (66–131 g/kg DM) and lower (P<0.001) concentrations of fibre fractions. Contrary to in vitro apparently degraded DM, in vitro truly degraded DM coefficient was greater (P<0.001) in diets (0.63–0.77) than in sole grasses (0.48–0.68). The PF was on average higher in diets than in sole grasses. The proportion of potentially degradable CP fractions (A1, B1, B2 and B3, based on the Cornell Net Carbohydrate and Protein System) in the diets ranged from 971 to 989 g/kg CP. Crude protein fractions, A and B2 were greater in diets but B1 and B3 fractions were less in diets than in sole grasses. A similar trend was also observed in the CHO fractions. Results showed that the nutritive value of the four grasses was improved when MPTS leaves were incorporated into the diet and this could ensure higher productivity of the animals.

A mouse model for X-linked adrenoleukodystrophy

X-linked adrenoleukodystrophy (X-ALD) is a peroxisomal disorder with impaired β-oxidation of very long chain fatty acids (VLCFAs) and reduced function of peroxisomal very long chain fatty acyl-CoA synthetase (VLCS) that leads to severe and progressive neurological disability. The X-ALD gene, identified by positional cloning, encodes a peroxisomal membrane protein (adrenoleukodystrophy protein; ALDP) that belongs to the ATP binding cassette transporter protein superfamily. Mutational analyses and functional studies of the X-ALD gene confirm that it and not VLCS is the gene responsible for X-ALD. Its role in the β-oxidation of VLCFAs and its effect on the function of VLCS are unclear. The complex pathology of X-ALD and the extreme variability of its clinical phenotypes are also unexplained. To facilitate understanding of X-ALD pathophysiology, we developed an X-ALD mouse model by gene targeting. The X-ALD mouse exhibits reduced β-oxidation of VLCFAs, resulting in significantly elevated levels of saturated VLCFAs in total lipids from all tissues measured and in cholesterol esters from adrenal glands. Lipid cleft inclusions were observed in adrenocortical cells of X-ALD mice under the electron microscope. No neurological involvement has been detected in X-ALD mice up to 6 months. We conclude that X-ALD mice exhibit biochemical defects equivalent to those found in human X-ALD and thus provide an experimental system for testing therapeutic intervention.

Study of the 3-Hydroxy Eicosanoyl-Coenzyme A Dehydratase and (E)-2,3 Enoyl-Coenzyme A Reductase Involved in Acyl-Coenzyme A Elongation in Etiolated Leek Seedlings1

(R,S)-[1-14C]3-Hydroxy eicosanoyl-coenzyme A (CoA) has been chemically synthesized to study the 3-hydroxy acyl-CoA dehydratase involved in the acyl-CoA elongase of etiolated leek (Allium porrum L.) seedling microsomes. 3-Hydroxy eicosanoyl-CoA (3-OH C20:0-CoA) dehydration led to the formation of (E)-2,3 eicosanoyl-CoA, which has been characterized. Our kinetic studies have determined the optimal conditions of the dehydration and also resolved the stereospecificity requirement of the dehydratase for (R)-3-OH C20:0-CoA. Isotopic dilution experiments showed that 3-hydroxy acyl-CoA dehydratase had a marked preference for (R)-3-OH C20:0-CoA. Moreover, the very-long-chain synthesis using (R)-3-OH C20:0-CoA isomer and [2-14C]malonyl-CoA was higher than that using the (S) isomer, whatever the malonyl-CoA and the 3-OH C20:0-CoA concentrations. We have also used [1-14C]3-OH C20:0-CoA to investigate the reductant requirement of the enoyl-CoA reductase of the acyl-CoA elongase complex. In the presence of NADPH, [1-14C]3-OH C20:0-CoA conversion was stimulated. Aside from the product of dehydration, i.e. (E)-2,3 eicosanoyl-CoA, we detected eicosanoyl-CoA resulting from the reduction of (E)-2,3 eicosanoyl-CoA. When we replaced NADPH with NADH, the eicosanoyl-CoA was 8- to 10-fold less abundant. Finally, in the presence of malonyl-CoA and NADPH or NADH, [1-14C]3-OH C20:0-CoA led to the synthesis of very-long-chain fatty acids. This synthesis was measured using [1-14C]3-OH C20:0-CoA and malonyl-CoA or (E)-2,3 eicosanoyl-CoA and [2-14C]malonyl-CoA. In both conditions and in the presence of NADPH, the acyl-CoA elongation activity was about 60 nmol mg−1 h−1, which is the highest ever reported for a plant system.

