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1.
J Sci Food Agric ; 100(5): 2236-2243, 2020 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-31917481

RESUMO

BACKGROUND: To study if the effect on fermentation of yeasts added in ruminant diets can be improved, the effect of adding dry (DY) or pre-activated (AY) Saccharomyces cerevisiae, compared with unsupplemented rumen fluid (CT), on barley grain or sugar beet pulp was evaluated under in vitro high-concentrate fermentative conditions. Yeasts were pre-activated by culturing aerobically at 30 °C for 24 h. RESULTS: In Experiment 1, AY showed a higher concentration than DY at 6 h incubation (6.83 versus 5.76 log cfu mL-1 ; P = 0.007), differences disappearing at 12 h. This was supported by higher gas production with AY, especially on sugar beet pulp. In Experiment 2, incubation pH was 6.24 and 6.31 respectively for barley and sugar beet pulp at 8 h (P < 0.05), but no effect was recorded at 24 h (6.00 and 5.96; P > 0.05). With sugar beet pulp, gas production promoted by AY was the highest (P < 0.05) in the first 8 h of incubation. However, differences with barley were lower and only detected between AY and CT at 12 h (P < 0.05). Total volatile fatty acids (VFAs) concentration at 8 h followed the same trend, but no differences were detected on molar VFAs profile or lactate concentration. Microbial diversity was more affected by the incubation series than by experimental treatments, and inocula including yeasts (AY, DY) did not differ from unsupplemented rumen liquid. CONCLUSIONS: When pre-activated, the concentration of S. cerevisiae was initially higher and resulted in higher gas volumes, and more on a fibrous (sugar beet pulp) than a starchy (barley) substrate. The response is apparently quantitative, since no major changes were detected on biodiversity or fermentation profile. © 2020 Society of Chemical Industry.


Assuntos
Fibras na Dieta/metabolismo , Fermentação , Rúmen/metabolismo , Saccharomyces cerevisiae/metabolismo , Amido/metabolismo , Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Animais , Beta vulgaris , Ácidos Graxos Voláteis/metabolismo , Hordeum , Ruminantes
2.
Int J Legal Med ; 129(5): 963-75, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26162598

RESUMO

DNA profiling is a key tool for forensic analysis; however, current methods identify a suspect either by direct comparison or from DNA database searches. In cases with unidentified suspects, prediction of visible physical traits e.g. pigmentation or hair distribution of the DNA donors can provide important probative information. This study aimed to explore single nucleotide polymorphism (SNP) variants for their effect on hair colour prediction. A discovery panel of 63 SNPs consisting of already established hair colour markers from the HIrisPlex hair colour phenotyping assay as well as additional markers for which associations to human pigmentation traits were previously identified was used to develop multiplex assays based on SNaPshot single-base extension technology. A genotyping study was performed on a range of European populations (n = 605). Hair colour phenotyping was accomplished by matching donor's hair to a graded colour category system of reference shades and photography. Since multiple SNPs in combination contribute in varying degrees to hair colour predictability in Europeans, we aimed to compile a compact marker set that could provide a reliable hair colour inference from the fewest SNPs. The predictive approach developed uses a naïve Bayes classifier to provide hair colour assignment probabilities for the SNP profiles of the key SNPs and was embedded into the Snipper online SNP classifier ( http://mathgene.usc.es/snipper/ ). Results indicate that red, blond, brown and black hair colours are predictable with informative probabilities in a high proportion of cases. Our study resulted in the identification of 12 most strongly associated SNPs to hair pigmentation variation in six genes.


