RHINOCEROS RESEARCH MASTERPLAN 2019cincinnatizoo.org/system/assets/uploads/2019/08/... · 2019 Rhino Taxon Advisory Group Research Masterplan 4 Primary Research Priorities I. Impact

2019 Rhino Taxon Advisory Group Research Masterplan

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RHINOCEROS RESEARCH MASTERPLAN

2019

Association of Zoos and Aquariums

Rhinoceros Taxon Advisory Group

Rhino Research Council


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Introduction Rhino Research Council

The Rhino Research Council is comprised of advisors with extensive rhino expertise in one of

the following: genetics, nutrition, health, reproduction, behavior & ecology, and management.

Each advisor selects several co-advisors to assist them with their area of responsibility. The

AZA Rhino TAG’s Research Council is charged with developing a five-year Masterplan that

encompasses research priorities for rhinos and prioritizes those challenges deemed most

important to address during the life of the Masterplan (typically five years).

Masterplan Goals

The Rhino Research Council Masterplan document serves several purposes. First, because much

of the information shared and discussed by the Rhino Research Council is from studies that have

yet to be published, the Masterplan provides a summary of the most recent progress in rhino

research and some of the newest theories about the challenges still facing rhinos. Second, the

Masterplan is an excellent source of reference when the Rhino TAG is asked to endorse

proposed research projects. Finally, the priorities in the Masterplan help guide the International

Rhino Foundation’s targeted request for proposals.

Over the years, the philosophy guiding the Rhino Research Masterplan has shifted. When first

formed, the Rhino Research Council focused only on research priorities for ex situ rhinos. The

rationale for this focused approach was that the Council was a part of the Rhino TAG and

therefore, its role should be to assist the TAG with ex situ rhino management issues. However,

during the development of the 2004 Masterplan, there was much discussion about the integration

of both ex situ and in situ rhino research priorities into the document. That year, studies on wild

rhinos that could shed light on problems in the ex situ population were included. During the 2009

Masterplan meeting, we took a step further and embraced research priorities for wild rhinos as part

of our responsibility realizing that what we were calling “wild” rhinos were really minimally

managed rhino populations since all rhinos are now managed to some extent. Finally, in the 2014

Masterplan, an attempt was made to fully integrate the research needs for minimally managed and

intensively managed rhino populations in Africa, Asia and our zoos. We attempted to do the same

for the 2019 Masterplan. In contrast to 2014, when the top research priorities were in situ based,

the 2019 votes were primarily for challenges impacting rhinos managed ex situ.


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2019 Rhino Research Council

Chair: Dr. Terri Roth, Cincinnati Zoo/CREW, OH

TAG Chair: Adam Eyres, Fossil Rim Wildlife Center, TX

Health (Veterinary Medicine)

Advisor: Dr. Michele Miller, Stellenbosch University, South Africa

Advisor: Dr. Eric Miller, St. Louis Zoo, MO

Co-Advisor: Dr. Benn Bryant, Taronga Conservation Society, Australia

Co-Advisor: Dr. Robin Radcliffe, Cornell University, NY

Co-Advisor: Dr. Beth Hammond, Lion Country Safari, FL

Nutrition

Advisor: Dr. Katie Sullivan, Disney’s Animals, Science and Environment, FL

Co-Advisor Dr. Marcus Clauss, University of Zurich, Switzerland

Co-Advisor Dr. Ellen Dierenfeld, Ellen S. Dierenfeld LLC, MO

Co-Advisor Kerrin Grant, The Wildlife Center, NM

Co-Advisor Barbara Henry, Cincinnati Zoo, OH

Co-Advisor Dr. Eduardo V. Valdes, Disney’s Animals, Science and Environment, FL

Genetics

Advisor: Dr. Peter J. van Coeverden de Groot, Queens University, Canada

Co-Advisor: Dr. Alfred Roca, University of Illinois, IL

Co-Advisor: Dr. James Austin, University of Florida, FL

Co-Advisor: Dr. Candace Scott, Queens University, Canada

Behavior and Ecology

Advisor: Dr. Lara Metrione, SE Zoo Alliance for Reproduction & Conservation, FL

Co-Advisor: Dr. Elizabeth Freeman, George Mason University, VA

Co-Advisor: Dr. Rachel Santymire, Lincoln Park Zoo, IL

Reproduction

Advisor: Dr. Monica Stoops, Cincinnati Zoo/CREW, OH

Co-Advisor: Dr. Justine O’Brien, Taronga Zoo, Australia

Co-Advisor: Dr. Linda Penfold, SE Zoo Alliance for Reproduction & Conservation, FL

Co-Advisor: Dr. Parker Pennington, San Diego Zoo Global/ICR, CA

Co-Advisor: Dr. Mandi Schook, Disney’s Animal Programs, FL

Co-Advisor: Dr. Jessye Wojtusik, Cincinnati Zoo/CREW, OH

Management

Advisor: Adam Eyres, Fossil Rim Wildlife Center, TX

Co-Advisor: Lance Aubrey, San Diego Zoo Global, CA (Black rhino, GOH)

Co-Advisor: Paul Reinhart, Cincinnati Zoo, OH (Sumatran rhino)

Co-Advisor: Randy Rieches, San Diego Zoo Global, CA (White rhino, GOH)

Co-Advisor: Lisa Smith, Buffalo Zoo, NY (Black rhino)

Co-Advisor: Clarice Brewer, White Oak Conservation Center, FL


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Primary Research Priorities

I. Impact of and control over body condition/weight

➢ Reproductive dysfunction

➢ Foot/joint problems

➢ Phytoestrogen relationship

➢ Diet composition and variety versus nutrient composition impact

➢ Health/disease impacts

➢ Overall effect on well-being

➢ How to best monitor/measure and alter to improve animal well-being

II. Iron overload in browsing rhinos

➢ Epidemiological review – what really is the significance/prevalence of IOD?

➢ Best biomarkers for detecting, monitoring and assessing condition or treatment

➢ Organ iron accumulation versus organ damage

➢ Association with other health issues (is it primary or secondary?)

➢ Interaction with other micro-nutrients

III. Understand/address early and late stage reproductive dysfunction

➢ Impact of over-conditioning

➢ Cause of stillbirths/pregnancy loss

➢ Why so much cyclic dysfunction (silent estrus, acyclicity, anovulation)?

IV. Investigate behavioral and environmental factors that affect rhino well-being

➢ Ex situ health, body condition, reproduction, socialization, enriched

environment

➢ In situ impact of factors like dehorning and traumatic injury recovery


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Introduction

It is important to note that these four research priorities are not ranked in order of importance.

They were simply the research topics receiving the four highest vote counts from RRC advisors

and SSP Chairs. It is also worth noting that there is a lot of overlap among these priorities.

I. Impact of and control over body condition/weight

As with most animals managed ex situ, rhinos typically receive more nutrient dense diets and

get less exercise than they do in situ. Not surprisingly, these conditions lead to over-

conditioning and obesity, which probably impact rhinos in the same way they do people and

pets. Just how much obesity accounts for health, reproductive and well-being issues and/or

what physiological pathways are involved are not known, so research in these areas would be

very enlightening and could contribute to future management/husbandry recommendations.

II. Iron overload in browsing rhinos

Research on the prevalence and significance of iron overload disorder (IOD) in browsing

rhinos continues to be a priority. It is generally accepted that many browsing rhinos exhibit

high levels of serum iron saturation and accumulate iron deposits in organ tissues

(hemosiderosis) that are noted post-mortem. However, the relationship between iron loading

and disease appears complex, as it is not yet known if hemosiderosis is directly related to

iron overload disease or is occurring in response to other disease states (i.e., dental disease,

metabolic disturbance) or vice versa. It is also unclear if the iron deposits are actually

damaging the organ tissue since iron overload is rarely identified as the primary cause of

death in black rhinos. Recent studies have brought to question some of the previous held

beliefs about monitoring IOD progression in rhinos, and new biomarkers/approaches may be

necessary for future studies. Further work is also needed in determining the bioavailability

and impact of dietary iron on IOD progression.

III. Understand/address early and late stage reproductive dysfunction

Although reproductive challenges in white and GOH rhinos continue to rise to the top of the

research priority list, there has been significant progress in recent years. There is a better

understanding of acyclicity in white rhinos and treatments for overcoming it. Additionally, a

dietary change that reduces phytoestrogen intake has been associated with resumed

reproductive success in some white rhinos. Furthermore, artificial insemination is proving to

be a useful tool for some rhinos that are not breeding naturally. However, the incidence of

anovulation in all rhino species is much higher than previously realized and the cause is

unknown. Early embryonic loss and stillbirths continue to plague rhino reproductive success.

IV. Investigate behavioral and environmental factors that affect rhino well-being

Animal welfare and well-being are of utmost importance to animal managers caring for

individuals in zoos, on private ranches or in sanctuaries/national parks. Although rhino

welfare issues differ among these environments, in all cases, the goal is to provide the best

possible conditions for the rhinos. Ex situ, the challenge is to provide an environment that is

as stimulating as that rhinos would experience in their native habitat during good times to the

greatest extent possible. In situ, the challenges range from identifying factors that impact

translocation success to understanding how to lessen the trauma and facilitate the recovery of

rhinos either suffering from poaching injuries or orphaned as calves so that they can resume a

productive, natural life with their counterparts in their native habitat. Identifying how to

achieve these well-being goals requires evidence-based scientific study.


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Detailed Areas in Need of Research Within Each Discipline

Health (Submitted by the Veterinary Advisory Group)

Goal: AZA Research initiatives of member institutions ideally should support global

conservation programs with a focus on preserving the species in their native habitats.

ASIAN RHINO HEALTH

1. Sumatran and Javan rhinos

1.1.Disease risk analyses

1.1.1. Translocations within and between range states, or from abroad present risks for

introduction of disease with or to introduced rhinos, especially if animals have

been held ex situ for some time. Research into diseases that present potential risk

to rhinos, logistically appropriate diagnostic techniques for screening, and

incorporation into protocols should be considered part of the risk analysis and

translocation process.

1.1.2. Disease prevention in and around sanctuaries and protected areas – particularly

the role of hemoparasites (trypanosomes) and tick-borne diseases (Babesia,

Theileria and Anaplasma) and other transmissible infectious diseases that can be

spread from domestic animals to sympatric rhinos. This should include

development of effective quarantine and biosecurity protocols to prevent, detect,

and treat these diseases. Vaccination for tetanus should also be evaluated for this

species. This topic was recently highlighted by the IUCN Asian Rhino Specialist

Group as a very important area of study given the small populations for both

Sumatran and Javan rhinos that are located in areas often completely surrounded

by humans and animal-based agriculture.


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1.2. Iron overload disorder in Asian browsing rhino species. There should be a thorough

pathology review of all ex situ rhino deaths for Sumatrans to better understand historical

extent and impact of confinement and stress in this syndrome. In addition, further

research on specific biomarkers for diagnosis are needed.

1.3.Nutrition of Asian rhino browsers

1.3.1 Comprehensive study of sanctuary rhino browse selections that have supported

rhino health and reproduction to be used as a template for other developing

sanctuaries to follow.

1.3.2 Are there any problems with ex situ diets due to low browse selection and

availability?

1.3.3 Studies on browse nutrient degradation related to varied collection, handling and

storage protocols employed at zoos and sanctuaries.

1.4.Protocol development

1.4.1. Animal movements among institutions (successful case examples are available

from work done in preparation for movement of two male rhinos from the U.S.

back to Indonesia; i.e. government requirements, vaccination strategies, etc.)

1.4.2. Capture and translocation protocols in situ

1.4.3. General anesthesia protocols in-situ (based on available drugs). Currently

available data within range countries should be compiled and reviewed to

continue improving protocols. Research on the use of short and long-acting

tranquilizers is also needed for Sumatran rhinos.

1.5. Establish normal biological and health parameters for ex situ and wild animals

(some work from SRS recently published, but need more normal data from truly

wild rhinos to better understand health of ex situ animals or those moved for

future translocation programs)

2. Greater one-horned (GOH) rhino

2.1 Obesity/body condition index – Overconditioning in ex situ rhinos (all species) leads to

a multitude of health problems including musculoskeletal, foot, and reproductive

problems. This has particularly been associated with pododermatitis along with ex situ

husbandry conditions in this species of rhino. Research examining methods of

standardizing body condition scores for all rhino species, along with improved nutritional

management should be a priority for ex situ health. Existing species-specific rhino body

condition scoring systems should be used as starting points and continue to be revised.

