Using 15N Dilution Method to Correct for Microbial Contamination When Assessing in situ Protein Degradability of Fresh Ryegrass

محورهای موضوعی : Camel

ر. طهماسبی ¹ ( Department of Animal Science, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran|School of Environmental and Rural Science, University of New England, New South Wales, Armidale, Australia )
ج. و. نولان ² ( School of Environmental and Rural Science, University of New England, New South Wales, Armidale, Australia )
ر. س. دوبس ³ ( Department of Primary Industries, Beef Industry Centre of Excellence, New South Wales, Armidale, Australia )

تاریخ دریافت : 1394/03/22 تاریخ پذیرش : 1394/10/10 تاریخ انتشار : 1395/03/12

کلید واژه: Degradation, ryegrass, Microbial contamination, labeled nitrogen, &lt, i&gt, in situ&lt, /i&gt, ,

چکیده مقاله :

This experiment was done to investigate microbial contamination and in situ disappearance rates of dry matter (DM), N and 15N of fresh labeled ryegrass. Perennial ryegrass (Lolium perenne) were labeled with 15N during growth in a glasshouse, harvested at 4th leaves stage and were incubated up to 34 h in situ in the rumen of 3 individually housed sheep. The animals were fed 800 g/d chopped alfalfa and had free access to drinking water. Six bags were placed in the rumen of each sheep simultaneously and removed after 0, 3, 7, 12, 21 and 33 h after incubation. The results were fitted to a model describing the degradation of DM and total N with time. It was found that residues from the washed zero time bags had lower 15N enrichments (7.7% vs. 8.3% enriched) than the original fresh samples. Under-estimation of effective degradability (ED) of protein in fresh forages by about 4% would have potential consequences for predictions of ruminally fermentable and escape protein and thus for dietary protein feeding management. However, because the correction assumes contaminating microbial N is unlabeled, but microbes attached to labeled ryegrass would become labeled to some extent, the true error and effective degradability may still be underestimated. Studies with two markers would help us to better understand the errors associated with the in situ technique.

چکیده انگلیسی:

این آزمایش برای تعیین میزان آلودگی میکروبی و نرخ‪های ناپدید شدن ماده خشک، نیتروژن و نیتروژن 15 در گیاه نشان‪دار شده رای‪گراس صورت گرفت. پرنیال رای‪گراس (لولیوم پرنه) با استفاده از محلول نیتروژن 15 در هنگام رشد در گلخانه نشان‪دار گردید و در مرحله چهار برگی برداشت شد. سپس به مدت 33 ساعت درون شکمبه سه رأس گوسفند فیستولا شده، که به صورت مجزا در قفس‪های متابولیکی نگهداری می‪شدند، قرار داده شد. گوسفندان روزانه 800 گرم یونجه خرد شده دریافت می‪کردند و به آب آشامیدنی تازه دسترسی داشتند. تعداد 6 کیسه به صورت همزمان درون شکمبه گوسفندان قرار داده شد و سپس در زمان‪های 0، 3، 7، 12، 21 و 33 ساعت پس از شکمبه‪گذاری برداشته شدند. نتایج با استفاده از یک مدل نشان دهنده تجزیه‪پذیری ماده خشک و کل نیتروژن در زمان تنظیم گردیدند. نتایج نشان داد که بقایای حاصل از شسته شدن کیسه‪های زمان صفر در مقایسه با نمونه‪های گیاه تازه، درصد غنی شدن پایینتری داشتند (7/7 درصد در مقایسه با 3/8 درصد). تخمین کمتر از حد تجزیه‪پذیری مؤثر پروتئین گیاه تازه در حدود 4 درصد می‪تواند در باره پیش‪بینی پروتئین عبوری و تجزیه‪پذیر در شکمبه و همینطور مدیریت تغذیه پروتئین پیامدهایی داشته باشد. بنابراین به دلیل این که تصحیح نمودن بر پایه این فرض استوار می‪باشدکه نیتروژن میکروبی نشان‪دار نمی‪باشد اما میکروب‪ها با چسبیدن به رای‪گراس تا اندازه‪ای نشان‪دار می‪شوند، خطای واقعی و تجزیه‪پذیری مؤثر کماکان ممکن است کمتر از حد تخمین زده شوند. انجام مطالعات با دو مارکر می‪تواند برای درک بهتر خطاهای مرتبط با تکنیک کیسه‪های نایلونی کمک شایانی نماید.

