Developing a Modified in vitro Gas Production Technique to Replace the Nylon Bag Method of Evaluating Protein Degradation of Alfalfa Hay in Ruminants
محورهای موضوعی : Camelا. آقاجانزاده گلشنی 1 , ن. ماهری سیس 2 , ر. سلامت دوست نوبر 3 , ی. ابراهیم نژاد 4 , ا. قربانی 5
1 - Department of Animal Science, Shabestar Branch, Islamic Azad University, Shabestar, Iran
2 - Department of Animal Science, Shabestar Branch, Islamic Azad University, Shabestar, Iran
3 - Department of Animal Science, Shabestar Branch, Islamic Azad University, Shabestar, Iran
4 - Department of Animal Science, Shabestar Branch, Islamic Azad University, Shabestar, Iran
5 - Department of Animal Science, Shabestar Branch, Islamic Azad University, Shabestar, Iran
کلید واژه: alfalfa, gas production, faeces, nylon bag,
چکیده مقاله :
The present study was conducted to investigate the possibility of using a modified in vitro gas production technique in place of the nylon bag method to estimate protein degradability of alfalfa hay in ruminants. In the in situ experiment dacron bags were filled with 3 g alfalfa hay. This was incubated in the rumen of three ruminally cannulated Ghezel rams for the periods of 0, 2, 4, 8, 12, 16, 24, 36 and 48 h. At the end of the experiment, dry matter, organic matter and crude protein degradability (CPD) were calculated. In the in vitro experiment, buffered rumen fluid was prepared in a solution of 19:1 artificial saliva to rumen fluid, which was pre-incubated by rapidly fermentable carbohydrates for 4 h. After pre-incubation, 30 mL of the buffered rumen fluid were added to 100 mL syringes, containing the alfalfa sample, with 7.5 mg N. The samples were incubated for 2, 4, 6, 8, 12, 16, 24, 36, 48, 60 and 72 hours, after which net gas production was computed. In the third experiment, 25 grams of faeces were mixed with 50 mL of artificial saliva, which was then made up to 1 liter by adding more artificial saliva and filtered. Then, the suspension was pre-incubated for 4 h. After pre-incubation, the same steps used to measure gas production were conducted. Results showed that there were significant differences between gas production with rumen liquor and faeces suspension at 2, 4, 24, 36 and 48 h of incubation, while no significant differences were found at other incubation times. There was a close relationship between crude protein degradation at different times and amount of gas production using rumen liquor [CPD= 58.93 + 0.32 gas (r2=0.76, n=18)] and fecal suspension [CPD= 58.38 + 0.27 gas (r2=0.60, n=18)]. The results indicated that faeces suspension can be used instead of rumen liquor as the medium in the gas production method. Development of the gas production technique could result in reducing the need to use fistulated animals in feed evaluation studies.
این مطالعه به منظور بررسی امکان استفاده از روش تولید گاز آزمایشگاهی تغییر یافته به جای روش کیسه‌های نایلونی برای برآورد تجزیهپذیری پروتئین یونجه خشک در نشخوارکنندگان به اجرا در آمد. در آزمایش کیسه‌های نایلونی مقدار سه گرم از نمونه یونجه داخل کیسه‌های داکرونی ریخته شد. کیسه‌ها به مدت صفر، 2، 4، 8، 12، 16، 24، 36 و 48 ساعت در شکمبه سه رأس گوسفند نر قزل کانوله‌دار انکوبه شدند. در پایان آزمایش تجزیهپذیری ماده خشک، ماده آلی و پروتئین محاسبه شدند. در آزمایش تولید گاز، شیرابه شکمبه بافری شده به نسبت 19 قسمت بزاق مصنوعی و یک قسمت شیرابه شکمبه تهیه و با کربوهیدرات سریع قابل تخمیر به مدت 4 ساعت پیش انکوبه شد. بعد از پیش انکوباسیون مقدار 30 میلی‌لیتر از این محلول داخل سرنگ 100 میلی‌لیتری که درون آن مقداری نمونه یونجه که دارای 5/7 میلی‌گرم نیتروژن بود، ریخته شد. نمونه‌ها در زمان‌های 2، 4، 6، 8، 12، 16، 24، 36، 48، 60 و 72 ساعت انکوبه شدند و بعد از آن گاز خالص تولیدی محاسبه شد. در سومین آزمایش 25 گرم از نمونه مدفوع با 50 میلی‌لیتر بزاق مصنوعی مخلوط شده و با اضافه کردن بزاق مصنوعی، حجم آن به یک لیتر رسانده شده و صاف گردید. سپس این سوسپانسیون به مدت 4 ساعت پیش انکوبه شد. پس از پیش انکوباسیون مراحل تولید گاز همانند روش معمول انجام شد. نتایج نشان داد که اختلاف معنی داری بین تولید گاز با شیرابه شکمبه و مدفوع در زمان‌های 2، 4، 24، 36 و 48 ساعت انکوباسیون وجود داشت در حالی‌که در بقیه ساعت‌های انکوباسیون اختلاف معنی داری وجود نداشت. رابطه نزدیکی بین تجزیهپذیری پروتئین خام در ساعت‌های مختلف انکوباسیون و مقدار گاز تولیدی با استفاده از شیرابه شکمبه (18= n و 76/0= r2وگاز 32/0+93/58=تجزیهپذیری پروتئین خام) و سوسپانسیون مدفوع (18= n و 60/0= r2وگاز 27/0+38/58=تجزیهپذیری پروتئین خام) وجود داشت. نتایج نشان داد که سوسپانسیون مدفوع می‌تواند به جای شیرابه شکمبه در روش تولید گاز آزمایشگاهی استفاده شود. توسعه این روش تولید گاز آزمایشگاهی می‌تواند باعث کاهش نیاز به دام‌های فیستوله‌گذاری شده در آزمایش‌های ارزیابی مواد خوراکی شود.