2.2 Gastrointestinal and cardiovascular problems – may be underreported in this species.

These were significant contributors to adult mortality in a recently reported review of

necropsy reports. Research to investigate the presence of health issues and contributing

factors (nutrition, management, stress, etc.) should be considered for this species.

2.3 Disease risk analyses

2.3.1 Translocations within and between range states, or from abroad present risks for

introduction of disease with or to introduced rhinos, especially if animals have

been held ex situ for some time. Research into diseases that present potential risk


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to rhinos, logistically appropriate diagnostic techniques for screening, and

incorporation into protocols should be considered part of the risk analysis and

translocation process.

2.3.2 Disease prevention in and around sanctuaries and protected areas – particularly

the role of transmissible infectious diseases that can spread from domestic

animals to sympatric rhinos including mycobacteriosis. A recent case of

Mycobacterium orygis was discovered in a free-ranging GAOH in Nepal. Since

GOH live in high burden TB countries, further research should also investigate

human-animal diseases. Similar to the Sumatran and Javan rhinos, GAOH rhinos

are located in areas often completely surrounded by humans and animal-based

agriculture. In addition, vaccination for tetanus and possibly other diseases that

may be associated with trauma during capture, translocation or in sanctuaries

should be investigated.

AFRICAN RHINO HEALTH

1. African Black rhino - Many of the disease issues of black rhinos have been well studied and

it is clear there are significant challenges to keeping browsing rhinos in captivity. Certainly,

better strategies can and should be established to help alleviate these concerns and research

will be required to address these issues (iron overload disorder, (IOD); idiopathic

hemorrhagic vasculopathy syndrome, (IHVS); nutrition, husbandry, infectious disease, etc.).

1.1 Nutrition- is the inclusion of cereal based concentrates in rhino diets detrimental (all

rhinos)?

1.2 Quantifying stress (especially chronic stress) in rhino - develop laboratory and

behavioral markers. What is the health impact, for instance, on development of GI

ulcers, increased iron stores, etc. in both ex situ and recently captured free-ranging rhinos

(all rhinos)? New ideas include evaluation of neutrophil function using portable

luminometer and the analyses of five markers in fecal samples that include thyroid

hormones.

1.3 Iron overload disorder – Does iron overload cause disease in ex situ browser rhinos or

not (i.e., what is the clinical significance)? Does it increase susceptibility to other

diseases, such as infections? Determine what is the most useful marker (or suite of

markers) for iron overload; e.g., is rhino specific ferritin an accurate marker? Research is

needed to determine whether dietary and therapeutic interventions are effective or even

necessary, and if other methods such as stress/management play a role. What are the

clinical/laboratory indications for therapy? Is there a link between excessive body

condition &/or chronic stress and the presence of chronic inflammatory mediators in ex

situ rhino (as in humans and domestic horses)? Is it possible that there is a ‘rhino

metabolic syndrome’ characterized by insulin resistance and the chronic elaboration of

inflammatory mediators by adipocytes that contributes to excessive iron uptake (and

other black rhino diseases)? Current research is underway to investigate the role of the GI

microbiome in diseases of black rhino, particularly IOD. This is a high priority research

area for the ex situ population.


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1.4 Oro-dental health – Factors in development of dental disease in black rhino (all

rhinos?) and methods for treating and preventing routine dental issues need further

research. In a recent survey, only 29% of black rhinos in North American zoos have ever

had an oral examination under general anesthesia; all of these individuals had orodental

disease ranging from enamel points to severe periodontal disease and tooth loss.

Association of renal disease with orodental disease has been shown in other species.

Therefore, more information is needed, not just for browsers, but all rhino species. What

is the prevalence of periodontal (or general orodental) disease? More information is

needed to describe orodental abnormalities and determine the pathogenesis. What is the

association between browse feeding and dental health? Is the problem simply caused by

high starch ex situ diets or abnormal tooth wear and periodontal trauma due to

chewing/grinding inappropriate forage?

1.5 In addition to the causes of black rhino mortality, a superficial necrolytic dermatitis

(SND)-like disease continues to plague ex situ black rhinos affecting both their

appearance and welfare, though not necessarily causing mortality. In the past, data

suggested that SND affected as many as 50% of ex situ black rhinos. In one study, the

hypothesis that SND was caused by hypoaminoacidemia was refuted. Furthermore, there

was no positive correlation between corticoid levels and animals with lesions vs. those

without lesions. Animals without lesions exhibited higher mean corticoid levels over

time than animals with lesions. Disease could be immune-mediated.

1.6 Renal disease – Necropsy reports on ex situ black rhinos in the U.S. have shown an

increased number of cases of renal disease as a cause of death with secondary tissue

mineralization. Does renal disease predispose to hypertension with secondary

arterio/atherosclerosis and possibly other health issues such as intermittent epistaxis? Is

there an age or dietary association with development of renal disease? Ante-mortem

bloodwork and urinalysis has not typically been useful diagnostically. Research to

investigate predisposing factors, including diet, and improved methods for early

diagnosis and intervention may benefit the ex situ population. In addition, relationship to

other health syndromes should be re-examined.

1.7 IHVS, idiopathic epistaxis – Cases are becoming rare, however, the death of a 4-year-

old black rhino was attributed to IVHS in 2014. When they occur, there is no obvious

etiology or consistently successful treatment. Idiopathic syndromes in black rhinos

should continue to receive research attention, including the role of stress, nutrition,

management, and infectious diseases.

1.8 Tuberculosis - Mycobacteriosis (TB) caused by Mycobacterium bovis has been

diagnosed in both free-ranging black rhinos and individuals on private game reserves in

South Africa. This has led to quarantine and restrictions on future translocations. Further

research is needed to develop accurate antemortem diagnostic tests, investigate the

epidemiology, perform risk assessments and create management and control plans.


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1.9 Management of poacher injured and orphaned rhinos in range countries - Recent

development of treatment options for poaching injuries have been implemented in South

Africa but additional research is needed to improve pain management, infection control,

and accelerate wound healing. Development of prognostic indicators would be very

useful to best utilize limited resources. In addition, research into orphan rhino care

including diet/formula and management (level of human contact, use of surrogate dams)

are needed to develop evidence-based protocols.

Ex situ Black Rhino Mortality (primary cause of death)

Note: does not represent co-morbidities

Australia 2000-2017 (n=11) North America 2008-2017 (n=28)

0% infectious disease 14% infectious disease – Clostridial

0% IHVS/hemolytic anemia 4% IHVS/hemolytic anemia

0% renal disease 28% renal disease

19% stillbirth/weak calf 11% stillbirth/weak calf/congenital anomalies

0% trauma 0% trauma

0% unknown 14% unknown

9% cardiovascular/vasculitis 4% cardiovascular/vasculitis

27% GI 7% GI – colonic rupture, splenic mass rupture

9% neoplasia (uterine adenocarcinoma) 7% neoplasia (uterine, thyroid adenocarcinoma)

9% liver disease 4% liver disease (iron overload disorder)

9% multisystemic disease 7% multisystemic disease

18% orodental disease 0% orodental disease

2. African white rhino

2.1 Idiopathic neurologic disease – Several white rhinos have exhibited episodic

tremors/ataxia in Australia and sporadic cases have been observed in white rhinos in

Kruger National Park. Survey of neurologic disease in white rhino would be worthwhile

to determine if this is an underreported condition.

2.2 Neosporosis – This infection has resulted in abortion and acute death in white rhinos. An

experimental ELISA is available. Research to determine seroprevalence may reveal if

this is a widespread issue for reproductive or potentially systemic health problems.

2.3 Gastrointestinal health issues – Investigation into potential factors such as

diet/nutrition, management, infectious diseases and stress to determine if

morbidity/mortality may be prevented. Anecdotal reports of Clostridial enterotoxemia

exist in white rhinos. Research into the predisposing factors, treatment and prevention by

use of available vaccines is needed.

2.4 Pharmacokinetics/dynamics of commonly used antibiotics and analgesics (all

rhinos) – Little scientific work has been done in rhinoceros species on therapeutic drugs,

especially on use of non-steroidal anti-inflammatory drugs. Empirical use may be


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inadequate, or potentially cause adverse effects. With the advent of husbandry training

and restraint devices, sample collection is now possible for these types of studies.

2.5 Management of poacher injured and orphaned rhinos in range countries - Similar to

the black rhino, there is an urgent need for research into effective management of

poaching injuries and orphan rhino care.


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Nutrition Compiled by the 2018 Rhino Research Council Nutrition Advisors

There have been about 36 peer-reviewed studies on rhino diets and nutrition related factors over

the last 5 years. Overall, rhino nutrition investigations have been geared towards understanding

dietary nutrients as they impact serum nutrients in relation to health, characterizing practical diet

needs under human care, characterizing consumption in wild areas, and how nutrients and diet

can impact wellness and ultimately welfare. The general consensus remains the same as in 2009

and 2014 - the highest priority nutritional studies would be those in association with potential

causes of rhino disease (in particular iron overload disorder) and/or reproductive problems.

1. Iron values (Iron Overload Disorder - IOD) – epidemiological information on diets, iron

parameters and health serum markers, and current health status would be useful to re-evaluate

the population since it has been many years since the original data were generated and many

dietary/management changes have been implemented.

What has been done: 14 publications

Current Need:

1.1 Identifying and improving diagnostics of IOD

1.2 Documenting diet ingredient impact on iron loading vs. inflammation (ie. alfalfa, forms

of phosphorus)

1.3 What is the upper limit for total dietary iron concentrations vs. pelleted diet iron? (< 300

ppm Fe currently recommended)

1.4 Understanding of antioxidant dietary impacts on iron loading (supplemented antioxidant /

anti-inflammatory in the form of vitamins, omega 3’s, etc.)

1.5 Potential treatment options – investigating safety and efficacy of natural chelators, as

well as long term large volume phlebotomy and their impacts on health

1.6 Iron loading in relation to pregnancy – potentially an epidemiological review


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Research being conducted / in progress:

A. Water contributions to iron load across institutions – Michigan State University (M.S.

study)

B. Iron related genetic mutations in relation to adenosine –San Diego Zoo Institute for

Conservation Research

C. Role of Gut Microbiota in Health and Disease Sensitivity of the Black Rhinoceros-

Smithsonian’s National Zoo, Center for Species Survival

D. Black rhino specific ferritin assay being tested - University of Florida

E. Examining oxidant stress in black rhino in relation to IOD potential measurements –

University of Florida

F. Labile plasma iron and miRNAs as potential diagnostic markers of IOD – Cincinnati

Zoo’s Center for Conservation and Research of Endangered Wildlife

2. Diet studies – several ongoing and relating to potential health hazards and impacts of diet

items / nutrient levels.

What has been done: 15 publications (including documentation of free-ranging animal nutrient

intakes in GOH, Sumatran and Javan species)

Current needs:

2.1 Continue monitoring serum versus dietary vitamin E (primarily in black rhinos, but

potentially in white rhinos as well due to obesity / reproduction concerns)

2.2 Establish serum vitamin E ranges in current population using appropriate commercial

laboratories (black rhinos and across species)

2.3 Investigate interaction of diet / nutrient content (macro or micro) and validated

inflammatory markers in each species

2.4 Establish more suitable minimum dietary nutrient concentrations for animals during

maintenance, gestation and breeding. Are horse requirements adequate / appropriate as

guidelines?

2.5 Develop science-based recommendations regarding the use of alfalfa for each rhino

species. The energy, protein, calcium, and bioavailable iron content of alfalfa, as well as

its overuse in diets, may be a factor in health including potential renal, iron and obesity

issues.

2.6 Investigate the use of phytase, versus presence of phytic acid, in conjunction with

phosphorus supplement, to combat hypophosphatemia. Ex situ black rhinos have

demonstrated serum hypophosphatemia and hypercalcemia, likely in relation to diet.

Currently high levels of NaPO4 supplementation are required to achieve low normal

serum P values, but there is a paucity of data regarding phosphorus digestibility.

