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      Effect of sugarcane fiber digestibility, conservation method and concentrate level on the ruminal ecosystem of beef cattle

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          Abstract

          The aim of this study was to investigate the effects of sugarcane neutral detergent fiber digestibility (NDFD), conservation method, and concentrate level on the ruminal microbial population of steers. Eight ruminal-cannulated Nellore steers were distributed in two contemporary 4 × 4 Latin Square design with a 2 × 2 factorial arrangement of treatments. Experiment 1: diets were formulated with 60% of concentrate level, and two sugarcane genotypes (high or low NDFD) either freshly cut or as silage. Experiment 2: diets were formulated with two levels of concentrate (60 or 80%), and two sugarcane genotypes (high or low NDFD) offered as freshly cut. Each experimental period lasted for 14 d, with the last 4 d used for ruminal fluid collection. Three cellulolytic bacteria ( Fibrobacter succinogenes, Ruminococcus albus, Ruminococcus flavefaciens), two amylolytic ( Streptococcus bovis, Ruminobacter amylophilus), and a lactate fermenting microorganism ( Megasphaera elsdenii) were quantified by qPCR. Experiment 1: diets with fresh sugarcane increased the population of S. bovis, and M. elsdenii. Sugarcane with high NDFD increased F. succinogenes population only when sugarcane was offered as freshly cut. Experiment 2: increasing concentrate in the diet decreased S. bovis population, and increased R. amylophilus. Sugarcane with high NDFD increased the population of cellulolytic bacteria only at the 60% concentrate diet. Providing sugarcane with high NDFD favored the growth of fibrolytic bacteria, and this effect were dependent on the conservation method and on diet concentrate level. In addition, sucrose appears to have great effect on the composition of ruminal microflora, especially S. bovis.

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          Statistical analysis of real-time PCR data

          Background Even though real-time PCR has been broadly applied in biomedical sciences, data processing procedures for the analysis of quantitative real-time PCR are still lacking; specifically in the realm of appropriate statistical treatment. Confidence interval and statistical significance considerations are not explicit in many of the current data analysis approaches. Based on the standard curve method and other useful data analysis methods, we present and compare four statistical approaches and models for the analysis of real-time PCR data. Results In the first approach, a multiple regression analysis model was developed to derive ΔΔCt from estimation of interaction of gene and treatment effects. In the second approach, an ANCOVA (analysis of covariance) model was proposed, and the ΔΔCt can be derived from analysis of effects of variables. The other two models involve calculation ΔCt followed by a two group t-test and non-parametric analogous Wilcoxon test. SAS programs were developed for all four models and data output for analysis of a sample set are presented. In addition, a data quality control model was developed and implemented using SAS. Conclusion Practical statistical solutions with SAS programs were developed for real-time PCR data and a sample dataset was analyzed with the SAS programs. The analysis using the various models and programs yielded similar results. Data quality control and analysis procedures presented here provide statistical elements for the estimation of the relative expression of genes using real-time PCR.
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            Factors that alter rumen microbial ecology.

            Ruminant animals and ruminal microorganisms have a symbiotic relationship that facilitates fiber digestion, but domestic ruminants in developed countries are often fed an abundance of grain and little fiber. When ruminants are fed fiber-deficient rations, physiological mechanisms of homeostasis are disrupted, ruminal pH declines, microbial ecology is altered, and the animal becomes more susceptible to metabolic disorders and, in some cases, infectious disease. Some disorders can be counteracted by feed additives (for example, antibiotics and buffers), but these additives can alter the composition of the ruminal ecosystem even further.
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              A grain-based subacute ruminal acidosis challenge causes translocation of lipopolysaccharide and triggers inflammation.

              The effects of a grain-based subacute ruminal acidosis (SARA) challenge on translocation of lipopolysaccharide (LPS) into the peripheral circulation, acute phase proteins in blood and milk, feed intake, milk production and composition, and blood metabolites were determined in 8 lactating Holstein cows. Between wk 1 and 5 of 2 successive 6-wk periods, cows received a total mixed ration ad libitum with a forage to concentrate (F:C) ratio of 50:50. In wk 6 of both periods, the SARA challenge was conducted by replacing 21% of the dry matter of the total mixed ration with pellets containing 50% wheat and 50% barley. Rumen pH was monitored continuously using indwelling pH probes in 4 rumen cannulated cows. Rumen fluid samples were collected 15 min before feed delivery and at 2, 4, 6, 12, 14, 16, 18, and 24 h after feed delivery for 2 d during wk 5 (control) and wk 6 (SARA). Peripheral blood samples were collected using jugular catheters 15 min before feeding and at 6 and 12 h after feeding at the same days of the rumen fluid collections. The SARA challenge significantly reduced average daily pH from 6.17 to 5.97 and increased the duration of rumen pH below pH 5.6 from 118 to 279 min/d. The challenge reduced dry matter intake (16.5 vs. 19 kg/d), milk yield (28.3 vs. 31.6 kg/d), and milk fat (2.93 vs. 3.30%, 0.85 vs. 0.97 kg/d), and tended to increase milk protein percentage (3.42 vs. 3.29%), without affecting milk protein yield (1.00 vs. 0.98 kg/d). The challenge also increased the concentration of free LPS in rumen fluid from 28,184 to 107,152 endotoxin units (EU)/mL. This was accompanied by an increase in LPS in peripheral blood plasma (0.52 vs. <0.05 EU/mL) with a peak at 12 h after feeding (0.81 EU/mL). Concentrations of the acute phase proteins serum amyloid A, haptoglobin, and LPS-binding protein (LBP) in peripheral blood as well as LBP concentration in milk increased (438.5 vs. 167.4, 475.6 vs. 0, 53.1 vs. 18.2, and 6.94 vs. 3.02 microg/mL, respectively) during SARA. The increase in LBP in combination with the increase in LPS in peripheral blood provides additional evidence of translocation of LPS. Results suggest that the grain-based SARA challenge resulted in translocation of LPS into the peripheral circulation, and that this translocation triggered a systemic inflammatory response.
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                Author and article information

                Contributors
                johnny_msouza@usp.br
                dannylo@usp.br
                brs.mesquita@gmail.com
                ligmesquita@hotmail.com
                (+55) 19-3565 4230 , lfpsilva@usp.br
                Journal
                AMB Express
                AMB Express
                AMB Express
                Springer Berlin Heidelberg (Berlin/Heidelberg )
                2191-0855
                6 March 2017
                6 March 2017
                2017
                : 7
                : 55
                Affiliations
                [1 ]ISNI 0000 0004 1937 0722, GRID grid.11899.38, Department of Animal Science, School of Veterinary Medicine, , Universidade de São Paulo, ; Pirassununga, São Paulo 13635-900 Brazil
                [2 ]Ouro Fino Saúde Animal, Cravinhos, São Paulo 14140-000 Brazil
                Author information
                http://orcid.org/0000-0003-3271-9864
                Article
                356
                10.1186/s13568-017-0356-7
                5339260
                28265975
                cc801824-21d5-4edf-9ec9-89990bd31a90
                © The Author(s) 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

                History
                : 20 December 2016
                : 24 February 2017
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100001807, Fundação de Amparo à Pesquisa do Estado de São Paulo;
                Award ID: 2013/22811-0
                Award ID: 2009/00976-2
                Award Recipient :
                Categories
                Original Article
                Custom metadata
                © The Author(s) 2017

                Biotechnology
                beef cattle,ruminal fermentation,ruminal microorganisms,qpcr,sugarcane
                Biotechnology
                beef cattle, ruminal fermentation, ruminal microorganisms, qpcr, sugarcane

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