Choosing the “best monkey” and training it for a complex

research project

Bushmitz Moshe DVM ,Yosi Shohat

BFC Israel

NHP in Diabetes research

• Diabetes mellitus, is a group of metabolic diseases in which a person has high blood sugar.

• As of 2012, an estimated 346 million people have type 2 diabetes.

• Animal models are important for determining the pathogenesis of and potential treatments for obesity and diabetes.

• Nonhuman primates (NHPs) are particularly useful for studying these disorders. As in humans, type 2 diabetes mellitus is the most common form of diabetes in NHPs and occurs more often in older & obese animals.

NHP in Diabetes research in Israel

• Research has been involved, for more than twenty years, with the subject of transplantation immunology and, more specifically, with questions relating to stem cell transplantation.

• The use of ‘mega dose’ transplants in leukemia patients, have already been translated into clinical achievements.

• The latter is already in advanced preclinical primate studies, suggesting potential cure of type 2 diabetes.

BFC - Israel

• Breeding facility for M. fascicularis established 1990

• All animals SPF Mauritian origin cynomolgus

Are all monkeys the same ?

Research success

Our experience is that 50% of the success is related to the personality of the monkey and 50% to the training process.

Choosing Training

Primate research evolution

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Goal of the study

• How to choose the”best animal” for a complicated research project which involves multiple interactions with the NHP & daily high No. of blood testing & injections ?

• To train the NHP to cooperate with multiple testing & injections in the pre and post operative phase.

• To be able to perform studies with NHP without forced action & with lower stress level & better animal welfare.

• To be able to perform studies with NHP without chemical restraint to avoid the risk thaat this could interfere with the diabetes research.

Choosing the right animal

• The vicinity of the breeding farm BFC to the research institute is the main advantage which allows to choose the animals which suit better the research goals.

• Chief behaviorist of Weitzmann inst. Yosi Shohat will first define the requirements for the specific research project.

• The Farm staff spot animals which show high level of curiosity and confidence with low level of fear .

• Then he comes to BFC & spend 3-4 hours over 2-3 visits , a couple of weeks apart to choose the right animal.

“Best animal” criteria

• Male , 3-5 kg ,2.5-4 years of age

• Physical and mental maturity.

• Not the Alpha male or the leader

• Highly curious.

• Highly self confident.

• Animal shows a stable level of fear.

• If 2 animals are selected ,they should be similar to avoid dependence.

• Will suit also the other animals in the facility.

• Selection is done by sitting out of the cage with 20-30 NHPs.

• Animals are offered food and items they never saw before.

• Curiosity is first evaluated and first selection done.

• Animal’s self confidence evaluated - how he interacts with humans.

• Animal’s level of fear evaluated - this is done now in standing position (of trainer) which is a major threat for the animals.

Choosing technic

• Confident NHP will show low level of fear in standing position & will not hesitate to take food with mouth to hand contact. This will also not change when trainer walks or moves along the cage. Less confident NHP will grab the food and run , or will show different behavior in standing position or when he walks along the cage.

• Animal’s social hierarchy in the group is evaluated.

• The chosen animals are signed with color spray.

Training general rules

• Training is done every day, 2-3 times in short sessions.

• Training is done when animal is in his normal 1.5x2x2.5 meter cage.

• During training other animals are moved to another cage in the same room so the trained animal is alone in his cage.

• Training is done with voice command or clicker or both.

• Reward is : voice reward + food.

Phase 1 - building human-animal trust

• Goal is to create a trust between trainer and animal

• Animal fed manually by trainer 4-5 times/d: hand to mouth.

• The most critical phase, and should follow by the book.

• Phase 1 = 2-4 weeks long

Phase 2 - building human-animal contact

• Goal = animal to be touched by the trainer in every area of his body with no fear.

• This phase starts after 1st week of phase 1

• Phase 2 = 2-4 weeks long

Phase 3 - animal offer his leg

• Goal = animal offers his leg outside the cage bars and is touched by the trainer .

• Trainer put his hand inside the cage and touches the leg

• Trainer pulls the leg out of the cage and holds it.

• Animal offers the leg voluntarily outside the cage bars.

• Phase 3 = 1 week long

Leg offering

Leg offering

Phase 4 - animal hold cage bars

• Goal = animal holds cage bars with both hands.

• Animal holds bars for different periods of time.

• Allow flexibility of location compare to “target”

• Animal learns not to interfere with trainer.

• When hold the bars also offer his leg.

• Phase 4 = 1 week long

Phase 5 - blood glucose testing

• Goal is to train the animal to sit quietly when trainer is taking blood sample from finger for glucose testing.

• First week is de-sensitization with glucose sampler without needle - animal gets use to the touch , noise & squeeze of finger.

• Animal holds bars with both hands offering his leg.

• When animal ready - use glucose sampler with needle and sample one drop of blood for glucose testing.

• Phase 5 - sometimes there is a regression but overall it takes up to 2 weeks

Phase 6 - injection of insulin

• Goal = train the animal to sit quietly when trainer is injecting insulin into the animal’s belly area.

• First week is de-sensitization with a syringe without needle - animal gets use to the touch .

• Animal holds bars with both hands offering his belly.

• Phase 6 - sometimes there is a regression but overall it takes up to 2 weeks.

Phase 7 - preparation for the transplantation

• 2 weeks before the transplantation , training starts to assimilate the post operative period.

• Blood sampling every 3 hours for 24 hours.

• Insulin injection - up to 3 per day

• Phase 7 = 1 week.

Phase 8 - post operative phase

• Start 3 hours after operation.

• Blood sampling every 3 hours for 24 hours.

• Insulin injection - up to 3 per day.

• Nutrition is 4 times a day and it is normal : 1 fruit , 2 vegetables, pellets , peanuts, sunflower, almonds.

• Phase 8 - takes up to 4 months.

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Training process

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Advantages of training

• NHP can be used where he is a valuable model.

• A trained NHP facilitates extremely the research as many tests and injections are required daily.

• NHP welfare is much better when animal is trained .

• Choosing the right animal is critical and saves time.

• Same system is implemented for choosing right NHP for brain research.

Problems and failure

• Low mental maturity will be expressed in loss of confidence and training failure.

• If animals are in pairs - one should not depend too much in the other.

• Good training is imitated by other animals but bad training or behavior is imitated & can regress training.

• One bad trainer can cause a failure of an animal training.

• Bad or wrong training can lead to an aggressive animal.

• Professional training guidance and supervision is essential.

Current

• In the last 6-7 years all animals in academic institutes in Israel are selected .

• Only a small % can fit this level of training , majority of animals go to short term studies.

• More breeders move into familiarization

• Contact between breeder and client must be strong to allow selection or pre-training

Future

• We at Bioculture - BFC start to do pre training according to client needs.

• This allow the holding time before the study starts at the CRO or research facility to be shorter.

• Animals already arrive with a certain level of training.

• animals which are difficult to adapt are excluded from study.

• Most of clients ask for entering transport box and oral dosing of drugs

Oral dosing of drugs

Thank you for your attention

• Bushmitz Mark Moshe - moshe@bushmitz.com

Scientific & research director at Bioculture group

• Yosi Shohat - shohat20@gmail.com