Assuntos
Cor de Cabelo/genética , Polimorfismo de Nucleotídeo Único , População Branca/genética , Europa (Continente) , Feminino , Marcadores Genéticos , Genótipo , Humanos , Funções Verossimilhança , Modelos Logísticos , Masculino , Fenótipo
3.
Transfus Med Hemother ; 42(6): 385-8, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26733770

RESUMO

BACKGROUND: DNA profiling with sets of highly polymorphic autosomal short tandem repeat (STR) markers has been applied in various aspects of human identification in forensic casework for nearly 20 years. However, in some cases of complex kinship investigation, the information provided by the conventionally used STR markers is not enough, often resulting in low likelihood ratio (LR) calculations. In these cases, it becomes necessary to increment the number of loci under analysis to reach adequate LRs. Recently, it has been proposed that single nucleotide polymorphisms (SNPs) could be used as a supportive tool to STR typing, eventually even replacing the methods/markers now employed. METHODS: In this work, we describe the results obtained in 7 revised complex paternity cases when applying a battery of STRs, as well as 52 human identification SNPs (SNPforID 52plex identification panel) using a SNaPshot methodology followed by capillary electrophoresis. RESULTS: Our results show that the analysis of SNPs, as complement to STR typing in forensic casework applications, would at least increase by a factor of 4 total PI values and correspondent Essen-Möller's W value. CONCLUSIONS: We demonstrated that SNP genotyping could be a key complement to STR information in challenging casework of disputed paternity, such as close relative individualization or complex pedigrees subject to endogamous relations.

4.
Electrophoresis ; 35(21-22): 3173-87, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24888494

RESUMO

The field of research and development of forensic STR genotyping remains active, innovative, and focused on continuous improvements. A series of recent developments including the introduction of a sixth dye have brought expanded STR multiplex sizes while maintaining sensitivity to typical forensic DNA. New supplementary kits complimenting the core STRs have also helped improve analysis of challenging identification cases such as distant pairwise relationships in deficient pedigrees. This article gives an overview of several recent key developments in forensic STR analysis: availability of expanded core STR kits and supplementary STRs, short-amplicon mini-STRs offering practical options for highly degraded DNA, Y-STR enhancements made from the identification of rapidly mutating loci, and enhanced analysis of genetic ancestry by analyzing 32-STR profiles with a Bayesian forensic classifier originally developed for SNP population data. As well as providing scope for genotyping larger numbers of STRs optimized for forensic applications, the launch of compact next-generation sequencing systems provides considerable potential for genotyping the sizeable proportion of nucleotide variation existing in forensic STRs, which currently escapes detection with CE.


Assuntos
Genética Forense/métodos , Repetições de Microssatélites/genética , Cromossomos Humanos Y/genética , Feminino , Humanos , Masculino , Mutação , Linhagem
5.
Int J Legal Med ; 128(1): 19-25, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23665814

RESUMO

Various strategies for analysing SNP markers and genotyping have been published with the goal of obtaining informative profiles from biological samples that contain only small amounts of template and/or degraded DNA. In this study, a multiplex assay of 52 autosomal single-nucleotide polymorphisms (SNPs) was used to analyse 438 individuals from urban populations from different regions of Colombia, as well as a sample of 50 Native American individuals of the Pastos ethnic group from Nariño. To determine if significant differences in these 52 SNPs exist between the distinct regions of Colombia, genetic distance and admixture analyses were performed based on the available data for 17 different Colombian population groups and for population groups from Africa, Europe and America. The results demonstrate significant differences between the populations from the Southwest Andean, Central-West Andean, Central-East Andean, Orinoquian and northern Colombian Pacific Coast regions. Most of the regions exhibited a European and Native American admixture. One exception is the population from the region of Chocó (on the northern Pacific Coast), which exhibits a high proportion of African admixture (54 %). From the observed genetic backgrounds, it is possible to conclude that a single reference database for the entire country would not be suitable for forensic purposes. The allele frequencies and the forensically relevant parameters were calculated for all of the markers in each Colombian region with significant values for the combined matching probability (power of discrimination ≥0.99999999999999990) and the combined probability of exclusion (≥0.9990) in trios that were obtained from all of the population groups.


