Abstracts of Scientific Papers 2016 Association of Primate Veterinarians Workshop.

Journal of the American Association for Laboratory Animal Science Copyright 2017 by the American Association for Laboratory Animal Science

Vol 56, No 2 March 2017 Pages 214–224

Abstracts of Scientific Papers

2016 Association of Primate Veterinarians Workshop Reported Analgesic and Anaesthetic Administration to Nonhuman Primates Undergoing Experimental Surgical Procedure: 2010-2015 H Bertrand*, C Sandersen, P Flecknell Newcastle University, Newcastle Upon Tyne, UK Anesthesia and analgesia techniques are directly tied to the principles of reduction and refinement established by Russell and Burch in 1959. Applied correctly in laboratory animals, use of modern anaesthetic and analgesic regimens should reduce mortality and morbidity and minimize the variation in the scientific data which is being obtained. Small numbers of nonhuman primates are still used every year in biomedical research since these species remain optimal models for some human diseases. This close relation to man enables many modern medical advances in anaesthesia and perioperative care to be applied to NHPs. However, an initial report showed a lack of detailed reporting of anaesthetic and analgesic techniques in larger laboratory species, including NHPs. In 2010, the ARRIVE guidelines were published with the aim of improving reporting of studies involving the use of laboratory animals. These guidelines are currently endorsed by more than 300 peer-reviewed journals. A literature review was conducted to examine the reporting of anaesthesia and analgesia methods used in nonhuman primate undergoing invasive procedures, with recovery from anaesthesia, for scientific purposes. The Pubmed database was consulted and 258 papers published between October 2010 and May 2015 were examined. Only 20.5% of paper reported the analgesic regimen used, with carprofen and buprenorphine as the two most widely used agents. A small survey conducted in parallel on a sample from the paper with no reported analgesia regimen, showed that postoperative analgesia was administered in the majority of studies. Reporting of the anaesthetic regimens was included in 49.7% of papers. Ketamine and isoflurane were most frequently used injectable and volatile anaesthetic agents. In conclusion, anaesthetic and analgesic regimens administered to NHPs remain poorly reported. This lack of detailed descriptions of protocols does little to reassure the public or regulatory authorities that appropriate high standards of perioperative care are employed. Pharmacokinetics of a Novel, Transdermal Fentanyl Solution in Rhesus Macaques (Macaca mulatta) GW Salyards*, HK Knych, AE Hill, KL Christe California National Primate Research Center, Davis, CA Rhesus macaques (Macaca mulatta) are the most commonly used nonhuman primate biomedical model, consisting of both research and clinical procedures requiring analgesia. Providing analgesia is imperative and opioids are a mainstay of analgesic therapy. A novel, transdermal fentanyl solution (TFS) was developed to be a long-acting, single-administration, topical opioid that delivers ≥4 d of therapeutic plasma concentrations as previously published in beagles (Canis familiaris). The TFS dose was applied to clipped dorsal skin under ketamine (10 mg/kg IM) sedation. We describe the pharmacokinetic profile of TFS in healthy, adult rhesus macaques (n = 6; 3M, 3F) in this 2-period, 2-treatment crossover study of a single topical administration of 1.3 (25) and 2.6 mg/kg (50 µL/kg) TFS. We hypothesized TFS in rhesus macaques would provide ≥4 d of therapeutic plasma concentrations (≥0.2 ng/mL) as compared to beagles. Plasma fentanyl concentrations were determined by

tandem liquid chromatography-mass spectrometry prior to drug administration and for up to 21 d post-administration (0, 0.5, 1, 2, 4, 8, 12, 24, 36, 48, 60, 72, 96, 120, 144, 168, 240, 336, 408, and 504 hours). Non-compartmental pharmacokinetic analysis was performed. For each dose (1.3 and 2.6 mg/kg), respectively, maximum plasma concentration was 1.95 ± 0.40 and 4.17 ± 0.70 ng/mL, occurring at 21.33 ± 4.09 and 30.67 ± 8.68 hours. The area under the curve was 227.25 ± 31.67 and 446.97 ± 49.08 h/ng/mL and the terminal elimination half-life was 93.65 ± 7.06 and 98.75 ± 5.40 hours. No adverse effects were noted after drug administration at either dose. Macaques maintained plasma fentanyl concentrations ≥0.2 ng/mL for at least 7 days after 1.3 mg/kg and at least 10 d after 2.6 mg/kg topical administration of TFS. A single TFS dose may provide efficacious therapy with less stress, discomfort, and risk to animals and personnel. Serological Screening of Tuberculosis in Nonhuman Primates using a Multiplex Antibody Diagnostic Assay RK Dhawan*, S Mehra, TW Foreman, NA Golden, KM Gentry, BA Bronson, ML Wunderlich, D Kaushal Charles River Laboratories, Wilmington, MA A rapid serological screening assay for tuberculosis (TB) in nonhuman primates has been developed using Luminex beadbased technology. Purified Mtb specific recombinant antigens were coupled to beads to create a 7-member multiplex (7-plex) panel. Sera from Indian rhesus macaques experimentally infected with Mtb strain CDC1551 via the aerosol route was used for sensitivity and specificity analyses. We used preimmune or preinfection sera as controls, and longitudinally collected samples at later time points, including at the end of the study. Samples derived from 3 different studies were used. Macaques exposed to high doses of Mtb via aerosol develop rapidly fulminating pulmonary TB, while a number of those exposed to low doses develop latent TB infection (LTBI), characterized by immunological reactivity to Mtb antigens sans any clinical signs of disease. In addition, we recently demonstrated that a Mtb mutant attenuated in stress response (MtbΔsigH) offers robust immune responses upon mucosal vaccination which protects against lethal challenge with Mtb. Both the immune responses and the magnitude of protection surpass that offered by a comparable BCG vaccination. Prior to vaccination/infection, all sera samples were negative on all multiplex assays while a majority of the sera exhibited reactivity to one or more of the peptides/assays postinfection with ESAT6 as the most reactive assay. Importantly, while positivity was detected in MtbΔsigH-vaccinated macaques (both the breadth and the magnitude of this reactivity was amplified following Mtb challenge), no reactivity was observed post-BCG vaccination. High specificity (>98%) of the multiplex assay was also observed when a large number of sera (n=740) were screened from historically known negative rhesus and cyno colonies. Our results have important implications for the development of diagnostic assays for TB in high burden (and thus high BCG vaccination prevalence) areas of the world. The data from these studies prove that this high throughput multiplex assay can be used for routine serological screening of Mtb in nonhuman primates. Cerebellar Dysfunction in a Captive-Bred Juvenile Rhesus Macaque (Macaca mulatta) Imported from China G Fleurie*, M Cottingham SNBL USA-SRC, Alice, TX

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Abstracts of scientific papers

