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ISSN 1479-5647Zoo Research GuidelinesNutrition and Diet Evaluation

Nutrition and Diet Evaluation British and Irish Association of Zoos and Aquariums 2009All rights reserved. No part of this publication my be reproduced or transmitted in anyform or by any means, electronic or mechanical, including photocopy, recording orany information storage and retrieval system, without permission in writing from thepublisher.Fidgett, A.L. and Plowman, A. (2009) Zoo Research Guidelines: Nutrition and dietevaluation. BIAZA, London, UK.First published 2009Published and printed by:BIAZAZoological Gardens,Regent’s Park,London NW1 4RY,United KingdomISSN 1479-5647

Nutrition and Diet EvaluationZoo Research Guidelines:Nutrition and Diet EvaluationAndrea L. Fidgett1 and Amy Plowman2North of England Zoological Society, Chester Zoo, Chester, CH2 1LH; 2Paignton ZooEnvironmental Park, Totnes Road, Paignton, Devon, TQ4 7EU1With contributions from and acknowledgements for valuable comments: Andy Beer,Ellen Dierenfeld, Anna Feistner and Stephanie Wehnelt; Sue Dow and Bristol Zoo forhosting this workshop at the BIAZA Research Meeting at Bristol Zoo (2003).Introduction to Nutrition and Diet Evaluation GuidelinesEffective management of species in zoos and aquariums requiresconsiderable knowledge of the biology of each species includingreproduction, behaviour, group dynamics, husbandry, nutrition, medicalneeds and so forth. Scientific investigations are the basis for understandingthe animals in a zoo and assessing the way they are cared for. Throughcareful observations and well-planned studies, much can be learned about,for example, reproductive and social behaviour, growth and development,and interactions with the physical environment.These guidelines form part of a series designed to clarify the series of stepsthat are usually involved in developing a research project in a zooenvironment. They discuss challenges which are characteristic for zooresearch projects which may differ from those encountered in laboratory orfield studies. Rather than repeat relevant advice offered elsewhere, theauthors suggest reading this document in conjunction with others from theseries, specifically: Project Planning and Behavioural Observations (Wehnelt etal., 2003); Research by questionnaire (Plowman et al., 2006); Research usingZoo Records (Fidgett et al., 2008).Non-invasive techniques of diet evaluation can be successfully employed inthe absence of specific nutritional expertise or laboratory analytical facilities,to provide information leading to the improvement of diets fed to zooanimals. These guidelines focus on methods that can be undertaken withoutspecialist knowledge or equipment as we assume that this will be thesituation for most readers, and are thus aimed at those who may be relativelynew to this field. The chapter ‘Feeding and Nutrition’ (Hosey et al., 2009) isalso useful background reading before any further investigation.-3-

Nutrition and Diet EvaluationThe Nutrition and Diet Evaluation Guidelines are divided into the followingheadings and it is useful to read all sections before beginning a project:1. What do we mean by Nutrition and Diet Evaluation?2. Dietary evaluation:a. by weighing feeds and remainsb. by focal animal observationsc. by survey and review3. Feed composition and nutrient calculation tools4. Chemical analysis of foodstuffs and faeces5. Dietary standards and recommendations6. Final considerations7. Further reading and other resourcesAppendix I - Conducting an Intake StudyAppendix II – Zoo Diet Questionnaire1. What do we mean by Nutrition and Diet Evaluation?Diet is the usual food of an animal, nutrition is the process by which a livingorganism assimilates that food and uses it for growth and for replacement oftissues. One of the challenges which zoological institutions face whenhousing and maintaining exotic animals is providing them with a dietadequate for achieving optimal nutrition.There are a number of factors that need to be taken into account whendesigning diets. These include: knowledge of the animal's diet compositionand foraging behaviour in the wild, knowledge of the functional anatomy ofits digestive tract, knowledge of dental morphology and adaptation to thenatural diet, knowledge of the bases on which animals select particular fooditems, and an understanding of the implications that foraging and dietselection have for social behaviour. This is a tall order, given our lack ofknowledge about many exotic species. It is compounded by the fact thateven when these factors are known, devising suitable substitutes for a wilddiet is hard. For example, we may know that 75% of a wild primate's diet isfruit, yet feeding 75% fruit in captivity may provide a very different intake interms of nutrients, since fruits produced commercially for consumption byhumans (and thus readily available as components of zoo diets) are sodifferent from wild fruits (see O’Brien et al., 1998).The goal of zoo feeding programmes must be to provide nutritional supportfor all stages of life, including egg development, gestation, lactation, andearly post-natal growth. Dietary deficiencies may result in increased-4-