Assuntos
Impressões Digitais de DNA/métodos , Etnicidade/genética , Genética Forense/métodos , Marcadores Genéticos/genética , Genética Populacional , Genótipo , Polimorfismo de Nucleotídeo Único/genética , Colômbia , Comparação Transcultural , Frequência do Gene , Variação Genética/genética , Humanos , Valor Preditivo dos Testes , Probabilidade
6.
Electrophoresis ; 34(8): 1151-62, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23400880

RESUMO

There is growing interest in developing additional DNA typing techniques to provide better investigative leads in forensic analysis. These include inference of genetic ancestry and prediction of common physical characteristics of DNA donors. To date, forensic ancestry analysis has centered on population-divergent SNPs but these binary loci cannot reliably detect DNA mixtures, common in forensic samples. Furthermore, STR genotypes, forming the principal DNA profiling system, are not routinely combined with forensic SNPs to strengthen frequency data available for ancestry inference. We report development of a 12-STR multiplex composed of ancestry informative marker STRs (AIM-STRs) selected from 434 tetranucleotide repeat loci. We adapted our online Bayesian classifier for AIM-SNPs: Snipper, to handle multiallele STR data using frequency-based training sets. We assessed the ability of the 12-plex AIM-STRs to differentiate CEPH Human Genome Diversity Panel populations, plus their informativeness combined with established forensic STRs and AIM-SNPs. We found combining STRs and SNPs improves the success rate of ancestry assignments while providing a reliable mixture detection system lacking from SNP analysis alone. As the 12 STRs generally show a broad range of alleles in all populations, they provide highly informative supplementary STRs for extended relationship testing and identification of missing persons with incomplete reference pedigrees. Lastly, mixed marker approaches (combining STRs with binary loci) for simple ancestry inference tests beyond forensic analysis bring advantages and we discuss the genotyping options available.


Assuntos
Genética Forense/métodos , Marcadores Genéticos/genética , Genética Populacional/métodos , Repetições de Microssatélites , Reação em Cadeia da Polimerase Multiplex/métodos , Grupos Raciais/genética , Teorema de Bayes , Análise por Conglomerados , Frequência do Gene/genética , Genômica/métodos , Humanos , Projetos de Pesquisa , Análise de Sequência de DNA/métodos
7.
Antonie Van Leeuwenhoek ; 103(5): 1057-68, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23354907

RESUMO

The effect of neutral detergent soluble fibre (NDSF) to neutral detergent fibre (NDF) dietary ratio (0.29, LR and 0.43, HR) on the caecal ecosystem of lactating does and their offspring was studied. From the 17th day of lactation, each diet was given to four does, allowing for free access to their litters. Does were sampled at 17 and 28 days of lactation, and also two pups per litter at 17 (milk-fed only), 28 (milk and solid fed) and 49 days of age. DGGE was used to study bacterial caecal biodiversity, and total bacterial concentration and relative proportions of Ruminococcus albus and Butyrivibrio fibrisolvens were quantified by real time PCR. In does, diet did not affect (P > 0.10) diversity indexes, total bacterial concentration or relative abundance of B. fibrisolvens, but at 28 days of lactation the proportion of R. albus was higher with LR (interaction Diet × Time, P = 0.037). Caecal communities of pups of 17 days were grouped by litter, but the influence of the mother was reduced at 28 days with solid feed intake, and at 49 days rabbits clustered by diet. Caecal biodiversity increased from 17 to 28 days, and was reduced at 49 days (Shannon index of 3.60, 3.71 and 3.57, respectively; P = 0.049). Total bacterial concentration and relative abundance of R. albus and B. fibrisolvens increased with solid feed intake from 17 to 28 days (P < 0.01), remaining unaffected thereafter. Access of pups to solid feed from 17 days of age modulates the development and composition of the caecal microbiota at weaning.


