Profil Fermentasi Rumen Sapi Pejantan Feedlot Dengan Administrasi Sikat Stimulator Mekanis Rumen (Rumenfibe ®).
RUMEN FERMENTATION PROFILE OF FEEDLOT STEER
ADMINISTERED WITH RUMEN MECHANICAL
STIMULATOR BRUSH (
Rumenfibe
®)
YULIANRI RIZKI YANZA
GRADUATE SCHOOL
BOGOR AGRICULTURAL UNIVERSITY
BOGOR
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DECLARATION
I do hereby declare that the thesis entitled "Rumen Fermentation Profile Of Feedlot Steer Administered With Rumen Mechanical Stimulator Brush (Rumenfibe®)" is my original work produced through the guidance of my academic advisors and that to the best of my knowledge, it has not been submitted for the award of any degree in any educational institution. All of the incorporated material originated from other published as well as unpublished papers are clearly stated in the text as well as in the references
Hereby I delegate the copy rights of this work to the Bogor Agricultural University.
Bogor, October 2015
Yulianri Rizki Yanza
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SUMMARY
YULIANRI RIZKI YANZA. Rumen Fermentation Profile of Feedlot Steer Administered With Rumen Mechanical Stimulator Brush (Rumenfibe®). Supervised by ANURAGA JAYANEGARA, DEWI APRI ASTUTI and JUNICHI TAKAHASHI.
In recent years concentrate feeding has been the main factor affecting production in feedlot cattle industry. The use of concentrate in feedlot is approximately 90% of the total feed consumed. Although such feeding practice is relevant in improving animal performance within a short period of time, however, its application may induce a metabolic disorder in the rumen, i.e. acidosis. Acidosis occurs when lactic acid and volatile fatty acid production rate are above their clearance rate through absorption and passage to the subsequent gastro-intestinal tract. Rumen mechanical stimulation (RMS) is a technology that may reduce such incidence although the results so far are varied. The aims of this study were to determine the effect of RMS (“RUMENFIBE®” or RF) on performance, rumen fermentation profile and to assess the economic feasibility of using RMS on feedlot steers
An experiment was conducted in Indonesia with twenty Brahman crossbreed steers (ca. 267+4 kg) randomly allocated in four pens and fed with a large proportion of concentrate, following PT Mitra Catur Taruma feeding management. Ten experimental steers in two pens were orally administered with three RMS into rumen of each animal that regarded as treatment group (RF) and another as control. Performances such average nutrients intake, body gain, and income over feed cost, and also by the rumen fermentation profile such pH, potential redox, ammonia, lactic acid, total and partial VFA, methane production and H2 recovery status were compared.
The result showed a similar performance between two treatments (with or without RMS), but RF treatment had to lowered income over feed cost (P< 0.05). The pH, potential redox (∆EH), ammonia, and lactic acid concentration gave a similar status on both treatment. Meanwhile, the total VFA and partial VFA such acetate and butyrate, methane production and H2 recovery status of RF group were higher than the control, although the propionate concentration, all partial VFA proportion (%) and methane relative (methane:total VFA) gave similar status on both treatments. Feedlot steer administered with rumen mechanical stimulation brush (Rumenfibe®) did not gave a negative effect in rumen fermentation profile but improved the rumen fermentation.
Keywords: feedlot steer, rumen fermentation profile, rumen mechanical stimulator rumenfibe.
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RINGKASAN
YULIANRI RIZKI YANZA. Profil Fermentasi Rumen Sapi Pejantan Feedlot Dengan Administrasi Sikat Stimulator Mekanis Rumen (Rumenfibe ®). Dibimbing oleh ANURAGA JAYANEGARA, DEWI APRI ASTUTI dan JUNICHI TAKAHASHI.
Pakan konsentrat sangat berperan dalam keberhasilan industri peternakan sapi pedaging. Penggunaannya bisa mencapai 90% bahkan lebih dari total konsumsi ransum sapi pedaging. Walaupun pakan konsentrat mampu meningkatkan performa sapi pedaging dalam pemeliharan jangka pendek, dalam aplikasinya justru berpotensi menyebabkan “metabolic-disorder” seperti asidosis. Asidosis terjadi bila produksi VFA dan asam laktat lebih tinggi dari kapasitas absorpsinya dan kemampuannya menuju jalur gastro-instentinal. Stimulan Mekanis Rumen (RMS) dengan administrasi sikat Rumenfibe® merupakan suatu teknologi yang mampu menurunkan resiko tersebut seperti asidosis walaupun hasilnya masih bervariasi. Tujuan penelitian ini adalah untuk mengetahui pengaruh RMS (“RUMENFIBE®” atau RF) terhadap performa, profil fermentasi rumen and perkiraan nilai ekonomis pemanfaatan RMS terhadap sapi pedaging feedlot.
Percobaan telah dilakukan di Indonesia terhadap dua puluh ekor sapi pejantan muda feedlot ( bobot badan 267+4 kg) yang diacak ke dalam empat kandang. Sapi perlakuan diberi proporsi konsentrat tinggi, sesuai dengan manajemen perusahaan PT. Mitra Catur Taruma. Sepuluh ekor sapi di administrasi tiga buah RMS Rumenfibe® ke dalam rumen menjadi perlakuan dan dikandangkan dalam dua buah pen sedangkan sepuluh ekor lainnya tidak diadministrasi sebagai kontrol. Parameter performa seperti rataan konsumsi nutrien, pertambahan bobot badan dan income over feed cost (IOFC), serta profil rumen seperti pH, potensi redoks (∆EH), kandungan ammonia, kandungan asam laktat, total VFA dan VFA parsial (asetat, propionat dan butirat), produksi methan dan potensi penyembuhan hidrogen dalam cairan rumen kemudian diamati.
Administrasi RMS dengan sikat Rumenfibe memberikan pengaruh yang sama terhadap perlakuan (RF) dan kontrol seperti pertambahan bobot badan dan rataan konsumsi nutrien, namun perlakuan RF memiliki nilai IOFC yang lebih rendah (P< 0.05). Sapi yang diadministrasi dan tidak diadministrasi RMS dengan sikat Rumenfibe juga memberikan pengaruh yang sama terhadap pH, potensi redoks, kandungan ammonia, kandungan asam laktat, dan asam propionat. Sedangkan asam asetat, butirat, prediksi methan, prediksi produksi dan pemanfaatan hidrogen dalam cairan rumen (P<0,05) pada perlakuan RF lebih baik daripada kontrol. Sapi yang diadministrasi sikat RMS (Rumenfibe®) tidak memberikan dampak yang buruk bagi performa dan fermentasi rumen, tetapi justru mampu meningkatkan fermentasi rumen.
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Copyright © 2015, Bogor Agricultural University.
All rights reserved.
No part or all of this work may be excerpted without inclusion or mentioning the sources. Exception only for research and education use, writing for scientific papers, reporting, critical writing or reviewing of a problem; and this exception does not inflict a financial loss in the proper interest of Bogor Agricultural University.
No part or this entire work may be transmitted and reproduced in any form without written permission from Bogor Agricultural University.
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RUMEN FERMENTATION PROFILE OF FEEDLOT STEER
ADMINISTERED WITH RUMEN MECHANICAL
STIMULATOR BRUSH (
Rumenfibe
®)
YULIANRI RIZKI YANZA
A Thesis submitted in partial fulfillment of the degree Master of Science
In
Nutrition and Feed Science
GRADUATE SCHOOL
BOGOR AGRICULTURAL UNIVERSITY
BOGOR
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PREFACE
There is no words except the grateful to Allah SWT, The One, The Creator, The Almighty and The Mercifull to His servants, and our nobleness prophet Muhammad SAW who has tought us the way of life to the Jannah, a place that He promised. I would like to express my deepest gratitude and thanks to the any lecturer of IPB who never gave up and led us for being a well educated men. This work could not be made possible without the supervision, support and constant guidance of my supervisors Prof. Dr. Dewi Apri Astuti and Dr. Anuraga Jayanegara who inspiring me in any moment. Thanks also to our Sensei, Prof. Junichi Takahashi, PhD, who guide this research without a doubt. Thanks to Meiwa Co. Ltd. that already supplied the RMS and funded this research and PT. Mitra Catur Taruma Feedlot that gave the opportunity in used their place and cattles since the beginning of this study. I am highly grateful to them for this kindness.