In April 2015, a 3-year-old, 2.77 kgs male rhesus macaque (Macaca mulatta), serologically negative for BV, SIV, SRV, and STLV was imported from China. Physical examination, at the time of the first TB testing of quarantine, was unremarkable. During the CDC quarantine, the animal was found to be uncoordinated, shaky, and losing balance. He was reported weak. By the time the animal was assessed, he was BAR, had normal hydration, and the iSTAT results were unremarkable (very mild BUN elevation). He was treated with primarily supportive care and he seemed to recover well post treatment. However, after close observations, it was found that the animal continued to exhibit the same symptoms. We observed central nervous symptoms such as repeated ataxia and missed occasional steps while jumping and climbing. He was exhibiting jerking movements and was uncoordinated. Appetite and hydration remained normal. Blood tests (CBC and chemistry) and X-rays were taken post quarantine and all results were within normal limits. A neurologic examination was conducted cage side and included evaluation of the mentation, behavior, posture, gait, strength, coordination, and cranial and spinal nerve function. Over time he seemed to improve slightly but continued to exhibit imbalance and uncoordination. All clinical findings pointed to a cerebellar dysfunction. The prognosis was poor for recovery and the animal was ultimately euthanized in September 2015. Final diagnostic was made on histopathology and will be discussed during the presentation. Sudden Paraplegia in a Moustached Tamarin (Saguinus mystax) AS Gozalo*, CR Michaud, R Herbert, WR Elkins Comparative Medicine Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD A wild-caught research naïve adult male tamarin (Saguinus mystax) presented a sudden onset of bilateral hind limb paresis. On physical examination, the tamarin had decreased musculature in hind legs with lack of movement, not moving the tail, no panniculus response, no anal tone, no palpable femoral pulse, no deep pain response, and both rear legs were cold to touch. Hydration was normal and mucous membranes were pink with a 2 second refill and there were no signs of trauma Differential diagnoses for sudden bilateral paraplegia include thoracolumbar spine trauma that compromises the innervations of the lower extremities; compression caused by a displaced intervertebral disk; neoplasia, either of the spinal cord or adjacent tissues, that exerts a mechanical impairment of the nerves; vascular and inflammatory disorders of the spinal cord; hypokalemia; saddle thrombus; dissecting aneurysm of the abdominal aorta. Due to a poor prognosis the tamarin was euthanized with sodium pentobarbital overdose. At necropsy, the left kidney presented a hemorrhagic infarct affecting the cortex, and a large well-adhered blood clot was visible at the trifurcation of the distal abdominal aorta extending towards the internal iliac arteries. Microscopically, the left kidney had a hemorrhagic infarct and both kidneys had interstitial lymphoplasmacytic infiltrates and expanded mesangial matrix with tubular proteinuria and proliferative arteriosclerosis. The abdominal aorta and iliac arteries had marked thrombosis characterized by focally extensive, severe, endothelial degeneration, sub-endothelial fibrin deposition and hemorrhage. Since no other lesions were found that might explain the clinical signs observed, sudden occlusion of the abdominal aorta was most likely the cause for the bilateral posterior paresis noted in this moustached tamarin. Clinical signs, pathophysiology, prospective treatments, and outcome of aortic thromboembolism in animals and humans will be discussed. Acute Paraplegia in an Aged Female Rhesus Macaque (Macaca mulatta) K Woodward*, R Sammak, R Keesler California National Primate Research Center, Davis, CA An 18-year-old female rhesus macaque (Macaca mulatta) presented

for acute paraplegia. Cageside neurologic exam showed normal mentation, normal cranial reflexes, and ability to urinate, but the flaccid paralysis of the hindlimbs with no response to deep pain. Decreased anal tone was noted after ketamine sedation. Spinal radiographs reveled severe thoracolumbar spondylosis. CT with myelogram was performed and a discrete compressive lesion could not be identified. Due to poor prognosis for recovery, the animal was euthanized the same day as presentation. No gross neurologic lesions were seen on necropsy, although a colonic adenocarcinoma was found incidentally. Within the thoracic and lumbar spinal grey matter, histopathology revealed arterioles surrounded by thin bands of highly eosinophilic fibrillar material with no lesions observed in peripheral nerves. Trichrome staining suggested that the fibrillar material was collagen and electron microscopy of the spinal cord confirmed irregular perivascular accumulation of collagen. Additionally, a discrete 4mm area of the proximal lumbar spinal cord had acute axonal swelling with no associated inflammation and no upstream or downstream histologic lesions. A discussion of the clinical signs, diagnostics, and differentials will be presented, along with further information on the disease processes. When Medical Intervention May Diminish Quality of Life DL Hasselschwert University of Louisiana, Lafayette New Iberia Research Center, New Iberia, LA Two novel chimpanzee cases are presented, that while intervention or corrective surgery was medically possible, doing so would have diminished quality of life for the animals. The phrases “first do no harm” and “consideration for quality of life” are philosophically integral to the medical and veterinary professions. The implication is that the medical professional should correct the problem; but what if intervention is detrimental to the animal? The first case was a chimpanzee born with complete unilateral cheiloschisis. Several factors were considered during assessment including surgical approach and staging, the extent of the defect, and quality of life. The defect was cosmetic and thought to have resulted from intrauterine trauma. Complications associated with cleft repairs such as selfmutilation were considered and lead to the decision not to repair the defect. The second case was an adult chimpanzee that presented with vitiligo. Depigmentation of the face and shoulders occurred at approximately age 12. Although thought to be autoimmune, treatment beyond homeopathic therapy was not considered due to the potential for immune suppression, thus comprising her quality of life. Both animals continue to thrive without complications. The uniqueness of the presenting problem may lead to the belief that intervention is necessary; however analysis of the quality of life for the animal may contraindicate some types of intervention. Clinical Management and Refinement Strategies for Cranial Implants in Common Marmosets (Callithrix jacchus) A Goodroe*, S Smith, R Shadmehr, X Wang, C Garrett Johns Hopkins University, Baltimore, MD An easily accessible auditory cortex and diverse vocal repertoire makes the common marmoset (Callithrix jacchus) an ideal model for studying the neural basis of auditory processing, perception, and vocal communication. Linking the neural basis of sound perception with behavior is facilitated through cranial implants, which allow the electrical activity of single or multiple neurons to be recorded. While 2 decades of investigator and veterinary collaboration have resulted in refinement of cranial implant placement and maintenance, 2 broad categories of ongoing clinical challenges exist. The first are challenges inherent to the traditional design of the implant. The second are challenges associated with novel modifications to the traditional structure and function of the implant. The traditional implant design features 2 regions of clinical importance: the accessible skin-dental cement margin and inaccessible dental cement and cortical bone interface. Complications associated with 215