Assuntos
Biodiversidade , Ceco/microbiologia , Dieta/métodos , Fibras na Dieta/metabolismo , Animais , Animais Recém-Nascidos , Carga Bacteriana , Butyrivibrio/isolamento & purificação , DNA Bacteriano/genética , Eletroforese em Gel de Gradiente Desnaturante , Detergentes , Lactação , Coelhos , Ruminococcus/isolamento & purificação
8.
Anaerobe ; 20: 50-7, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23403279

RESUMO

The effect of the level of neutral detergent fibre (NDF: 0.35, LI and 0.42, HI) and neutral detergent soluble fibre (NDSF: 0.14, LS and 0.17, HS) in the caecal ecosystem was studied in 24 weaned (28 days of age) rabbits, weighing 630 ± 80.2 g in a 2 × 2 factorial design. After 22 days, rabbits were slaughtered and their caecal contents sampled. The caecal pH (on average 6.2) and molar volatile fatty acids (VFA) proportions were not affected by dietary treatments, but total VFA concentration tended to be lower with NDF (84.7 vs. 74.1 mmol/l; P = 0.095). The amount of total bacteria tended (P = 0.075) to increase with NDSF, but only in diets with 0.35 NDF. The caecal proportions of Ruminococcus albus and Fibrobacter succinogenes were not affected by type or level of fibre, but Butyrivibrio fibrisolvens decreased (P = 0.055) with the NDF proportion in LS diets. Denaturing gradient gel electrophoresis (DGGE) analysis showed that bacterial communities clustered according to each combination of NDF and NDSF, but did not greatly differ among diets (similarity indexes between 0.67 and 0.70), nor biodiversity was affected (average Shannon and richness indexes 3.50 and 33.1; P > 0.10). Archaeal population revealed changes in the amount and composition that were particularly evident in HS diets, decreasing in concentration (from 4.37 to 4.12 log10 gene copy number/g) and biodiversity (Shannon index from 3.14 to 2.52 and richness index from 23.7 to 13.9) compared to LS. The type and level of dietary fibre had a minor impact on caecal fermentation traits or caecal bacterial community. However, the increase in NDSF from 0.14 to 0.17 reduced concentration and diversity of methanogenic archaea.


Assuntos
Archaea/genética , Bactérias/genética , Ceco/microbiologia , Fibras na Dieta/administração & dosagem , Fermentação , Coelhos/microbiologia , Animais , Archaea/classificação , Archaea/efeitos dos fármacos , Archaea/crescimento & desenvolvimento , Bactérias/classificação , Bactérias/efeitos dos fármacos , Bactérias/crescimento & desenvolvimento , Ceco/metabolismo , Eletroforese em Gel de Gradiente Desnaturante , Detergentes , Dieta/métodos , Ecossistema , Reação em Cadeia da Polimerase/métodos , Desmame
9.
Animals (Basel) ; 13(3)2023 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-36766232

RESUMO

The effect of the ensiling length (3, 6, or 9 months), and the processing by dehydration (D) or dehydration and pelleting (P) with respect to the fresh silages (F) were studied in vitro on three maize cultivars in three incubation runs to study the effect of these factors on the nutritive value of maize silage. Gas production pattern, in vitro true digestibility (IVTD), methane concentration (6 and 12 h), ammonia, and volatile fatty acid concentration (VFA) at 12 h were measured. The moisture and pH of F averaged 676 g/kg and 4.09, respectively, and were not affected by the ensiling length, but moisture was reduced, and the pH increased in D and P with respect to F (p < 0.05). The ensiling length did not affect the chemical composition, but differences among the processing forms were detected in a higher acid detergent insoluble nitrogen (ADIN) proportion in P than D, and D than F (p < 0.001). Silages opened at 9 months showed the lowest gas production (p < 0.05), and those that opened after 3 months showed the highest IVTD. The effects of processing on nutrient utilisation only manifested on 3 month silages, with the volume of gas production and IVTD being lower in D than F. However, processing tended (p = 0.064) to reduce the methane proportion at 12 h, indicating both a more efficient fermentation and a lower potential of greenhouse gas emissions compared to the fresh silages. Extending the length of ensiling to 9 months reduced the fermentation of maize silage. The processing increased the dry matter and buffered the feed as well as contributed to an increase in fermentation in 3 month silages.