2.7 Investigate rhino microbiomes by utilizing fecal DNA / RNA analyses as a means of

defining gut populations in rhinos on particular diets under human care versus wild diets.

2.8 Re-evaluate serum nutrient profiles in wild rhino species considering current

methodologies to update historical “reference” values for vitamins, minerals, etc.


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Research being conducted / in progress:

A. Vitamin E in serum and fecal samples on varied supplementation of Emcelle for white and

black rhinoceros species – Disney’s Animal Kingdom

B. Vitamin D in serum across seasons in black rhinos – Blank Park Zoo

C. Gut microbiome analysis in white and black rhinos – San Diego Zoo Global, Smithsonian

Center for Species Survival and Disney’s Animal Kingdom

3. Oro-dental issues – A research priority under rhino health since it can lead to oral lesions and

loss of body condition, but its cause could be related to diet (nutritional and/or mechanical).

There have been several cases of rhinos requiring repeated teeth floating procedures, and much

discussion on dental concerns across species. Potential evaluation of sugar content and amount of

enrichment items may also be warranted.

Current Need:

3.1 Investigate epidemiological correlation of dietary nutrients and dental disease

3.2 Impact of browse on dental health (quantity / type/ etc.)

3.3 Investigate use of alfalfa in relation to dental disease

What has been done: 1 publication

4. Fatty acids – Fatty acid ratios differ significantly between ex situ and wild populations of

black rhinos. Types of fatty acids available from diet could be associated with changes in

immune function and/or diseases such as SND.

Current Need:

4.1 What are the appropriate dietary ratios of n-3: n-6 fatty acid? Supplements are being

used to increase linolenic acid, but what is the minimum amount to offer to provide

benefits? Documentation of benefits would potentially lead to diet re-formulations.

4.2 Interaction between fatty acids / composition of diet and inflammation

5. Trace and macro minerals – Recent investigations are ongoing, including copper which is

an important anti-oxidant that interacts with iron. The recent reported incidence of renal disease

in the population calls calcium and phosphorus content of diets into question.

Current Need:

5.1 Appropriate dietary concentrations of zinc and copper, considering their interactions, and

potential binding with phytate and other secondary compounds. Zinc and copper are also

sometimes being supplemented for hoof health.

5.2 Investigate macro minerals in total diet in relation to renal health

Research being conducted:

A. Copper versus iron absorption using new in vitro methodology using rhino fecals –

University of Ghent

6. Body condition / body weight – Another research area that has also been noted under rhino

health. Body condition scoring (BCS) systems have been established for African and Asian


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rhino species and should be distributed/utilized by researchers and animal managers. These

scoring systems are very subjective at each institution, so using standardized protocols should be

helpful. Furthermore, efforts to control for bias by sending images of animals from different

institutions to one person for evaluation was equally as difficult because of the variation in

animal appearance based on photo angle/quality. Perhaps physiological values for leptin,

glucose, insulin, etc., could contribute to a continuum of data for a more solid method of

evaluating body condition, in addition to computerized assessments, body measurements

matched with weights and/or characterizing body types in the process. Tracking weight in some

context in relation to dietary nutrients consumed (fiber versus starch as energy sources in

pelleted diets for example), while challenging, is important for understanding nutritional impacts

on health. This is an important issue since body condition has been suggested as a factor

involved in potential iron loading, disease, historically skewed sex ratio of calves and

reproductive issues of F1 white rhinos.


Current Need:

6.1 Determining markers of body condition beyond BCS, correlated with body weight and

other health assessment

6.2 Investigating use and tracking of the variable nutrient impact of dietary enrichments on

BC/BW across institutions

7. Influence of dietary phytoestrogens on reproductive success of white rhinos

There is continued concern that phytoestrogens may be impacting white rhino reproductive

success. Work has evaluated the estrogen activity in rhino feeds; alfalfa and soy are predictably

high but Sudan grass also demonstrated high activity. West Coast Bermuda hay was found low in

activity but it was high in Bermuda hay grown on the East Coast. Therefore, both types of hay

and growing locations/condition likely affect phytoestrogen content. Initial diet changes which

lowered phytoestrogens, but also may have lowered energy intake and impacted body condition,

have been associated with two successful births at San Diego Zoo Global (unpublished)

warranting further investigation.


Need:

7.1 Test impact of low phytoestrogen diet and examine in relation to nutrient changes / body

weight and body condition, as well as reproductive success.

Research being conducted/ in progress:

A. Survey of white rhino holding institutions for diet changes versus reproductive success in

recent years - SDZG

8. Science-based browse recommendations – There are often requests for browse lists. In 2014,

seasonal browse analyses were encouraged to help institutions make informed decisions about

browse choices. Studies exist on the diversity of browse chosen by wild rhinos as well as the

nutritional components of the browse. However, the fact is that most zoos will feed what browse


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they can get locally and with the least amount of cost/effort. Regional browse studies on

nutritional value and palatability of local species are helpful as general guides and exist for some

areas. Recent papers have illustrated diversity of browse intake in wild areas for Greater One

Horned (GOH) and Javan rhinos as well. Developing a published browse database by region

could be helpful, but not a priority. Institutions with black rhino are encouraged to develop

browse farms to help provide better diets for their animals and these regional studies are a great

reference when initiating a browse farm.


9. Influence of diet on calf sex ratios – no research published in 5 years

This issue was a priority in the past, but the skewed sex ratio appeared spurious hence this is not

considered a current research need.

10. Neonatal growth, milk composition, and assisted-rearing

This issue was a priority in the past, and while existing information should be compiled for

easier access, there has been previous work published describing and documenting these

processes under human care.


Rhino Nutrition-related References 2013-2018 (see below for citations)

Prepared September 2018

Topics: # of references

General Nutrition/Diets 13

Iron Overload Disorder 14

Inflammatory Markers 2

Phytoestrogens 2

Body condition scoring 2

Tooth-wear / dental issues 1

Neonatal growth/assist-rearing/milk composition 2

Total 36


17

Reproduction

RRC Reproductive Discipline

Since the 2014 Masterplan, a total of 49 new peer reviewed manuscripts and conference abstracts

have been published in the area of rhino reproduction. As the wealth of research continues to

grow so have advancements in addressing previously established reproductive priorities for ex

situ, minimally managed and wild rhino populations. For example, mate compatibility, courtship

and aggression previously emerged as a high priority issue for Asian rhinos but has since been

effectively addressed through rhino manager/keeper/veterinary education regarding acceptable

behaviors and utilization of established ovarian biomarkers associated with successful breeding

in these species. Some priority issues still remain but have shifted research focus from

physiologic origin to that stemming from a management need. Ex situ African black rhino

populations were once plagued by skewed natal sex ratio of excess male births compared to that

of female calves. While there is no longer significant skew among any of our ex situ rhino

populations having an even birth sex ratio means that 50% of calves are males. For species like

the African white rhino which excels in large herds with one bull and several cows this presents

a population management challenge. Using novel approaches such as establishing protocols for

housing/exhibiting multi-male groups and/or integrating sex sorted sperm into assisted

reproductive techniques (ART) appears warranted for addressing this issue.

Emerging priorities in the reproductive discipline have and continue to be heavily focused on the

female side of the rhino breeding pair. Not many over-arching issues are associated with male

rhino reproduction. However, an area that continues to surface is a means by which to empower

African male rhinos in natural breeding circumstances. The innate female dominant social

dynamic of these species represents a particular research/management challenge. Reliable and

repeatable methods of collecting and cryopreserving rhino semen have been established with

significant advances in utilizing the biomaterial to provide proof of concept and applicability to


18

population management. There is continued need to strategically bank sperm from individual

males and establish cooperatively managed rhino genome resource banks among institutions.

While not a top priority, developing chemical induction for semen collection could be especially

useful in establishing advanced ART such as IVM/IVF. This methodology could also increase

the ability to bank sperm from more individual males (ie, institutions that do not want to use

general anesthesia or are not comfortable with administering best practice anesthetic drugs for

given species).

The reproductive discipline and the scientists involved in developing and conducting the research

needed to meet priority goals could not do so without the assistance and input from rhino

management and animal care teams. Operant conditioning for different procedures and overall

structured management is key for rhino reproductive research success. The substantial data

collected to date has helped understand the normative reproductive characteristics among the

different rhino species and identify early and late stage reproductive dysfunctions. However,

determining how to prevent these dysfunctions and identifying methods to treat underlying

causes are of primary concern moving forward. A multidisciplinary research approach will

likely be most effective in addressing select issues like over-conditioning (i.e., impact on

reproduction but extends to disciplines of health, nutrition, management and behavior).

Whereas, natural breeding is and continues to be the primary means of maintaining and growing

our rhino populations, the need for ART should not be overlooked and remains important for

those critically endangered species or doomed populations. Tremendous strides have been made

in developing ART for some of our managed rhino species. However, compared to the closest

domestic relative the horse, rhino ART is lagging. Therefore, a priority focus must be placed on

the advancement of available ART at our disposal to meet the genetic and demographic needs of

managed breeding programs for African and Asian rhinos. Optimizing ART for rhino species

will facilitate reproductive goals for each population and ease genetic as well as general

management. From a welfare perspective it is important to establish the types of reproductive

assessments that should be done and at what life stages to ensure optimal breeding

recommendations are met for each species.

The five reproductive priorities that emerged for 2019 Rhino Research Masterplan are as

follows:

1) Understand and address early and late stage reproductive dysfunction in African and

Asian rhinos

1.1 Impact of over-conditioning on reproductive dynamics

1.2 Stillbirth; while stillbirths have occurred in both African species it has predominately

been an issue in GOH rhinos. With up-to-date stats, there may emerge changes and/or

protocols for expectant females that minimize occurrence.

1.3 Silent estrus

1.4 Acyclicity

1.5 Anovulation

1.6 Early pregnancy loss


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2) Conduct studies to improve AI success across rhino species and develop the in vitro

laboratory techniques necessary to save doomed populations/genetically valuable

individuals.

2.1 AI timing

2.2 Application of lower sperm numbers (to help conserve genetic material and to aid in the

use of sorted sperm in which lower sperm concentration is mandated)

2.3 In vitro maturation/fertilization (northern white and Sumatran rhinos)

3) Develop novel biomarkers of follicular development to enhance natural and assisted

reproductive efforts in African and Asian rhinos.

3.1 Estrogen detection. With the exception of GOH rhinos, there has not been a reliable

method of measuring estrogen and having it reflect physiologic state.

3.2 Impending ovulation (biomarkers such as behavior, specific hormone milieu and/or

ultrasound)

3.3 Anovulation vs. ovulation

4) Determine best practices for reproductive management of male and female rhinos with

respect to age, prior reproductive performance and genetic contribution

5) Develop ideal test paradigms for operant conditioning for reproductive procedures in

African and Asian rhinos - An improved understanding of rhino behavior as it relates to

training for specific reproductive procedures and how reproductive status may influence

behavioral response. Framework of established training responses, timeframes for adaptation

and to reach steady state behavior.

5.1 Males; hand injection of anesthetic drugs, manual semen collection

5.2 Females; blood collection, ultrasound, artificial insemination


20

Genetics

The potential benefits of employing genetic tools for studying rhino conservation and population

genetics are tremendous, but considerable research and development is still needed in many areas

before direct, efficient, reliable application will be possible. Areas where conservation genetics

could be most valuable include studies on: genetic structure among and diversity within

surviving populations; dispersal; paternity; and censusing. The potential methodologies would

include analyses of: mtDNA, microsatellites, genetic sexing, single-nucleotide polymorphism

(SNPs), immunogenetics and the microbiome.

Over the next five years, there needs to be an emphasis on developing the appropriate

methodologies:

1. Microsatellite analysis - Microsatellites are powerful if they can be amplified from fecal

DNA and should be a priority. However, fecal samples in the tropics deteriorate very

rapidly.

2. High throughput sequencing (HTS) – Since fecal DNA typing is difficult and genetic

diversity in some rhinoceros species is low, next generation sequencing (NGS) may be of

great value in providing the initial screening for polymorphisms needed to develop

markers to examine rhino population genetics. These sequences are being used to

assemble the genomes of all rhinoceros species.


