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Journal of the American Association for Laboratory Animal Science Vol 55, No 5 Copyright 2016 September 2016 by the American Association for Laboratory Animal Science Pages 606–710 Abstracts of Scientific Presentations 2016 AALAS National Meeting Charlotte, North Carolina Poster Sessions blood collection method called the “one man.” The rat is manually restrained and bled by one person. In preparation for this technique, P1 A Novel Vascular Button Connection Using Combined Tech- rats are handled daily. The increase in handling reduces the amount nologies while Allowing for Social Enrichment by Pair Housing of stress the animal will have while being restrained. To restrain the animal is held in a vertical position and is grasped under each axilla, AJ Hehman*1, A Zuvich1, KA Adams2, D Shuey1 using the thumb and middle finger. The pad of the forefinger is used to pull the head back. To aid in visualizing the jugular vein, the 1Toxicology, Incyte Corporation, Wilmington, DE; 2Laboratory thoracic portion of the rat’s chest is shaved and wiped with an Animal Resources, Incyte Corporation, Wilmington, DE alcohol. Once the vein is located, a 25g needle attached to a 1 or 3mL syringe is inserted into the vessel and a blood sample obtained. After Traditional practice has been to single-house vessel-cannulated the appropriate amount of blood is collected, the needle is removed rodents post surgically to protect exposed exteriorized catheters. A and digital pressure is applied. A sample size of up to 2mLs can be port/protective cap model is available that allows for pair and group obtained. The restraint of the animal (tightness of the skin and housing of cannulated animals and is compatible with a harness/ alignment of the neck) is critical when learning this technique; swivel automated blood sampling system. Our goal was to develop a however, it can be learned easily and because this technique does not vessel-cannulated surgical model that could be socially housed and require any specialized equipment it can easily be implemented. would be compatible with response movement caging. To do this, we Hematomas have been seen on occasion with this method. One other modified the automated blood sample connection to be compatible risk of this procedure is death. This is rarely seen but can occur if the with the available port/protective cap system. A sterile custom vessel is not occluded properly. While both methods produce the connector was designed in collaboration with an industry vendor. same quality of sample; the refinement made to the commonly used Animals were dual jugular cannulated using the available port/ jugular vein blood collection method reduces cost in supplies, protective metal cap to allow for pair or group housing. The newly personnel needed, and the amount of stress on the animal. designed custom connector was primed and attached to the port after removal of the protective metal cap. The modified connector allows P3 Table Restraint Method for Working with Awake Nonhuman the standard extension line from the response movement caging Primates system to easily connect for blood sample withdraws. Studies were * conducted using our standard inhouse study design for pharmacoki- AM Storm , A Vinar, H Eric netic studies with multilple blood sample collections out to 24 hours post dose. All dosing and blood sample collections were performed NIH Division of Veterinary Resources, SoBran, Inc, Bethesda, MD in accordance with an IACUC-approved animal use protocol. The end result is a connector that allows for dosing and timed collections The need to use physical restraint for hands-on procedures involving of high-quality blood samples using a response movement caging nonhuman primates (NHPs) is common throughout the laboratory automated blood sampling system. The modified connector further animal community. Historically, our organization has used chemical allows for enhancement to the welfare of the animals by providing restraint for the vast majority of these procedures. After experiencing social enrichment through pair and group housing without compro- issues related to drug tolerance, anesthesia-related physiologic mising the integrity of the surgical model. complications, and the initiation of protocols requiring multiple procedures per day or multiple procedures over a period of P2 A Unique Blood Collection Method in a Conscious Rat successive days, our organization began a search for a new method of restraint which could increase efficiency, reduce or eliminate the need AM Walters*1, M Hirashima1, K Tsuyama1, S Wakamatsu2 for anesthesia, and improve overall animal welfare. There is an abundance of information related to the various restraint devices and 1SNBL USA Ltd, Everett, WA; 2Shin Nippon Biomedical Laboratories, protocols in the literature. Some of the more common methods LTD (SNBL) Japan, Kagoshima , Japan include chair restraint with pole and collar and the use of squeeze cages. Our organization adopted the use of a lesser known restraint Pharmakokinetics and toxicokinetics are important components of device, the primate restraint table, in combination with cooperative drug discovery and development process. The removal of blood at behavioral training for a variety of hands-on procedures including multiple time points from an animal is required for these evaluations. blood collection and veterinary evaluations. Animals were trained to Blood sampling can be stressful for animals because of the handling, use this device by applying positive reinforcement and desensitiza- restraint, anaesthesia, or discomfort associated with a particular tion techniques allowing the NHPs to participate in research technique. The jugular vein is one of a few collection sites which can procedures while fully awake. This was accomplished by acclimat- be used for multiple blood collections in a conscious rat. Blood ing the NHP to the table, then progressing through a series of steps parameters, supplies, and number of personal required, along with to shape the desired behavior of