Cloning of human acetyl-CoA carboxylase-beta and its unique features.

Acetyl-CoA carboxylase, which has a molecular mass of 265 kDa (ACC-alpha), catalyzes the rate-limiting step in the biosynthesis of long-chain fatty acids. In this study we report the complete amino acid sequence and unique features of an isoform of ACC with a molecular mass of 275 kDa (ACC-beta), which is primarily expressed in heart and skeletal muscles. In these tissues, ACC-beta may be involved in the regulation of fatty acid oxidation, rather than fatty acid biosynthesis. ACC-beta contains an amino acid sequence at the N terminus which is about 200 amino acids long and may be uniquely related to the role of ACC-beta in controlling carnitine palmitoyltransferase I activity and fatty acid oxidation by mitochondria. If we exclude this unique sequence at the N terminus the two forms of ACC show about 75% amino acid identity. All of the known functional domains of ACC are found in the homologous regions. Human ACC-beta cDNA has an open reading frame of 7,343 bases, encoding a protein of 2,458 amino acids, with a calculated molecular mass of 276,638 Da. The mRNA size of human ACC-beta is approximately 10 kb and is primarily expressed in heart and skeletal muscle tissues, whereas ACC-alpha mRNA is detected in all tissues tested. A fragment of ACC-beta cDNA was expressed in Escherichia coli and antibodies against the peptide were generated to establish that the cDNA sequence that we cloned is that for ACC-beta.

Fluidity of the lipid domain of cell wall from Mycobacterium chelonae.

The mycobacterial cell wall contains large amounts of unusual lipids, including mycolic acids that are covalently linked to the underlying arabinogalactan-peptidoglycan complex. Hydrocarbon chains of much of these lipids have been shown to be packed in a direction perpendicular to the plane of the cell surface. In this study, we examined the dynamic properties of the organized lipid domains in the cell wall isolated from Mycobacterium chelonae grown at 30 degrees C. Differential scanning calorimetry showed that much of the lipids underwent major thermal transitions between 30 degree C and 65 degrees C, that is at temperatures above the growth temperature, a result suggesting that a significant portion of the lipids existed in a structure of extremely low fluidity in the growing cells. Spin-labeled fatty acid probes were successfully inserted into the more fluid part of the cell wall. Our model of the cell wall suggests that this domain corresponds to the outermost leaflet, a conclusion reinforced by the observation that labeling of intact cells produced electron spin resonance spectra similar to those of the isolated cell wall. Use of stearate labeled at different positions showed that the fluidity within the outer leaflet increased only slightly as the nitroxide group was placed farther away from the surface. These results are consistent with the model of mycobacterial cell wall containing an asymmetric lipid bilayer, with an internal, less fluid mycolic acid leaflet and an external, more fluid leaflet composed of lipids containing shorter chain fatty acids. The presence of the low-fluidity layer will lower the permeability of the cell wall to lipophilic antibiotics and chemotherapeutic agents and may contribute to the well-known intrinsic resistance of mycobacteria to such compounds.

PXA1, a possible Saccharomyces cerevisiae ortholog of the human adrenoleukodystrophy gene.