10.
Animals (Basel) ; 13(5)2023 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-36899762

RESUMO

The aim of this study was to evaluate the effect of a phytomelatonin-rich diet, including by-products from the food industry, on ram sperm quality and seminal plasma composition. Melatonin content in several by-products before and after in vitro ruminal and abomasal digestion was determined by HPLC-ESI-MS/MS. Finally, 20% of a mix of grape pulp with pomegranate and tomato pomaces was included in the rams' diet, constituting the phytomelatonin-rich diet. Feeding the rams with this diet resulted in an increase in seminal plasma melatonin levels compared with the control group (commercial diet) in the third month of the study. In addition, percentages higher than those in the control group of morphologically normal viable spermatozoa with a low content of reactive oxygen species were observed from the second month onwards. However, the antioxidant effect does not seem to be exerted through the modulation of the antioxidant enzymes since the analysis of the activities of catalase, glutathione reductase and glutathione peroxidase in seminal plasma revealed no significant differences between the two experimental groups. In conclusion, this study reveals, for the first time, that a phytomelatonin-rich diet can improve seminal characteristics in rams.

11.
Anaerobe ; 18(6): 602-7, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23123831

RESUMO

To verify if non-invasive collection of soft faeces (SF) from rabbits can be used as an index of bacterial biodiversity in caecal contents (CC), 24 weaned rabbits were given diets with low (LI) and high (HI) levels of insoluble fibre (neutral detergent fibre, NDF) and low (LS) and high (HS) levels of soluble fibre (neutral detergent soluble fibre, NDSF). After 21 days, animals were fitted with neck collars for SF collection. Two days later, animals were slaughtered and CC sampled. Total bacterial concentration quantified by real time PCR (log(10) ng DNA/mg DM) was higher in SF than CC (2.615 vs. 2.383). Among diets, in CC it was (P = 0.059) lowest in LILS diet, whereas in SF it decreased (P = 0.025) with the NDF level. DGGE profiles showed that structure of bacterial communities of SF was close to that of CC; however, similarity was higher in LI than HI diets (0.82 vs. 0.74). Diversity indexes in CC decreased with NDSF (P < 0.05), whereas the effect of NDF (P < 0.05) was also appreciated in SF. Soft faeces can be an alternative to surgery or slaughter techniques to monitor changes in caecal bacterial community; however, high dietary NDF may decrease similarity between both communities.


Assuntos
Bactérias/classificação , Bactérias/genética , Biodiversidade , Ceco/microbiologia , Dieta/métodos , Fibras na Dieta/administração & dosagem , Fezes/microbiologia , Animais , Carga Bacteriana , Eletroforese em Gel de Gradiente Desnaturante , Coelhos
12.
Animals (Basel) ; 12(3)2022 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-35158667

RESUMO

Interest in using clays in the diets of ruminants to improve health and performance is increasing. The microbial fermentation of 65:35 (HC) or 35:65 (HF) concentrate:forage feeds, alone or with zeolite (Z), bentonite (B), or sepiolite (S), was studied in an in vitro semicontinuous culture system. The medium pH was allowed to drop for the first 6 h and was gradually buffered thereafter. For the HC diet, the medium pH was higher with Z throughout incubation (p < 0.05). Similar results were observed for the HF diet, but with lower differences between the additives. Throughout incubation, the volume of gas produced was higher with HC than HF (p < 0.05). The gas volume with S was the lowest (p < 0.05) for HC, whereas for HF it was lowest with B from 8 h onwards (p < 0.05). No treatment differences (p > 0.05) were observed in dry matter disappearance, microbial mass, or volatile fatty acids. However, the inclusion of B in HC reduced the ammonia concentration at 6 and 12 h with respect to C (p < 0.05). The inclusion of zeolite as an additive in the diets of ruminants stabilizes the rumen environment during the first stages of fermentation in terms of pH and ammonia concentration, especially in high-concentrate diets. The buffering effect of bentonite and sepiolite was lower, and both might reduce ruminal microbial fermentation, depending on the concentrate proportion.