21

3. Standardization – different labs using various loci are reporting different results. There

needs to be some consistency, or the techniques may wrongly be considered unreliable

for answering conservation/population questions.

4. Shorter amplicons (75-150 bp) need to be targeted by genetic markers and made

available for fecal DNA studies since degradation can occur rapidly in fecal material.

5. Fecal microbiome analysis may be useful in some cases as an alternative to analysis of

an individual’s own genetic material. Microbial DNA is more plentiful and in better

condition when excreted in the feces and each individual has their own microbial profile.

This was the methodology employed to determine that the fecal samples from the Viet

Nam Javan rhino were all from the same individual.

6. Immunogenetic variability and its relationship to disease resistance and mate choice

would be interesting to investigate. If pre-testing for mate compatibility or interest could

occur prior to animal transport, it would be very valuable to animal managers in zoos and

managed reserves/ranches. It may also be a means of ensuring that immune system

diversity is maintained in small populations.

7. Environmental DNA – eDNA is an emerging tool in genetic studies but may be more

challenging under tropical conditions just as fecal DNA is more challenging in samples

from tropical forests.

Ongoing Priority Genetic Studies

➢ Immunogenetic variability – analyses of the major histocompatibility complex and of

innate immune system genes such as Toll-like receptors are being planned and once

established will help significantly when analyzing populations for variation in genes that

have been under selection and may provide greater fitness when their diversity is maintained.

This is particularly important information for species that appear to have low neutral genetic

variation, for example the GOH and white rhinoceros.

➢ Fecal DNA analyses – Some work has been done, but the method needs to be optimized so

that individual genotypes from fecals can be used for census work and assessing and studying

breeding strategies, dispersal, etc. in reserves and parks. The analyses are especially needed

for the Javan and Sumatran rhinos since it is the only material available for analyzing

populations and learning more about animal numbers, locations, sex, relatedness, etc.

Progress has been made towards this goal, and newly developed markers have been used to

determine population structure in Sumatran rhinos, while the use of these markers on fecal

samples is being implemented in Indonesia for both species. A project to optimize non-

invasive genotyping from fecals as a tool for estimating numbers and sex of white and black

rhinos from the Kruger National Park has been initiated. Finally, a project optimizing fecal

genotyping of D. b. bicornis for census of the Etosha population is being launched.

➢ Study of disease history and parasite load - A critical emerging arena to which genetics

can contribute is disease risk assessment of different rhino populations. This is particularly


22

relevant in light of the tragic loss of five ex situ Sumatran rhino in Malaysia. A particular

avenue worth pursuing is PCR techniques that can determine presence and load of different

pathogens in wild populations from tissue collections and fecals. A major study investigating

these possibilities is proposed for the Kruger National Park for their C.s.simum and

D.b.minor populations.

➢ Social aggregation, dispersal, variance in reproductive success in extant D.b.bicornis

(Namibia) - Nearly 200 samples have been collected from D. b. bicornis in Etosha National

Park and Damaraland D.b. bicornis. With > 500 animals this is the largest population of

endemic black rhinos. The characterization of their genetic structure using 9 microsatellite

loci has been completed. Although there are suggestions that these animals cluster when

numbers are higher - which is counter to the popular notion that these animals are solitary -

genetic analyses at 9 microsatellite loci do not support any significant structure in this

population. This may be because so little time has passed since the numbers of this

population started to increase and a dispersal /genetic structure equilibrium may not have

been reached. Using more NGS-derived genetic markers, another look at this population 15

years hence would be instructive with respect to emerging genetic structure in this, the

largest black rhino population.

➢ Individual genetic variation – Such information should be examined and compared to help

guide breeding recommendations. Such genetic studies can verify assumptions made

regarding founder animals. Genetic markers can also be used to verify pedigree-based

analyses and identify genetically valuable individuals.


23

Behavior & Ecology

Priority Issue: Investigating behavioral and environmental factors that affect rhino

wellbeing

1. Developing and validating methods to assess wellbeing: Documentation and evaluation of

animal wellbeing is now a requirement of all AZA zoos, and WAZA encourages

considerations for wellbeing even among conservation activities executed in situ. Wellbeing

involves the promotion of positive states within five broad domains relevant to an animal’s

life, including nutrition, physical health, environment, behavior, and mental state. Within the

context of environment, behavior, and mental state, metrics for gauging whether an animal is

in a more positive state or a more negative state need to be developed, and the validity of

their application to rhinos and their appropriate interpretation in the context of rhino biology

and husbandry require investigation. Some research has already evaluated potential

associations between behavior, zoo environment, and successful (or unsuccessful)

reproduction. Evidence supporting a relationship between other endpoints of wellbeing (i.e.,

good (or poor) health or nutrition, a positive (or negative) mental state and specific

behaviors, environments, or measurable physiological parameters associated with

psychological states (e.g., inflammatory markers, acute phase proteins) needs to be built.

One assessment tool, WelfareTrak (by Brookfield Zoo), has been tested for its informative

ability regarding black rhino welfare.

2. Specific factors that might affect wellbeing that are a priority for research

2.1 Behavioral effects of dehorning – Dehorning rhinos is one option being used to curtail

the poaching epidemic. While dehorning itself is not painful and glucocorticoid

concentrations appear to return to pre-procedure values within a few days, the rhino’s

response to the sudden absence of the horn and how the rhino adapts behaviorally to that

physical change in the short- and long-term are not understood, especially related to


24

overall wellbeing. Sparring behavior, territorial defense, breeding success, and predator

defense are all key areas where the absence of a horn might alter wellbeing outcomes,

either positively or negatively.

2.2 Behavioral response to intensive medical care following traumatic injury and the re-

release process – Following traumatic injury, such as poaching attempts, some wild

rhinos are left in a grisly condition that requires many months of intensive medical care

and rehabilitation. During this time, the rhinos are exposed to stimuli they have never

experienced before and are isolated from natural stimuli and choices to which they were

accustomed. Although treatment is necessary for the rhino to survive and metrics of

health are monitored closely to determine the progress of recovery, how the rhino

responds behaviorally to this dramatic change in life conditions also should be monitored.

Behaviors that might be associated with distress or depressed mental state v. recovery

need to be described. Once rehabilitated and deemed healthy for release, the rhino’s

behavioral response to the release process needs to be monitored and evaluated to

determine its relevance to positive or negative states of wellbeing.

2.3 Behavioral response to management in bomas and during transport – During

translocation/reintroduction or quarantine for shipment, rhinos are kept in bomas.

Behavioral and physiological measures of wellbeing for rhinos in transition should be

developed. A scoring system has recently been developed for white rhinos in bomas

(Miller et al., 2016) and additional study would be useful, especially for the other species.

Non-invasive measures of the stress response other than glucocorticoids would be

valuable.

2.4 Ex situ meta-analyses across management practices, environments, etc. – The effects

of various demographic (e.g., age-sex ratios), social (e.g., group size and density), and

environmental characteristics of zoos (e.g., climate, enclosure characteristics) on

behavior, especially features that differ from what the species would experience in range

country and those that involve human interactions (e.g., enrichment, training and

education/guest-relations programs), need to be assessed from a wellbeing perspective for

rhino populations in managed care. Beyond population-level data, the meta-analysis

approach should make it possible to track changes in an individual rhino’s indices of

wellbeing as it moves from one facility to another, and to subsequently design controlled

experiments based on observed trends.

Additional Behavior and Ecology Research Needs

1. Understanding and managing small populations in the face of habitat fragmentation,

poaching, and climate change: In the shrinking habitat that exists for rhinos, whether a

viable population can sustain itself there is a concern, especially as environmental conditions

change over time and the poaching crisis continues.

1.1 Population viability and habitat suitability – This is a very active area of research for

all rhino species; much has been learned, but many questions need additional study.

Assessments in each subpopulation and habitat pocket are needed to determine which


25

subpopulations are sources and which are sinks. New research in this area can take

advantage of techniques developed in other fields, including dung DNA analysis and

fecal hormone metabolite analysis for determination of pregnancy status, to improve

survey methods. Advancements in the use of recently-developed technologies, including

drones and on-animal accelerometers, might improve managers’ abilities to monitor rhino

populations. How the source versus sink fate of a given subpopulation or how the long-

term viability of a habitat pocket will affect the status of the larger metapopulation is

crucial to understand. The reasons for subpopulation growth or decline over time and for

changes in habitat suitability across seasons need to be better understood in order to

manipulate the outcome. In addition, a deeper understanding of how reproduction and

population growth are affected by population density and spatial distribution, limitations

on sub-adult dispersal, social constraints such as mate choice and dominance, and

environmental variability and climate shifts are needed. Key factors that promote adult

and sub-adult survival to maximize fecundity also need to be identified.

1.2 Corridor habitat – Assessments to determine key positive and negative attributes that

affect rhino dispersal between subpopulations are needed. Habitat and climate changes

over time need to be modeled to determine where corridors will be needed in the future.

1.3 Habitat restoration and management – Habitat restoration and management can

improve conservation outcomes, but the techniques themselves require further research.

How to control and eliminate the invasive plant Mikania micrantha in GOH rhino habitat,

and how to prevent or mitigate desertification and habitat water loss in Africa are of

particular concern. The effects of fire and changes in megaherbivore densities, including

the potential loss of rhino populations, on savanna ecosystems need additional study.

Recent studies have looked at the potential effects of competition between rhinos and

other megaherbivores, such as elephants and the Javan bull (banteng), on habitat quality,

plant food availability, and habitat carrying capacity. Additional studies of inter-species

interactions, especially in marginal and human-modified habitats, would be very useful.

2. Determining behavioral and ecological factors that result in successful

translocation/reintroduction: While many of the challenges noted above for small

populations apply to situations in which groups of rhinos or individuals are

translocated/reintroduced, there are some additional considerations specific to this situation

that require further study.

2.1 Meta-analyses of data on translocation attempts and outcomes – As a first step,

accessibility to this type of data needs to be improved in such a way that the safety of the

rhinos is not compromised. Meta-analysis could provide insights about how individual

rhino characteristics and specific release area characteristics might influence the outcome

of the attempt.

2.2 Dispersal – How much and where rhinos move immediately after release, and what

influences that movement are important areas of study. Recent research looking at what

types of information rhinos communicate to each other through olfaction and vocalization


26

might provide new opportunities for manipulating and improving translocation and

reintroduction.

2.3 Territoriality and breeding success – Factors that influence which bulls establish a

territory and secure reproductive opportunities need to be better understood. This area of

study might benefit from developing methods for small reserve managers to document

social interactions.

2.4 Sub-adults – As more calves are orphaned by poaching, successful reintroduction

techniques for sub-adults after hand-rearing are essential. Important areas of study

include the impact of boma location, the effects of age upon release, predator and

territorial bull avoidance, and social and environmental factors that influence early home

range establishment.


27

Management

Priority Issue: Determine management aspects that can enhance areas of rhino conservation

within the ex situ facilities and potential benefits to wild rhinos.

1. Optimize/Standardize Body Condition Scoring - Whereas, this is a veterinary/nutrition

focus, the caretakers of the animals should all be aware of the different scales and what the

scores mean in relation to their charges. Management will be affected by low and high

scores, especially in regard to breeding success. There are several systems—mainly based on

a five-point scale, for rhinos.

2. Determine factors contributing to success or failure of male/male housing or bachelor

groups - There are more and more opportunities to house two young males together in

traditional zoo settings, and larger groups of males in larger landscape facilities. The

International Rhino Keepers Association (IRKA) is interested in helping to do the research

with respect to ages, size of exhibit, exhibit design/layout, factors that have been known to

help, issues to avoid, etc. to determine if/when this can work and for how long we should

expect successful groupings to remain in-tact.

3. Determine success/failure in pairing older animals with younger animals - There are

many situations in which one of two companion animals has died and the SSP wants to pair a

younger animal with that lone animal. The IRKA is also interested in looking into the

success of those introductions over the past years since this will most likely need to continue

into the future as pairs become lone animals. Ultimately, when the other older animal dies, it

would then make sense to put another compatibly aged animal with the lone animal.