منابع و مأخذ:

AFRC. (1992). Agricultural and Food Research Council. AFRC Technical Committee on Responses to Nutrients. Nutritive Requirements of Ruminant Animals: Protein. Nutr. Abstr. Rev. 62, 787-835.

Arroyo J.M., González J., Rodríguez C.A., Alvir M.R. and Ouarti M. (2009). In situ evaluation of the protein value of wheat grain corrected for ruminal microbial contamination. J. Sci. Food Agric. 89, 731-734.

Barrell L.G., Burke J.L., Waghorn G.C., Attwood G.T. and Brookes I.M. (2000). Preparation of fresh forages for incubation and prediction of nutritive value. Pp. 5-8 in Proc. Soc. Anim. Prod. New Zealand.

Beckers Y., Thewis A., Maudoux B. and Francois E. (1995). Studies on the in situ nitrogen degradability corrected for bacterial contamination of concentrate feed in steers. J. Anim. Sci. 73, 220-227.

Broadbent F.E. and Carlton A.B. (1980). Methodology for field trials with nitrogen 15 depleted nitrogen. J. Environ. Qual. 9, 236-242.

Broderick G.A., Huhtanen P., Ahvenjarvi S., Reynal S.M. and Shingfield K.J. (2010). Quantifying ruminal nitrogen metabolism using the omasal sampling technique in cattle-a meta-analysis. J. Dairy Sci. 93, 3216-3230.

Chen X.B. (1997). Neway Excel, a Utility for Processing Data of Feed Degradability and in vitro Gas Production. Rowett Research Institute, Aberdeen, UK.

CohenD.C. and Doyle P.T. (2001). Effect of sample preparation on in situ estimates of protein degradability for white clover herbages. Australian J. Exp. Agric. 41, 619-624.

Colombini S. and Broderick G.A. (2008). In vitro ruminal protein degradation and microbial protein formation of seed legumes. J. Dairy Sci. 91(1), 43-51.

Danso S.K.A., Hardarson G. and Zapata F. (1988). Dinitrogen fixation estimates in alfalfa-ryegrass swards using different nitrogen-15 labeling methods. Crop. Sci. 28, 106-110.

Deinum B. and Maassen A. (1994). Effects of drying temperature on chemical composition and in vitro digestibility of forages. Anim. Feed Sci. Technol. 46, 75-86.

González J., Paria-Marrnol J., Rodriguez C.A. and Martinez A. (2007). Effects of ensiling on ruminal degrada- ability and intestinal digestibility of Italian rye-grass. Anim. Feed Sci. Technol. 136, 38-50.

González J., Centeno C., Morujo N., Faría-Mármol J. and Martínez A. (2009). In situ ruminal amino acid degradability of green and ensiled Italian rye-grass according to particle transit model and microbial contamination correction. Livest. Sci. 123, 209-214.

Harris D.C. and Hesterman O.B. (1990). Quantifying the nitrogen contribution from alfalfa to soil and two succeeding crops using nitrogen-15. Agron. J. 82, 129-134.

Hoffman P.C., Sievert S.J., Shaver R.D., Welch D.A. and Combs D.K. (1993). In situ dry matter, protein and fiber degradation of perennial forages. J. Dairy Sci. 76, 2632-2643.

Hristov A.N. and Broderick G.A. (1996). Synthesis of microbial protein in ruminally cannulated cows fed alfalfa silage, alfalfa hay, or corn silage. J. Dairy Sci. 79, 1627-1637.

Hristov A.N., Huhtanen P., Rode L.M., Acharya S.N. and McAllister T.A. (2001). Comparison of the ruminal metabolism of nitrogen from 15n labeled alfalfa preserved as hay or as silage. J. Dairy Sci. 84, 2738-2750.

Ipharraguerre I.R., Reynal S.M., Lineiro M. and Broderick G.A. (2007). A comparison of sampling sites, digesta and microbial markers and microbial references for assessing the post ruminal supple of nutrients in dairy cows. J. Dairy Sci. 90, 1904-1919.