This master study would not succeeded without the firm support and prayers of my beloved parents Zakir Has and Nurbayani, and my beloved brothers Farhan, and Ises. Thanks a lot of the support from huge family of H. Ramli; Nurzakiah, Wiwit, Inel, Cecep, Desi, Sian, Neneng, and special for Asmadi that always thought me wise thinkings. I am also grateful to my classmates and others such, Ali, Ridho, Dila, Rossy, Tekad, Andre, Putri (Alm.), Nur, Mega, Teguh Wahono, Kaleem Saleem and to all of them whose names are not mentioned here. A special thanks to Devide Maric Hersade who always accompany me in any moment since studying in IPB. I am very grateful to Gita Swara Pascasarjana IPB and those members such Dr. James Unitly, F. Ochieng (Alm.), Silvia, Weni, Donal, Lista, Merry, Janet, Dedi, Desi, Iswahyudi, Ella, and many others that gave me spirit in a voice.
I extremely indebted to HMI and KAHMI Cabang Bogor, Forum Wacana IPB, Kedai IPOK, Sayogyo Institute ca. SAINS (ie. Eko Cahyono and especially to Dr. HC. Gunawan Wiradi, that tought a lot about philosophy, history, sciences and reality), The Nastari Consultant (such Said Abdullah, Wahono, M. Nur Amin, and Kaboul). Thanks to P4W IPB such Dr. Hadi, Prof. Iman Supriyatna, Dr. Iwan Prihantoro, Mr. Agustinus and Dani by took me in any agendas. Thanks to Dendi, Mr Abimanyu, Mr. Yusuf and and many others below the PT. Mitra Catur Taruma. Thanks to excentric lecturers, Prof. Komang G. Wiryawan and Prof. Asep Syaifudin. Thanks to all civities and alumnous of SMAN PLUS Provinsi Riau. Thanks to KEDAI IPOK members such Iham the philosopher, Dito, Riza, Chenko, Imam, Mega, Heru, Mubarok, Arfi, Ulfah, Afifah, Iyuth and many others who also helped me and brought any benefecial discussion in social issues, philosophy and nationalism. Special thanks to my dearest, Aulia Nur Rahma, who always struggle and put her faith on me and in any unforgatten moment.
Finally, I am extremely thankful to IPB itself, I have experienced living in one of several historical college, with wonderful and friendly people. Last but not the least by borrowing a sentence from my friend Kalim Saleem from Pakistan, Indonesia is no doubt a piece of paradise on earth.
Bogor, October 2015
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TABLE OF CONTENTS
SUMMARY... ix
RINGKASAN... x
FOREWORDS... xx
TABLE OF CONTENT... xi
TABLE OF FIGURES... xii
TABLE OF APPENDICES... xiii
1. INTRODUCTION... 1.
Background... 1.
Research Objectives... 2.
Research Hypotheses... 2.
2. METHODOLOGY... 3.
Experimental Design... 3.
In vivo study... 3.
Rumen Fermentation analysis... 5.
a. pH and ORP (∆EH) ... 5.
b. Ammonia... 6.
c. Lactic acid content... 6.
d. Partial VFA... 6.
e. Methane and Hydrogen prediction... 7.
Statistical Analyses... 7.
3. RESULTS AND DISCUSSION... 8.
Nutrient Intake and Feedlot Steer Performance... 8.
RMS Influence Into Rumen Fermentation Profile Of Feedlot Cattle... 9.
4. CUNCLUSIONS... 15.
5. LIST OF REFERENCES... 16.
6. APPENDICES... 18.
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LIST OF FIGURES
1. Figure 1. Rumen Mechanical Stimulator by Smith study (1969; left) and Rumenfibe® by Meiwa Co. Ltd. (present; right)... 4.
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LIST OF TABLES
1. Table 1. Nutrient Composition of Feeds Source by Proximate
Analysis... 3. 2. Table 2. Nutrient Composition of Formulated Rations (100g-1)...
4. 3. Table 3. Total Nutrient Intake of Feedlot Steer Administered With
(RF) and Without Rumen Mechanical Stimulator (Control) in short-time rearing...
8. 4. Table 4. Performance, gain:feed ratio and income over feed cost
(IOFC) of feedlot steers with (RF) and without rumen mechanical stimulator administration (control) in short-time rearing...
8. 5. Table 5. Rumen Fermentation Profile of Feedlot Steer Administered
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LIST OF APPENDICES
1. 1 st Appendix. Mean Difference of Dry Matter Intake in 95% Confidents
Level (P<0.05)... 18. 2. 2 nd Appendix. Mean Difference of Crude Fiber Intake in 95%
Confidents Level (P<0.05)... 18. 3. 3 rd Appendix. Mean Difference of Crude Protein Intake in 95%
Confidents Level (P<0.05)... 18. 4. 4 th Appendix. Mean Difference of Ether Extract in 95% Confidents
Level (P<0.05)... 18. 5. 5 th Appendix. Mean Difference of NFE in 95% Confidents Level
(P<0.05) ... 18. 6. 6 th Appendix. Mean Difference of TDN Intake in 95% Confidents Level
(P<0.05) ... 19. 7. 7 th Appendix. Mean Difference of Concentrate : Forage Ratio Intake in
95% Confidents Level (P<0.05)... 19. 8. 8 th Appendix. Mean Difference of GE Intake in 95% Confidents Level
(P<0.05)... 19. 9. 9 th Appendix. Mean Difference of Total Gain in 95% Confidents Level
(P<0.05)... 19. 10. 10 th Appendix. Mean Difference of Average Daily Gain (ADG) in 95%
Confidents Level (P<0.05)... 19. 11. 11 th Appendix. Mean Difference of Gain : Feed Ratio in 95% Confidents
Level (P<0.05)... 20. 12. 12 th Appendix. Mean Difference of IOFC in 95% Confidents Level
(P<0.05)... 20. 13. 13 th Appendix. Mean Difference of pH in 95% Confidents Level
(P<0.05)... 20. 14. 14 th Appendix. Mean Difference of ∆EH in 95% Confidents Level
(P<0.05)... 20. 15. 15 th Appendix. Mean Difference of Ammonia in 95% Confidents Level
(P<0.05)... 20. 16. 16 th Appendix. Mean Difference of Lactic acid in 95% Confidents Level
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17. 17 th Appendix. Mean Difference of Acetate in 95% Confidents Level
(P<0.05)... 21. 18. 18 th Appendix. Mean Difference of Propionate in 95% Confidents Level
(P<0.05)... 21. 19. 19 th Appendix. Mean Difference of Butyrate in 95% Confidents Level
(P<0.05)... 21. 20. 20 th Appendix. Mean Difference of total VFA in 95% Confidents Level
(P<0.05)... 21. 21. 21 th Appendix. Mean Difference of Acetate : Propionate Ratio in 95%
Confidents Level (P<0.05)... 22. 22. 22 th Appendix. Mean Difference of Methane in 95% Confidents Level
(P<0.05)... 22. 23. 23 th Appendix. Mean Difference of Methane/VFA in 95% Confidents
Level (P<0.05)... 22. 24. 24 th Appendix. Mean Difference of H2 prodiction in 95% Confidents
Level (P<0.05)... 22. 25. 25 th Appendix. Mean Difference of H2 utilised in 95% Confidents
Level (P<0.05)... 22. 26. 26 th Appendix. Mean Difference of Acetate Proportion in 95%
Confidents Level (P<0.05)... 23. 27. 27 th Appendix. Mean Difference of Propionate Proportion in 95%
Confidents Level (P<0.05)... 23. 28. 28 th Appendix. Mean Difference of Butyrate Proportion in 95%
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1
1. INTRODUCTION
Background
Concentrate feeding has been a primary practice in ruminant livestock industries in several decades. In contrary, it has been suggested that a certain amount of roughage in rations is important to maintain the rumen functions (van Houtert, 1983). This condition was influenced by high population of cattle that need intensive feeding, but roughage availability is limited. This limitation believed as the influence of wet and dry season. Althought may lowering cost production, roughage also considered less effective to improve beef cattle performance than the concentrate, certainly for rearing in short time period (Orskorv, 2001). Moreover, tropical forages in Indonesia has low quality of nutrient was believed did not mutual to feedlot cattle that dominated by Australian breed, i.e. Brahman Cross. These factors was drove the feedlot industry to intensified the concentrate feeding which based on grains and cropwaste by-product.
