DOCSLIB.ORG

EFFECT of ENERGY SOURCE, TIMING of PROVISION, and DAYS on FEED on FEED EFFICIENCY of FINISHING BEEF CATTLE a Thesis Submitted T

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

EFFECT OF ENERGY SOURCE, TIMING OF PROVISION, AND DAYS ON FEED ON FEED EFFICIENCY OF FINISHING BEEF CATTLE A Thesis Submitted to the College of Graduate and Postdoctoral Studies in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy in the Department of Animal and Poultry Science University of Saskatchewan Saskatoon, Saskatchewan By Faustin Joy ©Copyright Faustin Joy, December 2019, All rights reserved PERMISSION TO USE STATEMENT In presenting this thesis in partial fulfillment of the requirements for a Postgraduate degree from the University of Saskatchewan, I agree that the Libraries of this University may make it freely available for inspection. I further agree that permission for copying of this thesis in any manner, in whole or in part, for scholarly purposes may be granted by the professor or professors who supervised my thesis work or, in their absence, by the Head of the Department or the Dean of the College in which my thesis work was done. It is understood that any copying or publication or use of this thesis or parts thereof for financial gain shall not be allowed without my written permission. It is also understood that due recognition shall be given to me and to the University of Saskatchewan in any scholarly use which may be made of any material in my thesis. DISCLAIMER This report was exclusively created to meet the thesis requirements for the degree of Doctor of Philosophy at the University of Saskatchewan. Reference in this thesis to any specific commercial products, process, or service by trade name, trademark, manufacturer, or otherwise, does not constitute or imply its endorsement, recommendation, or favoring by the University of Saskatchewan. The views and opinions of the author expressed herein do not state or reflect those of the University of Saskatchewan, and shall not be used for advertising or product endorsement purposes. Requests for permission to copy or to make other uses of materials in this thesis in whole or part should be addressed to: Head of the Department of Animal and Poultry Science 51 Campus Drive University of Saskatchewan Saskatoon, Saskatchewan S7N 5A8 Canada OR Dean College of Graduate and Postdoctoral Studies University of Saskatchewan 116 Thorvaldson Building, 110 Science Place Saskatoon, Saskatchewan S7N 5C9 Canada i ABSTRACT The objective of this research was to evaluate if the decline in gain-to-feed ratio (G:F) during finishing in beef cattle could be due to a reduction in nutrient digestion, short chain fatty acid absorption, or post-absorptive nutrient utilization, and those responses were influenced by the dietary energy source or timing of provision. A high-lipid byproduct pellet (HLP) was used as a partial replacement for barley grain in a high concentrate finishing diet to partially replace starch with lipid as the energy source. Yearling steers were used for performance evaluations and ruminally cannulated Hereford crossbred heifers were used for nutrient metabolism studies. The dry matter (DM), organic matter (OM), crude protein (CP) and neutral detergent fibre (NDF) digestibility of HLP diets were lower than the barley-based control diet (CON). Utilizing a phase- feeding strategy and partially replacing barley grain and canola meal with a high-fibre high-lipid byproduct pellet in the latter part of the finishing period may improve carcass yield grade without affecting average daily gain and G:F. With advancing days on feed, diet DM digestibility (P = 0.02) and insulin resistance (P = 0.04) increase without changes in ruminal pH and plasma metabolite clearance rates. A marginal increase in forage inclusion with the HLP diet increased ADG (P = 0.04). Increasing dietary lipid supply up to 6% of DM using HLP did not affect the digesta flow and rumen fermentation parameters, therefore, increased lipid content is not associated with reduced feed efficiency of the HLP diet. In conclusion, decreasing feed efficiency in the later stages of finishing in beef cattle is most likely due to changes in the post absorptive nutrient metabolism, and these changes are not influenced by the dietary energy source. The small particle size of feed ingredients within the byproduct pellet may be the factor associated with decreased feed efficiency of HLP pellet rather than lipid content. ii ACKNOWLEDGEMENTS I thank Almighty God for all his blessings and giving this opportunity to work with a bunch of wonderful people at the Department of Animal Science, University of Saskatchewan as part of my Doctoral program. I would like to extend my heartfelt gratitude to my supervisor Dr. Greg Penner for all his amazing support, guidance and encouragement throughout my study. I will always cherish his encouragement and support as an invaluable experience in my academic career. I thank my committee members Dr. John McKinnon, Dr. Steve Hendrick, Dr. Luis Burciaga and the graduate committee chair Dr. Fiona Buchanan for their continued assistance with my research project and thesis writing. My sincere appreciation to Dr. Pawel Górka, who patiently devoted his time for numerous discussions regarding this project. A special thanks to Team Rumen, Gillian Gratton, Kasia, Coral, Katie, Dan, Silvia, Keshia, Rodrigo, Larissa, Jordan, Brittney, Liam, Tyler, Brittany, JK, Brittney Schurmann and numerous others for all their dedication and assistance with this research project. I would also like to extend my sincere thanks to the Manager and staff of Livestock Research Building and Beef Cattle Research and Teaching Unit at the University of Saskatchewan for assisting me with the sample collection. I also extend my gratitude to the Alberta Crop Industry Development Fund Ltd. (Lacombe, AB, Canada), Agriculture Development Fund administered through the Ministry of Agriculture in Saskatchewan (Regina, SK, Canada), and the Beef Cattle Research Council of Canada (Calgary, AB, Canada) for funding my projects. I thank Natalia, lab manager for her guidance with the laboratory work of this project. I dedicate this work to my parents, Joy and Annamma, and my dear sisters for their constant motivation and strength in my academic career. I thank my dear wife Divya for always being there to support me in all the ups and downs of life, and instilling in me the confidence to go ahead and succeed in life. I also thank my son Enric for cooperating with my studies and writing and making my world more meaningful and beautiful each day. iii TABLE OF CONTENTS PERMISSION TO USE STATEMENT .......................................................................................... i ABSTRACT .................................................................................................................................... ii LIST OF TABLES ........................................................................................................................ vii LIST OF FIGURES ........................................................................................................................ x LIST OF ABBREVIATIONS ........................................................................................................ xi 1.0 INTRODUCTION .................................................................................................................... 1 2.0 LITERATURE REVIEW ......................................................................................................... 3 2.1 Feeding management in North America ............................................................................... 3 2.2 Feed efficiency in feedlot finishing diets .............................................................................. 5 2.2.1 Patterns of feed efficiency .............................................................................................. 6 2.2.2 Importance of feed efficiency (Sustainability) ............................................................... 9 2.2.3 Physiological factors affecting feed efficiency ............................................................ 13 2.2.4 Strategies to increase feed efficiency ........................................................................... 27 2.3 Alternative diets in feedlot cattle ........................................................................................ 29 2.3.1 Single byproduct substitution- Advantages .................................................................. 30 2.3.2 Disadvantages of byproducts in feedlot diets ............................................................... 30 2.3.3 Blended byproducts ...................................................................................................... 31 2.3.4 Rumen fermentation and feed efficiency with byproduct diets .................................... 32 2.4 Global hypothesis and objectives ........................................................................................ 34 3.0 EVALUATION OF A PHASE-FEEDING STRATEGY UTILIZING A HIGH- LIPID HIGH-FIBRE BYPRODUCT PELLET IN DIETS FOR FEEDLOT STEERS ............... 36 3.1. Introduction ........................................................................................................................ 37 3.2. Materials and methods ....................................................................................................... 38 3.2.1 Study 1 .......................................................................................................................... 38 3.2.2 Study 2 .........................................................................................................................
Recommended publications
  • WAGYU BREEDERS HANDBOOK an Introduction to Wagyu