28

4. Conditioning recommendations to help with other RRC needs.

4.1 Ultrasonography, phlebotomy, oral exams, etc. These behaviors have already been

conditioned in many rhinos throughout the US and Europe. Perhaps the Rhino Research

Council (in conjunction with the TAG and SSP Coordinators) should encourssage a more

formalized training/conditioning program that is focused on the aspects that each of the

RRC disciplines might require. A compilation of best practices could be useful.

4.2 Are there other training opportunities that don’t fall under ‘medical, scientific research’

headings? Blood collection will be valuable for the genetics discipline, as well as

nutrition and reproduction. Ultrasound conditioning is obviously beneficial to the

reproductive research. Collection of urine and saliva could be beneficial for various

research projects, and of course, fecal collections which require no training. Scale

training, especially for those with portable scales, could be important for BCS

monitoring.

5. Can research be done in captivity that helps the wild populations? There has been some

opportunity for collecting measurements on ex situ rhinos to help with collar fitting in the

wild. Is there more opportunity for this type of work?

6. Management decisions that affect the potential to be a release candidate - This was

something discussed in 2013, regarding the ex situ animals being an ‘assurance’ population

for potential reintroduction. It is doubtful there will be large scale reintroductions in the near

future, but could managers participate in determining what makes a ‘good’ candidate for

reintroduction? This would go hand in hand with some of the needs described in the

behavior and ecology section.

7. Collection management styles

7.1 Exhibit strengths and weaknesses for non-breeding animals

7.2 Exhibit strengths and weaknesses for breeding groups

7.3 Conditioning for moves—crate training, facility design, factors that improve moves

7.4 Determine stressors and levels of stress in animals before, during and after moves (in

conjunction with medical and behavior disciplines)


29

In situ Research Priorities

A request for input regarding in situ rhino research priorities was distributed to a few

rhino field researchers/conservationists. The topics received that are not already

represented elsewhere in the document are summarized below.

African Rhinos

1.) Objective determination of conservation value of different rhino populations - This

issue becomes important to influence more appropriate metapopulation management of

rhinos by highlighting the fact that the conservation value of rhino populations drops

disproportionately as numbers decline, an issue that is really important for countries

that are not making appropriate policy decisions on the biological management of

rhinos (i.e., 25% of rhinos are on private land but that 25% does not equate to 25% of

the conservation value of all rhinos in Africa). What is happening in terms of genetic

exchange between small, privately-owned groups of rhinos in South Africa? What are

the effects of any age/sex skewing, derivation of trend data on inbreeding coefficients

to help provide guidelines for the IUCN/SSC African Rhino Specialist Group to

example current classifications (“Key”, “Important”, to also include “Marginally

viable,” “Non-viable”, etc.

2.) Rhino parentage analysis to determine effects of management practices – These

data are needed to determine what husbandry practices (translocations, dehorning, etc.)

may be impacting breeding dynamics.

3.) Exploration of alternative low-power radio frequency systems to track rhinos and

to integrate anti-poaching data – There is the need for a software programming

expert to work on a simple “platform” that integrates primary digital data from devices

in the field (digital radios that transmit GPS positions, Sigfox devices sending


30

locational information from tagged rhinos and from Sigfox-linked devices such as

vehicle trackers, gunshot detectors, etc., so that this information can be displayed in a

customized way on a computer screen in an ops room.

4.) Economic analysis of rhino conservation - What economic values do rhinos

contribute to national economies? How much do rhinos serve as proxy indicators for

ecosystem processes and other components of natural capital and what are the

economic implications arising from this? How much does rhino conservation cost over

and above the basic per area protection costs that need to be met for a typical spectrum

of other wildlife species in protected areas (both private and state) in Africa? What

implications does this have for international performance-based financial support for

productive rhino breeding situations? What economic stimuli can be applied to achieve

new range expansion options in South Africa, which depends upon small land units

being induced to amalgamate into larger ones?

Asian Rhinos

1.) Desk study documenting information on Sumatran rhino captures and

translocations in the 1980s. The ecology and sociobiology of Asian rhinos differs

vastly from that of the African species. Only anecdotal data are available on Sumatran

rhino captures from the 1980s. Two young females, Ratu and Rosa, wandered out of the

forest and were successfully transferred to the Sumatran Rhino Sanctuary in 2005, and a

capture attempt in 2016 failed. A desk study documenting information and lessons

learned from the captures in the 1980s would be a useful first step to lay the groundwork

for future efforts.

2.) Genetic studies of Javan rhinos - The only existing population of Javan rhinos in Ujung

Kulon National park has fluctuated in size over the years, but has never been estimated to

be over ~85 animals and currently stands at ~68 individuals. Because there is no other

wild population of this rhino species, the infusion of genetic diversity is not possible.

Regardless, there is considerable concern about the extent of inbreeding within this

population and a desire to conduct a population genetic study to determine relatedness

and a parentage analysis of the existing animals.

3.) Nutritional analysis of food plants most frequently fed to Sumatran rhinos at the

Sumatran Rhino Sanctuary in Way Kambas National Park - There is a large

collaborative effort underway to capture some of the last wild Sumatran rhinos for

intensive management in breeding centers. Only one native breeding center in SE Asia

has succeeded in producing calves, and therefore any new center should be built in its

likeness and staff should adopt similar husbandry and management practices. Because

browse availability will likely vary in different locations, it would be helpful to conduct a

nutrient analysis of the current diets of the rhinos at the Sumatran rhino sanctuary so that

nutrient value can be replicated at new facilities with different browse.


31

Addendum

Recent (2013-2018) Relevant Rhino Research Papers

Published Literature on Rhino Nutrition

Bapodra P, Dierenfeld E, Wolfe BA. Evaluation of season‐related dietary changes on the serum

profiles of fat‐soluble vitamins, mineral, fatty acids, and lipids in the captive greater one horned rhinoceros (Rhinoceros unicornis). Zoo Biol. 2014; 33: 314-319.

Dutta DK, Bora PJ, Mahanta R, et al. Seasonal variations in food plant preferences of

reintroduced Rhinos Rhinoceros unicornis (Mammalia: Perrissodactyla: Rhinocerotidae) in

Manas National Park, Assam, India. J Threatened Taxa. 2016; 8: 9525-36.

Edwards KL, Shultz S, Pilgrim M, et al. Irregular ovarian activity, body condition and

behavioural differences are associated with reproductive success in female eastern black

rhinoceros (Diceros bicornis michaeli). Gen Comp Endocrinol. 2015; 214: 186-94.

Gimmel A, Hoby S, Deillon L, von Houwald F, Schweizer R, Kölln M, Ratert C, Liesegang A.

Milk composition of Indian rhinoceros (Rhinoceros unicornis) and changes over lactation.

Journal of Zoo and Wildlife Medicine. 2018;49(3):704-14.

Hariyadi AR, Sajuthi D, Astuti DA, Alikodra HS, Maheshwari H. Analysis of nutrition quality

and food digestibility in male Javan rhinoceros (Rhinoceros sondaicus) in Ujung Kulon

National Park. Pachyderm. 2016 Jul 19(57):86-96.

Heidegger EM, von Houwald F, Steck B, Clauss M. Body condition scoring system for greater

one‐horned rhino (Rhinoceros unicornis): Development and application. Zoo Biology. 2016

Sep 1;35(5):432-43.

Huntley NF, Naumann HD, Kenny AL, Kerley MS. Black rhinoceros (Diceros bicornis) and

domestic horse (Equus caballus) hindgut microflora demonstrate similar fermentation

responses to grape seed extract supplementation in vitro. J Anim Physiol Anim Nutr. 2016;

1- 15.

Lavin SR, Sullivan KE, Wooley SC, Stone K, Russell S, Valdes EV. Near infrared reflectance

spectroscopy (NIRS) analyses of nutrient composition and condensed tannin concentrations

in Carolina willow (Salix caroliniana). Zoo Biology. 2015 Nov 1;34(6):576-82.

Lavin SR, Sullivan KE, Wooley SC, Robinson R, Singh S, Stone K, Russell S, Valdes EV.

Nutrient and plant secondary compound composition and iron‐binding capacity in leaves

and green stems of commonly used plant browse (Carolina willow; Salix caroliniana) fed to

zoo‐managed browsing herbivores. Zoo Biology. 2015 Nov 1;34(6):565-75.

Mbatha KR, Bakare AG, Browse silage as potential feed for captive wild ungulates in southern

Africa, Animal Nutrition Journal (2018), doi: 10.1016/j.aninu.2017.12.003.


32

Miller M, Chavey PS, Hofmeyr J, et al. Evaluation of serum ferritin and serum iron in free-

ranging black rhinoceros (Diceros bicornis) as a tool to understand factors affecting iron-

overload disorder. J Zoo Wildl Med. 2016; 47: 820-6.

Mukhlisi M, Atmoko T, Yassir I, Setiawan R, Kusuma AD. Abundance and nutrient content of

some food plants in Sumatran rhino habitat in the forest of Kutai Barat, East Kalimantan,

Indonesia. Pachyderm. 2017 Oct 2(58):77-87.

Oltman W, Olds J, Makowski AJ, et al. Seasonal variation of 25-hydroxy-vitamin D in two

captive Eastern black rhinoceros (Diceros bicornis michaeli). National Merial Veterinary

Scholars Symposium, The Ohio State University. 2016.

Roth TL, Reinhart PR, Kroll JL. Serum Ferritin Concentration is Not a Reliable Biomarker of

Iron Overload Disorder Progression or Hemochromatosis in the Sumatran Rhinoceros

(Dicerorhinus sumatrensis). Journal of Zoo and Wildlife Medicine. 2017;48(3):645-58.

Salyer, Lorien, Iron Metabolism Genes in Browsing and Grazing Rhinoceroses: Implications For

Iron Overload Disorder. 2017. Honors Thesis Projects. 54.

Schook MW, Wildt DE, Raghanti MA, et al. Increased inflammation and decreased insulin

sensitivity indicate metabolic disturbances in zoo-managed compared to free-ranging black

rhinoceros (Diceros bicornis). Gen Comp Endocr. 2015; 217: 10-19.

Sullivan KE, Lavin SR, Livingston SE, Knutson M, Valdes EV, Warren LK. Comparative

digestibility of dry matter, protein, and fiber between the horse and black rhinoceros.

Journal of Equine Veterinary Science. 2015 May 1;35(5):405.

Sullivan KE, Valdes EV. Update on Rhinoceros Nutrition. In: Miller RE, Lamberski N, Calle P,

editors. Miller-Fowler's Zoo and Wild Animal Medicine Current Therapy, Volume 9.

Elsevier Health Sciences; 2018. 699- 706.

Taylor LA, Müller DW, Schwitzer C, et al. Tooth wear in captive rhinoceroses (Diceros,

Rhinoceros, Ceratotherium, Perissodactyla) differs from that of free-ranging conspecifics.

Contributions to Zoology. 2014; 83.

Thakur S, Upreti CR, Jha K. Nutrient analysis of grass species consumed by greater one-horned

rhinoceros (Rhinoceros unicornis) in Chitwan national park, Nepal. International Journal of

Applied Sciences and Biotechnology. 2014 Dec 24;2(4):402-8.

Tubbs CW, Durrant BS, Milnes MR. Reconsidering the use of soy and alfalfa in southern white

rhinoceros diets. Pachyderm. 2017 Oct 2(58):135-9.

Tubbs CW, Moley LA, Ivy JA, et al. Estrogenicity of captive southern white rhinoceros diets and

their association with fertility. Gen Comp Endocr. 2016; 238: 32-38.

Watanabe M, Roth TL, Bauer SJ, Lane A, Romick-Rosendale LE. Feasibility Study of NMR

based serum metabolomic profiling to animal health monitoring: a case study on iron

storage disease in captive Sumatran rhinoceros (Dicerorhinus sumatrensis). PloS one.

2016;11(5):e0156318.

Wojtusik J, Roth TL. Investigation of Factors Potentially Associated with Serum Ferritin

Concentrations in the Black Rhinoceros (Diceros Bicornis) Using a Validated Rhinoceros-

Specific Assay. Journal of Zoo and Wildlife Medicine. 2018;49(2):297-306.

Published Conference Abstracts on Rhino Nutrition

Brooks M, Lee B, Pera J. Case Study: Winos for rhinos: feeding grape pomace to black

rhinoceros (Diceros bicornis) as a method for mitigating iron storage disease. 2017.