High concentrate diet in feedlot industries improved performance in short-term period. Meanwhile, ruminant actually need appropiate proportion of fibrous in order to mantain the rumen motility and to stimulate microbial activity (Chiba, 2014). Practically, concentrate is fed approximately 85% of total feed consumed (Ngadiyono, 1995) or even more. Gonzalez et al., (2012) states that grain and forage characteristics, and feed additives affect feeding behavior such feed intake, chewing behaviors, which has a great influence on ruminal fluid acid–base balance. However, high concentrate diet with low roughage may decrease rumen motility and may further cause a metabolic disorder such as acidosis.
Digestion process in ruminant involves the rumen and its associated microbes. Through the actions of rumen microbes, fibers (cellulose and hemi-cellulose) or other carbohydrates (such as starch and sugar) are fermented into volatile fatty acids (VFA), methane (CH4) and ammonia (NH3). The cattle which fed predominantly roughage diets high in cellulose intermediated in soluble sugars with lower starch, cellulose then hydrolysed primarily to acetate and methane via anaerobic glycolysis, where methane is the electron sink product (Hungate 1966). Propionate amounts produced on cellulose predominant diets are 15% to 20% of the total VFA pool. When cattle fed high concentrate that predominantly with starch diets, amylolytic bacteria fermenting this substrate produce a higher proportion of propionate (35% to 45% of total VFA pool). In comparison to propionate and acetate, butyrate is quantitatively the least important of the three major VFAs in the rumen fluid of animals fed roughages (van Houtert 1983). Most of VFA then absorbed through the rumen wall and further utilized as energy source. High starch diet will need the increased of buffer concentration (bycarbonates; naturally from from saliva) to maintain rumen status from acidity condition. It is according to starch fermentation that also produce a higher lactic acid and led to acidic rumen condition (RAGFAR, 2007). Such feeding practice (i.e. high concentrte diet) is characterized with a lower rumen pH, increased methane and lactic acid concentration, decreased microbes population, decreased nutrient absorption, and acidosis may occur (Cerrilla & Martinez, 2003; RAGFAR, 2007). Therefore, it is necessary to maintain good rumen status.
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2
Scientists have tried to anticipate this problem, for instance, by administering brushes called Rumen Mechanical Stimulator (RMS) into the rumen as an artificial fiber (Horiguchi & Takahashi 2000). This method has been developed since 1960s with similar circumstances where concentrate is a primary in feedlot cattle diet (Smith, 1969). Recently, an RMS i.e. Rumenfibe® (RF, Meiwa- Sangyo, Kyoto, Japan) has been developed in purpose to stimulate the rumen mucosa and ecology of beef cattle mechanically. It is considered that this artificial fiber is indigestible and functioning as scrubber by stimulating the rumen wall) and may promote microbial activities in the rumen (Horiguchi & Takahashi 2000). Despite its potentiality, RMS has not been previously introduced in beef cattle production in Indonesia.
Research Objectives
The aims of the present study were: (1) to determine the effect of RMS (“RUMENFIBE®” or RF) on performance of feedlot steers, (2) to investigate the effect of RMS on rumen fermentation profile of feedlot steers, and (3) to assess the economic feasibility of using RMS on feedlot steers.
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3
2. METHODOLOGY
Experimental Design
This research had been conducted since 5 October 2014 to 5 March 2015. In vivo study was held at PT. Mitra Catur Taruma feedlot company within three months of rearing (31 days a month), according to consortium term of reference between Meiwa Co. Ltd. and PT. Mitra Catur Taruma. This study was then continued with rumen fermentation analysis in which rumen liqour of each steer was stored and collected after being slaughtered, then sample was analyzed for several parameters. In vivo study
In vivo experiment was performed at the PT. Mitra Catur Taruma Feedlot, located at Cariu-Jonggol, Bogor-West Java. Experiment was done at the end of dry seasons, i.e. temperature about 27-340C and humidity about 49-82% where forage supply was limited. Twenty Australian steers (ca. 267 kg steer-1) were randomly allocated in four pens in which five steers were installed in each pen. All cattle were fed as usual feed which routinely fed into steers in the feedlot. Feed proportion consisted of formulated concentrates and roughages such green cut king grass, rice straw and corn cob waste on various dry matter basis, following PT Mitra Catur Taruma feeding management (Table 1).
Table 1. Nutrient Composition of Feeds Used in the Experiment Nutrient
Composition (%)
Concentrate Roughage
GR FN KG CC RS
---%---
Dry Matter 88.51 86.19 13.31 19.30 49.47
Organic Matter 85.29 90.45 88.38 92.52 89.49
Crude Fiber 18.18 16.79 20.11 27.47 27.43
Crude Protein 13.63 14.43 14.57 6.43 6.28
Ether Extract 4.15 4.71 2.54 1.41 2.13
Ash 14.71 9.55 11.62 7.48 10.51
NFE 49.33 54.52 51.16 57.21 53.65
TDN* 63.68 69.91 62.21 51.58 52.30
GEI (Kal kg-1 DM) 23.02 25.27 22.49 18.65 18.91
Notes: GR= Growing Feed; FN=Finishing feed; KG=King grass; CC= corn cob waste ; RS: Rice Straw; NFE=Nitrogen free extract ;TDN=Total Digestible Nutrient; GEI (Gross Energy Intake); * = National Research Council (1984).
Ten experimental steers in two pens were orally administered with three RMS brush (Rumenfibe; Figure. 1) per steer into the rumen through mouth at cattle crush dock. Furthermore, steers fed by dinamics composition of ration (Table 2) twice per day at 07.00 a.m. and 03.00 p.m. where forages were given about 30-60 minutes after the concentrates which offered concentrates:forages with 90:10 ratio. After three months of rearing, cattle performance such as final body weight and income over feed cost were evaluated then continued to slaughter at the Bubulak Slaughter House, Bogor, where the rumen liquor was sampled for further analyses.
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Figure 1. Rumen Mechanical Stimulator by Smith study (1969; left) and Rumenfibe® by Meiwa Co. Ltd. (present; right).
Table 2. Nutrient Composition of Formulated Rations Nutrient
Composition (%) GW1 GW2 GW3 FN1 FN2 FN3
Dry Matter 81.60 81.85 87.61 79.35 79.60 85.36
Organic Matter 83.76 83.27 90.26 88.32 87.84 94.83
Crude Fiber 17.94 18.21 20.41 16.65 16.92 19.11
Crude Protein 13.42 12.81 13.30 14.12 13.52 14.01
Ether Extract 3.96 3.87 4.08 4.46 4.37 4.58
TDN* 62.28 61.13 65.36 67.85 66.70 70.93
GEI (Kal kg-1 DM) 22.52 22.10 23.63 24.53 24.12 25.65
Notes: GW1= Grower feed + King Grass; GW2= Grower feed + Green cut corn; GW3= Grower feed + Rice Straw; FN1=Finishing feed + King Grass; FN2= Finishing feed + Green cut corn; FN3= Finishing feed + Rice Straw; NFE=Nitrogen free extract ;TDN=Total Digestible Nutrient; GEI (Gross Energy Intake); * = National Research Council (1984).
Nutrient intake was analyzed based on dry matter (DM) in feed rations. Steer was weighed each month (31 days a month) with weighing machine since the beginning of this study. The performance parameters such as nutrient intake, final body weight and average daily gain of each steer were then calculated. Feed conversion ratio (FCR) and income over feed cost (IOFC) were also calculated.
a. Nutrient intake evaluation
Nutrient intake was quantified by difference between offered feed and feed residual in each pen. The model to quatified dry matter intake was :
Nutrient Intake (kg head-1day-1) = (Σ intake x % nutrient in ration)
93 days
The nutrientintake was calculated since the beginning of this research after RF installment. Nutrient intake such as dry matter, organic matter, crude fiber, crude protein, ether extract, nitrogen free extract, total digestable nutrient (TDN) and gross energy intake (GEI; Kal kg-1day-1) were also calculated with the respective model.
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5 b. Performance evaluation
Performance of feedlot steers that quantified in this study was total gain and average daily gain (ADG). The model used to quatify the total gain:
Total Gain (kg head-1) = Final weight (kg head-1) – Initial weight (kg head-1) Meanwhile, for the average daily gain was according to that model divided by total days of rearing (93 days).
c. Gain : Feed evaluation
The Gain : Feed evaluation in this study was calculated by total DM intake (kg head-1day-1) per total gain (kg head-1). The model used to quatify feed efficiency was :
Feed efficiency = total DM intake (kg head-1)
Total Gain (kg head-1)
d. Income over feed cost (IOFC)
Income over feed cost was calculated after steers were sold to describe the bruto income by the difference on total price of bodyweight (BW) gain and total feed cost in Rupiah. The model used to quantify feed efficiency was :
IOFC (Rp head-1) = Total price of BW gain – Total feed cost
Rumen Fermentation Study
Each item of the rumen fermentation profile was measured in order to determine the feedlot steer rumen fermentation whether good or poor with and without administered the RMS brush fed high concentrate diet. Rumen fermentation profile of experimented steer was measured by its biochemical content analysis such pH,
oxidation reduction potential (∆EH), ammonia concentration, lactic acid concentration,
volatile fatty acid concentration (totally and partially), methane production and hydrogen recovered potencies of each steer.