13.
Int J Mol Sci ; 12(9): 5815-27, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22016628

RESUMO

AIMS: The relationship between variants in SLCO1B1 and SLCO2B1 genes and lipid-lowering response to atorvastatin was investigated. MATERIAL AND METHODS: One-hundred-thirty-six unrelated individuals with hypercholesterolemia were selected and treated with atorvastatin (10 mg/day/4 weeks). They were genotyped with a panel of ancestry informative markers for individual African component of ancestry (ACA) estimation by SNaPshot(®) and SLCO1B1 (c.388A>G, c.463C>A and c.521T>C) and SLCO2B1 (-71T>C) gene polymorphisms were identified by TaqMan(®) Real-time PCR. RESULTS: Subjects carrying SLCO1B1 c.388GG genotype exhibited significantly high low-density lipoprotein (LDL) cholesterol reduction relative to c.388AA+c.388AG carriers (41 vs. 37%, p = 0.034). Haplotype analysis revealed that homozygous of SLCO1B1*15 (c.521C and c.388G) variant had similar response to statin relative to heterozygous and non-carriers. A multivariate logistic regression analysis confirmed that c.388GG genotype was associated with higher LDL cholesterol reduction in the study population (OR: 3.2, CI95%:1.3-8.0, p < 0.05). CONCLUSION: SLCO1B1 c.388A>G polymorphism causes significant increase in atorvastatin response and may be an important marker for predicting efficacy of lipid-lowering therapy.


Assuntos
Ácidos Heptanoicos/uso terapêutico , Hipercolesterolemia/tratamento farmacológico , Hipercolesterolemia/genética , Transportadores de Ânions Orgânicos/genética , Polimorfismo de Nucleotídeo Único , Pirróis/uso terapêutico , Idoso , Anticolesterolemiantes/uso terapêutico , Atorvastatina , Feminino , Frequência do Gene , Genótipo , Haplótipos , Humanos , Desequilíbrio de Ligação , Transportador 1 de Ânion Orgânico Específico do Fígado , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Farmacogenética/métodos , Resultado do Tratamento
14.
Animals (Basel) ; 10(10)2020 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-33096765

RESUMO

In vitro rumen fermentation systems are often adapted to forage feeding conditions, with pH values ranging in a range close to neutrality (between 6.5 and 7.0). Several attempts using different buffers have been made to control incubation pH in order to evaluate microbial fermentation under conditions simulating high concentrate feeding, but results have not been completely successful because of rapid exhaustion of buffering capacity. Recently, a modification of bicarbonate ion concentration in the buffer of incubation solution has been proposed, which, together with using rumen inoculum from donor ruminants given high-concentrate diets, allows for mimicking such conditions in vitro. It is important to consider that the gas volume recorded is in part directly produced from microbial fermentation of substrates, but also indirectly from the buffering capacity of the medium. Thus, the contribution of each (direct and indirect) gas source to the overall production should be estimated. Another major factor affecting fermentation is the rate of passage, but closed batch systems cannot be adapted to its consideration. Therefore, a simple semicontinuous incubation system has been developed, which studies the rate and extent of fermentation by gas production at the time it allows for controlling medium pH and rate of passage by manual replacement of incubation medium by fresh saliva without including rumen inoculum. The application of this system to studies using high concentrate feeding conditions will also be reviewed here.

15.
Animals (Basel) ; 10(4)2020 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-32340162

RESUMO

The fermentation patterns of six fiber sources, soybean hulls (SH), sugarbeet pulp (BP), palm kernel cake (PK), oat hulls (OH), dehydrated alfalfa meal (DA), and barley straw (BS) were evaluated for this study on the effect of their presentation form (non-processed, NP and ground, GR). Substrates were tested in a conventional in vitro batch system, using rumen fluid obtained from ewes fed 0.5 alfalfa hay and 0.5 barley straw. All substrates rendered a higher gas production in GR form (p < 0.05) except for BS but ranked similarly irrespective of the presentation form. Among the substrates, when incubated NP, the highest volume of gas was recorded with BP from 8 h onwards (p < 0.05), whereas OH and BS resulted in the lowest gas volume (p < 0.05). During the first half of the incubation period, methane production was higher in GR than NP (p < 0.05). Among substrates, despite NP or GR, methane production with BP was the highest (p < 0.05). Similarly, the presentation form did not qualitatively affect fermentation, as no differences were observed in volatile fatty acids proportions. The effect of particle size of fibrous substrates does not have a major impact on the rate and extent of the rumen microbial fermentation.