Proceedings of the Twelfth Conference on Zoo and Wildlife Nutrition, AZA Nutrition

Advisory Group, Frisco, TX.


33

Ellis KB, Whisnant S, Fellner V, Koutsos E, Ange-van Heugten. 2013. The interaction of diet

and fecal cortisol in the Southern white rhinoceros (Ceratotherium simum simum). In Ward

A, Coslik A, Mahan K, Treiber K, Reppert A, Maslanka M, Eds. Proceedings of the Tenth

Conference on Zoo and Wildlife Nutrition, AZA Nutrition Advisory Group, Salt Lake City,

UT.

Koutsos E, Clauss M, Valdes E. Designing iron controlled diets for exotic hoofstock- variability

in raw materials and manufacturing contributions to total dietary iron. Comparative

Nutrition Society Symposium, Rio Grande, PR. 2016.

Mimiko J, Stringer E, Parsons J. 2017. Case Study: Iron In Black Rhinoceros Diets: The Impact

of Pasture. Proceedings of the Twelfth Conference on Zoo and Wildlife Nutrition, AZA

Nutrition Advisory Group, Friscoe, TX.

Murtaugh K, Power M, Ward A. Macronutrient composition of milk from an Asian rhinoceros.

2017. Proceedings of the Twelfth Conference on Zoo and Wildlife Nutrition, AZA Nutrition

Advisory Group, Frisco, TX.

Paglia D. Human Medical Experience Provides Paradigms Relevant To Captive Breeding of

Endangered Wildlife: Rationale for Prevention and Therapy of Hemolytic and Iron

Overload Propensities In Browser Rhinoceroses, Tapirs and Other Susceptible Species.

American Association of Zoo Veterinarian Conference. 2017, Frisco, TX.

Ricketts V, C Sauer, ES Dierenfeld and K Whitehouse-Tedd. 2018. Feed intake and dietary

content of iron (Fe) copper (Cu), tannic acid and vitamin E of five captive black rhinoceros

(Diceros bicornis) in a UK collection. Marwell Wildlife Nutrition Conference, Colden

Common, Winchester, UK.

Sullivan KE, Valdes EV, Livingston SE, et al. Use of a novel iron chelator (HBED) in black

rhinoceros. In Bissell H, Brooks M Eds. Proceedings of the Eleventh Conference on Zoo

and Wildlife Nutrition, AZA Nutrition Advisory Group, Portland, OR. 2015.

Sullivan KE, Coffey R, Lavin SR, et al. Sequencing the Black Rhino L-ferritin Gene: How

Accurate is our Testing? Proceedings of the Nutrition Advisory Group to the AZA bi-

annual conference, Frisco, TX. 2017.

Sullivan KE, Lavin SR, Livingston SE, Knutson M, Valdes EV, Warren L. Safety and efficacy

of a novel iron chelator in equine as a model for black rhinoceros. Flat Rock, NC:

Comparative Nutrition Society. 2014.

Sullivan KE, Livingston S, Williams S, Mylniczenko N, Rodriguez C, Pye G, Valdes EV. 2016.

Iron Overload Disorder in Browsing Rhinos 2016 Workshop: A Review of Current Goals

and Practical Action Planning. Comparative Nutrition Society 11th Biennial Symposium.

Rio Grande, Puerto Rico.

Sullivan KE, Mylniczenko ND, Emerson JA, Hall NH, Fontenot D, De Voe R, Nolan E, Stacy N,

Livingston SE, Lavin SR, Valdes EV, Pye GW. 2015. Case study: a hemolytic event in an

iron overloaded black rhinoceros (Diceros bicornis) in association with cessation of

chelation therapy. In Bissell H, Brooks M Eds. Proceedings of the Eleventh Conference on

Zoo and Wildlife Nutrition, AZA Nutrition Advisory Group, Portland, OR.

Sullivan KE, Ardente A, Livingston S, Williams S, Valdes EV. 2018. Impact of differing levels

of oral supplementation of vitamin E for white (Ceratotherium simum simum) and black

rhinoceros (Diceros bicornis) on serum levels and fecal losses. Comparative Nutrition

Society 12th Biennial Symposium. Quebec City, Canada.


34

Publications on Rhino Reproduction

Compilation of published rhino 'reproductive' research since last Masterplan

*highlighted= references abstract/conference proceedings

2018

Gomez, M. C., Cates, Y., Stansfield, D. B., Young, C., Klee, R., & Durrant, B. (2018). 187

Morphological Appearance and Expression of Spermatogonial Stem Cell Markers in White

Rhinoceros Testicular Tissue. Reproduction, Fertility and Development, 30, 233-234.

Hermes, R., Hildebrandt, T.B., Goritz, F. 2018. Cryopreservation in rhinoceros- setting a new

benchmark for sperm cryosurvival. PLoS One 13(7):e0200154. doi:

10.1371/journal.pone.0200154

Hildebrandt, T.B., Hermes, R., Colleoni, S., Diecke, S., Holtze, S., Renfree, M.B., Stejakal, J.,

Hayashi, K., Drukker, M., Loi, P., Goritz, F, Lazzari, G., Galli, C. 2018. Embryos and

embryonic stem cells from the white rhinoceros. Nat Commun 9(1):2589. doi:

10.1038/s41467-018-04959-2

Pennington, P.M., Durrant, B.S. (2018). Assisted reproductive technologies in captive

rhinoceroses. Mammal Review https://doi.org/10.1111/mam.12138

Stoops, M.A., Winget, G.D., DeChant, C.J., Ball, R.L., Roth, T.L. (2018). Early fetal sexing in

the rhinoceros by detection of male-specific genes in maternal serum. Molecular

Reproduction & Development, 85 (3) 197-204. https://doi.org/10.1002/mrd.22953.

Wojtusik, J., Stoops, M.A., Roth, T.L. (2018). Comparison of soy lecithin, coconut water, and

coconut milk as animal protein-free alternatives to egg-yolk in extender for semen

cryopreservation of the African black rhinoceros (Diceros bicornis) and Indian rhinoceros

(Rhinoceros unicornis). Theriogenology, 121: 72-77.

https://doi.org/10.1016/j.theriogenology.2018.07.042

2017

Bickley, S.M., Pollock, K.E., Stoops, M.A. (2017). Biological and behavioral reasons for

unsuccessful breeding in captive female African black rhinos: olfactory stimulation as a

method for improvement. In Proceedings: the 6th International Society of Wildlife

Endocrinology, Orlando, FL, USA Aug 14-16. P75.

Korody, M. L., Pivaroff, C., Nguyen, T. D., Peterson, S. E., Ryder, O. A., & Loring, J. F. (2017).

Four new induced pluripotent stem cell lines produced from northern white rhinoceros with

non-integrating reprogramming factors. bioRxiv, 202499.

Kottwitz, J., Stoops, M., Reeves, J., Harmon, R., Wilborn, R., Edmonson, M., Boothe, D. (2017).

The negative effects of analgesic and anesthetic drugs on sperm motility: implications for

assisted breeding in captive rhino. In Proceedings: the 49th Annual American Association of

Zoological Veterinarians Conference, Dallas, TX USA Sept 22-29. P107-108.

Roth, T. L., Schook, M. W., & Stoops, M. A. (2017). Monitoring and controlling ovarian

function in the rhinoceros. Theriogenology. DOI: 10.1016/j.theriogenology.2017.12.007

Schwarzenberger, F. (2017). Comparative evaluation of gestation in three rhinoceros species

(Diceros bicornis; Ceratotherium simum, Rhinoceros unicornis). In Proceedings:

International Society of Wildlife Endocrinology, Orlando, FL USA Aug 14-16. P69.

Stoops, M.A., O'Brien, J.K., Niederlander, J., Metrione, L., Pootoolol, J., Delk, K., Niemuller,

C., Hagen, D., Douglas, R.H., Proudfoot, J. (2017). Administration of biorelease

progesterone and estradiol or GnRH to induce estrous cycles and breeding in anestrous


35

African white rhinoceroses (Certatotherium simum simum). In Proceedings: International

Society of Wildlife Endocrinology, Orlando, FL USA Aug 14-16. P111.

Tubbs, C. W., Durrant, B. S., & Milnes, M. R. (2017). Reconsidering the use of soy and alfalfa

in southern white rhinoceros diets. Pachyderm, 58, 135-139.

Ververs, C., Hostens, M., van Zijll Langhout, M., Otto, M., Govaere, J., & Van Soom, A. (2017).

109 REPRODUCTIVE PERFORMANCE PARAMETERS IN A LARGE HERD OF

CONFINED FREE-ROAMING WHITE RHINOCEROSES (CERATOTHERIUM

SIMUM). Reproduction, Fertility and Development, 29(1), 163-163.

Wojtusik, J., Stoops, M.A., Roth, T.L. (2017). Use of animal-protein free extenders as

alternatives to standard egg-yolk based extender in semen cryopreservation of the black

(Diceros bicornis) and Indian (Rhinoceros unicornis) rhino. In Proceedings: 11th

International Conference on Behaviour, Physiology, and Genetics of Wildlife, Berlin,

Germany Oct 4-7th. P135.

2016

Galli, C., Hermes, R., Goeritz, F., Colleoni, S., Diecke, S., Drukker, M., Hayashi, K., Holtze, S.,

Lazzari, G., Payne, J., Sos, E., Steiskal, J., Wiesner, M., Zainuddin, Z.A., Hildebrandt, T.B.

2016. First results of oocyte maturation and in-vitro-fertilisation (IVF) in Sumatran and

northern white rhinoceroses. In: Proceedings

15th International Elephant & Rhino Conservation & Research Symposium, Singapore Zoo,

Singapore Nov. 13-18. 51.

Hermes, R., Schwarzenberger, F., Göritz, F., Oh, S., Fernandes, T., Bernardino, R., ... &

Saragusty, J. (2016). Ovarian down regulation by GnRF vaccination decreases reproductive

tract tumour size in female white and greater one-horned rhinoceroses. PloS one, 11(7),

e0157963.

O'Brien, J.K., Stoops, M.A., Roth, T.L., Ball, R.L., Montano, G.A., Stenman, K.L., Posy, J.L.,

Saiers, R., Ramer, J.C., Love, C.C., Robeck T.R. (2016). Progress in sperm sorting and

cryopreservation technologies for modifying population sex ratio and preserving genetic

diversity in the rhinoceros and elephant. In: Proceedings 15th International Elephant &

Rhino Conservation & Research Symposium, Singapore Zoo, Singapore Nov. 13-18. P48.

Roth, T. L., Stoops, M. A., Robeck, T. R., & O’Brien, J. K. (2016). Factors impacting the

success of post-mortem sperm rescue in the rhinoceros. Animal Reproduction Science, 167,

22-30.

Roth, T. L., Stoops, M. A., Robeck, T. R., & O’Brien, J. K. (2016). 116 FACTORS

IMPACTING THE SUCCESS OF POSTMORTEM SPERM RECOVERY AND

CRYOPRESERVATION IN THE RHINOCEROS. Reproduction, Fertility and

Development, 28, 188-188.

Santymire, R.M., Misek, S., Gossett, J., Kamhout, M., Chatroop, E., Rafacz, M. (2016). Male

behaviours signal the female's reproductive state in a pair of black rhinoceros housed at

Lincoln Park Zoo. Journal of Zoo and Aquarium Research, 4, 30-37.

Saragusty, J., Diecke, S., Drukker, M., Durrant, B., Friedrich, B-N. I., Galli, C., Goritz, F.,

Hayashi, K., Hermes, R., Holtze, S., Johnson, S., Lazzari, G., Loi, P., Loring, J.F., Okita, K.,

Renfree, M.B., Seet, S., Voracek, T., Stejskal, J., Ryder, O.A., Hildebrandt, T.B. (2016).

Rewinding the process of mammalian extinction. Zoo Biology, 35, 280-292.

Stoops, M. A., Campbell, M. K., DeChant, C. J., Hauser, J., Kottwitz, J., Pairan, R. D.,

Shaffstall, W., Volle, K., Roth, T. L. (2016). Enhancing captive Indian rhinoceros genetics


36

via artificial insemination of cryopreserved sperm. Animal Reproduction Science, 172, 60-

75.