Jarrige R. (1987). Ingestion et digestion des aliments. Pp. 29-56 in Alimentation des Bovins, Ovins et Caprins. R. Jarrige, Ed. INRA Publications, Paris, France.

Kamoun M., Ammar H., Théwis A., Beckers Y., France J. and López S. (2014). Comparison of three ¹⁵N methods to correct for microbial contamination when assessing in situ protein degradability of fresh forages. J. Anim. Sci. 92, 5053-5062.

Kamoun M., López S., Beckers Y., Lecomte P., Francois E. and Théwis A. (2007). Effect of feed ¹⁵N incorporation into solid-associated bacteria on the in situ nitrogen degradability of ¹⁵N labeled Italian ryegrass. Anim. Feed Sci. Technol. 135, 353-361.

Krawielitzki K., Schmidt T., Voigt J., Kowalczyk J. and Gabel M. (2006). Dynamics of microbial contamination of protein during ruminal in situ incubation of feedstuffs. J. Anim. Feed Sci. 15, 313-328.

Ma T., Deng K.D., Tu Y., Zhang N.F., Jiang C.G., Liu J., Zhao Y.G. and Diao Q.Y. (2014). Effect of dietary forage-to-concentrate ratios on urinary excretion of purine derivatives and microbial nitrogen yields in the rumen of Doper cross bred sheep. Livest. Sci. 160, 37-44.

Machado P.A.S., Valadares Filho S.C., Detmann E., SantosS.A., Valadares R.F.D., Ducatti C., Rotta P.P. and Costa e Silva L.F. (2013). Development of equations to estimate microbial contamination in ruminal incubation residues of forage produced under tropical conditions using N as a label. J. Anim. Sci. 91, 3836-3846.

Mathers J.C. and Aitchison E.M. (1981). Direct estimation of the extent of contamination of food residues by microbial matter after incubation within synthetic fiber bags in the rumen. J. Agric. Sci. 96, 691-693.

Mitchell R.B., Redfearn D.D., Moser L.E., Grant R.J., Moore K.J. and Kirch B.H. (1997). Relationships between in situ protein degradability and grass developmental morphology. J. Dairy Sci. 80, 1143-1149.

Noceck J.E. and Grant A.L. (1987). Characterization of in situ nitrogen and fiber digestion and bacterial nitrogen contamination of hay crop forages preserved at different dry matter percentages. J. Anim. Sci. 64, 552-564.

Olubobokun J.A., Craig W.M. and Pond K.R. (1990). Effects of mastication and microbial contamination on ruminal in situ forage disappearance. J. Anim. Sci. 68, 3371-3381.

Orskov E.R. and McDonaldI. (1979). The estimation of protein degradability in the rumen from incubation measurements weighted according to rate of passage. J. Agric. Sci. 92, 499-503.

Rodríguez C.A. and González J. (2006). In situ study of the relevance of bacterial adherence to feed particles for the contamination and accuracy of rumen degradability estimates for feeds vegetable origin. Br. J. Nutr. 96, 316-325.

SAS Institute. (2003). SAS^®/STAT Software, Release 9.1. SAS Institute, Inc., Cary, NC. USA.

Ulyatt M., Dellow D., John A., Reid C. and Waghorn G. (1986). Contribution of chewing during eating and rumination to the clearance of digesta from the ruminoreticulum. Pp. 498-515 in Control of Digestion and Metabolism in Ruminants. L.P. Milligan, W.L. Grovum and A. Dobson, Eds. Prentice Hall Publication, Englewood Cliffs, New Jersey, USA.

Van Bruchem J., Voigt J., Lammers-Wienhoven T.S.C.W., Schonhusen U., Ketelaars J.J.M.H. and Tamminga S. (1997). Secretion and reabsorption of endogenous protein along the small intestine of sheep: estimates derived from ¹⁵N dilution of plasma non-protein-N. Br. J. Nutr. 77, 273-286.

Varvikko T. and Lindberg J.E. (1985). Estimation of microbial nitrogen in nylon-bag residues by feed ¹⁵N dilution. Br. J. Nutr. 54, 473-481.

Waghorn G.C., Shelton I.D. and Thomas V.J. (1989). Particle breakdown and rumen digestion of fresh ryegrass (Lolium perenne) and lucerne (Medicago sativa) fed to cows during a restricted feeding period. Br. J. Nutr. 61, 409-423.