    WAGYU BREEDERS HANDBOOK an Introduction to Wagyu

    2018 WAGYU BREEDERS HANDBOOK An introduction to Wagyu. Pamela Armstrong, LVT Maple Row Stock Farm [email protected] www.Wagyupedia.com FOREWARD Many people consider Wagyu beef to be the most tender and flavorful beef in the World. The cattle used to make this beef are docile with good temperaments, and they are known for their intense intramuscular marbling, high fertility rates and calving ease traits. Why wouldn’t a cattle farmer want to raise Wagyu? The internet is flush with information about Wagyu, some of it is accurate and some of it is misleading. This handbook is designed to help breeders decide whether or not raising this breed is the right choice for them. Peer-reviewed journals and academic textbooks were used to create this handbook, and world-renowned Wagyu experts were consulted. There are good opportunities for producers who are informed, careful and realistic. There are many variances within the Wagyu breeds and bloodlines; as well as differences in short and long-fed animals, and results of feeding protocols. Wagyu are very special animals, they are considered a national treasure in Japan. I hope you enjoy and appreciate them as much as I do. Pam Armstrong, LVT © 2018 Pamela Armstrong, LVT Page 2 Table of Contents FOREWARD ................................................................................................................................................... 2 ORIGIN OF WAGYU ......................................................................................................................................
  • Japanese Beef Cattle - Wa Means Japan[Ese] and Gyu Means Cow