Rumen liquor of each steer was storaged into thermos bottles (2 bottles head-1; 2 litres bottle-1) by squeezed the rumens content with sterylized hand. The rumen liquor then filtered and separated into small bottles at Feed Quality Testing Laboratory, IPB’s Faculty of Animal Science as soon as possible after storaged to partially analyzed in several laboratory with difference sample bottle. Feed resources were analyzed by proximate analysis at IPBs General Laboratory (PAU). Rumen fermentation profile analysis was measured partially into several indicators, below:
a. pH and ORP (∆EH)
The pH and reduction and oxidation potential/ redox potential (∆EH) were analyzed with digital pH and ORP kit analysis used electrodes and callibration test solutions at Land Chemistry and Fertility Laboratory, Faculty of Agriculture of IPB. pH status was measured as soon as possible after storaged. The portable electrodes then pulled in a small bottle. Normal pH indicates around + 7,00 and normal ∆EH of the rumen measures about -330 to -220 mV (Wang et al., 2012).
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6
b. Ammonia
Ammonia (NH3) was analyzed with conway micro-diffusion method (General Laboratory Procedures, 1996) used the conway cup. Conway cup was smeared with vaseline on both lips. one ml supernatant of rumen liquor then dropped in the left side of Conway cup and saturated with an indicator of Na2C03 solution at another side of chamber. Before mixed, one ml of boric acid indicator was dropped into the middle of the conway cup. The conway cup then sealed in order to prevent oxidation. Furthermore, the Na2CO3 then saturated with the supernatant and mixed by slow-shaking the Conway cup. Sample then left at normal room temperature (+25oC) until boric acid were showed a blue color, then opened. In the next 24 hours, cup were opened then titrated into the midle of chamber with 0.06 N HCI until showed a pink color (as same as boric acid indicator). NH3 concentration (mM) then calculated by the formula:
Amount of NH 3( mM) = ∆H2SO4 titrated (ml) x N H2SO4 (mM) x 1000
c. Lactic acid concentration
Lactic acid concentration in the rumen liquor was determined by Baker and
Summerson’s method (1941) with spectrophotometer analyzer. Rumen liquor sample plus 10% TCA were filtered and putted in the centrifuge tube in order to get 10 ml filtrate. Then, sample was centrifuged at 2750 rpm speed for 15 minutes. The supernatant was poured and aligned round and round with one ml TCA 10%. About 20% CuSO4 solution was added into the tube and was diluted with 10 ml distilled water. Then, 2 grams Ca(OH) powder was added. All tubes were closed with Parafilm (in practical case was closed with marbles) then homogenized for more than 30 minutes, then centrifuged again. The supernatant was taken and pulled into a tube test, then were dried and cleaned (φ 18-23 mm). Each tube was diluted and added with 0.05 ml of CuSO4 4%, then was added with six ml of H2SO4 burette. Tube was heated about 5 minutes (> 80°C) then refrigerated in a few minutes (<200C). At the next step, 0.1 ml of reagent p-hydroxyphenyle was added drop by drop until shown the white color. Tube then homogenized with mixer before pulled into the water bath (300C) for more than 30 minutes then reinserted into boiling water for 90 seconds before cooled the tube. While liquid in tube turned into a violet color, the tube was moved into the cuvet in order determined the wavelength each liquids with OD spectrophotometer valued at 20 amplitude and 560 nm of wavelength (Good reading transmition around 20-80 amplitude). After wavelength was measured in spectrophotometer machine, lactic acid content was determined with the formula:
Lactic acid content (mM) = 0,006347 + 15,266 OD Molecular weight (g) d. Partial VFA
Partial VFA was analyzed at the Nutritional Laboratory of Technology Agriculture Gadjah Mada University with gas chromatography machine (GC 8-Schmidzu: 10%GP-SP-1200/1% H3PO4 on Chromosorb WAW 80/100). One ml of each sample standard VFA liquid indicator and one ml of rumen liquid were inserted
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7 into an Eppendorf tube before added with 0.003 g of 5-dihydrate sulfo salicylic acid. The mixed liquor was shaked then centrifuged for 10 minutes at 12000 rpm and 70C temperature, then sample was injected into the gas chromatograph sample (GC). The GC Machine specification used 2 kg cm2 -1 gas flow pressure of N2 with 2 metr column range and 150oC column temperature (3mm φ).
The separation system was based on the separation properties of the partition, absorption of two stationary phase (column) and the mobile phase (gas). The big difference in the partition or absorption in the second phase was led to the peak on the monitor screen. By reading the chromatogram VFA standard references, VFA concentration of each sample was measured by formula:
VFA (mM) = VFA sample area x standard VFA content x 1000 Standard VFA Area X VFA molecular weight e. Methane and Hydrogen prediction
Methane emission was measured by estimating stoichiometrically based on VFA concentration profile that adjusted by Moss et al. (2000) equation:
CH4(mM) = 0.45 C2 – 0.275 C3 + 0.40 C4
Hydrogen utilised and hydrogen produced were calculated follows : H2produced (mM) = 2 C2 + C3 + 4 C4
H2utilised (mM) = 4 CH4 + 2 C3 + 2 C4
Since the amount of methane emission is influenced by hydrogen recovery (Jayanegara
et al., 2013), adjustment was made by considering a hydrogen recovery of 90% (Jayanegara et al., 2015).
Statistical Analyses
Data were analyzed with SPSS software version 16.0. Comparison between control group (without RF administration) and treatment group (with RF administration) was performed by t-test procedure.
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8
3. RESULTS AND DISCUSSION
1. Nutrient Intake and Feedlot Steer Performance
The RF and control treatments gave similar average nutrient intake during the experimental period (Table 3). Daily intake of each steer was approximately 9.5 kg day-1 at dry matter (DM) basis. The proportion of concentrate and forage intake was 91.5% and 8.5%, respectively. Similar pattern to that of DM intake, crude fiber intake, crude protein intake, ether extract intake, nitrogen free extract intake, total digestable nutrients (TDN) intake and gross energy intake (GEI) also gave similar results on both treatments.
Table 3. Total nutrient intake of feedlot steer administered with (RF) and without rumen mechanical stimulator (Control) in short-time rearing.
Item Treatment SEM P-value
Control RF
Average Intake of Nutrient
Dry Matter (kg h-1 d-1) 9.589 + 0.007 9.590 + 0.025 0.002 0.876
Crude Fiber (kg h-1 d-1) 1.760 + 0.002 1.759 + 0.005 0.000 0.852
Crude Protein (kg h-1 d-1) 1.292 + 0.001 1.293 + 0.003 0.000 0.572
Ether Extract (kg h-1 d-1) 0.397 + 0.001 0.397 + 0.001 0.000 0.423
NFE (kg h-1 d-1) 4.904 + 0.004 4.905 + 0.014 0.001 0.902
TDN£ (kg h-1 d-1) 6.208 + 0.005 6.210 + 0.016 0.001 0.729
GEI (MJ h-1 d-1) 16.472 + 0.013 16.475 + 0.045 0.003 0.811
Concentrate : Roughages (%) 91.5 : 8.5 91.5 : 8.5
Notes: NFE=Nitrogen free extract ;TDN=Total Digestible Nutrient. £ = National Research Council,
1984; GEI=Gross Energy Intake; Different upper case letters within rows indicate differences
among treatments (P≤0.05).
Nutrient intake is influenced by the chemical composition within the concentrate and forage. This result indicated that RF treatment was unable to increase feed intake of steers when reared in short-time period. Previous studies also reported that RMS administration resulted in a similar nutrient intake compared to control such as in Thai native steers (Bos indicus) (Angthong et al., 2011), lactating Holstein (Matsumoto et al., 2011) and Holstein steers (Horiguchi & Takahashi, 2004).
Table 4. Performance, gain:feed ratio and income over feed cost (IOFC) of feedlot steers with (RF) and without rumen mechanical stimulator administration (control) in short-time rearing.