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Vol 56, No 2 Journal of the American Association for Laboratory Animal Science March 2017

the margin include localized inflammation (often associated with commensal skin bacteria) and proliferative granulation tissue. Complications associated with the interface, while less frequent due to margin maintenance regimens, are far more serious and involve the centripetal migration of specific opportunistic bacteria, fungi, or chronic, proliferative granulation tissue between the cortical bone and dental cement. Complications include loss of bone or dental cement integrity, cranial implant dislodgement, and meningitis or meningoencephalitis, as well as systemic pathology associated with chronic inflammation. More recently, modifications in the traditional implant design have presented new clinical challenges. Examples include the use of artificial dura, addition of a cochlear implant, and a reduced profile titanium cranial implant. While similar principles of cranial implantation are widely used across nonhuman primate neuroscience models, our institution was the first to apply and continue optimizing these techniques in marmosets. Case examples highlighting refined implant maintenance practices, concepts critical to the margin and interface management, and unique implant design and function will be presented. Dynamic Contrast Computed Tomographic Imaging as a Method of Determining Liver Function in Nonhuman Primates in Ebolavirus Research M St Claire*, S Chefer, RA Byrum, R Reeder, RF Johnson National Institutes of Health, Frederick, MD In the aftermath of the 2014 Ebolavirus (EBOV) outbreak in West Africa, major efforts have been undertaken to develop innovative methods for the study of EBOV in animal models. Terminal stages of EBOV disease are characterized by multiorgan system failure, including the liver as one of the primarily targets of infection. Vascular damage to the organ is expected to result in alterations of tissue perfusion, eventually leading to liver failure. Abnormalities in liver function during disease progression can be assessed by clinical imaging which is used in the monitoring, staging, and treatment of human patients; but has not been examined in nonhuman primates, which are the gold standard animal models for pathogenesis, treatment, and vaccine research. In the past, liver function has been difficult to assess in animal models. We recently evaluated an application of dynamic contrast-enhanced (DCE) computed tomography (CT) imaging in NHPs with a clinical scanner as a method for staging liver disease during EBOV infection. Tissue alterations may be assessed comparing perfusion rates pre- and post-EBOV exposure. This method requires an injection of contrast media in defined small quantities at high flow rates to obtain a short and well-defined intravenous bolus. Since a clinical scanner has less sensitivity and resolution as compared with a micro-CT, to meet the requirements for DCE application the contrast media injection strategy (dose, volume, rate) was adjusted and evaluated in rhesus macaques (Macaca mulatta) and common marmosets (Callithrix jacchus). In addition, the impact of injection location (tail or leg veins versus arm vein) on the general distribution of contrast material was assessed. The results indicate that while the contrast-enhanced image quality was acceptable for application of this method in large NHPs such as Rhesus macaques, it has limited application in marmosets due to artifacts associated with respiration and limitations with site of contrast injection. The Clinical Management of an Outbreak of Yersinia enterocolitica Infection in a Colony of African Green Monkeys (Chlorocebus aethiops sabeus) Q Wilson*, A LeGrande, AR Hutchison, G Balamayooran, RN Andrews, HM Atkins, KT Michalson, ND Kock, D Caudell, MK Gee Wake Forest School of Medicine, Winston-Salem, NC A 64-day-old male African green monkey (Chlorocebus aethiops sabeus) was found dead in his enclosure. This animal was housed in 1 of 16 indoor-outdoor breeding pens, each holding a group of 11-30 individuals. Gross necropsy findings revealed severe bilateral

pneumonia which was positive for Yersinia enterocolitica on bacterial culture. Increased visual surveillance of this pen was implemented, but concerns not identified. One month later, a monkey from a different group died with severe necrotizing typhlocolitis which was also confirmed to be Y. enterocolitica via bacterial culture and histopathology. Colony-wide surveillance, increased sanitation, and quarantine procedures were implemented. Animals showing even subtle abnormalities, including mild ataxia or depression, were examined and treated with antibiotics and supportive care. Over the subsequent 6 months, 115 animals had become infected with Y. enterocolitica, with 23 mortalities. This led to a strategy for colonywide quarantine and relocation of individual pens into rack-caging for treatment. Screening was conducted by fecal PCR specific for Y. enterocolitica, before returning the animals testing negative to the indoor-outdoor enclosures. When managing a colony outbreak of yersiniosis, early identification of infected individuals, colonywide treatment and intensified sanitation are critical for successful treatment and reduction of the microbial burden. Computed Tomographic Diagnosis of Multiple Bullae and Pneumothorax and Treatment of Pneumothorax in a Rhesus Macaque (Macaca mulatta) JM Kim*, JS Shin, BH Min, WY Jeong, GE Lee, MS Kim, JE Kim, CG Park Xenotransplantation Research Center, Seoul National University Hospital, Seoul, Korea Bullae and pneumothorax have various aetiologies in veterinary medicine. We observed multiple bullae and pneumothorax in a transplanted porcine islet and 9-year-old male rhesus monkey (Macaca mulatta). Ascites was observed at 5 days after transplantation (TPL). While the computed tomography (CT) was performed to evaluate the liver, bullae and pneumothorax were observed on the CT on day 8 after TPL. No findings related to pneumothorax were seen on the chest X-ray before TPL. It was thought that bullae were secondarily induced by lung mites and bullae ruptured during positive ventilation at the TPL. Although the monkey had moderate pneumothorax, dyspnea was not observed. The serial chest X-rays were performed to monitor the pneumothorax. As the pneumothorax worsened, the air was removed by thoracocentesis at 13 days after TPL. A thoracotomy between the left 3rd and 4th rib was performed to fixate the ruptured lung at 14 days after TPL. Air leakage was found in the middle of the left cranial lung lobe parenchyma, and the ruptured parenchyma was sutured with a purse string suture pattern using 4-0 absorbable material. The monkey had no complications after surgery. To the best of our knowledge, this is the first reported treatment of pneumothorax caused by ruptured bullae in nonhuman primates. Oral Squamous Cell Carcinoma in a Sub-Adult Rhesus Macaque (Macaca mulatta) H Sidener*, C Bishop, O Slayden Oregon National Primate Research Center, Beaverton, OR A 4-year-old female rhesus macaque presented for gingivitis, halitosis, and gingival hyperplasia of the left maxillary and mandibular dental arcade. Oral gingival cultures were negative for bacterial pathogens, and the hyperplastic tissues were trimmed and submitted for histopathology. All tissues showed abundant dysplasia, but the mandibular tissues were diagnosed as an oral squamous cell carcinoma (OSCC). The presence of viral cytopathology (koilocytosis, chromatin margination, and intranuclear inclusions) suggested papilloma virus infection as an underlying etiology. This animal is enrolled in an IACUC-approved research project that includes placement of a testosterone implants at the time of menarche (age 2.5 years) to maintain an elevated serum testosterone at 3-4 fold normal (1-1.5 ng/ml). The tumor progressed only minimally in the 6 months following the diagnosis, and the animal has shown no adverse clinical consequences outside of mi-