Tubbs, C. W., Moley, L. A., Ivy, J. A., Metrione, L. C., LaClaire, S., Felton, R. G., Durrant,

B.S., Milnes, M. R. (2016). Estrogenicity of captive southern white rhinoceros diets and

their association with fertility. General and Comparative Endocrinology, 238, 32-38.

von Houwald, F. (2016). Husbandry, management and breeding of the Greater one-horned

rhinoceros Rhinoceros unicornis at Zoo Basel. International Zoo Yearbook, 50, 203-214.

2015

DeCourcy, K., Campbell, M., Hauser, J., Kottwitz, J., Levens, G., Pairan, R., Volle, K, Stoops,

M.A. (2015). Using urinary hormone analysis to predict gender and assess fetal viability in

the Indian rhinoceros (Rhinoceros unicornis). In: Proceedings Annual Conference of the

American Association of Zoo Veterinarians. Portland, OR USA Sept25-Oct 2. P82-83.

Edwards, K. L., Shultz, S., Pilgrim, M., & Walker, S. L. (2015). Irregular ovarian activity, body

condition and behavioural differences are associated with reproductive success in female

eastern black rhinoceros (Diceros bicornis michaeli). General and Comparative

Endocrinology, 214, 186-194.

Edwards, K. L., Shultz, S., Pilgrim, M., & Walker, S. L. (2015). Male reproductive success is

correlated with testosterone in the eastern black rhinoceros (Diceros bicornis

michaeli). General and Comparative Endocrinology, 213, 40-49.

Edwards, K. L., Walker, S. L., Dunham, A. E., Pilgrim, M., Okita-Ouma, B., & Shultz, S.

(2015). Low birth rates and reproductive skew limit the viability of Europe’s captive eastern

black rhinoceros, Diceros bicornis michaeli. Biodiversity and Conservation, 24, 2831-2852.

Gener, S., Schwarz, C., Grothmann, P., Bernhard, A., Eulenberger, K., Einspanier, A., &

Gottschalk, J. (2015). Oestrus and pregnancy detection in the black rhinoceros (Diceros

bicornis) by faecal hormone analysis. Reproduction in Domestic Animals, 50, 33-34.

Moresco, A., Larsen, R.S., Boon, D., O'Brien, J.K., Stoops, M.A. (2015). Semen collection in

black rhinoceros (Diceros bicornis) via urethral catheterization. In: Proceedings Annual

Conference of the American Association of Zoo Veterinarians. Portland OR USA Sept25-

Oct2. P170-171.

O’Brien, J. K., Roth, T. L., Stoops, M. A., Ball, R. L., Steinman, K. J., Montano, G. A., Love,

C.C., Robeck, T. R. (2015). Sperm sex-sorting and preservation for managing the sex ratio

and genetic diversity of the southern white rhinoceros (Ceratotherium simum

simum). Animal Reproduction Science, 152, 137-153.

Van der Goot, A. C., Martin, G. B., Millar, R. P., Paris, M. C. J., & Ganswindt, A. (2015).

Profiling patterns of fecal 20-oxopregnane concentrations during ovarian cycles in free-

ranging southern white rhinoceros (Ceratotherium simum simum). Animal Reproduction

Science, 161, 89-95.

Ververs, C., van Zijl Langhout, M., Govaere, J., & Van Soom, A. (2015). Features of

reproduction and assisted reproduction in the white (Ceratotherium simum) and black

(Diceros bicornis) rhinoceros. Vlaams Diergeneeskundig Tijdschrift, 84, 175-187.

2014

Cain, B., Wandera, A.B., Shawcross, S.G., Edwin, H.W., Stevens-Wood, B., Kemp, S.J., Okita-

Ouma, B., Watts, P.C. (2014). Sex-biased inbreeding effects on reproductive success and

home range size of the critically endangered black rhinoceros. Conservation Biology, 28,

594-603.


37

Capiro, J. M., Stoops, M. A., Freeman, E. W., Clawson, D., & Schook, M. W. (2014). Effects of

management strategies on glucocorticoids and behavior in Indian rhinoceros (Rhinoceros

unicornis): translocation and operant conditioning. Zoo Biology, 33, 131-143.

Edwards, K. L., McArthur, H. M., Liddicoat, T., & Walker, S. L. (2014). A practical field

extraction method for non-invasive monitoring of hormone activity in the black

rhinoceros. Conservation Physiology, 2(1).

Freeman, E.W., Meyer, J.M., Bird, J., Adendorff, J., Schulte, B.A., Santymire, R.M. (2014).

Impacts of environmental pressures on the reproductive physiology of subpopulations of

black rhinoceros (Diceros bicornis bicornis) in Addo Elphant National Park, South Africa.

Conservation Physiology, 2 doi:10.1093/consphys/cot034.

Hermes, R., Göritz, F., Saragusty, J., Stoops, M. A., & Hildebrandt, T. B. (2014). Reproductive

tract tumours: the scourge of woman reproduction ails Indian rhinoceroses. PloS one, 9(3),

e92595.

Metrione, L., & Eyres, A. (2014). Rhino Husbandry Manual. Fort Worth (TX): International

Rhino Foundation.

Penfold, L.M., Powell, D., Traylor-Holzer, K., Asa, C.S. (2014). "Use it or lose it":

characterization, implications, and mitigation of female infertility in captive wildlife. Zoo

Biology, 33, 20-28.

Schwwarz, C., Grothmann, P., Gottschalk, J., Eulenberger, K., Einspainer, A. (2014). Breeding

management of black rhinos (Diceros bicornis michaeli) in Magdeburg zoo. Tierarztliche

Praxis Ausgabe G Grosstiere/Nuztiere, 42, 150-155.

Stoops, M. A., West, G. D., Roth, T. L., & Lung, N. P. (2014). Use of urinary biomarkers of

ovarian function and altrenogest supplementation to enhance captive breeding success in the

Indian rhinoceros (Rhinoceros unicornis). Zoo Biology, 33, 83-88.

Tubbs, C., McDonough, C.E., Felton, R., Milnes, M.R. (2014). Advances in conservation

endocrinology: the application of molecular approaches to the conservation of endangered

species. General and Comparative Endocrinology, 203, 29-34.

2013

Benco, A., Campbell, M., Barthel, M., Pinto, C., MacKinnon, K., Stoops, M. (2013). Urinary

hormone concentrations and pharmacokinetics/pharmacodynamics of haloperidol in a

female Indian rhinoceros (Rhinoceros unicornis). In: Proceedings International Elephant and

Rhino Conservation and Research Symposium. Pittsburgh, PA USA Aug 26-30. P11-12.

Nau, M., Pairan, P., Pinto, C., Sims, R., MacKinnon, K., Stoops, M. (2013). Relationship of

salivary hormone concentrations to urinary hormone excretion profiles in the Indian

rhinoceros (Rhinoceros unicornis). In: Proceedings International Elephant and Rhino

Conservation and Research Symposium. Pittsburgh, PA USA Aug 26-30. P9.

Van der Goot, A., Martin, G., Metrione, L., Paris, M., Schook, M., Penfold, L. (2013). Attempt

to control estrus and ovulation in white rhinoceroses using a synthetic progestagen and slow

release GnRH analogue. In: Proceedings International Elephant and Rhino Conservation

and Research Symposium. Pittsburgh, PA USA Aug26-30. P9.

Roth, T.L., Reinhart, P.R., Romo, J.S., Candra, D., Suhaery, A., Stoops, M.A. (2013). Sexual

maturation in the Sumatran rhinoceros. Zoo Biology, 32, 549-555.

Van der Goot, A.C., Dalerum, F., Ganswindt, A., Martin, G.B., Millar, R.P., Paris, M.C. (2013).

Faecal progestagen profiles in wild southern white rhinoceros (Ceratotherium simum

simum). African Zoology, 48, 143-151.


38

Publications on Rhino Behavior and Ecology

Adhikari, K. 2015. Ecology, demography, conservation and management of greater one horned

rhinoceros (Rhinoceros unicornis) in Chitwan Natioal Park, Nepal. PhD Dissertation.

Saurashtra University, Rajkot, India.

Aldila, D., A.J. Hutchinson, M. Woolway, N. Owen-Smith, and E. Soewono. 2015. A

mathematical model of black rhino translocation strategy. Journal of ssMathematical and

Fundamental Sciences 47(1):104-115.

Anderson, T.M., P.M. Ngoti, M.L. Nzunda, D.M. Griffith, J.D.M. Speed, F. Fossøy, E. Røskaft,

and B.J. Graae. 2018. The burning question: does fire affect habitat selection and forage

preference of black rhinos (Diceros bicornis) in East African savannas? Oryx 1-10

doi:10.1017/S0030605318000388.

Badenhorst, M., M. Otto, A.C. van der Goot, and A. Ganswindt. 2016. Stress steroid levels and

the short-term impact of routine dehorning in female southern white rhinoceros

(Ceratotherium simum simum). African Zoology 51(4):211-215.

Barman, R., B. Choudhury, N.V.K. Ashraf, and V. Menon. 2014. Rehabilitation of greater one-

horned rhinoceros calves in Manas National Park, a World Heritage Site in India.

Pachyderm 55:78-88.

Bhattacharya, A., and K. Chakraborty. 2017. Why do Indian rhinos eat elephant grasses?

International Journal of Trend in Research and Development 4(5):355-357.

Cain, B., A.B. Wandera, S.G. Shawcross, W.E. Harris, B. Stevens-Wood, S.J. Kemp, and B.

Okita-Ouma. 2014. Sex-biased inbreeding effects on reproductive success and home range

size of the critically endangered black rhinoceros. Conservation Biology 28(2):594-603.

Capiro, J.M., M.A. Stoops, E.W. Freeman, D. Clawson, and M.W. Schook. 2014. Effects of

management strategies on glucocorticoids and behavior in Indian rhinoceros (Rhinoceros

unicornis): translocation and operant conditioning. Zoo Biology 33(2):131-143.

Christie, K.S., S.L. Gilbert, C.L. Brown, M. Hatfield, and L. Hanson. 2016. Unmanned aircraft

systems in wildlife research: current and future applications of a transformative technology.

Frontiers in Ecology and the Environment 14(5):241-251.

Cinková, I., and R. Policht. 2014. Contact calls of the northern and southern white rhinoceros

allow for individual and species identification. PLoS ONE 9(6):e98475.

Cinková, I., and R. Policht. 2015. Discrimination of familiarity and sex from chemical cues in

the dung by wild southern white rhinoceros. Animal Cognition 18(1):385-392.

Clark, J.H. 2013. Habitat use analysis of a reintroduced black rhino (Diceros bicornis)

population. Honors Thesis. Western Kentucky University, Bowling Green, USA.

Cromsigt, J.P.G.M., and M. te Beest. 2014. Restoration of a megaherbivore: landscape-level

impacts of white rhinoceros in Kruger National Park, South Africa. Journal of Ecology

102(3):566-575.

D’Amen, M., N.E. Zimmermann, and P.B. Pearman. 2013. Conservation of phylogeographic

lineages under climate change. Global Ecology Biogeography 22(1):93-104.

Deka, R.J., and N.K. Sarma. 2015. Studies on feeding behavior and daily activities of Rhinoceros

unicornis in natural and captive condition of Assam. Indian Journal of Animal Research

49(4):542-545.

Di Minin, E., J. Laitila, F. Montesino-Pouzols, N. Leader-Williams, R. Slotow, P.S. Goodman,

A.J. Conway, and A. Moilanen. 2015. Identification of policies for a sustainable legal trade


39

in rhinoceros horn based on population projection and socioeconomic models. Conservation

Biology 29(2):545-555.

Donaldson, J.E., S. Archibald, N. Govender, D. Pollard, Z. Luhdo, and C.L. Parr. 2018.

Ecological engineering through fire-herbivory feedbacks drives the formation of savanna

grazing lawns. Journal of Applied Ecology 55(1):225-235.

Dutta, D.K., and R. Mahanta. 2015. A study on the behavior and colonization of translocated

greater one-horned rhinos Rhinoceros unicornis (Mammalia: Perissodactyla:

Rhinocerotidae) during 90 days from their release at Manas National Park, Assam India.

Journal of Threatened Taxa 7(2):6864-6877.