The adrenoleukodystrophy protein (ALDp) is an ATP-binding cassette (ABC) transporter in the human peroxisome membrane. It is defective in X chromosome-linked adrenoleukodystrophy (ALD), a neurodegenerative disorder with impaired peroxisomal oxidation of very long chain fatty acids. We report cloning and characterization of PXA1, a yeast gene encoding a protein (Pxa1p) exhibiting high similarity to ALDp. Disruption of PXA1 results in impaired growth on oleic acid and reduced ability to oxidize oleate. Pxa1p is peroxisome associated; however, in the PXA1 mutant yeast, as in ALD cells, peroxisomes are morphologically intact. Disruption of a second yeast gene, YKL741, which encodes a more distantly related ALDp homolog (Yk174p), in either wild-type or PXA1 mutant yeast, results in a growth phenotype identical to that of the PXA1 mutant. This result suggests that Yk1741p and Pxa1p may be subunits of the same transporter. Sequence analysis of Pxa1p, ALDp, and related ABC transporters reveals a possible fatty acid binding domain and a 14-amino acid EAA-like motif, previously described only in prokaryotes. Because of the similarities in sequence and function, we propose that Pxa1p is the Saccharomyces cerevisiae ortholog of ALDp.

Análise da microbiota intestinal em adultos com hábitos alimentares distintos e de associações com a inflamação e resistência à insulina

Introdução: A microbiota intestinal possui grande diversidade de bactérias, predominantemente dos filos Bacteroidetes e Firmicutes, com múltiplas funções. A alimentação pode alterar sua composição e função. Alto teor de gordura saturada altera a permeabilidade intestinal, eleva os lipopolissacarídeos e predispõe à inflamação subclínica crônica. Dieta rica em fibras, como a vegetariana, induz elevação de ácidos graxos de cadeia curta e benefícios metabólicos. Objetivos: Para analisar a composição da microbiota intestinal de adventistas com diferentes hábitos alimentares e associá-los à inflamação subclínica e resistência à insulina, esta tese incluiu: 1) revisão dos mecanismos que associam a alimentação à microbiota intestinal e ao risco cardiometabólico; 2) verificação da composição da microbiota intestinal segundo diferentes hábitos alimentares e de associações com biomarcadores de doenças cardiometabólicas; 3) avaliação da associação entre a abundância de Akkermansia muciniphila e o metabolismo da glicose; 4) análise da presença de enterótipos e de associações com características clínicas. Métodos: Este estudo transversal incluiu 295 adventistas estratificados segundo hábitos alimentares (vegetariano estrito, ovo-lacto-vegetariano e onívoro). Foram avaliadas associações com dados clínicos, bioquímicos e inflamatórios. O perfil da microbiota foi obtido por sequenciamento do gene 16S rRNA (Illumina® Miseq). Resultados: 1) Há evidências de que as relações entre dieta, inflamação, resistência à insulina e risco cardiometabólico são em parte mediadas pela composição da microbiota intestinal. 2) Vegetarianos apresentaram melhor perfil clínico quando comparados aos onívoros. Confirmou-se maior abundância de Firmicutes e Bacteroidetes, que não diferiram segundo a adiposidade corporal. Entretanto, vegetarianos estritos apresentaram mais Bacteroidetes, menos Firmicutes e maior abundância do gênero Prevotella quando comparados aos outros dois grupos de hábitos alimentares. Entre os ovo-lactovegetarianos verificou-se maior proporção de Firmicutes especialmente do gênero Faecalibacterium. Nos onívoros, houve super-representação do filo Proteobacteria (Succinivibrio e Halomonas) comparados aos vegetarianos. 3) Indivíduos normoglicêmicos apresentaram maior abundância de Akkermansia muciniphila que aqueles com glicemia alterada. A abundância desta bactéria correlacionou-se inversamente à glicemia e hemoglobina glicosilada. 4) Foram identificados três enterótipos (Bacteroides, Prevotella e Ruminococcaceae), similares àqueles previamente descritos. As concentrações de LDL-C foram menores no enterótipo 2, no qual houve maior frequência de vegetarianos estritos. Discussão: 1) Conhecimentos sobre participação da microbiota na fisiopatologia de doenças poderão reverter em estratégias para manipulá-la para promover saúde. 2) Apoia-se a hipótese de que hábitos alimentares se associam favorável ou desfavoravelmente a características metabólicas e inflamatórias do hospedeiro via alterações na composição da microbiota intestinal. Sugerimos que a exposição a alimentos de origem animal possa impactar negativamente nas proporções de comunidades bacterianas. 3) Sugerimos que a abundância da Akkermansia muciniphila possa participar do metabolismo da glicose. 4) Reforçamos que a existência de três enterótipos não deva ser específica de certas populações/continentes. Apesar de desconhecido o significado biológico destes agrupamentos, as correlações com o perfil lipídico podem sugerir sua utilidade na avaliação do risco cardiometabólico. Conclusões: Nossos achados fortalecem a ideia de que a composição da microbiota intestinal se altera mediante diferentes hábitos alimentares, que, por sua vez, estão associados a alterações nos perfis metabólicos e inflamatórios. Estudos prospectivos deverão investigar o potencial da dieta na prevenção de distúrbios cardiometabólicos mediados pela microbiota.