cooperative restraint and limb stress of the animal were evaluated using 2 different jugular vein extension. Training success was measured by the number of fully puncture methods. The first method requires the use of a jugular acclimated animals to the restraint table prior to the initiation of board for the blood draw and requires minimally 2 people (bleeder medical and research procedures. The implementation of this and holder). The rat is restrained in an unnatural position, which can technique has reduced the labor involved with these procedures by cause stress. The ties used to hold the forepaws can cause damage to eliminating the time needed for anesthesia induction, monitoring the limbs and nerves in this area, causing lameness. Tilting the head and recovery, and the complications associated with the use of of the rats in the head cap can cause haemorrhage to the ears. The rat anesthesia. We recommend this restraint device for laboratories can also lose consciousness. This method is commonly used for serial interested in alternative restraint methods in their research and blood collections in rats. The second method evaluated is a unique medical procedures. 606 Abstracts of scientific papers 2016 AALAS National Meeting P4 Strategies to Increase Time Usage and Accessibility of Environ- P6 Optimization of in Vivo Blood Sampling for Pharmacokinetic mental Enrichment Provided to Socially Housed Primates to Studies Improve Animal Welfare and Wellbeing ST Ulufatu, CK McCaughey* CA Carrier* Safety Assessment, Genentech, San Francisco, CA Covance Research Products, Alice, TX Currently blood collection for preclinical mouse pharmacokinetic Group housing is the standard method for housing nonhuman studies has been refined from rotated retro-orbital (RO) eye bleeds to primates at our facility. Delivering enrichment to all social group a microsampling technique via the “whole blood” or “tail prick” members can be difficult due to the social hierarchies. Often the method. This technique in conjunction with an automated, miniatur- dominant animal will guard and chase off other group members when ized immunoassay platform is now primarily employed such that we enrichment is thrown in. While training can be done to combat some of may obtain a full serum-concentration time profile from each individ- this dominant behavior, we looked at other changes that could be ual mouse. By using this combination, we were able to produce a made to improve our environmental enrichment delivery methods to more robust data set than using the RO method and ELISA. The tail large groups of primates. Simple refinements in our methods were prick method involves warming the tail of a mouse and then using a tried to improve animals’ access to offered enrichment. The novel 27g needle which is injected into the tail vein and quickly taken out. strategies discussed here are piñatas, game feeders on timers, and Approximately 20ul of whole blood is collected from the tail vein via larger frozen forage blocks. Experience with primates reveals a hematocrit glass tube and spun down after 30 minutes of clotting. ingestible items encourage foraging and destructible items prove most The serum is collected by cutting the glass hematocrit and pipetting interesting. As a result, selected new strategies targeted these “likes.” at least 10ul of serum. The serum is then processed with an automat- The most successful enrichment strategies provided the group a longer ed, miniaturized immunoassay. Previous methods used staggered time of use, gave them something to do, and provided foraging RO blood collection which combines data across animals to produce opportunities. The longer it took to empty the item, the sooner the a full pharmacokinetic profile. The limiting factor with RO bleeding dominant animal moved on. This allowed other animals in the group a is the amount (~150ul) and frequency blood is able to be collected. chance to forage. The new methods also provided more accessibility to The microsampling technique offers us the opportunity to take a group-housed primates especially the lower ranking members who smaller amount of blood (~20ul) at every time-point from the same typically only had access to leftovers.
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  • (Gekko Gecko LINNAEUS, 1758) Saliva on Angiogenesis

    (Gekko Gecko LINNAEUS, 1758) Saliva on Angiogenesis

    Jurnal Biologi Indonesia 13(2): 253-260 (2017) Effect of Tokay Gecko (Gekko gecko LINNAEUS, 1758) Saliva on Angiogenesis During Wound Healing Phase of Autotomized Tail in Common Sun Skink (Eutropis multifasciata KUHL, 1820) (Pengaruh Saliva Tokek (Gekko gecko, LINNEAUS 1758) Terhadap Angiogenesis Pada Fase Penyembuhan Luka Ekor Kadal Kebun (Eutropis multifasciata KUHL, 1820) Setelah Autotomi) Nurul Inayah1,2, Nyoman Puniawati Soesilo2 & Rarastoeti Pratiwi3 (1)Zoology Division, Research Center for Biology-Indonesian Institute of Sciences (LIPI) (2,3) Faculty of Biology, Gadjah Mada University Email: [email protected] Received: December 2016, Accepted: June 2017 ABSTRACT The purpose of this study was to investigate the effect of Tokay gecko saliva on morphology and angiogenesis response on the healing process of skink tail wound and also to characterize the protein profile of Gecko saliva. Twelve skinks were autotomized and wound surface of tail smeared by young gecko saliva, adult gecko saliva, and human’s saliva twice per day and control. The morphological changes of the wound surface were observed. The angiogenesis response was observed in vitro using Chorioallantois Membrane (CAM) of the ninth day's chick embryos. Protein profile of gecko saliva analyzed with SDS-PAGE. Generally, treated wound showed a better healing. Young gecko saliva able to stimulate angiogenesis in wound healing stage of sun skink tail after autotomy. Saliva protein of young and adult Gecko differences was not only in the size (or density) but also in the number of the bands. The young and adult Gecko revealed a striking consistency of protein patterns, indicating a profound physiological stability of the whole saliva.