Nutrition and Diet Evaluationsusceptibility to disease, reduced fertility, reduced neonate viability, suboptimal yolk or milk production, retarded growth and physical deformities.Deficiencies in diet presentation may result in obesity, stereotypies, orincreased aggression. It is clear that an understanding of the nutrition ofexotic animals is vital to their well-being and our ability to maintain themsuccessfully in captivity, yet animal nutrition is a new and relativelyunexplored field. Part of the problem is a lack of facilities in zoologicalinstitutions and a lack of expertise. There is, thus, a strong need to developnutritional studies and departments in zoological institutions.Research on nutrition is carried out both as a problem-solving exercise (inrelation to ill-health or apparent infertility) and as part of ongoingprogrammes to improve animal husbandry. Studies carried out to solveproblems tend to focus on assessing the nutrient quality of the diet, whereasthe husbandry work is more oriented to food presentation methods andbehavioural and environmental enrichment involving food, and feeding andforaging techniques. The aims are to enhance nutritional quality andpalatability and to stimulate natural foraging behaviour. The emphasis is onnon-invasive research methods. The ideal situation for diet evaluation wouldbe to monitor the intake of individual animals. However, for the evaluation tobe valid it is essential that the animals are behaving normally and separatinganimals for the purpose of a study would likely cause stress, which in turnmay affect appetite and hence intake. Therefore, animals should not beseparated from their usual social groupings, normal routines disturbed aslittle as possible, and most assessments can be done indirectly throughweighing food remains or analysing faeces.2. Dietary evaluationCurrently in the UK, the Secretary of State’s Standards for Modern ZooPractice (2004) are designed to ensure that the welfare of animals in zoos isprotected, that zoos are safe places for the public to visit and, that zoosparticipate in appropriate conservation and public education measures. TheStandards recommend best practice by which zoos are inspected and grantedlicenses by local authorities. The Standards consider food and water as basicneeds and recommends the frequency of feeds and their nutritional balancebe taken into account. It states “Food should be presented in a manner andfrequency commensurate with the natural behaviour of the species, as well asits nutritional requirements, which may vary according to season.” Thisstatement is expanded to a series of more specific recommendations,including “A record of all diets must be maintained.”-5-

Nutrition and Diet EvaluationFor many zoos, diets are recorded on kitchen whiteboards, a practicalsolution to keep all keepers informed and easily allow for changes as speciesor numbers of animals change. However useful, they only form a startingpoint since they often lack sufficient detail about the feeds and quantitiesused, data which are necessary for properly evaluating the efficacy of a diet.a. Diet evaluation by weighing feeds and remainsThe most commonly used means of diet evaluation involves weighing feedsand remains. Also termed an intake study, it is relatively simple but can betime-consuming. The purpose is to determine the actual amount of food fedto and consumed by an animal or group of animals in an enclosure. Becausesome diet components may be offered but not consumed, an evaluation ofthe feeds actually consumed can provide considerably more information andthus allow for a more accurate assessment of the diet than simple evaluationof the diet offered. There is a step-by-step guide to conducting an intakestudy in Appendix I.Intake studies are run over a period of time, during which all the food itemsoffered to the study animals are weighed at the time of preparation. All foodremaining at the end of a feeding period is carefully collected and alsoweighed. Some means of adjusting for changes in moisture content of theuneaten food should be used, and this is achieved either by drying feed andremains samples to a constant weight in a drying oven or through the use ofa dummy or control dish to assess changes in moisture content in the actualclimatic conditions experienced. Although the first method is more accurate,it can often prove impractical for large volumes of food. Adjusted weights ofremains, subtracted from original weights fed, produce an estimate of feedintake. These values are then averaged over the length of the diet trial. Thenutritional composition of the diet is typically estimated from data on thenutrient composition of individual items in the diet and is therefore only asaccurate as the data upon which the estimate is based. Hence from weighingfeeds and collection of remains and the use of food tables/diet managementsoftware it is possible to arrive at an estimate of nutrient composition.Care must be taken to ensure the correct units are used, particularly whenthe use of a conversion factor is required. Comparisons between the intakesof different groups are best made on the basis of total group body weight,rather than by the number of animals in each group.-6-