    Japanese Beef Cattle - Wa Means Japan[Ese] and Gyu Means Cow

    Definition and breed makeup Origins Meat Quality Breed Register and its Structure Wagyu is the name of Japanese beef cattle - wa means Japan[ese] and gyu means cow. There are four(4) types of Wagyu Cattle in Japan; Japanese Black Japanese Brown or Red (also known as Akaushi) Japanese Shorthorn Japanese Poll Japanese Beef cattle Japanese Black Japanese Brown (Wagyu) (Red Wagyu) Native Cattle Japanese Shorthorn Japanese Poll These breed have been established by cross between Japanese native cattle and western breeds. The Japanese Black, called “Wagyu”, is the most popular beef cattle in Japan, and this breed was established by cross between Japanese native cattle and several European breeds, including Brown Swiss, Devon, and Ayrshire Japanese Brown, known as “Red Wagyu”, was established by cross with Korean originated cattle and Simmental breed. We have also Japanese shorthorn and Japanese polled, but the population of these two breeds has recently significantly reduced in recent times • For centuries Buddhism was predominant in Japan and meat consumption was prohibited. The utilization of animal products did not become popular until the Meiji restoration in 1868 In the Meiji era many foreign breeds were introduced to Japan and initially crossbred with the native cattle under the direction of the government. Through this, the gene pool for Japanese breed had diluted but the population had expanded. After the initial frenzy of crossbreeding was over, cattle breeders began to improve and promote their own breeds without further crossbreeding within the prefectures. Meat has been consumed in Japan for only about 140 years. Meat eating has only reached widespread popularity in the last 40 years after the “Law for Improvement and Increased Livestock Production” was enacted in 1950s.
  • Wagyu and the Factors Contributing to Its Beef Quality: a Japanese Industry Overview

    Wagyu and the Factors Contributing to Its Beef Quality: a Japanese Industry Overview

    Meat Science 120 (2016) 10–18 Contents lists available at ScienceDirect Meat Science journal homepage: www.elsevier.com/locate/meatsci Wagyu and the factors contributing to its beef quality: A Japanese industry overview Michiyo Motoyama a,b,⁎, Keisuke Sasaki a, Akira Watanabe a a Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization (NARO), Japan b UR370 Qualité des Produits Animaux, Institute Nationale de Recherche Agronomique (INRA), France article info abstract Article history: Wagyu cattle are originated from native Japanese breeds, which have evolved by adapting to the unique climate Received 7 February 2016 and environment of Japan. Since the modern beef-eating culture started to flourish in Japan in the 1860s, Wagyu Received in revised form 17 April 2016 has been improved for higher quality beef to satisfy the taste preferences of Japanese consumers. The most no- Accepted 19 April 2016 ticeable characteristic of Wagyu beef is its intense marbling. The high intramuscular fat (IMF) content improves Available online 20 April 2016 the texture, juiciness and thereby the overall palatability. In addition, the composition of the fat in Wagyu is con- Keywords: siderably different from that in other beef breeds. Characteristic Wagyu beef aroma gives sweet and fatty sensa- Wagyu tion. Wagyu beef is also valued for its high traceability and uniformity guaranteed because of the nationwide Japanese Black standards for beef carcass and trading. Although Wagyu producers are currently facing issues regarding calf pro- Marbling duction, Wagyu beef is increasingly being exported to the global market and creating new market value as one of Intramuscular fat the world's most luxury food products.
  • Wagyu Breeder Handbook