Efeito das farinhas de jatobá-do-cerrado (Hymenaea stigonocarpa Mart.) in natura e extrusada no metabolismo lipídico e parâmetros fermentativos em hamsters e resposta glicêmica em humanos após a extrusão

Introdução: A polpa farinácea do jatobá-do-cerrado (Hymenaea stigonocarpa Mart.) apresenta alto teor de fibra alimentar, em média 60 g/100 g, que são importantes para a redução do risco e controle de doenças crônicas não transmissíveis (DCNT). A extrusão termoplástica neutraliza aromas intensos, proporciona a formação de amido resistente, aumenta a fibra alimentar solúvel e melhora a textura do produto final. Objetivo: Estudar o efeito das farinhas de jatobá-do-cerrado in natura (FIN) e extrusada (FE) no metabolismo lipídico e parâmetros fermentativos em hamsters, bem como verificar a resposta glicêmica em humanos após a extrusão. Métodos: Processo de extrusão: velocidade de 200 rpm; matriz com 4 mm de diâmetro; taxa de compressão 3:1; alimentação constante de 70 gramas/minuto; temperatura de 150 °C; proporção farinha de jatobá-do-cerrado e amido de milho: 70:30 por cento e umidade a 25 por cento . Foi realizado um experimento animal com hamsters durante 21 dias, em que se analisou alguns parâmetros do metabolismo lipídico e colônico (fermentativos) dos animais, divididos em quatro grupos experimentais, se diferenciando pela dieta. As dietas controle (GC), in natura (GFI) e extrusada (GFE) eram hipercolesterolemizantes (13,5 por cento de gordura de coco e 0,1 por cento de colesterol) e a dieta referência (GR) com óleo de soja como fonte lipídica, não. Todas as dietas apresentavam 15 por cento de fibra alimentar, sendo que as dietas GR e GC tinham como fonte de fibra a celulose, e as dietas GFI e GFE tiveram as próprias fibras como fonte. A resposta glicêmica em humanos foi verificada por meio do ensaio do índice glicêmico e carga glicêmica da FE, com dez voluntários saudáveis que consumiram 25 gramas de carboidratos disponíveis do alimento teste (farinha extrusada) ou do pão branco como alimento controle. Resultados: Não foi observada diferença significativa entre o peso final, ingestão diária média e total, ganho de peso e CEA entre os animais dos quatro grupos. A concentração de triglicerídeos foi menor em 41 por cento e 38 por cento nos animais que receberam as dietas GFI e GFE, em relação aqueles que receberam a dieta GC, assim como também para o colesterol total (55 por cento e 47 por cento ), LDL-c (70 por cento e 53 por cento ) e não-HDL-c (63 por cento e 49 por cento ) séricos, lipídeos totais hepáticos (39 por cento e 45 por cento ) e o peso dos fígados dos animais também foi menor (21 por cento em ambos os grupos). Não houve diferença no colesterol hepático e excretado nas fezes dos animais dos quatro grupos. Os animais do GFE excretaram 57 por cento mais ácidos biliares nas fezes que os animais do GC. Com relação aos parâmetros fermentativos, observou-se maior excreção de fibras (1,24 ± 0,08 e 1,52 ± 0,09 gramas) nos animais dos grupos GR e GC respectivamente, em relação aos do GFI e GFE (0,50 e 0,48 gramas), porém o escore fecal (3,50 ± 0,19 e 3,38 ± 0,18) e o grau de fermentação (54 e 52 por cento ) foi maior nos animais dos grupos GFI e GFE. Houve uma maior produção de AGCC no ceco dos animais dos grupos GFI e GFE (80 e 57,5 µmol/g de ceco respectivamente) e maior diminuição do pH no conteúdo cecal nos animais do grupo GFI (7,49 ± 0,10), em relação ao GC (8,06 ± 0,13). Os ácidos acético e propiônico, estiveram presentes em maior quantidade no ceco dos animais dos grupos GFI (58,5 e 6,1 µmol/g de ceco) e GFE (42,5 e 6,6 µmol/g de ceco) e os animais do GFI produziram mais ácido butírico (15 µmol/g de ceco), em relação aos demais grupos. Quanto à resposta glicêmica da farinha pós extrusão, não houve diferença entre a área de resposta glicêmica da farinha extrusada e do pão branco, o índice glicêmico da farinha extrusada (glicose como controle) foi classificado como moderado, e a carga glicêmica (na porção de 30 gramas), baixa. Conclusão: As FIN e FE favoreceram a redução do colesterol total, LDL-c, não-HDL-c e dos triglicerídeos séricos, além da diminuição do acúmulo de lipídeos hepáticos. Foi observado também aumento expressivo na formação de AGCC e no grau de fermentação. A FE proporcionou um aumento na excreção de ácidos biliares nas fezes e apresentou índice glicêmico moderado e baixa carga glicêmica.