Item Treatment SEM P-value
Control RF
Performance
Total Gain (kg h-1 d-1) 184.30 + 26.4 171.00 + 27.7 2.721 0.286
ADG (kg h-1 d-1) 1.98 + 0.28 1.83 + 0.29 0.029 0.287
Gain : Feed ratio 0.21 + 0.03 0.19 + 0.03 0.003 0.288 IOFC (Rp h-1) 4,248,709 + 950,414b 2,768,625 + 998,981a 121,535 0.003
Notes: ADG=Average Daily Gain; IOFC = Income Over Feed Cost; Different upper case letters within
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9 The RF and control treatment showed similar performance status on total body weight, gained about 171-184.3 kg. Steers were boosted about 1.84 - 1.98 kg day-1 (Table 4). Lallman (1990) adjusted that the yearling beef cattle requires about 8-11 kg day-1 of DM intake which contained 60-70% TDN, to gain at least 1.8 kg day-1 of steer cattle body weigh. Meanwhile, Addah et al. (2014) studied that feedlot steer fed with inoculation and chop-length of whole -crop barley silage about 11 kg day-1 DM capable to gaining the steer weigh up to 1.9-2.0 kg day-1. Veracini et al. (2012) also reported that Angus beef steer fed around 8.7-10.1 kg day-1 DM and gained the body weight about 1.6 kg day-1 in early 84 days finishing phase fed with shelled corn and soybean meal that was replaced with distillers grain. Ngadiyono (1995) achieved a lower result on BX cattle, about 0.8 to 1.2 kg day-1. Compared with Lallman (1990) and Addah et
al. (2014), the present study resulted appropriate performance. In comparison to Ngadiyono (1995) and Veracini et al. (2012), the average bodyweight gain in this study was obtained better.
The results of gain : feed ratio also gave similar value on both treatment, about 0.192-0.207. This result represent that each kg of feed intake was capabled to boosting 0.19-0.20 kg of body weigh (Table 4). High concentrate in ration might gained the steer performance effectively, but the RMS administration did not improve body weight gain better than the Control. Angthong et al. (2011) also resulted similar performance by studied Thai native steers with and without RMS administration.
Administering the RMS brush into the feedlot steer through mouth at the weighing dock gave an earlier stress that might dropped steers body weight in several days. After the stressed period, the compensatory feeding expectedly could boost steer weight gain but did not gave significant improve than the control. It was believed the RMS administration was unsuitable to improve feedlot steer bodyweigh in short-time period of rearing till the slaughtering limits. Similar performance on both groups also believed as the influence of nutrient intake such crude fibre and crude protein. Althought believed to promote utilised the energy better, the RF group not showed a better result than the control according to fibrous and protein intake on both treatments that achieved above the minimum expectation of steer requirement, 12% and 7% of DM respectively. (Lazarini et al., 2009) states that the microbial growth rate was improved at CP levels better than 7% and improve digestibility of low-quality forage. However, the RMS brush administration in this study resulted a similar bodyweight gain and gave the unpoor performance of feedlot steer.
With regard to the economy beneficial measured as income over feed cost (IOFC), RMS administration was more costly than control (P<0.05). The RF treatment in each steer had lower IOFC (Rp 1,470,000) than the control. This lower IOFC was due to the cost to purchase RMS, i.e. as high as Rp.1,000,000.-, for each cattle.
2. RMS Influence Into Rumen Fermentation Profile Of Feedlot Cattle
The pH status of RF and Control treatment were showed a similar value, about 6.045. The pH status in this study obviously still at a tollerance value, althought below than the minimum standar of minor acidic status catagoryzed. RAGFAR (2007) confirmed that minor acidic status of cattle if pH status of the rumen liquor below than
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10
6.2 and could reduced the fiber digestion, especially while cattle fed high concentrate diet. Chiba (2014) also affirmed the normal pH status of the rumen liquor normally marked about 6 to 7, but the rumen ecology could depressed below than 5.5 while cattle fed with mostly grains ration, then brought to acid intolerance status that promotes protozoa population. Horiguchi & Takahashi (2004) also found with and without RMS administration through brush administration gave a normal pH status even below 6 at 3 hours after feeding fed with high concentrates diet to Holstein steer, then raised up to 7 after 18 hours. These results explained that the RMS brush administration in this present study was no decreased the ruminal pH of feedlot steer fed high concentrate diet.
Table 5. Rumen fermentation profile of feedlot steer administered with (RF) and without rumen mechanical stimulator (Control).
Item Treatment SEM P-value
Control RF
Rumen Profile
pH 6.045 + 0.49 6.045 + 0.44a 0.048 1.000 ∆ EH (mV) -179.760 + 112.52 -132.844 + 74.66a 10.199 0.296 Ammonia (mM) 14.472 + 5.85 12.420 + 3.55a 0.514 0.365 Lactic acid (mM) 6.512 + 1.64 6.352 + 0.96a 0.140 0.796 Total VFA (mM) 117.906 + 12.08a 142.842 + 27.08b 2.780 0.039
Partial VFA
Acetate (mM) 73.247 + 8.96a 86.100 + 15.05b 1.592 0.046
Propionate (mM) 25.582 + 4.17 32.452 + 11.52a 1.047 0.146 Butyrate (mM) 19.078 + 3.94a 24.290 + 4.18b 0.559 0.019
VFA Proportion
Acetate % 62.07 + 3.32 60.56 + 3.99a 0.078 0.410 Propionate % 21.65 + 2.32 22.22 + 5.27a 0.693 0.770 Butyrate % 16.29 + 3.62 17.21 + 2.55a 0.003 0.556 Acetate : Propionate Ratio 2.90 + 0.37 2.912 + 0.92a 0.004 0.969 Methane (mM) 33.56 + 3.61a 39.54 + 6.54b 0.005 0.032
Methane / VFA % 28.49 + 1.61 28.03 + 3.735a 0.004 0.738 H2 Production (mM) 248.39 + 24.12a 301.81 + 51.69b 5.537 0.014
H2 Utilised (mM) 223.55 + 21.71a 271.63 + 46.52b 4.984 0.014
Notes: Different upper case letters within rows indicate differences among treatments (P≤0.05). During fermentation, a reducing equivalent ions is produced, which is accompanied by the release and transfer of protons and electrons (Wang et al., 2012). This process presenting the effectiveness of fermentation and microbial activity that measured by redox potential (∆EH; Julien et al., 2009). In this study, the redox potential of RF and Control treatment were measured -132,84 and -179.76 mV in the rumen liquor, respectively. This result showed a similar value between RF and Control treatment, and also were not found the outlier data that might influenced the results although had width deviation on both treatments. The redox potential of RF and Control treatments resulted below than usually found in the rumen liquor, normally about 300 mV (Moss et al., 2000), or accurately about -323mV by in vitro and about -213 mV by in vivo study (Julien et al., 2009). Wang et al. (2012) also found that normal potential redox status in the rumen liquor before electrocuted about -300 to -200 mV. Both
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11 treatments might in a tolerance rate by ∆EH condition compared with those standard even showed below than the usual. This low rate was notified as the effect of open air
analysis while putted in the ∆EH electrodes then rumen liquor was oxidized that might decreased the anaerob fermentation status.
Ammonia was a ruminal fermentation product that absorbed in cattle rumen as proteolytic microbe acts where amino acid deaminated (Chiba, 2014). The concentration of ammonia was found in a similar value, resulted about 12.42-14.472 mM in the rumen liquor. This results was approximate with the usually found that reported by Patra (2013) on meta-analysis study, quantified at 12.9 mM in the rumen liquor of dairy and steer cattle. But Horiguchi & Takahashi (2003) found a lower result than the present study i.e. about 5.3-7.1 mM. Meanwhile in contrary, Matsuyama et al.
(2002) assured that the cattle administered with RMS had a lower ammonia than the control (10 mmol dL-1 vs 14 mmol dL-1) although its nitrogen retention was improved. The ammonia concentration in the present study resulted in a similar with previous studies althought there were resulted various. This result may assure that the RMS administration did not influence proteolytic microbes activity in produced ammonia.
There were found difference results according to total and partial VFA concentration (acetate and butyrate) between RF and Control treatment. The RF treatment resulted a higher VFA than the control, i.e 142.842 mM vs 117.906 mM, influnced by the partial VFA accumulation (p<0.05). In partially, the acetate and butyrate concetration in RF group were higher than the control treatment, i.e. 86.1 vs 73.247 mM and 24.29 vs 19.1 mM in the rumen liquor, respectively (p<0.05). Instead from the acetate and butyrate, The RF treatment unable to increase the propionate concentration, resulted 24.3-32.4 mM as similar as the control. But the acetate, propionate and butyrate of the RF and control treatment had a similar proportion in percentages. Therefore, the RF and control group achieved a similar quantities on Acetic:Propionic ratio, pointed out about 2.89-2.91.