AOAC. (1990). Official Methods of Analysis. Vol. I. 15th Ed. Association of Official Analytical Chemists, Arlington, VA, USA.
Aghajanzadeh-Golshani A., Maheri-Sis N., Baradaran-Hasanzadeh A., Asadi-Dizaji A. and Mirzaei-Aghsaghali A. (2012). Determining nutrients degradation kinetics of chickpea (Cicer arietinum) straw using nylon bag technique in sheep. Open Vet. J. 2, 54-57.
Alvir M.R., González J. and Rodríguez C.A. (1999). Relationship between in situ rumen protein degradability and chemical composition of alfalfa hays. Ann. Zootech. 48, 447-456.
Balfe B. (1985). The development of a two-stage technique for the in vitro digestion of hay using ovine faeces (instead of rumen liquor) as a source of microorganisms. MS Thisis. University of Wales, Bangor.
Bani P., Iamartino N. and Orskov E.R. (1999). In vitro gas production from ruminal or faecal inoculum and comparison with in situ degradability. Pp. 357-359 in Proc. A.S.P.A. 13th Cong. Piacenza, Italy.
Biagi G., Piva A., Ligabue M. and Formigoni A. (2005). Evaluation of in vitro ruminal degradation of alfalfa. Biotechnol. Anim. Husband. 21(6), 259-264.
Borba A.E.S., Correia P.J.A., Fernandes J.M.M. and Borba A.F.R.S. (2001). Comparison of three sources of inocula for predicting apparent digestibility of ruminant feedstuffs. Anim. Res. 50, 265-273.
Bueno I.C.S., Cabral Filho S.L.S., Godoy P.B. and Abdalla A.L. (2010). Comparison of in situ and in vitro dry matter rumen degradability of three distinct quality hays in sheep. Trop. Subtrop. Agroecosyst. 12, 321-332.
Chen X.B. (1995). "Fitcurve" macro, IFRU, the Macaulay Institute, Aberdeen, UK.
Cone J.W., Rodrigues M.A.M., Guedes C.M. and Blok M.C. (2009). Comparison of protein fermentation characteristics in rumen fluid determined with the gas production technique and the nylon bag technique. Anim. Feed Sci. Technol. 153, 28-38.
El-Shaer H.M., Omed H.M., Chamberlain A.G. and Axford R.F.E. (1987). Use of faecal organisms from sheep for the in vitro determination of digestibility. J. Agric. Sci. Camb. 109, 257-259.
England M.L., Broderick G.A., Shaver R.D. and Combs D.K. (1997). Comparison of in situ and in vitro techniques for measuring ruminal degradation of animal by-product proteins. J. Dairy Sci. 80, 2925-2931.
Fon F.N. and Nsahlai I.V. (2012). Laboratory cultured faecal inoculum as a substitute for fresh rumen inoculum for in vitro feed evaluation. African J. Agric. Res. 7(49), 6595-6604.
Getachew G., Makkar H.P.S. and Becker K. (1998). The in vitro gas coupled with ammonia measurement for evaluation of nitrogen degradability in low quality roughages using incubation medium of different buffering capacity. J. Sci. Food Agric. 77, 87-95.
Gressley T.F., Hall M.B. and Armentano L.E. (2011). Ruminant nutrition symposium: productivity, digestion and health responses to hindgut acidosis in ruminants. J. Anim. Sci. 89, 1120-1130.
Jahani-Azizabadi H., Danesh Mesgaran M., Valizadeh R. and Nasiri Moghadam H. (2009). Comparison of in vivo with in situ mobile bag and three step enzymatic procedures to evaluate protein disappearance of alfalfa hay and barley grain. Iranian J. Vet. Res. 10(3), 260-266.
Jalilvand G., Naserian A., Kebreab E., Odongo N.E., Valizadeh R., Eftekhar Shahroodi F., Lopez S. and France J. (2008). Rumen degradation kinetics of alfalfa hay, maize silage and wheat straw treated with fibrolytic enzymes. Arch. Zootech. 57(218), 155-164.
Jiri T., Miroslav S. and Tomas K. (1996). The influence of a nylon bag carrier on alfalfa crude protein degradability. Anim. Feed Sci. Technol. 57, 129-137.
Julier B., Guines F., Emile J.C. and huyghe C. (2003). Variation in protein degradability in dried forage legumes. Anim. Res. 52, 401-412.