Organic carbon and biomarker record from the Laptev Sea continental margin

In order to understand the processes controlling organic carbon deposition (i.e., primary productivity vs. terrigenous supply) and their paleoceanographic significance, three sediment cores (PS2471, PS2474. and PS2476) from the Laptev Sea continental margin were investigated for their content and composition of organic carbon. The characterization of organic matter indudes the determination of buk parameters (hydrogen index values and C/N ratios) and the analysis of specific biomarkers (n-alaknes, fatty acids, alkenones, and pigments). Total organic carbon (TOC) values vary between 0.3 and 2%. In general, the organic matter from the Laptev Sea continental margin is dominated by terrigenous matter throughout. However. significant amounts of marine organic carbon occur. The turbidites, according to a still preliminary stratigraphy probably deposited during glacial Oxygen Isotope Stages 2 and 4, are characterized by maximum amounts of organic carbon of terrigenous origin. Marine organic carbon appears to show enhanced relative abundances in the Termination I (?) and early Holocene time intervals, as indicated by maximum amounts of short chain n-alkanes, short-chain fatty acids, and alkenones. The increased amounts of faity acids, however, may also have a freshwater origin due to increased river discharge at that time. The occurrence of alkenones is suggested to indicate an intensification of Atlantic water inflow along the Eurasian continental margin starting at that time. Oxygen Isotope Stage l accumutation rates of total organic carhon are 0.3, 0.17, and 0.02 C/cm**2/ky in cores PS2476, PS2474, and PS2471, respectively.