Dutta, D.K., A. Sharma, R. Mahanta, and A. Swargowari. 2017. Behaviour of post released

translocated greater one-horned rhinoceros (Rhinoceros unicornis) at Manas National Park,

Assam, India. Pachyderm 58:58-66.

Edwards, K.L. 2013. Investigating population performance and factors that influence

reproductive success in the eastern black rhinoceros (Diceros bicornis michaeli). PhD

Dissertation. University of Liverpool, Liverpool, United Kingdom.

Eyres, A., J. Capiro, and J. Ivy. 2017. Population analysis and breeding and transfer plan

southern white rhinoceros (Ceratotherium simum simum) AZA species survival plan yellow

program. Silver Spring, MD: Association of Zoos and Aquariums Population Management

Center. 43 p.

Ferreira, S.M., C. Greaver, G.A. Knight, M.H. Knight, I.P.J. Smit, and D. Pienaar. 2015.

Disruption of rhino demography by poachers may lead to population declines in Kruger

National Park, South Africa. PLoS ONE 10(6):e0127783.

Freeman, E.W., J.M. Meyer, J. Adendorff, B.A. Schulte, and R.M. Santymire. 2014. Scraping

behavior of black rhinoceros is related to age and fecal gonadal metabolite concentrations.

Journal of Mammalogy 95(2):340-348.

Freeman, E.W., J.M. Meyer, J. Bird, J. Adendorff, B.A. Schulte, and R.M. Santymire. 2014.

Impacts of environmental pressures on the reproductive physiology of subpopulations of

black rhinoceros (Diceros bicornis bicornis) in Addo Elephant National Park, South Africa.

Conservation Physiology 2(1):cot034.

Gedir, J.V., P.R. Law, P. du Preez, and W.L. Linklater. 2018. Effects of age and sex ratios on

offspring recruitment rates in translocated black rhinoceros. Conservation Biology

32(3):628-637.

Greaver, C., S. Ferreira, and R. Slotow. 2014. Density-dependent regulation of the critically

endangered black rhinoceros population in Ithala Game Reserve, South Africa. Austral

Ecology 39(4):437-447.

Gyöngyi, K., and M. Elmeros. 2017. Forage choice of the reintroduced black rhino and the

availability of selected browse species at Majete Wildlife Reserve, Malawi. Pachyderm

58:40-50.

Hariyadi, A.R. 2015. Assessment of Halimun-Salak National Park as a potential site for

establishing the second population of Javan rhinoceros (Rhinoceros sondaicus).

http://www.rhinoresourcecenter.com/pdf_files/146/1460994396.pdf

Harjanto, E. 2017. On a territorial competition between Rhinoceros sondaicus and Bos javanicus

at Ujung Kulon National Park. Communication in Biomathematical Sciences 1(1):46-53.

Kafley, H., M.E. Gompper, M. Khadka, M. Sharma, R. Maharjan, and B.P. Thapaliya. 2015.

Analysis of rhino (Rhinoceros unicornis) population viability in Nepal: impact assessment

of antipoaching and translocation strategies. Zoology and Ecology 25(4):288-294.


40

Kruger, M. 2017. Capture and boma stress responses in the white rhinoceros (Ceratotherium

simum). PhD Dissertation. University of the Witwatersrand, Johannesburg, South Africa.

Landman, M., and G.I. Kerley. 2014. Elephant both increase and decrease availability of browse

resources for black rhinoceros. Biotropica 46(1):42-49.

Landman, M., D.S. Schoeman, and G.I. Kerley. 2013. Shift in black rhinoceros diet in the

presence of elephant: evidence for competition? PLoS ONE 8(7):e69771.

Law, P.R., B. Fike, and P.C. Lent. 2013. Mortality and female fecundity in an expanding black

rhinoceros (Diceros bicornis minor) population. European Journal of Wildlife Research

59(4):477-485.

Law, P.R., B. Fike, and P.C. Lent. 2014. Birth sex in an expanding black rhinoceros (Diceros

bicornis minor) population. Journal of Mammalogy 95(2):349-356.

Law, P.R., B. Fike, P.C. Lent. 2015. Dynamics of an expanding black rhinoceros (Diceros

bicornis minor) population. European Journal of Wildlife Research 61(4):601-609.

Law, P.R., and W.L. Linklater. 2014. Black rhinoceros demography should be stage, not age,

based. African Journal of Ecology 52(4):571-573.

le Roux, S.P., J. Marias, R. Wolhuter, and T. Niesler. 2017. Animal-borne behavior classification

for sheep (Dohne merino) and rhinoceros (Ceratotherium simum and Diceros bicornis).

Animal Biotelemetry 5:25.

Linklater, W.L., K. Mayer, and R.R. Swaisgood. 2013. Chemical signals of age, sex and identity

in black rhinoceros. Animal Behaviour 85(3):671-677.

Linn, S.N., M. Boeer, and M. Scheumann. 2018. First insights into the vocal repertoire of infant

and juvenile southern white rhinoceros. PLoS ONE 13(3):e0192166.

Marneweck, C., A. Jurgens, and A.M. Shrader. 2017. Dung odours signal sex, age, territorial and

oestrous state in white rhinos. Proceedings of the Royal Society of London B

284(1846):20162376.

Mays, H.L. Jr., C. Hung, P. Shaner, J. Denvir, M. Justice, S. Yang, T.L. Roth, D.A. Oehler, J.

Fan, S. Rekulapally, and D.A. Primerano. 2018. Genomic analysis of demographic history

and ecological niche modeling in the endangered Sumatran rhinoceros Dicerorhinus

sumatrensis. Current Biology 28(1):70-76.

Menargues, A., V. Urios, and R. Limiñana. 2013. Seasonal pattern of salivary cortisol secretion

in the greater one-horned rhinos. Animal Welfare 22(4):467-472.

Miller, M., M. Kruger, M. Kruger, F. Olea-Popelka, and P. Buss. 2016. A scoring system to

improve decision making and outcomes in the adaptation of recently captured white

rhinoceroses (Ceratotherium simum) to captivity. Journal of Wildlife Diseases 52(2):S78-

S85.

Mulero-Pázmány, M., R. Stolper, L.D. van Essen, J.J. Negro, and T. Sassen. 2014. Remotely

piloted aircraft systems as a rhinoceros anti-poaching tool in Africa. PLoS ONE

9(1):e83873.

Murphy, S.T., N. Subedi, S.R. Jnawali, B.R. Lamichhane, G.P. Upadhyay, R. Kock, and R.

Amin. 2013. Invasive mikania in Chitwan National Park, Nepal: the threat to the greater

one-horned rhinoceros Rhinoceros unicornis and factors driving the invasion. Oryx

47(3):361-368.

Ngoti, P.M. 2017. The feeding ecology of eastern black rhinocoeroses (Diceros bicornis

michaeli) in southern Serengeti National Park, Tanzania. Master’s Thesis. Norwegian

University of Science and Technology, Trondheim, Norway.


41

Odendaal-Holmes, K., J.P. Marshal, and F. Parrini. 2014. Disturbance and habitat factors in a

small reserve: space use by establishing black rhinoceros (Diceros bicornis). South African

Journal of Wildlife Research 44(2):148-160.

Ojah, S., A. Saikia, and P. Sivakumar. 2015. Habitat suitability of Laokhowa Burhachapori

wildlife sanctuary complex of Assam, India for Rhinoceros unicornis Linn. The Clarion

4(2):39-47.

Olsson, S. 2015. A survey of the available browse for the black rhinoceros (Diceros bicornis

Linnaeus, 1758) in a farmland area in the Kunene region, Namibia. Bachelor’s Thesis.

Halmstad University, Halmstad, Sweden.

Plotz, R.D. 2014. The interspecific relationships of black rhinoceros (Diceros bicornis) in

Hluhluwe-iMfolozi Park. PhD Dissertation. Victoria University of Wellington, Wellington,

New Zealand.

Plotz, R.D., W.J. Grecian, G.I.H. Kerley, and W.L. Linklater. 2016. Standardising home range

studies for improved management of the critically endangered black rhinoceros. PLoS ONE

11(3):e0150571.

Pluháček, J., B.L. Steck, S.P. Sinha, and F. von Houwald. 2017. Interbirth intervals are

associated with age of the mother, but not with infant mortality in Indian rhinoceros. Current

Zoology 63(3):229-235.

Prinsloo, D. 2017. Impact of African elephant feeding on white rhinoceros foraging

opportunities. Master’s Thesis. Nelson Mandela University, Port Elizabeth, South Africa.

Pusparini, W. 2014. Ecology and conservation of endangered species in Sumatra: smaller cats

and the Sumatran rhinoceros (Dicerorhinus sumatrensis) as case studies. Master’s Thesis.

University of Massachusetts, Amherst, USA.

Pusparini, W., P.R. Sievert, T.K. Fuller, T.O. Randhir, and N. Andayani. 2015. Rhinos in the

parks: an island-wide survey of the last wild population of the Sumatran rhinoceros. PLoS

ONE 10(9):e0136643.

Pusparini, W., and H.T. Wibisono. 2013. Landscape-level assessment of the distribution of the

Sumatran rhinoceros in Bukit Barisan Selatan National Park, Sumatra. Pachyderm 53:59-65.

Schwabe, F., T. Göttert, N. Starik, S.R. Levick, and U. Zeller. 2015. A study on the postrelease

behavior and habitat preferences of black rhino (Diceros bicornis) reintroduced into a

fenced reserve in Namibia. African Journal of Ecology 53(4):531-539.

Soka, G.E., A.A. Rija, and A. Owino. 2014. Modeling black rhinoceros (Diceros bicornis)

population performance in East Africa: the case of Lake Nakuru National Park, Kenya.

Biodiversity & Endangered Species 2(3):1-6.

Subedi, N., S.J. Jnawali, M. Dhakal, N.M.B. Pradhan, B.R. Lamichhane, S. Malia, R. Amin, and

Y.V. Jhala. 2013. Population status, structure and distribution of the greater one-horned

rhinoceros Rhinoceros unicornis in Nepal. Oryx 47(3):352-360.

Subedi, N., B.R. Lamichhane, R. Amin, S.R. Jnawali, and Y.V. Jhala. 2017. Demography and

viability of the largest population of greater one-horned rhinoceros in Nepal. Global

Ecology and Conservation 12:241-252.

Thapa, K., S. Nepal, G. Thapa, S.R. Bhatta, and E. Wikramanayake. 2013. Past, present and

future conservation of the greater one-horned rhinoceros Rhinoceros unicornis in Nepal.

Oryx 47(3):345-351.

Thompson, S., T. Avent, and L.S. Doughty. 2016. Range analysis and terrain preference of adult

southern white rhinoceros (Ceratotherium simum) in a South African private game reserve:

insights into carrying capacity and future management. PLoS ONE 11(9):e0161724.


42

Thuo, D.N., J.O. Junga, J.M. Kamau, J.O. Amimo, F.M. Kibegwa, and K.E. Githui. 2015.

Population viability analysis of black rhinoceros (Diceros bicornis michaeli) in Lake

Nakuru National Park, Kenya. Biodiversity & Endangered Species 3(1):1-5.

Tripathi, A.K. 2013. Social and reproductive behavior of great Indian one-horned rhino,

Rhinoceros unicornis in Dudhwa National Park, U.P., India. International Journal of

Pharmacy and Life Sciences 4(11):3116-3121.

Veldhuis, M.P., M.I. Gommers, H. Olff, and M.P. Berg. 2018. Spatial redistribution of nutrients

by large herbivores and dung beetles in a savanna ecosystem. Journal of Ecology

106(1):422-433.

Ververs, C., M. van Zijll Langhout, M. Hostens, M. Otto, J. Govaere, B. Durrant, and A. Van

Soom. 2017. Reproductive performance parameters in a large population of game-ranched

white rhinoceroses (Ceratotherium simum simum). PLoS ONE 12(12):e0187751.

Whitham, J.C., and L. Miller. 2016. Using technology to monitor and improve zoo animal

welfare. Animal Welfare 25(4):395-409.

Yadav, P.K., K. Sarma, and S. Dookia. 2013. The review of biodiversity and conservation study

in India using geospatial technology. International Journal of Remote Sensing and GIS

2(1):1-10.

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