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nor gingivitis and periodontal disease. She exhibits normal mense and growth patterns, and has no observable difficulties eating or conducting normal species specific behaviors. A second animal was diagnosed with gingival hyperplasia but biopsy did not reveal evidence of papillomavirus infection, and neoplasia was not diagnosed. Papillomaviruses have been connected with cervical, genital, and oral squamous cell carcinomas in older macaques, but this is the first report of OSCC in a rhesus macaque of this age, with or without underlying viral etiology. Androgen receptors have been shown to be present on 60% of examined specimens of head and neck squamous cell carcinomas in one clinical study, and it has been suggested that the expression of androgen receptors is critical for OSCC growth, survival, and tumorigenic activity. Some types of carcinomas with androgen receptors have potentially shown treatment promise using androgen deprivation therapy. Nonhuman Primate Models of Traumatic Hemorrhagic Shock MV Campagna*, CA Koeller* Naval Medical Research Unit San Antonio Hemorrhage is a major cause of mortality among both humans and animals sustaining traumatic wounds. In addition, it is the leading potentially survivable cause of death among military personnel injured on the battlefield. Multiple animal models of traumatic hemorrhagic shock have been developed in order to better understand the associated pathophysiology. Nonhuman primates have surfaced as an invaluable asset in the evaluation of potential therapies for soldiers injured in combat due to their similarities to humans in physiologic and immunologic responses and their decreased likelihood of xenogenic reactions to human blood products. Here, we describe and compare historical, current, and novel models of hemorrhagic shock in order to define their specific advantages, disadvantages, and clinical translatability. Particular focus will be on novel rhesus macaque models of traumatic hemorrhagic shock. Clinical Relevance of Nonhuman Primate Models of Traumatic Hemorrhagic Shock to Both Human and Nonhuman Primate Patients MV Campagna*, CA Koeller* Naval Medical Research Unit San Antonio The goal of present studies is to use novel rhesus macaque models of pressure-targeted, poly-traumatic hemorrhagic shock to assess multiple parameters, including indicators of physiology, hematology, coagulation, immunological response, and clinical outcome. These studies have been designed to help identify potential treatments for traumatic hemorrhage sustained by combat casualties in austere environments. However, their clinical relevance can spread beyond their original intentions. As traumatic hemorrhage is also a potential cause of morbidity and mortality among nonhuman primates, particularly socially housed males, the pathophysiology and clinical outcomes observed in these models are relevant to veterinary medicine as well. Data obtained from these models may assist primate veterinarians in their evaluation and clinical management of fight-wound or trauma-induced hemorrhage, by identifying indicators of more severe physiologic compromise, and potentially elucidating ideal therapies for use in critical cases. Development of a Process to Enhance Trio Formation in Adult Male Rhesus Macaques (Macaca mulatta) A Ruhde*, KC Baker, JL Blanchard, RP Bohm, K Russell-Lodrigue Tulane National Primate Research Center, Covington, LA The purpose of this study is to evaluate a methodology to enhance and to systematically assess outcomes of indoor trio housing adult male rhesus macaques. The formation of trios can significantly improve animal welfare by providing an additional indoor social

housing option. The addition of trio housing may reduce the necessity to singly house animals when experimental groups are designed with an odd number of animals. When only pair housing is pursued, some animals must be housed alone. Our study reports potential predictors of successful trio housing based on data collected from adult males (n = 37, ages 4.7-12.9 yrs.) during assessment and initial introduction phases. Behavioral data (6-8 hours) in single housing was collected prior to the start of introductions. The composition of the trios was predetermined from a limited pool of subjects. Each subject was introduced separately to both potential trio-mates for 48 hours in protected contact (PC). There were significant behavioral differences in single housing between animals that had at least one successful PC introduction (“successful males”) and those that failed all the PC attempts (“unsuccessful males”). Unsuccessful males exhibited 3 times more aggressive displaying directed outside their cage and spent twice as much time self-grooming in comparison to the successful males. Successful males spent almost twice as much time resting versus the unsuccessful males. We found that a greater weight disparity between potential partners yielded a higher success rate for the PC introduction outcomes. Only 34% of the PC introductions were successful when the weight difference between potential partners was ≤1 kg, while 72% of the PC introductions were successful when the weight difference was >1 kg. Successful group housing of adult male rhesus macaques may be improved by taking such data into consideration during the preliminary trio formation processes. Characterization of Multidrug-Resistant Enterococcus faecalis Isolated from Research Macaques MT Lieberman*, SE Woods, F Lebreton, MS Gilmore, JG Fox Massachusetts Institute of Technology, Division of Comparative Medicine, Cambridge, MA Multi-drug resistant (MDR) Enterococcus faecalis is a common and serious cause of nosocomial infections. Previous characterization of 14 E. faecalis isolates from cephalic recording chambers of macaques used in neuroscience research revealed 2 lineages with marked multi-drug resistance. Lineage A (LA) isolates (n = 7) showed differing susceptibilities to gentamicin, with 4/7 isolates displaying high-level gentamicin resistance, while Lineage B (LB) isolates (n = 7) displayedwi resistance to neomycin. All isolates from Lineages A and B displayed marked streptomycin resistance. Resistance to tetracycline, chloramphenicol, erythromycin, and trimethoprimsulfamethoxazole was also noted. DNA was extracted from 2 LA and 1 LB isolate and sequenced on a single SMRT cell on an RS2 sequencer. Genomes were assembled and FASTA sequences were analyzed to confirm sequence type (ST) and identify genes of interest. Two isolates from LA were confirmed to be ST4 and the isolate from LB was confirmed to be ST55. All isolates had a unique antimicrobial resistance gene profile with the lsa(A) gene encoding intrinsic resistance to lincosamides and streptogramins A as the only common gene between all 3 isolates. Additional macrolide resistance encoded by erm(B) was identified in 2 isolates. Four genes encoding aminoglycoside resistance were identified: str and 3 aminoglycoside-modifying enzymes: aph(3’)-III, aac(6’)-aph(2”), and ant(6)-Ia. Genes encoding tetracycline resistance included mechanisms for both efflux pumps (tetL) and ribosomal protection (tetS and tetM). Phenicol resistance was conferred by the cat gene and was present in ST4 isolates, but not the ST55 isolate. Trimethoprim resistance encoded by dfrG was noted in 1 ST4 isolate. Other virulence factors identified included cytolysin, enterococcal surface protein, aggregation substance, gelatinase, collagen adhesion precursor, and endocarditis antigen. E. faecalis isolates from cephalically implanted macaques display genetic similarities to isolates associated with human nosocomial infections. Macaques represent a unique research model to study nosocomial infections due to their long-term residence in a healthcare setting and intermittent antimicrobial exposure.

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Sex Skin in a Chimpanzee

A 49-year-old female chimpanzee (Pan troglodytes) presented in January 2014 with a nonhealing lesion to her sex skin. This case report will detail the case progression, diagnostic tests, biopsy results, and various administered therapies. There will be opportunities for the audience to weigh in on differential diagnoses, special tests, and treatment options. The case will conclude with a summary of results and progression to the present day, including recommendations for case management.

gonadotropin, which causes a decidual reaction within the uterus, and is the cause of the uterine enlargement. Chorionic gonadotropin is also responsible for the luteinization noted within the ovary. Ovarian choriocarcinoma is extremely unusual in a nulliparturient juvenile primate. However, it is a germ cell tumor, and it has been reported in children and chinchillas (who have similar placentation). Germ cell tumors represent 2-4% of human pediatric cancers. Due to metastasis, the prognosis for ovarian choriocarcinoma is poor. Diagnosis is achieved antemortem via a combination of imaging, a high index of suspicion, and elevated circulating chorionic gonadotropin levels. The chief differential diagnosis in this case is granulosa cell tumor, but characteristic Call-Exner bodies were not present, and granulosa cell tumors are not biphasic. There was no evidence of granulomatous disease such as tuberculosis in this young monkey.