Electrolyte transport in the mammalian colon: mechanisms and implications for disease Kunzelmann and Marcus Mall

Electrolyte Transport in the Mammalian Colon: Mechanisms and Implications for Disease. Physiol. Rev. 82: 245-289, 2002.The colonic epithelium has both absorptive and secretory functions. The transport is characterized by a net absorption of NaCl, short-chain fatty acids (SCFA), and water, allowing extrusion of a feces with very little water and salt content. In addition, the epithelium does secret mucus, bicarbonate, and KCl. Polarized distribution of transport proteins in both luminal and basolateral membranes enables efficient salt transport in both directions, probably even within an individual cell. Meanwhile, most of the participating transport proteins have been identified, and their function has been studied in detail. Absorption of NaCl is a rather steady process that is controlled by steroid hormones regulating the expression of epithelial Na+ channels (ENaC), the Na+-K+-ATPase, and additional modulating factors such as the serum- and glucocorticoid-regulated kinase SGK. Acute regulation of absorption may occur by a Na+ feedback mechanism and the cystic fibrosis transmembrane conductance regulator (CFTR). Cl- secretion in the adult colon relies on luminal CFTR, which is a cAMP-regulated Cl- channel and a regulator of other transport proteins. As a consequence, mutations in CFTR result in both impaired Cl- secretion and enhanced Na+ absorption in the colon of cystic fibrosis (CF) patients. Ca2+- and cAMP-activated basolateral K+ channels support both secretion and absorption of electrolytes and work in concert with additional regulatory proteins, which determine their functional and pharmacological profile. Knowledge of the mechanisms of electrolyte transport in the colon enables the development of new strategies for the treatment of CF and secretory diarrhea. It will also lead to a better understanding of the pathophysiological events during inflammatory bowel disease and development of colonic carcinoma.

Dysfonctions mitochondriales associées à l’acidose lactique du Saguenay-Lac-Saint-Jean révélées par l’étude d’un nouveau modèle murin de la maladie

Le syndrome de Leigh version canadienne-française (LSFC) est une maladie autosomale récessive causée par une mutation du gène LRPPRC, encodant une protéine du même nom. LRPPRC est impliquée dans la traduction des gènes mitochondriaux qui encodent certains complexes de la chaine respiratoire. Les répercussions biochimiques incluent un déficit tissu spécifique de la cytochrome c oxydase (COX), principalement dans le foie et le cerveau, et la survenue de crises d’acidose fatales chez 80 % des enfants atteints avant l’âge de 3-4 ans. L’identification d’options thérapeutiques demeure encore un défi de taille et ceci est en partie relié au manque de connaissances des fonctions biologiques de LRPPRC et des mécanismes impliqués dans la pathogenèse du LSFC, au niveau des dysfonctions mitochondriales résultantes. Afin d’étudier ces mécanismes, le consortium de l’acidose lactique, dont fait partie notre laboratoire, a récemment développé un modèle murin portant une ablation de LRPPRC spécifique au foie (souris H-Lrpprc-/-). L’objectif principal est de déterminer si ce modèle reproduit le phénotype pathologique observé dans les cultures de fibroblastes humains issus de biopsies de peau de patients LSFC. Dans le cadre des travaux de ce mémoire, nous avons amorcé la caractérisation de ce nouveau modèle, en examinant le phénotype général, l’histopathologie hépatique et les fonctions mitochondriales, et en nous focalisant principalement sur les fonctions respiratoires et la capacité à oxyder divers types de substrats. Nous avons observé un retard de croissance, une hépatomégalie ainsi que plusieurs anomalies histologiques du foie chez la souris HLrpprc-/-. De plus, l’ablation de LRPPRC induit un déficit du complexe IV, mais aussi de l’ATP synthase, et affecte l’oxydation des acides gras à longues chaines. À la lumière de ces résultats, nous croyons que le modèle murin H-Lrpprc-/- contribuera à l’avancement des connaissances générales sur LRPPRC, nous permettant de mieux comprendre l’influence de la protéine sur les fonctions mitochondriales.