    Wagyu Breeder Handbook

    2018 WAGYU BREEDER HANDBOOK An introduction to Wagyu. Pamela Armstrong, LVT Maple Row Stock Farm [email protected] www.Wagyupedia.com FOREWARD Many people consider Wagyu beef to be the most tender and flavorful beef in the World. The cattle used to produce this beef are docile with good temperaments, and they are known for their intense intramuscular marbling, high fertility rates and ease of calving. Why wouldn’t a cattle farmer want to raise Wagyu? The internet is flush with information about Wagyu, some of it is accurate and some of it is misleading. This handbook is designed to help breeders decide whether or not raising this breed is the right choice for them. Peer-reviewed journals and academic textbooks were used to create this handbook, and world-renowned Wagyu experts were consulted. There are good opportunities for producers who are informed, careful and realistic. There are many variances within the Wagyu breeds and bloodlines; as well as differences between short and long-fed animals, and results of different feeding protocols. Wagyu are very special animals, they are considered a national treasure in Japan. I hope you enjoy and appreciate them as much as I do. Pam Armstrong, LVT (Pam and MRS Kitaguni Shigatsu “April”). © 2018 Pamela Armstrong, LVT Page 2 Table of Contents ORIGIN OF WAGYU ....................................................................................................................................... 5 What are Wagyu? ....................................................................................................................................
  • The Japanese Wagyu Beef Industry: Current Situation and Future Prospects — a Review

    The Japanese Wagyu Beef Industry: Current Situation and Future Prospects — a Review

    Open Access Asian-Australas J Anim Sci Vol. 31, No. 7:933-950 July 2018 https://doi.org/10.5713/ajas.18.0333 pISSN 1011-2367 eISSN 1976-5517 The Japanese Wagyu beef industry: current situation and future prospects — A review Takafumi Gotoh1,2,*, Takanori Nishimura3, Keigo Kuchida4, and Hideyuki Mannen5 * Corresponding Author: Takafumi Gotoh Abstract: In Japan, Wagyu cattle include four Japanese breeds; Black, Brown, Shorthorn, and Tel: +81-99-285-8699, Fax: +81-99-285-8699, E-mail: [email protected] Polled. Today, the renowned brand name Wagyu includes not only cattle produced in Japan, but also cattle produced in countries such as Australia and the United States. In recent years, 1 Department of Agricultural Sciences and Natural the intramuscular fat percentage in beef (longissimus muscle) from Japanese Black cattle has Resources, Faculty of Agriculture, Kagoshima University, Kagoshima, 890-0065, Japan increased to be greater than 30%. The Japanese Black breed is genetically predisposed to pro­ 2 Kuju Agricultural Research Center, Faculty of ducing carcass lipids containing higher concentrations of monounsaturated fatty acids than Agriculture, Kyushu University, Takeda 878-0201, other breeds. However, there are numerous problems with the management of this breed Japan 3 Muscle Biology and Meat Science laboratory, including high production costs, disposal of untreated excrement, the requirement for im­ Research Faculty of Agriculture, Hokkaido University, ported feed, and food security risks resulting from various viral diseases introduced by im­ Sapporo 060-8589, Japan ported feed. The feeding system needs to shift to one that is more efficient, and improves 4 Obihiro University of Agriculture and Veterinary Medicine, Obihiro 080-8555, Japan management for farmers, food security for consumers, and the health environment for resi­ 5 Laboratory of Animal Breeding and Genetics, dents of Japan.
  • The Difference Between Kobe and Wagyu Beef by John Cecala, Buedel Fine Meats, Published in Food Industry News

    The Difference Between Kobe and Wagyu Beef by John Cecala, Buedel Fine Meats, Published in Food Industry News