Hepatomegaly, Weight Loss, and a Fetus in Distress

Abdominal Mass in a Female Cebus paella

A Haertel*

J Steinbach*, M Klinger , T Albers

Oregon National Primate Research Center, Beaverton, OR

New York University, New York, NY

A 9-year-old pregnant female rhesus macaque (Macaca mulatta) socially housed in an outdoor shelter presented while sedated at a semi-annual physical exam with severe hepatomegaly, alopecia, BCS of 2/5, and 23% weight loss. The animal was transported to the clinic hospital still sedated, and a mild tachypnea was observed. Abnormal lung sounds were not detected on auscultation, and sedation recovery was reasoned as the cause of tachypnea. Results of a CBC demonstrated leukocytosis with a mature neutrophilia, a mild anemia, and thrombocytosis. Blood chemistry results showed hypoalbuminemia and elevation of alkaline phosphatase and gamma-glutamyl transferase. Abdominal ultrasound examination revealed hepatomegaly, a strikingly hypoechoic liver parenchyma and a second trimester fetus with marked tachycardia. Hepatic amyloidosis and imminent fetal demise were strongly suspected. Besides diarrhea, the patient was stable upon recovery from sedation. Sulfasalazine and s-adenosylmethionine were started. Ultrasound assessment performed 6 days after presentation was consistent with placental abruption. Because of this potentially grave complication of pregnancy in a compromised animal, euthanasia was performed, and a postmortem evaluation was conducted. The gross lesions included uterine hemorrhage subjacent to the placenta, hepatomegaly with waxy and friable parenchyma, typhlocolitis, a gastric trichobezoar, lung abscessation, pleuritis, and pericarditis. The differential diagnosis of the lung lesions included Klebsiella pneumoniae, Corynebacterium spp., Streptococcus pneumoniae, Staphylococcus aureus, Escherichia coli, Yersinia pseudotuberculosis/enterocolitica, Nocardia spp., and Mannheimia spp.

An approximately 15-year-old clinically healthy female Cebus apella weighing 3 kg was euthanized followed by perfusion at the end of an IACUC-approved operant behavioral study. Blood work was completed within the previous 3 month period as part of normal quality health assurance for the Cebus colony. Serology indicated negative antibodies to H. tamarinus and H. samarinus and positive antibodies to measles and CMV. GI culture was negative for Salmonella, Shigella, Campylobacter, and Yersinia. CBC and chemistry values were all within normal limits. During necropsy, gross examination revealed a mass connected to the cranial aspect of the uterus. All other organs and tissues were observed to be within normal limits. The mass was filled with dark fluid. Possible rule outs include endometriosis, adenomyosis, endometritis, endometrial hyperplasia, adenocarcinoma, endometrial polyps, endometrial cyst, endometrial fibroid, leiomyomas, ovarian cysts, and ovarian neoplasia. Tissues were submitted for histopathology. Histopathological diagnosis confirmed a uterine cyst. Spontaneous diseases of the female nonhuman primate reproductive tract have been the subject of several reviews. A uterine cyst was reported in a female Rhesus. Several reports have identified ovarian cysts and endometrial polyps in Old World primates and there has been at least 1 report of an endometrial polyp in a spider monkey. To the author’s knowledge this is the first reported diagnosis of uterine cyst in Cebus apella.

A Beck, E Magden*, S Buchl, W Baze Albert Einstein College of Medicine, New York, NY

Pulmonary and Hepatic Masses in a Young African Green Monkey (Chlorocebus sabaeu) DJ Schwahn*, J A Smith, DL Hasselschwert DJ Schwahn Pathology, Madison, WI A 1-year-old female African green monkey presented for lethargy. Physical examination revealed hepatomegaly and a firm lower abdominal mass. A foreign body was suspected. Exploratory laparotomy identified an irregular, friable, 4 x 2 cm mass engulfing the left ovary and oviduct, as well as numerous 2-3 mm diameter, firm, solid, white nodules suspected to be granulomas within the 4-times-larger-than-expected liver. Euthanasia was elected. Similar 2-3 mm white nodules were identified within the lungs. The overall impression was of disseminated granulomatous disease. Additionally, the uterus 5 five times larger than expected with a greatly thickened wall and mucosa. Histopathology revealed the presence of a biphasic ovarian choriocarcinoma with metastases to the liver and lungs. The tumor consists of 2 populations of cells—an outer rim of syncytiotrophoblasts with many nuclei and sparse cytoplasm surrounding an inner ring of more plentiful, typically uninucleate cells with abundant cytoplasm. Inside both rings is a central core of necrotic cellular debris. Ovarian choriocarcinomas secrete chorionic

What’s Your Diagnosis? DM Sergio, B Ogden*, R Bunte, MM Varela, E Pena Singapore Health Services Pte Ltd , Singapore A 4-year-old male cynomolgus macaque that had appeared normal 1 hour before was found recumbent in its cage with tremors of the head and neck. Following treatment for shock, the tremors ceased. Radiographs showed mild dilatation of the stomach and marked gas distention of the bowels. The animal was returned to the cage for observation and fluid therapy continued. Tremors were again observed 6 hours later, but ceased after additional treatments and the animal appeared to rest comfortably. Lab results showed mild increases in the WBC, monocytes, neutrophils, RBC, HGB, and HCT, prompting addition of antibiotic to the treatment regimen. Serum biochemistry showed creatinine, ALP, ALT, AST, LDH, GGT, and CPK were all remarkably high. Because the animal was still weak the next morning and had labored breathing, it was sedated and placed under light isoflurane anesthesia via an endotracheal tube. Orogastric intubation revealed a mixture of gas and a coffee ground appearing fluid from the stomach. An exploratory laparotomy was performed, confirming marked gas dilatation of the small and large bowel, but minimal gas in the stomach. No other abnormalities were noted. During closure of the abdomen, the heart rate became erratic and the animal went into respiratory arrest. When emergency treatments failed to improve the condition, the animal was euthanized.

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Necropsy revealed a mixture of partially digested and fresh blood in the stomach and a 3 cm diameter reddened area of the mucosa in the pyloric region. Gas distention of the intestines was noted, but the appearance of all other organs was unremarkable. Anaerobic culture of the stomach contents was positive for Clostridium bifermentans. Morphologic diagnosis was severe focal gastric hemorrhage and histopathology showed severe intramucosal pyloric hemorrhage.

the femur with a butterfly fragment. Due to the complexity of the case, a board-certified surgeon was consulted on a treatment plan. This surgeon volunteered to collaborate with veterinarians to repair the fracture with a plate-rod technique (an IM pin combined with a bone plate). Thanks to these combined efforts, the patient recovered successfully and all of the vets involved gained valuable experience.

Critical Masses: What’s Your Diagnosis?

Proximal Tibial Osteosarcoma in an Adult Rhesus Macaque (Macaca mulatta)

SN Poulle*

E Van Bebber*, DL Hasselschwert

Bioculture Group, Riviere des Anguilles, Mauritius

University of Louisian Lafayette New Iberia Research Center New Iberia, LA

Three cases of unusual masses were detected in M.fascicularis. In the first case, abdominal distention was noted in a 3-year old male animal. X-ray and ultrasonography revealed a fluid-filled mass in the right upper quadrant of the abdomen. The animal was BAR and eating normally. In the second case a hard, radio-dense 2 x 2cm mass was detected in the right hip of a 3-year old male macaque. The mass was fairly mobile and did not seem to be associated with any of the adjacent skeletal structure. The animal was bright, alert, and responsive and with only slightly impaired locomotion. In case three, the sudden onset of ataxia and head tilt in a 2-year old female was observed. Pupils and other neurologic parameters were unaltered. Euthanasia was elected and upon necropsy a tumor-like mass was found on left side of brain case, next to the cerebellum and not associated with the brain.