Nutrition and Diet EvaluationThe data provide estimates of what is fed, and more importantly, what isbeing consumed. Comparing these data sets may reveal whether nutritionalinadequacies apparent in the diet consumed are due to the animals' choice offood items, or because the diet provided is inadequate. A simple index ofpreference is also useful, whereby food items are categorized into low,neutral or high palatability according to the percentage eaten.b. Diet evaluation by focal animal observationsWhile attending to the animals' welfare, evaluating the diets of species intheir natural groupings is an unreliable means of estimating their individualintakes. It relies on the assumption that every animal in a group eats anidentical amount and that no single animal eats all of one food item. Factorssuch as age, sex, grouping, dominance status and breeding origin may affectfood choices made by individuals and direct behavioural observations madeat feeding times can provide valuable information about these choices.To calculate individual feed intake through direct observation it is necessaryto know the average piece size of each food type and then to count howmany pieces of each food are consumed. This is relatively straightforward forsome species e.g. macaques that are fed discrete food items, such as pelletsand pieces of fruit or vegetables, which can be easily weighed (e.g. seePlowman, 2008). However, for other animals that are provided with large ornon-discrete items, such as browse or exudates, or have access to paddockgrazing/browsing in their enclosure it is necessary to estimate bite size. Forbrowsers, average bite size can be determined by providing a known-weightof browse, counting the number of bites taken then removing the browse andre-weighing. This is likely to be affected by type of browse, relative leaf/twigratio (e.g. Shipley et al.,1999) and individual animal (especially size) soaverage bite size needs to be determined over a range of these conditionsand preferably for each individual animal if feasible. For grazers, bite sizedetermination is also affected by sward type and structure (e.g. Burlinson etal., 1991) and ideally should be estimated in a similar fashion for browsers(above) using a sward sample in a movable container. If this is not feasible itcan be estimated by the hand-plucking method (e.g. Vries, 1995).This type of study provides much more detailed information about individualfood preferences and consumption than can normally be obtained by anintake study (a. above). However it is extremely time-consuming; observingonly one animal per feeding session will require a long period of time over-7-

Nutrition and Diet Evaluationwhich to establish a reliable data set if group size is large. Nevertheless it isimportant to consider this type of diet evaluation in situations where theremay be strong social effects in group-living species, leading tomonopolization of palatable food items by one or a small number ofdominant individuals. Attempts to ensure equal access of all individuals toall food types include feeding groups more food than is actually required(these can lead to dominant individuals becoming overweight), and providingfood in more than one location. However, short of separating animals atfeeding times it is extremely difficult to guarantee that every animal willobtain its 'fair' share.Evaluation by this method also allows the determination of the contributionof enclosure vegetation to the diet. Zoo enclosures are becoming increasinglymore naturalistic and contain more vegetation than in the past. Some of thisvegetation may have been chosen deliberately because it is palatable to theanimals and some deliberately because it is not palatable. If animals eatsignificant amounts of enclosure vegetation this will have an impact on thenutrient composition of their diet and also needs to be taken into account.There is no way to do this other than by focal animal observation and eventhen it is still difficult to reliably estimate weight of food eaten due tovariation in bite size (see above).c. Diet evaluation by survey and reviewThis technique is increasingly used by EEPs/TAGs to gather diet data for asingle species across a range of zoos. If done well it can provide goodinformation on nutrient contents of diets which can be compared withbreeding and health records across institutions. Given the lack of data onwild diets for many exotic species this is often the only way to determinedietary recommendations (see section 5 below) However, this method hasmajor limitations in that the quality of data returned is usually not goodenough to do anything with at all. This is largely due to poor survey methodswhich do not make clear what information is required; but also because toprovide the level of information needed requires quite a lot of effort on thepart of the respondent.To be of any real use, diet survey respondents must give the weight of eachindividual food item offered to the animal/group of animals each day over aperiod of time. This period should be long enough to incorporate all theregular variations in the daily diet – typically a week in many zoos. Other-8-