16.
Animals (Basel) ; 10(2)2020 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-32041290

RESUMO

The fermentation pattern of several carbohydrate sources and their interaction with the nature of microbial inoculum was studied. Barley (B), maize (M), sorghum, (S), sugarbeet pulp (BP), citrus pulp (CP) and wheat bran (WB) were tested in an in vitro semicontinuous system maintaining poorly buffered conditions from 0 to 6 h, and being gradually buffered to 6.5 from 8 to 24 h to simulate the rumen pH pattern. Rumen fluid inoculum was obtained from lambs fed with either concentrate and barley straw (CI) or alfalfa hay (FI). The extent of fermentation was higher with CI than FI throughout the incubation (p < 0.05). Among the substrates, S, BP and M maintained the highest pH (p < 0.05), whereas CP recorded the lowest pH with both inocula. Similarly, CP recorded the highest gas volume throughout the incubation, followed by WB and B, and S recorded the lowest volume (p < 0.05). On average, the total volatile fatty acid (VFA), as well as lactic acid concentration, was higher with CP than in the other substrates (p < 0.05). The microbial structure was more affected by the animal donor of inoculum than by the substrate. The in vitro semicontinuous system allows for the study of the rumen environment acidification and substrate microbial fermentation under intensive feeding conditions.

17.
Br J Nutr ; 100(5): 977-83, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18334044

RESUMO

The contribution of microbial amino acids through caecotrophy to tissue protein metabolism was investigated in lactating does. Attempts were made to vary microbial supply through a dietary antibiotic, Zn bacitracin, and to vary tissue demand through manipulation of litter size. Three groups of eight New Zealand does were fed different experimental diets from day 28 of pregnancy to day 26 of lactation. The control group received the basal diet formulated to meet requirements with grass hay, wheat, soybean meal and barley grain. The second (no antibiotic) group and the third (bacitracin; BAC) group ingested the basal diet supplemented with ammonium sulfate (5 g/kg), initially unlabelled (day 1 to day 8) then labelled with 15N (day 9 to day 30), while the BAC diet was also supplemented throughout with antibiotic (Zn bacitracin; 100 mg/kg). From just after birth each group of does was subdivided into two groups, each of four females, with the litter size either five (LS5) or nine (LS9) pups. The 15N enrichment in liver, milk and caecal bacteria amino acids was determined by GC-combustion-isotope ratio MS. All amino acids in bacterial protein were enriched with the (15 NH 4)2SO4 treatment, with lysine 15N enrichment significantly greater in caecal bacteria (0.23 (SE 0.0063) atom % excess (ape)) than in liver (0.04 (SE 0.0004) ape) or milk protein (0.05 (SE 0.0018) ape), confirming the double origin (bacterial and dietary) of tissue lysine. The contribution of microbes to tissue lysine was 0.23 (SE 0.006) when milk protein was used as reference.


Assuntos
Aminoácidos/biossíntese , Bactérias/metabolismo , Lactação/fisiologia , Fígado/metabolismo , Lisina/metabolismo , Leite/metabolismo , Aminoácidos/análise , Animais , Antibacterianos/farmacologia , Ceco/microbiologia , Feminino , Marcação por Isótopo , Tamanho da Ninhada de Vivíparos , Gravidez , Coelhos
18.
PLoS One ; 13(10): e0205572, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30321211