A 5-year-old Rhesus macaque (Macaca mulatta) presented with left leg lameness. Upon physical examination, a large, palpable mass was present at the level of the proximal tibia. The animal appeared otherwise healthy. Radiographs revealed a large, lytic lesion at the level of the proximal tibia indicative of osteosarcoma. The animal was euthanized, and grossly, an osteolytic mass invaded the proximal tibia and surrounding soft tissues. Animal presentation, physical examination, radiographic findings and ultimately histopathology confirmed a diagnosis of osteosarcoma. Prevalence of Flavivirus Antibodies to Zika Virus, Dengue Virus, and West Nile Virus in North American Nonhuman Primate Colonies

Alphaxalone Use in Rhesus Macaques

RK Dhawan*, ML Wunderlich, B Bronson, L Campbell, C Wang

D Kempf, J Johnston*, D Levesque, L Howell

Charles River Laboratories, Wilmington, MA

Yerkes National Primate Research Center, Atlanta, GA

The ability to detect Zika virus (ZIKV) infections has become a recent health concern in NHP colonies due to implications with fetus development. Identification of animals infected with ZIKV in a timely manner is therefore critical. A Luminex bead-based multiplex assay was developed using conventional whole virus lysate and recombinant (rNS1 and rENV) ZIKV proteins. Test sera were also analyzed by ZIKV, Dengue virus ((DENV) and West Nile virus (WNV) ELISA kits to investigate possible serological cross-reactivity between these flavivirus agents. Assay specificity and serological cross-reactivity between flaviviruses was assessed using DENV and WNV human seroconversion panels. The DENV samples scored positive on DENV (homologous) ELISA, ZIKV multiplex and ELISA assays and WNV ELISA assay. Similar cross reactivity was seen with the WNV panel on DENV ELISA but not on ZIKV assays. Mauritian and Chinese cynomolgus macaques from Charles River indoor colonies showed 0/200 positives (100% specificity) tested by ZIKV, DENV, and WNV assays. Similar analysis of sera samples (n = 300) from 3 outdoor North American NHP colonies showed 1-2% positives for ZIKV, 10-15% Dengue and 2-7% WNV positives for individual colonies. Same samples gave positive scores on two or all three assays with lowest scores on ZIKV assays. These results indicate possibility of DENV infection with limited serological cross reactivity to WNV and ZIKV assays. Thus serology assays for these agents can’t distinguish between different flavivirus NHP antibodies and individual flavivirus infections can only be confirmed by agent specific PCR assays. Therefore, we have developed a ZIKV RT-qPCR assay based on ~90 ZIKV sequences in the NCBI database. Inactivated ZIKV was acquired and tested to confirm that the process from RNA isolation to PCR testing was functional. We evaluated 45 NHP blood samples obtained from 3 different institutions and all were determined to be negative (0% positive). This confirms that PCR can be used to routinely monitor for active infection of ZIKV in NHP research colonies, and may also be considered for use in ZIKV research studies.

Alphaxalone is a neuroactive steroid anesthetic that is poorly water soluble. The original formulation used Cremaphor EL, a nonionic surfactant, to increase solubility. However, the additive caused hypersensitivity and histamine release. These side effects led to the eventual withdrawal of the anesthetic from the market. Recently, the FDA approved the use of a new, water soluble formulation of alphaxalone for intramuscular (IM) and intravenous (IV) use in dogs and cats in the United States. This study aimed to determine the physiologic effects of alphaxalone on heart rate, respiratory rate, blood pressure, oxygen saturation, end-tidal carbon dioxide, body temperature, and depth of anesthesia using a behavior scoring system in rhesus macaques (Macaca mulatta, n = 4). In the first experiment, alphaxalone was administered IM at 5 mg/kg. The induction time was rapid (5.0 ± 0.02 min) and provided a 42 ± 6.7 minutes period of sedation. Cardiovascular and respiratory parameters remained stable. Adequate sedation was achieved; however, the single injection did not provide a surgical plane of anesthesia as evidenced by only a slightly decreased jaw tone and maintenance of the medial palpebral reflex. In the second experiment, alphaxalone (5 mg/kg IM) was administered and anesthesia was maintained using a continuous rate infusion (CRI, 0.125-0.150 mg/kg/min IV) for a 1-hour period. Again, cardiovascular and respiratory parameters remained stable, although, in this experiment subjects did reach a surgical plane of anesthesia. The recovery time was 19 ± 0.05 min following termination of the CRI. In conclusion, these results suggest that alphaxalone may represent an attractive alternative or adjunct to commonly used anesthetic protocols in nonhuman primates. It provides consistent induction and sedation while maintaining cardiovascular and respiratory parameters. Femoral Fracture Repair in a Juvenile Chimp E Romero*, DL Hasselschwert, J V Coutin New Iberia Research Center, New Iberia, LA A 6-year-old male chimpanzee presented with acute lameness and swelling in a thigh. Radiographs revealed a spiral fracture of

One Is a Party, Two is a Crowd: The Case of a Squirrel Monkey Twin C-Section S Milligan*, J Ruiz 219


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Texas A&M University, College Station, TX The squirrel monkey is a popular and well-known model for cardiovascular research, pharmacologic studies and pelvic organ prolapse. We maintains a breeding colony of both species of squirrel monkey used in biomedical research, Saimiri boliviensis and Saimiri sciureus. At 64 days gestation, a 4-year-old female member of the Saimiri sciureus breeding colony presented on ultrasound with a twin pregnancy following 3 previous pregnancies, one of which was an abortion that occurred during this same breeding season. Unfortunately, twin pregnancies in squirrel monkeys often result in either intra-uterine death of the fetuses or dystocia resulting death of the dam and offspring. Due to the high-risk nature of this pregnancy, which appeared to be progressing normally, a scheduled cesarean section was performed at 146 days gestation (normal gestation is 152 days). This case presentation aims to highlight the surgical approach for the successful delivery of the one male, one female live twins as well as postoperative management of the dam and offspring. Unfortunately, the male twin was euthanized at 3 days old due to failure to thrive. This presentation also aims to describe his postmortem gross and histopathologic lesions. Peritonitis and Cholangiohepatitis in an SIV-Infected Rhesus Macaque (Macaca mulatta) C Cullin, J Stanton*, A Lewis Oregon National Primate Research Center, Beaverton, OR A 12-year-old, 9.8-kg male rhesus macaque was reported for gradual weight loss, intermittent partial anorexia, and loose stool 14 months after protocol-associated infection with simian immunodeficiency virus (SIV). Fecal culture revealed normal enteric flora; fecal parasitology was positive for Balantidium sp. Initial treatment included metronidazole, oral fluids, caloric support, and probiotic and fiber supplementation. Bloodwork changes included mild lymphopenia, moderate hypoalbuminemia, and marked elevation in liver enzymes. Mildly concentrated urine and bilirubinuria were present on urinalysis. Despite treatment, the monkey continued to decline and was euthanized at clinical endpoints. At necropsy, the serosal surfaces and omentum were mildly thickened and hyperemic, consistent with mild peritonitis. The gallbladder wall was thickened; the cystic and common bile ducts were markedly dilated. The intrahepatic bile ducts were prominent on sectioned surface of the liver, and the hepatic parenchyma was discolored yellowish tan. On microscopic evaluation, there was proliferation of the biliary epithelium in the liver, gallbladder, and extrahepatic bile ducts accompanied by lymphocytic, eosinophilic, and histiocytic inflammation. Similar proliferative and inflammatory changes were present in the omentum and intestinal serosal. Within the gallbladder mucosa, there were numerous individual epithelial cells extruded into the lumen. Immunohistochemical staining for Enterocytozoon revealed numerous intracellular perinuclear organisms consistent with Enterocytozoon bieneusi in the biliary epithelium, as well as in the mesothelium of the omentum and duodenal enterocytes. Also found in human AIDS patients, E. bieneusi is an important opportunistic infection observed in SIV-induced AIDS in macaques. In macaques, it must be differentiated from SIV-induced pathology and other opportunistic infections, including Cryptosporidium parvum, a frequent cause of cholangiohepatitis. E. bieneusi has been increasingly identified in immunocompromised macaques and remains underreported in this population. Though fumagillin treatment may be used in combination with antiretroviral therapy in human patients, treatment in macaques often remains unrewarding. Strategies to Increase Time Usage and Accessibility of Environmental Enrichment Provided to Socially Housed Primates to Improve Animal Welfare and Wellbeing C Carrier* Covance, Alice, TX