    The Difference Between Kobe and Wagyu Beef by John Cecala, Buedel Fine Meats, published in Food Industry News Wagyu beef is intensely marbled with softer fat, has higher percentages of monounsaturated fats, omega-3 and omega-6 fatty acids and is lower in cholesterol than commodity beef. The combinations of these fats deliver a distinctive rich and tender flavor compared to other beef. The most exclusive Wagyu in the world comes from Kobe, Japan. People use the terms Kobe and Wagyu beef interchangeably in the U.S. thinking it refers to the same premium imported Japanese beef, when it does not. All Kobe is Wagyu, but not all Wagyu is Kobe Many restaurant menus feature “Kobe Burgers” or “Kobe Steaks”. The internet is flooded with on-line companies offering Kobe Beef, Kobe Burgers, Kobe-Style Beef, and Wagyu beef. The truth is authentic Kobe beef is very rarely seen on restaurant menus in the USA. Legitimate Kobe beef is priced around $200 per portion for a steak, and $50 for a burger. If you see something on a menu referred to as Kobe priced less than that, it is most likely domestic or imported Wagyu. How can you tell the difference? Key Terms & Definitions Kobe A city in Japan and the capital city of the Hyōgo Prefecture. Kobe is also considered a region of Japan like Champaign is a region in France, and Parma is a region in Italy. Wa Japanese or Japanese Style Gyu The Japanese word for a Cow or Cattle Wagyu Japanese or Japanese Style Cattle. Japanese cattle consist of four breeds: Japanese Black, Japanese Brown, Japanese Poll, and Japanese Shorthorn.
  • Explore Night Time Hyogo Experience History・Culture Manners & Basic Knowledge Nature・Landscape HYOGO Sports・Activity the Origin of Japan Gourmet・Onsen Climate Food

    Explore Night Time Hyogo Experience History・Culture Manners & Basic Knowledge Nature・Landscape HYOGO Sports・Activity the Origin of Japan Gourmet・Onsen Climate Food

    Tips for enjoying Explore Night time Hyogo experience History・Culture Manners & basic knowledge Nature・Landscape HYOGO Sports・Activity The Origin of Japan Gourmet・Onsen Climate Food The climate differs greatly between the north and Many hotels and restaurants in Japan are unable south of Hyogo prefecture. The north (Tajima, to provide special vegetarian or halal menus. Tamba) gets a lot of snow in winter (Dec-Feb) and Please check with your destination ahead of time temperatures may fall below freezing. if you have special dietary needs. ONSEN Manners (hot spring) Wearing a bathing suit in the ONSEN is not Japanese people have great respect for propriety. allowed. In some cases, you may be refused if It’s important to behave politely and handle tools you have tattoos. (In Japan, people with tattoos with care when taking part in a tourist experi- are generally assumed to be gangsters.) ence. Also, please speak quietly and remove hats and sunglasses when entering a temple or shrine. Few Japanese people speak English, compared to Language the rest of Asia, and may not be able to under- stand even simple things in English. 5-10-1 Shimoyamatedori,Chuo-ku,Kobe,Hyogo 650-8567 E-mail：[email protected] Explore Hyogo, The Origin of Japan E125° E135° E145° N45° Hyogo, the heart of Japan Hyogo Prefecture is located in the center of Japan at N35° 35 degrees north latitude and 135 degrees east longitude. Close to the ancient Japanese capital of Kyoto, Hyogo has a rich cultural background. Due to its convenient location, Hyogo is well-connected to N25° other major cities by railroads and airlines.
  • Meat Produced by Japanese Black Cattle and Wagyu Report

    Meat Produced by Japanese Black Cattle and Wagyu Report

    Meat produced by Japanese Black cattle and Wagyu T. Gotoh,* H. Takahashi,* T. Nishimura,† K. Kuchida,‡ and H. Mannen§ *Kuju Agricultural Research Center, Faculty of Agriculture, Kyushu University 8780201, Japan †Muscle Biology and Meat Science laboratory, Research Faculty of Agriculture, Hokkaido University, 060-8589, Japan ‡Obihiro University of Agriculture and Veterinary Medicine, 080-8555, Japan §Laboratory of Animal Breeding and Genetics, Graduate School of Agricultural Science, Kobe University, 657-8501, Japan Black cattle produced in foreign countries such as Australia or the USA. Implications There are numerous studies investigating the meat quality, quantity, and • In Japan, Wagyu cattle include four types of Japanese cattle: the muscle physiology of crossbreed Wagyu (Japanese Black) in foreign Black, Brown, Shorthorn, and Polled breeds. The famous brand countries (May et al., 1993; Cafe et al., 2006, 2009; Greenwood et al., name Wagyu includes not only Japanese Black cattle produced in 2006, 2009). In this review, Wagyu will be referred to as the Japanese Japan, but also animals or even crossbred Japanese Black cattle Black cattle because of their predominant numbers. produced in foreign countries such as Australia or the USA. Mishima cattle, which are thought to be a Japanese native, are bred on Mishima Island in the Yamaguchi prefecture. They seem to have main- • In recent years, the intramuscular fat percentage of beef from tained the native breed characteristics, and the cattle are expected to im- Japanese Black cattle has been greater than 30%. Japanese Black prove meat quality when crossed with other breeds. The Mishima steers is genetically predisposed to producing carcass lipids containing averaged 517 ± 20 kg (n = 4) of body weight at 35.8 ± 0.5 months of age greater concentrations of monounsaturated fatty acids than other (Morita et al., 2000).
  • Taste Excellence AWA NEWSLETTER