The increased of total VFA status in RF treatment was resulted by acetate and butyrate concentration (P<0.05), even the propionate concentration tended to be higher than the control. This present study achieved better result compared with Horiguchi and Takahashi (2004) study between with and without RMS administration to Holstein steer that gave a similar quantitiy of acetate, propionate and butyrate concentration, resulted about 40 mM; 24 mM; and 26 mM in the urmen liquor. Matsuyama et al. (2002;2004) found the propionic and butyric concentration of RMS treatment increased than the control i.e. 34.3 and 20 mmol dL-1 vs 27.1 and 16.6 mmol dL-1, respectively. But by the acetate and total VFA in RF treatment had lowered than the control by 24 hours 50 vs 58 mmol dL-1 and 100 vs 125 mM while Holstein steers fed mainly rolled barley.
In the contrary, Horiguchi & Takahashi (2002) found that the RMS treatment had a lower total VFA propotion than the control ( 61.8 vs 87.9 mmol dL-1 after 4 hours feeding and 67.3 vs 78.3 mmol dL-1 after 24 hours feeding), but produced higher total VFA proportion than the Control 89.3 vs45 mM) while fed long rice straw and high concentrate diet to Holstein steers. These previous studies were generally unidentical according to present result by their total and partial VFA concentration, such acetate,
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12
propionate and butyrate concentration. Instead, the proportion of partial VFA in the present study with and without RMS administration resulted in similar percentage.
Stimulating the rumen wall with brush administration may increasing microbes activity in mechanical way, functioning as an artificial fiber. The RMS brush existency in the rumen might improve rumen motility and made the best use of nutrient fermentation into short-chain VFA as the major end product and promote the cellulolytic bacteria activity to digest fibrous. It was showed by higher production of acetate and butyrate in RF treatment, although the propionate had similar value with control. Horiguchi and Takahashi (2002) said that rumination is linked to rumen contraction and reticulorumen motility is a major factor in regulating the turnover of rumen digesta. They assumed may affect the passage rate of digesta through the ruminoreticulum that influence reticulorumen motility.
Lactic acid was produced by amylolytic bacteria during the degradation of starch, furthermore in low concentrations, transformed by another amylolytic bacteria to produce propionate (Nagaraja, 2012). The lactic acid concentration in the rumen liquor was found about 6.29-6.45 mM and gave a similar value between RF and control treatment. The control group potentially produced higher lactic acid concentration than the RF group, althought had a similar result with RF. Suspectedly crude fiber in feed ration intake reached the minimum requirement of feedlot steer that mantained the selulolytic bacteria activity. Chiba (2014) states that few part of the roughage component as long as can replaced by grain based concentrates which have lower potential energy than roughage, about 7.5 vs 5 mol VFA kg DM-1 were digested respectively, concentrate can compensate by the greater digestibility about 75-90% tthan roughage (45-70%), and higher fermentation rates via amylolytic than the cellulolytic pathways.
Increased the utilization of lactic acid in rumen occured while lactic microbes fermenters growth was promoted that indicated with this acidic status. The lower pH is often associated with higher lactate concentration in the ruminal liquor, thus this occurrence requiring a higher ruminal pH that favoring increased fiber digestibility (Nagaraja et al., 2012). The buffering process was able from saliva (naturally) or other buffers in a ration (bicarbonate) to maintaining good rumen status. Because fed high grains shall decrease the rumen motility and pH fall very quickly. Promoting lactic acid hydrogenation was stronger acid than the VFA, then it was need transformed the lactic acid into propionate, thus ruminal pH tends to normal (RAGFAR, 2007). Cerrilla and Martinez (2003) confirmed high concentrate diet made the high rate of total VFA, but also increased lactic acid concentration and took to acidity status in the rumen.
Chiba (2014) states that only 10% of lactic acid was changes into VFA, and the common compound is metabolized to pyruvate (en route to glucose and glycogen) by the liver. The RMS brush may stimulate the physical function of rumen mucosa and mantain the rumen ecology from unnature digestion in fed high concentrate (Horiguchi and Takahashi, 2002). The RMS brush might increased chewing activity of steer that forced a lot of saliva as the effect of excessive mastication, then rumination might increased. Horiguchi and Takahashi (2002) confirmed that the increased rumination duration time of the reticular contractions might be affect the increased passage rates
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13 might triggered the buffering capacity in the rumen. It was also noted that the present study had lower lactic acid concentration and normal pH status from adjusment of acidosis may occur.
Furthermore, steers administrated with and without RMS brush in this study was never assigned into acute acidosis indicated by upper than 50mM of lactic acid pH values lower than 5.0 (Nagaraja et al., 1985; Castrillo et al., 2013). The RMS brush might stimulated short chain degradable microbes and synthesized the lactic acid compound into short chain VFA, even unable to promoted amilolytic bacteria activity better according to propionate concentration, but kept the rumen motility from acidic status with high VFA.
Meanwhile, better VFA production in RF treatment had not gave a better performance of feedlot steer. Melo et al. (2013) reported that the increased VFA absorptive capacity may allow the use of energy-dense diets without excessive VFA accumulation, thereby reducing the incidence and severity of ruminal acidosis (Melo
et al., 2013). that better nutrient digestion such VFA would improve cattle performance but in this present study resulted a similar bodyweight gain. Suspiciously, the early stressed happens since installation of RF by used devices and affected cattles phisiologies condition. The adaptation period after that pressure slowed down the weight gain because lot of metabolism energy loosest by stressed condition (Yanza et al., 2015). Pond et al. (2005) confirmed that nutrient intake positively related with the growth rate and the body composition during the development. But compensatory feeding in some period unable boosted the cattle performance.
Methane emission was associated with microbes fermentative species and H2-users that may facilitating interspecies electrons transfer in the rumen. Moss et al.
(2000) states that the formation of methane is the major way of hydrogen elimination through the following reaction. This process brought a benefit by predicting the methane production from partial VFA concentration by adjusted models which accurate with a high coefficient determination by in-vitro method (Jayanegara et al., 2015). This present study resulted higher methane concentration The RF than the control treatment, resulted about 39.54 vs 33.6 mM (P<0.05). But the methane production per total VFA gave a similar proportion, about 28 mM. Furthermore, The H2 production and H2 utilised status resulted an improvement, where the RF treatment was higher than the Control by their H2 production (301.8 mM vs 248.4 mM) and H2 utilised (271.6 mM vs 223.5 mM) (P<0.05). These quantities were accorded to 90% H2 recovery that was stoichiometrically measured based on adjusted formula (Jayanegara et al., 2015).
This high quantity of methane in RF treatment was resulted by the accumulation of higher acetate and butyrate concentration. Meanwhile, the methane concentration per total VFA (which called relative methane) which achieved similar proportion with the control due to total VFA certainly reflected as methane production rather than the factual methane.
Higher H2 production and ionisation at the RF treatment may lead the rumen fermentation into methanogenesis since fermentation of glucose and protein to acetic and butyric yields H2, a main compound of methane formation (Jayanegara et al., 2015). But, some others bacteria iniciated oxaloacetate and lactic acid to produced propionate
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14
by attached the H2 ions. This synthesized may lowering the methane production, but in this study did not showed a better result. Propionate bacteria was unable inhibited the methane production since the cellulolytic bacteria was stimulated to produce higher acetate and butyrate while digesting fibrous by celulolitic microbes activity that yields H2 ions. However, the relative methane production could reduced since the total VFA concentration increased, although the factual methane concentration significantly tended to be higher than the control.
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15
4. CONCLUSIONS
This study suggests that feedlot steers administered with rumen mechanical stimulator (RMS) brush: 1) did not improve performance of feedlot steer; 2) improved the rumen fermentation by the total and partial VFA produced in the rumen, such as the acetate and butyrate concentration, but not the propionate, and 3) increased the production cost as indicated by its lower income over feed cost.