Group housing is the standard method for housing nonhuman primates at our facility. Delivering enrichment to all social group members can be difficult due to the social hierarchies. Often the dominant animal will guard and chase off other group members when enrichment is thrown in. While training can be done to combat some of this dominant behavior, we looked at other changes that could be made to improve our environmental enrichment delivery methods to large groups of primates. Simple refinements in our methods were tried to improve animals’ access to offered enrichment. The novel strategies discussed here are piñatas, game feeders on timers, and larger frozen forage blocks. Experience with primates reveals ingestible items encourage foraging and destructible items prove most interesting. As a result, selected new strategies targeted these “likes.” The most successful enrichment strategies provided the group a longer time of use, gave them something to do, and provided foraging opportunities. The longer it took to empty the item, the sooner the dominant animal moved on. This allowed other animals in the group a chance to forage. The new methods also provided more accessibility to group-housed primates especially the lower ranking members who typically only had access to leftovers. These simple and practical to employ refinements, increased duration of use, and allowed for more animals to access the environmental enrichment being offered in the group setting. As such, these methods should increase the welfare and wellbeing for low ranking animals in a group setting. Rare Uterine Angioleiomyoma in an African Green Monkey (Chlorocebus aethiops sabeus) MJ Valentine, A Beierschmitt*, JJ Callanan Behavioral Science Foundation, Estridge Estate, St Kitts, West Indies A uterine neoplasm was observed, as an incidental finding, during postmortem examination of a 26-year-old female multiparous African Green Monkey (Chlorocebus aethiops sabeus). The intramural, expansile, 2-3 cm well-demarcated, dark-red, nodular neoplasm located on the anterior uterine body (corpus) wall was confirmed as a cavernous uterine angioleiomyoma (syn. vascular leiomyoma) characterised by abundant intratumoral vasculature lined by Factor VIII positive endothelial cells and surrounded by smooth muscle actin positive cell proliferations. Human uterine angioleiomyoma, observed in middle age, can be single or multiple and occur submucosally, intramurally, or subserosally and are further classified based on their vascular component as cavernous, venous, or solid (capillary). Angioleiomyoma sharing the characteristics of intramural human cavernous uterine angioleiomyoma should be considered in the differential of uterine tumours in nonhuman primates. The Combination of Internal and External Bone Fixation for a Comminuted Fracture in a Rhesus Macaque (Macaca mulatta) A Haertel*, T Hobbs, K Prongay Oregon National Primate Research Center, Beaverton, OR Comminuted fractures in animals are caused by high-energy trauma. The inherent lack of stability of these fractures makes them difficult to treat. A closed comminuted fracture of the distal third diaphysis of the left tibia occurred in an 11-year-old, third-trimester pregnant female rhesus macaque socially housed in an outdoor shelter. A fixation system consisting of an intramedullary pin, cerclage wire, and a type IIb external skeletal fixator (ESF) stabilized the fracture. The patient displayed weight-bearing lameness initially, and, within a week of fixator stabilization, lameness resolved. Parturition occurred at 9 days post op, and the patient nursed and cared for her infant henceforth. Cover bandages were impetuously removed by the patient; however, unexpectedly, the ESF was welltolerated. Radiographs taken every 2 weeks provided monitoring of the fracture and fixator as well as a cleaning opportunity. The type IIb ESF was converted to less-rigid type IIa and then to a type Ia which promoted limb mobility and exuberant periosteal callus formation. The patient was encouraged to continue loading the

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limb in an exercise tunnel attached continuously to the home cage. There were 2 complications with our fixation system. First, the IM pin migrated into the stifle joint and required surgical correction. Second, radiolucent cortical bone developed at ESF cross pins. We suspect calcium mobilization associated with pregnancy and lactation may have partly impeded fracture healing. Despite these setbacks, fracture healing progressed throughout evaluations, and the external fixator was removed successfully. Our fracture fixation system provided rigidity in early stages of fracture healing, followed by the gradual reduction of support to allow for an incremental increase in load-bearing on the bone to promote healing. Some internal fixation devices do not overcome forces imposed on comminuted fractures to allow for stabilization or cause pressure necrosis of overlying tissue when placed in the distal third diaphysis of the tibia. Contrary to our expectations, this NHP tolerated the ESF throughout pregnancy, delivery, and infant care. The combination of internal and external bone fixation provided excellent stability and adaptability to a healing comminuted tibia fracture.

and treated upon discovery; however clinical signs of lower bowel disease did not improve. As gluten sensitivity has been described in rhesus macaques, the monkey was placed on a trial of nutritionally complete, wheat free, nonhuman primate diet. Cessation of soft stool and diarrhea was observed almost immediately upon diet change, resulting in a favorable weight gain of 3kg in a few short months. This progress was short lived, and within months new clinical signs of intermittent vomiting with blood and melena were appreciated. Next, endoscopy was performed on the stomach, duodenum, and colon revealing additional information relevant to this case study.