RESUMO

This study investigated the effect of the feeding behaviour on growth performance, and carcass and meat characteristics of 96 barrows fed ad libitum or restrictively with high or low amino acids (AA) diets according to a 2 × 2 factorial design. The feeding behaviour traits were measured with automated feeders. From 86 kg BW, half of the pigs were given feeds with high indispensable (AA) contents, while the other half received feeds with indispensable AA contents reduced by 9% in early finishing (86-118 kg BW) and by 18% in late finishing (118-145 kg BW). Body lipid and protein retentions were estimated from BW and backfat depth measures recorded at the beginning and end of each period. Pigs were slaughtered at 145 kg BW and carcass and meat quality data were recorded. Phenotypic correlations among feeding behaviours, growth performances, and carcass and meat traits were computed from all the data after adjustment for the effects of feeding treatments. As feeding rate was the behavioural trait most highly correlated with performance and carcass traits, the records of each pig were classified into feeding rate tertiles. Then, the data were statistically analysed using a mixed model, which included feed restriction (FR), AA reduction (AAR), the FR × AAR interaction and the feeding rate tertile as fixed factors, and pen as a random factor. Pigs eating faster (52.1 to 118.9 g/min) had significantly greater final body weights (16%), average daily weight gains (27%), estimated protein gains (22%), estimated lipid retention (46%), carcass weights (16%), weights of lean cuts (14%), weights of fat cuts (21%), proportions of fat in the carcass (14%), and 4% lower proportions of carcass lean cuts than pigs eating slowly (12.6 to 38.2 g/min). Manipulating the eating rate, through management or genetic strategies, could affect feed intake and subsequent growth performance, hence carcass quality, but have little influence on feed efficiency.


Assuntos
Composição Corporal , Peso Corporal , Comportamento Alimentar , Sus scrofa/anatomia & histologia , Sus scrofa/crescimento & desenvolvimento , Ração Animal , Animais , Automação , Gorduras na Dieta , Proteínas Alimentares , Carne , Sus scrofa/psicologia , Fatores de Tempo
19.
FEMS Microbiol Lett ; 272(2): 144-53, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-17498210

RESUMO

The effect feeding antibiotics has on the bacterial population of the rabbit caecum was investigated. No changes in total volatile fatty acid production or total bacterial counts were observed compared with nonantibiotic treated controls. However, treatment with chlortetracycline resulted in an increase of propionate at the apparent cost of butyrate (P<0.05). Denaturing gradient gel electrophoresis analysis indicated that the two antibiotics that inhibit protein synthesis (chlortetracycline and tiamulin) exerted the most similar changes on the bacterial population structure, decreasing the diversity of the profiles. Sequence analysis of DNA from excised denaturing gradient gel electrophoresis bands was carried out. The majority of the sequences observed were most similar to bacterial sequences previously described in other gut environments, with 11% being most similar to those previously reported from the rabbit, and 95% of the sequences having 95% or greater identity to sequences already in GenBank.


Assuntos
Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Ceco/microbiologia , Animais , Bactérias/classificação , Bactérias/isolamento & purificação , Biodiversidade , Butiratos/análise , Contagem de Colônia Microbiana , DNA Bacteriano/análise , DNA Bacteriano/genética , Eletroforese em Gel de Poliacrilamida , Ácidos Graxos Voláteis/biossíntese , Fezes/química , Fezes/microbiologia , Modelos Animais , Dados de Sequência Molecular , Desnaturação de Ácido Nucleico , Filogenia , Reação em Cadeia da Polimerase , Propionatos/análise , Coelhos , Análise de Sequência de DNA
20.
Methods Mol Biol ; 1420: 233-53, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27259744

RESUMO

An expanding choice of ancestry-informative marker single nucleotide polymorphisms (AIM-SNPs) is becoming available for the forensic user in the form of sensitive SNaPshot-based tests or in alternative single-base extension genotyping systems (e.g., Sequenom iPLEX) that can be adapted for analysis with SNaPshot. In addition, alternative ancestry-informative variation: Indels and STRs can be analyzed using direct PCR-to-CE techniques that offer the possibility to detect mixed profiles. We review the current forensically viable AIM panels, their optimized PCR multiplexes, and the population differentiation power they offer. We also describe how improved population divergence balance can be achieved with the enlarged multiplex scales of next-generation sequencing approaches to enable analysis of admixed individuals without biased estimation of co-ancestry proportions.


Assuntos
Genética Forense/métodos , Marcadores Genéticos , Eletroforese Capilar/métodos , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Repetições de Microssatélites , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Nucleotídeo Único
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