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16
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APPENDICES
1 st Appendix. Mean Difference of Dry Matter Intake in 95% Confidents Level (P<0.05) Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Low er Upp er Equal variances
assumed -0.18 18 0.86 0.00 0.01 -0.02 0.02 Equal variances
not assumed -0.18 10.31 0.86 0.00 0.01 -0.02 0.02 2 nd Appendix. Mean Difference of Crude Fiber Intake in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed
7.757E+ 13
3.6183
E-115 0.29 18 0.78 0.00 0.00 0.00 0.00 Equal variances
not assumed 0.29 10.61 0.78 0.00 0.00 0.00 0.00
3 rd Appendix. Mean Difference of Crude Protein Intake in 95% Confidents Level (P<0.05) Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed
8.8971E +13
1.0532
E-115 -0.95 18 0.36 0.00 0.00 0.00 0.00 Equal variances
not assumed -0.95 10.98 0.36 0.00 0.00 0.00 0.00 4 th Appendix. Mean Difference of Ether Extract in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed . . -1.34 18 0.20 0.00 0.00 0.00 0.00 Equal variances
not assumed -1.34 13.24 0.20 0.00 0.00 0.00 0.00 5 th Appendix. Mean Difference of NFE in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr> F Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed
9.198E +13
7.807E
-116 -0.11 18 0.92 0.00 0.00 -0.01 0.01 Equal variances
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19 6 th Appendix. Mean Difference of TDN Intake in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t Df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed
8.0415 E+14
2.6163
E-124 -0.37 18 0.71 0.00 0.01 -0.01 0.01 Equal variances
not assumed -0.37 10.51 0.72 0.00 0.01 -0.01 0.01 7 th Appendix. Mean Difference of Concentrate : Forage Ratio Intake in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed . . -2.15 18 0.05 -0.09 0.04 -0.19 0.00 Equal variances
not assumed -2.15 10.28 0.06 -0.09 0.04 -0.19 0.00 8 th Appendix. Mean Difference of GE Intake in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed
2.0638E +14
5.4157E-119 -0.20 18 0.84 0.00 0.01 -0.03 0.03 Equal variances
not assumed -0.20 10.39 0.84 0.00 0.01 -0.04 0.03 9 th Appendix. Mean Difference of Total Gain in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed 0.012 0.912 1.099 18 0.286 13.3 12.10 -12.12 38.72 Equal variances
not assumed 1.099 17.96 0.286 13.3 12.10 -12.13 38.73 10 th Appendix. Mean Difference of Average Daily Gain (ADG) in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed 0.013 0.911 1.097 18 0.287 0.14 0.13 -0.13 0.42 Equal variances
(36)
20
11 th Appendix. Mean Difference of Gain : Feed Ratio in 95% Confidents Level (P<0.05) Analysis Assumed F Sig. t df Pr>F Mean
Diff.
Std. Error Diff.
95% CID Lower Upper Equal variances
assumed 0.008 1.095 18.00 0.288 0.015 0.014 -0.014 0.043 0.41 Equal variances
not assumed 0.931 1.095 17.95 0.288 0.015 0.014 -0.014 0.044 0.41 12 th Appendix. Mean Difference of IOFC in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed 0.012 0.913 3.394 18 0.003
1,480,084 436,033 564,013 2,396,156 Equal variances
not assumed 3.394 17.96 0.003
1,480,084 436,033 563,850 2,396,319 13 th Appendix. Mean Difference of pH in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed 0.13 0.73 -5.16946E-05 17 1.00 0.00 0.215 -0.453 0.453 Equal variances
not assumed -5.19906E-05 17 1.00 0.00 0.214 -0.451 0.451 14 thAppendix. Mean Difference of ∆EH in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr> F Mean Diff. Std. Error Diff. 95% CID Lower Uppe
r Equal variances
assumed 2.86 0.11 -1.06 17 0.31 -46.92 44.37 -140.53 46.70 Equal variances
not assumed -1.08 15.73 0.30 -46.92 43.42 -139.09 45.26 15 th Appendix. Mean Difference of Ammonia in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Uppe
r Equal variances
assumed 1.66 0.22 0.91 17 0.375 2.052 2.254 -2.703 6.807 Equal variances
(37)
21 16 th Appendix. Mean Difference of Lactic acid in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed 4.32 0.05 0.26 17 0.801 0.161 0.627 -1.161 1.483 Equal variances
not assumed 0.26 14.75 0.796 0.161 0.610 -1.141 1.463 17 th Appendix. Mean Difference of Acetate in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed 2.56 0.13 -2.17 15 0.046 -12.853 5.923 -25.479 -0.228 Equal variances
not assumed -2.11 11.14 0.059 -12.853 6.104 -26.267 0.560 18 th Appendix. Mean Difference of Propionate in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Uppe
r Equal variances
assumed 9.08 0.01 -1.68 15 0.115 -6.870 4.101 -15.611 1.870 Equal variances
not assumed -1.60 8.63 0.146 -6.870 4.304 -16.671 2.931 19 th Appendix. Mean Difference of Butyrate in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed 0.12 0.73 -2.65 15 0.018 -5.212 1.969 -9.409 -1.016 Equal variances
not assumed -2.64 14.51 0.019 -5.212 1.976 -9.437 -0.988 20 th Appendix. Mean Difference of total VFA in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed 5.987 0.027 -2.504 15 0.024 -24.94 9.96 -46.16 -3.71 Equal variances
(38)
22
21 th Appendix. Mean Difference of Acetate : Propionate Ratio in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed 6.81 0.02 -0.04 15 0.969 -0.013 0.331 -0.718 0.692 Equal variances
not assumed -0.04 8.96 0.971 -0.013 0.346 -0.796 0.770 22 th Appendix. Mean Difference of Methane in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed 4.21 0.06 -2.37 15 0.032 -5.979 2.521 -11.354 -0.605 Equal variances
not assumed -2.29 10.61 0.043 -5.979 2.608 -11.744 -0.214 23 th Appendix. Mean Difference of Methane/VFA in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed 6.86 0.02 0.34 15 0.738 0.464 1.364 -2.444 3.373 Equal variances
not assumed 0.33 9.27 0.752 0.464 1.425 -2.745 3.673 24 th Appendix. Mean Difference of H2 prodiction in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Uppe
r Equal variances
assumed 1.00 0.33 0.85 15 0.410 1.502 1.772 -2.275 5.279 Equal variances
not assumed 0.84 13.71 0.417 1.502 1.793 -2.350 5.354 25 th Appendix. Mean Difference of H2 utilised in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff. Std. Error Diff. 95% CID Lower Upper Equal variances
assumed 6.28 0.02 -0.30 15 0.770 -0.577 1.933 -4.696 3.543 Equal variances
(39)
23 26 th Appendix. Mean Difference of Acetate Proportion in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error
Diff.
95% CID Lower Upper Equal variances
assumed 5.58 0.03 -2.79 15 0.014 -53.43 19.17 -94.30 -12.56 Equal variances
not assumed -2.68 9.66 0.024 -53.43 19.97 -98.13 -8.73 27 th Appendix. Mean Difference of Propionate Proportion in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Upper Equal variances
assumed 5.58 0.03 -2.79 15 0.014 -48.08 17.26 -84.87 -11.30 Equal variances
not assumed -2.68 9.66 0.024 -48.08 17.97 -88.32 -7.85 28 th Appendix. Mean Difference of Butyrate Proportion in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Upper Equal variances
assumed 1.91 0.19 -0.60 15 0.556 -0.93 1.54 -4.20 2.35 Equal variances
(40)
24
CURRICULUM VITAE
The author was born at 20th of July, 1989 in Pekanbaru-Riau with the atmosphere of Malayan life. Author is the middle son of Zakir Has and Nurbayani who bear the Mandailing clan of malay ethnic. The author has the eldest brother Sesfrika Yanza and the youngest Farhan Ramdhani Yanza. Academic track of the author start at SDN 020 Simpang Tiga in elementary school (1995-1996), then continued at MTsN Pekanbaru which equal with junior high school (2001-2004). The author then attended the senior high school the SMAN Plus
of Riau province (2004-2006) then graduate at SMAN 1 Pekanbaru (2007). The author’s bachelor degree was achieved at Bogor Agricultural University, Faculty of Animal Science, Nutrition Science and Feed Technology Mayor (IPB; 2007-2012). The author continued his master degree in Nutrition and Feed Science and finished at 2015. The author was an active person and joined several organisations since studied in IPB. Since studied in bachelor degree, the author had been a general secretary of HMI Komisariat Fakultas Peternakan IPB (2010-2012) and vice secretary of “Cadre Guidience Division” of HMI Cabang Bogor (2012). The author was also member of the HIMASITER (2010-2011). Since studied in master degree, the author was nominated as a chairman candidate of Forum Wacana IPB Election (2013-2014) at IPB Graduate Student Conference (2013) and trusted being a Coordinator of Public Policy Discussion Forum of Forum Wacana IPB at 2013-2014. The author also was the member of Gita Swara Pascasarjana Choir of IPB (2012-present). The author hobbies were reading, arts and sports such music, futsal and basketball and achieved several honours. The author was an assistent at laboratorium of agrostology under the Animal Science Faculty (2010-2012) and was an auxillary teacher of SMK Agri Insani Vocational High School (2012-2014) for Mathematics and Physics lessons. The author, local government and several native figures had concepted and established the Vocational High Scool of Agriculture, Major: Livestock Science, Plantation Science and Food Technology Science at Kampar, then named as SMKN 1 Kuok-Kampar Riau (2012-2014). The author also was a research assistant of Medium-term Development Planning Investment of P4W IPB and Kampar Riau Consortium Project (2013) and was an expert assistant of CSR Funds Evaluation Project of Joint Operating Body of Pertamina and PT. Talisman-Indonesia at Ogan Komering Ulu, South Sumatera (2014-2015). The author has an interest in being a lecturer and willingness to serve his hometown, Riau-Indonesia.