Kamalak A., Canbolat O., Gurbuz Y. and Ozay O. (2005). Comparison of in vitro gas production technique with in situ nylon bag technique to estimate dry matter degradation. Czech J. Anim. Sci. 50(2), 60-67.
Karlsson L., Hetta M., Uden P. and Martinsson K. (2009). New methodology for estimating rumen protein degradation using the in vitro gas production technique. Anim. Feed Sci. Technol. 153, 193-202.
Maheri-Sis N., Eghbali-Vaighan M., Mirzaaghazadeh A. and Ebrahimnezhad Y. (2012). Effect of microwave irradiation on the fermentation characteristics and nutritive value of tomato pomace for ruminants using in vitro gas production technique. J. BioSci. Biotechnol. 1(3), 261-266.
Maheri-Sis N., Eghbali-Vaighan M., Mirza-Aghazadeh A., Ahmadzadeh A.R., Aghajanzadeh-Golshani A., Mirzaei-Aghsaghali A. and Shaddel-Telli A.A. (2011). Effects of microwave irradiation on ruminal dry matter degradation of tomato pomace. Curr. Res. J. Biol. Sci. 3, 268-272.
Mauricio R.M., Owen E., Mould F.L., Givens I., Theodorou M.K., France J., Davies D.R. and Dhanoa M.S. (2001). Comparison of bovine rumen liquor and bovine faeces as inoculum for an in vitro gas production technique for evaluating forages. Anim. Feed Sci. Technol. 89, 33-48.
Menke K.H., Raab L., Salewski A., Steingass H., Fritz D. and Schneider W. (1979). The estimation of digestibility and metabolisable energy content of ruminant feedstuffs from the gas production when they incubated with rumen liquor in vitro. J. Agric. Sci. 93, 217-222.
Michalet-Doreau B. and Ould-Bah M.Y. (1992). In vitro and in sacco methods for the estimation of dietary nitrogen degradability in the rumen: a review. Anim. Feed Sci. Technol. 40, 57-86.
Mirzaei-Aghsaghali A., Maheri-Sis N., Mirza Aghazadeh A., Ebrahimnezhad Y., Dastoori M.R. and Aghajanzade-Golshani A. (2008). Estimation of methane production in sheep using nutrient composition of the diet. J. Anim. Vet. Adv. 7(6), 765-770.
Mirzaei-Aghsaghali A., Maheri-Sis N., Mansouri H., Ebrahim Razeghi M., Safaei A.R. Aghajanzadeh-Golshani A. and Alipoor K. (2011). Estimation of the nutritive value of tomato pomace for ruminant using in vitro gas production technique. Afr. J. Biotechnol. 10(33), 6251-6256.
NRC. (2001). Nutrient Requirements of Dairy Cattle. 7th Ed. National Academy Press, Washington, DC, USA.
NRC. (2007). Nutrient Requirements of Small Ruminants: Sheep, Goats, Cervids and New World Camelids. National Academy Press, Washington, DC, USA.
Nikkhah A. and Mahdavi A. (2006). Comparison of in situ and gas test method in determination of feedstuff nutritive value. Iranian J. Agric. Sci. 37(2), 281-292.
Omed H.M., Lvett D.K. and Axford R.F.E. (2000). Faeces as a source of microbial enzymes for estimating digestibility. Pp. 135-154 in Forage Evaluation in Ruminant Nutrition. D.I. Givens, E. Owen, R.F.E. Axford and H.M. Omed, Eds. CABInternational, Wallingford.
Orskov E.R. and Macdonald I. (1979). The estimation of protein degradability in the rumen from incubation measurement weight according to rate of passage. J. Agric. Sci. 92, 499-503.
Parand E. and Taghizadeh A. (2009). Examination of digestibility of processed barley grain with different methods, using gas production technique with two sources of inocola. J. Anim. Sci. Res. 2(4), 1-13.
Paya H., Taghizadeh A., Janamohamadi H. and Moghadam G.A. (2008). Ruminal dry matter and crude protein degradability of some tropical (Iranian) feeds used in ruminant diets estimated using the in situ and in vitro techniques. J. Biol. Sci. 3(7), 720-725.
SAS Institute. (2001). SAS®/STAT Software, Release 8.02. SAS Institute, Inc., Cary, NC. USA.
Tilley J.M.A. and Terry R.A. (1963). A two stage technique for in vitro digestion of forage crops. J. Br. Grass Soc. 18, 104-111.
Varadyova Z., Baran M. and Zelenak I. (2005). Comparison of two in vitro fermentation gas production methods using both rumen fluid and faecal inoculum from sheep. Anim. Feed Sci. Technol. 123, 81-94.
Van Soest P.J., Robertson J.B. and Lewis B.A. (1991). Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J. Dairy Sci. 74, 3583-3597.
Williams W.L., Tedeschi L.O., Kononoff P.J., Callaway T.R., Dowd S.E., Karges K. and Gibson M.L. (2010). Evaluation of in vitro gas production and rumen bacterial populations fermenting corn milling (co) products. J. Dairy Sci. 93, 4735-4743.