Supplementation of cattle fed tropical grasses with microalgae increases microbial protein production and average daily gain

A series of 3 experiments were conducted to evaluate the use of microalgae as supplements for ruminants consuming low-CP tropical grasses. In Exp. 1, the chemical composition and in vitro protein degradability of 9 algae species and 4 protein supplements were determined. In Exp. 2, rumen function and microbial protein (MCP) production were determined in Bos indicus steers fed speargrass hay alone or supplemented with Spirulina platensis, Chlorella pyrenoidosa, Dunaliella salina, or cottonseed meal (CSM). In Exp. 3, DMI and ADG were determined in B. indicus steers fed speargrass hay alone or supplemented with increasing amounts of NPN (urea combined with ammonia sulfate), CSM, or S. platensis. In Exp. 1, the CP content of S. platensis and C. pyrenoidosa (675 and 580 g/kg DM) was highest among the algae species and higher than the other protein supplements evaluated, and Schizochytrium sp. had the highest crude lipid (CL) content (198 g/kg DM). In Exp. 2, S. platensis supplementation increased speargrass hay intake, the efficiency of MCP production, the fractional outflow rate of digesta from the rumen, the concentration of NH3N, and the molar proportion of branched-chain fatty acids in the rumen fluid of steers above all other treatments. Dunaliella salina acceptance by steers was low and this resulted in no significant difference to unsupplemented steers for all parameters measured for this algae supplement. In Exp. 3, ADG linearly increased with increasing supplementary N intake from both S. platensis and NPN, with no difference between the 2 supplements. In contrast, ADG quadratically increased with increasing supplementary N intake from CSM. It was concluded that S. platensis and C. pyrenoidosa may potentially be used as protein sources for cattle grazing low-CP pastures.

Effects of feeding carnitine and ractopamine on rainbow trout (Oncorhynchus mykiss, Walbaum 1972)

L-carnitine is required for the transfer of long-chain fatty acids from the cytosol to the mitochondrial matrix for 13-oxidation of them and ractopamine, beta adrenergic agonists, have potential stimulating lipolysis and altering rates of protein degradation and synthesis. Present study was carried out to improve lipid body oxidation and protein-sparing action of fish through addition of L-carnitine and ractopamine to diet of rainbow trout, Oncorhynchus mykiss, Walbaum 1972. An eight-week feeding trial was carried out to evaluate the effects of supplementation of tree levels of L-carnitine tartrate (0, 1 and 2 g/kg) and two levels of ractopamine hydrochloride (0 and 10 ppm) on growth performance, fillet muscle fatty acid compositions and blood biochemical parameters in 288 juvenile rainbow trout (130 g) at 3X2 factorial experimental design. Ractopamine and 1 g/kg carnitine improved the specific growth rate, feed conversion ratio, protein efficiency ratio and weight gain at the end of experiment. The protein and lipid contents of fillet muscle were affected by the inclusion of 10 mg/kg ractopamine in the diet, increasing crude protein and reducing crude fat (P<0.05) of fish fillet muscle. The highest protein and lowest fat contents of fish fillet were observed in diet that contains 2 g/kg carnitine plus ractopamine. Ractopamine and carnitine increased levels of albumin, total protein and globulin in fish blood serum, but carnitine increased triacylglycerol and cholesterol. Fatty acids compositions of fish fillet were also affected by ractopamine and carnitine. All fatty acids except for eicosapentaenoic acid and docosahexaenoic acid, were increased by dietary supplementation of ractopamine. Total saturated fatty acids were not affected by carnitine. Supplementation (P>0.05). However, total n-3 poly unsaturated fatty acids were reduced by carnitine supplementation. A significant interaction was observed between ractopamine and carnitine supplementation regarding the saturated (P<0.01) and n-3 poly unsaturated fatty acid (P<0.001) of fish fillet. This study shows that supplementation of 1 g/kg carnitine and 10 ppm ractopamine could improve performance of juvenile rainbow trout and their combination in diet results in protein increment, fat reduction and change in profile of fatty acids in fillet muscle.