Nutrition and Diet Evaluationinformation required includes the numbers, sexes and ages of animals in thegroup and their weights if known. The manufacturer and product name ofany manufactured feeds must be included. To ensure that all this informationis received the surveyor must ensure that the questionnaire they send out isvery precise, clear, and easy to complete (see Zoo Research Guidelines:Questionnaires for further advice on how to conduct research byquestionnaire). An example of a good diet survey questionnaire is included inAppendix II.Even if a large number of institutions respond to a survey with excellentinformation it must be remembered that, unlike the above methods, thistechnique only tells us the diet actually offered - not what is actuallyconsumed. As previously stated, these can differ quite substantially but it isunrealistic to expect many zoos to provide accurate actual consumption data.The best way to acquire these data from many zoos is to actually visit themand collect it yourself.3. Feed composition and nutrient calculation toolsHaving recorded the ingredients in a diet, along with quantities fed andconsumed, these data must then be combined with information about thenutrient composition of each ingredient to provide a nutritional summary ofthe diet. Various food tables are available in print and online, from which therelevant nutrient values can be extracted and copied into a spreadsheet toperform the calculations. Alternatively, Zootrition software is acomprehensive electronic database with the facility to compare nutritionalcontent of specific food items and calculate overall nutritional composition ofdiets.-9-

Nutrition and Diet EvaluationTable 1 Common zoo feed categories and the best sources ofcomposition informationFruit, Veg,Nuts, SeedsAZA’s Nutrition AdvisoryGroup (NAG) websiteForageWholeprey9192Manufacturer’s GuaranteedAnalysisPrepared feeds/ supplements9McCance & Widdowsons’Composition of Foods9Peer-reviewed literature9USDA Nutrient Database9ZOOTRITION 993999399See NAG Technical Paper 6: Hay and pellet ratios: considerations in feeding ungulates. 2SeeUSDA Publication “Nutrient composition of whole vertebrate prey (excluding fish) fed inzoos”. 3Contains data for meat, fish and shellfish products as used for human consumption(e.g. fish fillets, rather than whole fish).1The Royal Society of Chemistry publishes “McCance and Widdowsons’ TheComposition of Foods (2002), now in its 6th Summary Edition. This is arevised and updated set of official UK food tables, covering more than 1100different types of food. The nutrient coverage of this edition has beenextended to include selenium, manganese, iodine and fatty acids. The textalso covers proximates, vitamins, inorganics and lists scientific names forfoods.The United States Department of Agriculture (USDA) are responsible forcompiling equivalent food tables in North America and their NutrientDatabase is available to search online. The web address is for the home pageof their Agriculture Research Service Nutrient Data Laboratory, which hasother useful information in addition to the Nutrient Database link.(www.nal.usda.gov/fnic/foodcomp/)ZOOTRITION features: Over 3000 feedstuffs with published nutrient values, many unique to zooand wildlife species. Comprehensive information covering nutrient recommendations for USdomestic and zoo species, the latter largely based on AZA Species SurvivalPlan and Taxon Advisory Group nutrient specifications and- 10 -

Nutrition and Diet Evaluationrecommendations for several species are from international husbandryguidelines. Energetics calculator to estimate energy needs based on animaltaxonomy, food habits, physiological stage, and activity levels. Only data from peer-reviewed sources are considered for inclusion in the‘locked’ global database embedded within Zootrition. However, most ofthe information on feedstuffs and nutrient recommendations is still ofNorth American origin.When using either published tables or software programmes as nutrientcalculation tools, be aware the final nutrient values are calculated, notmeasured. In all instances, missing data (indicated by a dash ‘-‘) do notindicate the nutrient is not present. It simply means the specific content forthe feed was not assayed and there are no available data. Thus totals for adiet or combination of feeds should b

successfully in captivity, yet animal nutrition is a new and relatively unexplored field. Part of the problem is a lack of facilities in zoological institutions and a lack of expertise. There is, thus, a strong need to develop nutritional studies and departments in zoological institutions. Research on nutrition is carried out both as a problem-solving exercise (in relation to ill-health or .

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