(1)
6
thAppendix. Mean Difference of TDN Intake in 95% Confidents Level (P<0.05)
AnalysisAssumed F Sig. t Df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Upper Equal variances
assumed
8.0415 E+14
2.6163
E-124 -0.37 18 0.71 0.00 0.01 -0.01 0.01 Equal variances
not assumed -0.37 10.51 0.72 0.00 0.01 -0.01 0.01
7
thAppendix. Mean Difference of Concentrate : Forage Ratio Intake in 95% Confidents
Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Upper Equal variances
assumed . . -2.15 18 0.05 -0.09 0.04 -0.19 0.00 Equal variances
not assumed -2.15 10.28 0.06 -0.09 0.04 -0.19 0.00
8
thAppendix. Mean Difference of GE Intake in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error
Diff.
95% CID Lower Upper Equal variances
assumed
2.0638E +14
5.4157E-119 -0.20 18 0.84 0.00 0.01 -0.03 0.03 Equal variances
not assumed -0.20 10.39 0.84 0.00 0.01 -0.04 0.03
9
thAppendix. Mean Difference of Total Gain in 95% Confidents Level (P<0.05)
AnalysisAssumed F Sig. t df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Upper Equal variances
assumed 0.012 0.912 1.099 18 0.286 13.3 12.10 -12.12 38.72 Equal variances
not assumed 1.099 17.96 0.286 13.3 12.10 -12.13 38.73
10
thAppendix. Mean Difference of Average Daily Gain (ADG) in 95% Confidents
Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error
Diff.
95% CID Lower Upper Equal variances
assumed 0.013 0.911 1.097 18 0.287 0.14 0.13 -0.13 0.42 Equal variances
(2)
11
thAppendix. Mean Difference of Gain : Feed Ratio in 95% Confidents Level (P<0.05)
Analysis Assumed F Sig. t df Pr>F Mean Diff.
Std. Error Diff.
95% CID Lower Upper Equal variances
assumed 0.008 1.095 18.00 0.288 0.015 0.014 -0.014 0.043 0.41 Equal variances
not assumed 0.931 1.095 17.95 0.288 0.015 0.014 -0.014 0.044 0.41
12
thAppendix. Mean Difference of IOFC in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Upper Equal variances
assumed 0.012 0.913 3.394 18 0.003
1,480,084
436,033
564,013
2,396,156 Equal variances
not assumed 3.394 17.96 0.003
1,480,084
436,033
563,850
2,396,319
13
thAppendix. Mean Difference of pH in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error
Diff.
95% CID Lower Upper Equal variances
assumed 0.13 0.73 -5.16946E-05 17 1.00 0.00 0.215 -0.453 0.453 Equal variances
not assumed -5.19906E-05 17 1.00 0.00 0.214 -0.451 0.451
14
thAppendix. Mean Difference of ∆EH in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>
F
Mean Diff.
Std. Error Diff.
95% CID Lower Uppe
r Equal variances
assumed 2.86 0.11 -1.06 17 0.31 -46.92 44.37 -140.53 46.70 Equal variances
not assumed -1.08 15.73 0.30 -46.92 43.42 -139.09 45.26
15
thAppendix. Mean Difference of Ammonia in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Uppe
r Equal variances
assumed 1.66 0.22 0.91 17 0.375 2.052 2.254 -2.703 6.807 Equal variances
(3)
16
thAppendix. Mean Difference of Lactic acid in 95% Confidents Level (P<0.05)
AnalysisAssumed F Sig. t df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Upper Equal variances
assumed 4.32 0.05 0.26 17 0.801 0.161 0.627 -1.161 1.483 Equal variances
not assumed 0.26 14.75 0.796 0.161 0.610 -1.141 1.463
17
thAppendix. Mean Difference of Acetate in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error
Diff.
95% CID Lower Upper Equal variances
assumed 2.56 0.13 -2.17 15 0.046 -12.853 5.923 -25.479 -0.228 Equal variances
not assumed -2.11 11.14 0.059 -12.853 6.104 -26.267 0.560
18
thAppendix. Mean Difference of Propionate in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Uppe
r Equal variances
assumed 9.08 0.01 -1.68 15 0.115 -6.870 4.101 -15.611 1.870 Equal variances
not assumed -1.60 8.63 0.146 -6.870 4.304 -16.671 2.931
19
thAppendix. Mean Difference of Butyrate in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error
Diff.
95% CID Lower Upper Equal variances
assumed 0.12 0.73 -2.65 15 0.018 -5.212 1.969 -9.409 -1.016 Equal variances
not assumed -2.64 14.51 0.019 -5.212 1.976 -9.437 -0.988
20
thAppendix. Mean Difference of total VFA in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error
Diff.
95% CID Lower Upper Equal variances
assumed 5.987 0.027 -2.504 15 0.024 -24.94 9.96 -46.16 -3.71 Equal variances
(4)
21
thAppendix. Mean Difference of Acetate : Propionate Ratio in 95% Confidents
Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Upper Equal variances
assumed 6.81 0.02 -0.04 15 0.969 -0.013 0.331 -0.718 0.692 Equal variances
not assumed -0.04 8.96 0.971 -0.013 0.346 -0.796 0.770
22
thAppendix. Mean Difference of Methane in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Upper Equal variances
assumed 4.21 0.06 -2.37 15 0.032 -5.979 2.521 -11.354 -0.605 Equal variances
not assumed -2.29 10.61 0.043 -5.979 2.608 -11.744 -0.214
23
thAppendix. Mean Difference of Methane/VFA in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Upper Equal variances
assumed 6.86 0.02 0.34 15 0.738 0.464 1.364 -2.444 3.373 Equal variances
not assumed 0.33 9.27 0.752 0.464 1.425 -2.745 3.673
24
thAppendix. Mean Difference of H2 prodiction in 95% Confidents Level (P<0.05)
AnalysisAssumed F Sig. t df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Uppe
r Equal variances
assumed 1.00 0.33 0.85 15 0.410 1.502 1.772 -2.275 5.279 Equal variances
not assumed 0.84 13.71 0.417 1.502 1.793 -2.350 5.354
25
thAppendix. Mean Difference of H2 utilised in 95% Confidents Level (P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error
Diff.
95% CID Lower Upper Equal variances
assumed 6.28 0.02 -0.30 15 0.770 -0.577 1.933 -4.696 3.543 Equal variances
(5)
26
thAppendix. Mean Difference of Acetate Proportion in 95% Confidents Level
(P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error
Diff.
95% CID Lower Upper Equal variances
assumed 5.58 0.03 -2.79 15 0.014 -53.43 19.17 -94.30 -12.56 Equal variances
not assumed -2.68 9.66 0.024 -53.43 19.97 -98.13 -8.73
27
thAppendix. Mean Difference of Propionate Proportion in 95% Confidents Level
(P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Upper Equal variances
assumed 5.58 0.03 -2.79 15 0.014 -48.08 17.26 -84.87 -11.30 Equal variances
not assumed -2.68 9.66 0.024 -48.08 17.97 -88.32 -7.85
28
thAppendix. Mean Difference of Butyrate Proportion in 95% Confidents Level
(P<0.05)
Analysis
Assumed F Sig. t df Pr>F
Mean Diff.
Std. Error Diff.
95% CID Lower Upper Equal variances
assumed 1.91 0.19 -0.60 15 0.556 -0.93 1.54 -4.20 2.35 Equal variances
(6)