Developing an index of dietary estimation of fermentable carbohydrate to allow dietary analysis for epidemiological studies

It has been proposed that long-term consumption of diets rich in non-digestible carbohydrates (NDCs), such as cereals, fruit and vegetables might protect against several chronic diseases, however, it has been difficult to fully establish their impact on health in epidemiology studies. The wide range properties of the different NDCs may dilution their impact when they are combined in one category for statistical comparisons in correlations or multivariate analysis. Several mechanisms have been suggested to explain the protective effects of NDCs, including increased stool bulk, dilution of carcinogens in the colonic lumen, reduced transit time, lowering pH, and bacterial fermentation to short chain fatty acids (SCFA) in the colon. However, it is very difficult to measure SCFA in humans in vivo with any accuracy, so epidemiological studies on the impact of SCFA are not feasible. Most studies use dietary fibre (DF) or Non-Starch Polysaccharides (NSP) intake to estimate the levels, but not all fibres or NSP are equally fermentable. It has been proposed that long-term consumption of diets rich in non-digestible carbohydrates (NDCs), such as cereals, fruit and vegetables might protect against several chronic diseases, however, it has been difficult to fully establish their impact on health in epidemiology studies. The wide range properties of the different NDCs may dilution their impact when they are combined in one category for statistical comparisons in correlations or multivariate analysis. Several mechanisms have been suggested to explain the protective effects of NDCs, including increased stool bulk, dilution of carcinogens in the colonic lumen, reduced transit time, lowering pH, and bacterial fermentation to short chain fatty acids (SCFA) in the colon. However, it is very difficult to measure SCFA in humans in vivo with any accuracy, so epidemiological studies on the impact of SCFA are not feasible. Most studies use dietary fibre (DF) or Non-Starch Polysaccharides (NSP) intake to estimate the levels, but not all fibres or NSP are equally fermentable. The first aim of this thesis was the development of the equations used to estimate the amount of FC that reaches the human colon and is fermented fully to SCFA by the colonic bacteria. Therefore, several studies were examined for evidence to determine the different percentages of each type of NDCs that should be included in the final model, based on how much NDCs entered the colon intact and also to what extent they were fermented to SCFA in vivo. Our model equations are FC-DF or NSP$ 1: 100 % Soluble + 10 % insoluble + 100 % NDOs¥ + 5 % TS** FC-DF or NSP 2: 100 % Soluble + 50 % insoluble + 100 % NDOs + 5 % TS FC-DF* or NSP 3: 100 % Soluble + 10 % insoluble + 100 % NDOs + 10 % TS FC-DF or NSP 4: 100 % Soluble + 50 % insoluble + 100 % NDOs + 10 % TS *DF: Dietary fibre; **TS: Total starch; $NSP: non-starch polysaccharide; ¥NDOs: non-digestible oligosaccharide The second study of this thesis aimed to examine all four predicted FC-DF and FC-NSP equations developed, to estimate FC from dietary records against urinary colonic NDCs fermentation biomarkers. The main finding of a cross-sectional comparison of habitual diet with urinary excretion of SCFA products, showed weak but significant correlation between the 24 h urinary excretion of SCFA and acetate with the estimated FC-DF 4 and FC-NSP 4 when considering all of the study participants (n = 122). Similar correlations were observed with the data for valid participants (n = 78). It was also observed that FC-DF and FC-NSP had positive correlations with 24 h urinary acetate and SCFA compared with DF and NSP alone. Hence, it could be hypothesised that using the developed index to estimate FC in the diet form dietary records, might predict SCFA production in the colon in vivo in humans. The next study in this thesis aimed to validate the FC equations developed using in vitro models of small intestinal digestion and human colon fermentation. The main findings in these in vitro studies were that there were several strong agreements between the amounts of SCFA produced after actual in vitro fermentation of single fibre and different mixtures of NDCs, and those predicted by the estimated FC from our developed equation FC-DF 4. These results which demonstrated a strong relationship between SCFA production in vitro from a range of fermentations of single fibres and mixtures of NDCs and that from the predicted FC equation, support the use of the FC equation for estimation of FC from dietary records. Therefore, we can conclude that the newly developed predicted equations have been deemed a valid and practical tool to assess SCFA productions for in vitro fermentation.