    Taste Excellence AWA NEWSLETTER

    eNEWS Official Publication SALE RESULTS | SELLING THAT STEAK | WAGYU EVERYWHERE | NEW BOARD MEMBERS BEEF CATTLE FEED EFFICIENCY | CARE AND CLEANING | INCREASE MARBLING | CATTLEFAX Taste Excellence AWA NEWSLETTER JAN - FEB 208-262-8100 www.wagyu.org 2017 AWA NEWSLETTER JAN - FEB 2017 2017 MILE HIGH WAGYU EXPERIENCE SALE RESULTS 2017 Mile High Wagyu Experience Sale Top Selling Semen Lot: January 18th, 2017 – Denver, Colorado Lot 42- $1100 per unit – MT FUJI, 5 units; consigned by Auctioneer: C.D. Butch Booker Crescent Harbor Ranch, purchased by Theresa Olson of Manager: JDA, Inc. North Branch, MN. Averages Donation Lot: Females: $7858 7 Day South African Safari - $2300 – donated by Wagyu Bulls: $5750 Breeder.Net, purchased by Troy Gabriel, Gabriel Family Embryos: $786/Embryos Farms of Ashville, OH. All proceeds go to the A.W.A. Semen: $348/Unit A beautiful day in Denver set the tone for a wonderful TOPS: evening. The Mile High Wagyu Experience Sale followed Top Selling Live Lot: the Wagyu Show at the 2017 National Western Stock Lot XX – $40,500 - CMC Genie D3, 3-01-2016, sired by Show and was hosted at the Cowboy Lounge in Down- GENESIS; consigned by CMC Livestock. 1/3 interest town Denver, CO. A large crowd filled the Cowboy purchased by BAR V Wagyu of Abilene, KS. Lounge to enjoy an array of good eats and participate is an exciting Wagyu auction. The added lots of a few Top Selling Embryo Lot: of the day’s champions helped to fuel the anticipation Lot 24 – $1500 per embryo – Rueshaw X LAG B0589N, which brought in over 70 registered buyers to the event.
  • Canadian Inspection Failed to Meet U.S. Standards Natural Programs

    Canadian Inspection Failed to Meet U.S. Standards Natural Programs

    The National Livestock Weekly January 16, 2006 • Vol. 85, No. 14 “The Industry’s Largest Weekly Circulation” www.wlj.net • E-mail: [email protected] • [email protected] • [email protected] A Crow Publication Canadian inspection failed to meet U.S. standards In November 2003, U.S. food safety officials warned that Canadian meat and poultry inspec- tions failed to meet minimum U.S. standards, yet R-CALF pushes USDAdid nothing to stop the flow of imports from Canada, a recently released study by USDA’s Of- fice of the Inspector General (OIG) found. Canadian Since the alarm was raised, more than 4.4 bil- lion pounds of processed meat has made its way border suit to U.S. consumers. USDA, in response to the report, said it had ad- R-CALF United Stockgrowers of America (R- dressed numerous violations at multiple Cana- CALF) is pressing their case against USDA and dian plants, some of which lost their export cer- the agency’s decision to open the Canadian bor- tification following inspections which noted the der to cattle under 30 months of age, referred to deficiencies. in the motion as USDA’s minimal risk rule (Final “In no instance was public health placed at Rule). R-CALF was granted a preliminary injunc- risk,” said Richard Raymond, undersecretary for tion in Montana federal district court in March food safety. 2005 which halted the movement of Canadian Since USDA inspectors decertified the Cana- cattle and specific beef products into the U.S. as dian processors, the Canadian government has a result of R-CALF concerns about the prevalence been working with industry officials in that coun- of bovine spongiform encephalopathy (BSE).