Infant Diarrhea Reduces Growth and Survival Rates of Rhesus Macaques

Between 2014 and 2015, a group of rhesus macaques due to sudden death were suspected to be caused by Encephalomyocarditis virus (EMCV) infection based on gross examination and histopathology findings. Two sets of published primers targeting 3Dpol gene of EMCV were used to detect EMCV RNA extracted from heart tissue samples by RT-PCR. Results showed that the difference of testing sensitivity between these two RT-PCR assays was up to 90%, which implied that the sequence of EMCV 3Dpol gene tested might be highly divergent from known EMCV strains. EMCV isolation was therefore performed and viral genome sequences were analyzed. Of the 14 necropsied animals, EMCV was successfully isolated from a variety of tissues collected from 10 animals. Phylogenetic analysis of the entire open reading frame (ORF) showed that our isolates were grouped into a new lineage other than published lineage A, B, C, D of EMCV-1 serotype, Cardiovirus A species, Cardiovirus genus. This new lineage is more closely related to EMCV Mengovirus strain (rhesus monkey/Uganda/1946), but only with 79% nucleotide identity in the ORF area. Seroprevalence study by testing representing 67 rhesus monkeys from the facility showed a 7.5% EMCV neutralizing antibody positive rate (5/67), which suggests that some of infected animals might be asymptomatic. It is not clear how this novel EMCV strain is transmitted, virus isolation from trapped rodents will be further investigated since rodents are usually considered as the EMCV spreading source.

A Haertel, K Prongay* Oregon National Primate Research Center, Beaverton, OR Weight is a commonly collected metric used to assess normal growth, nutritional status, and health. Infant rhesus macaques experience linear weight gain during the first year of life. We hypothesized that animals who develop diarrhea during the first year of life would experience poor weight gain and a higher risk of death relative to healthy infants, and that the difference in weights could allow early identification and intervention of infants with diarrhea prior to death. Using a retrospective cohort design, daily weight gain through the first year of life was determined by sex and housing type (small-group shelters or large-group corrals). From a dataset of 2,636 animals and 9,345 weight records, standard weight curves for healthy male and female infant rhesus were developed for corral and shelter animals. Healthy infant weight curves were compared with animals in the same social environment who develop diarrhea during the first year of life. Diarrhea significantly stunted shelter male, shelter female, and corral females, reducing daily weight gain by as much as 32% over the first year of life. Survival analysis revealed infants with diarrhea in the first year of life also had a 4.2 times greater chance of death than healthy infants, and death was 2.6 times more likely in shelter infants than corral infants. Sex or any cofactor with diarrhea did not contribute as a significant risk of death; however, visual inspection of survival curves demonstrated age periods that differed between corral and shelters animals with diarrhea where mortality rates accelerate rather than remain static. Shelter animals with diarrhea appear to have a dramatic mortality spike between 90 and 250 days of age; whereas mortality rates of corral animals with diarrhea increase between 90 and 150 days of age. Optimal cut-points of weight for age-sex-housing-type from standard weight curves could identify at-risk infants. Additional morphometric data would allow a more complete picture of stunting growth beyond weight gain. An Interesting Case of Chronic Gastrointestinal Disease in a Macaque J Dewar*, L Shankle, F Sentz, J Ascher FDA/Center for Biologics Evaluation and Research/OM/DVS, Silver Spring, MD Over a span of 10 years, a male rhesus macaque presented with chronic soft stool and episodic diarrhea. The animal displayed chronic gastrointestinal upset different from his group on study. He was treated with various enteric therapies with waxing and waning progress. This rhesus maintained lean, yet overall favorable body condition, and a good appetite. Early on he tested positive for several known enteric pathogens which were quickly identified

A Novel Encephalomyocarditis Virus Lineage Closely Related To Mengovirus Discovered in Rhesus Macaques (Macaca mulatta) C Zao*, Y Yang, A Cooke, R Berger, OD Gonzalez Velez VRL, San Antonio, TX

Comparison of Direct and Indirect Methods of Arterial Blood Pressure Measurement in Healthy Adult Male Rhesus Macaques (Macaca mulatta) LK France*, C Garrett Johns Hopkins University, Baltimore, MD Blood pressure is a critical parameter for evaluating the overall health of an animal, assessing the effects of drugs and procedures, monitoring the physiologic status during anesthesia, and making clinical decisions. The placement of an arterial catheter for direct measurement of blood pressure remains the gold standard in both human and veterinary medicine for accurately measuring blood pressure; however, this procedure is invasive, technically challenging, and often impractical during brief anesthetized examinations. To date, noninvasive methods for measuring blood pressure have not been rigorously evaluated nor compared to direct arterial catheter measurements in nonhuman primates. The objective of this study was to determine which method of indirect, noninvasive blood pressure monitoring was most accurate when compared to direct, invasive arterial catheterization. The indirect, noninvasive methods evaluated were ultrasonic Doppler flow detection and oscillometry. The specific aims of this study were to determine the relative accuracy of each indirect method at a given body location and to assess whether the accuracy of each indirect method was dependent on body location. Nine, healthy 11- to 13-year-old male rhesus macaques ranging 10 to 16 kgs were anesthetized with ketamine (20 mg/kg IM), intubated, and maintained within a stable 221


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surgical plane under general inhalant anesthesia using isoflurance. Blood pressure measurements were taken via direct arterial catherization of the saphenous artery and compared to ultrasonic Doppler flow detection and oscillometric measurements at 3 body locations (forearm, distal leg, and tail base). Each measurement was repeated 3 times at 2-minute time intervals. Initial results from this ongoing study indicate that blood pressure measurements taken from the arm and tail base using oscillometry most closely approximate blood pressure measurements obtained via direct arterial catheterization. Blood pressure measurements taken from the leg using oscillometry were the least accurate when compared to blood pressure measurements obtained by a direct arterial catheter. Results of this study will provide guidelines for the use of noninvasive methods to accurately monitor blood pressure in anesthetized rhesus macaques across a variety of clinical or experimental settings. Reference Growth Curve For Long-Tailed Macaques (Macaca fascicularis) from Mauritius S Naiken* Bioculture Group, Riviere des Anguilles, Mauritius The long–tailed macaque (Macaca fascicularis) of Mauritian origin is widely used in biomedical research. However, there is little information on its growth pattern. The purpose of this study is to provide weight for age and sex reference standards for this important animal model. Reference values were derived from retrospective data over a 10-year period for 37,105 male and 35,454 female captive-bred monkeys aging from 0 to 84 months. The smooth curves of weight for age and sex were created using the Generalised Additive Models for Location Scale and Shape (GAMLSS) package under R.3.1.2 software and included 244,809 and 223,432 weight observations for males and females, respectively. It was found that males not only grow faster than females, but they also continue to grow well after females have attained their peak body mass at around the age of 48 months. Male and females growth curves will be tested for statistical difference. It is expected that when males go through an adolescent growth spurt at around the age of 30 months, females will continue to grow at a constant rate. The results from this study establish ageweight reference ranges which can be useful for assessing growth in long-tailed macaques of Mauritian origin. Successful Group Formation of Mature Male Long-Tailed Macaques (Macaca fascicularis) T Andrianjazalahatra, P Honess* Bioculture Group, Riviere des Anguilles, Mauritius Forming groups of mature male macaques in captivity without potentially fatal aggression has long been a challenge. With roughly equal birth sex ratios in breeding facilities where breeding groups are single-/two-male, bachelor groups are a necessity for efficient use of caging. We aimed to reduce aggression and improve welfare when creating groups of long-tailed macaque males over 4 years old, by reducing the number of existing coalitions between introduced males through minimizing previous experience of each other. It was hypothesised that minimizing coalitions would reduce aggression. In a retrospective analysis of colony records (Jan 2010 – Nov 2015) we compared aggression levels for 30 days following mergers using the new reduced coalition system (43 groups) and the old system (36 groups) (overall mean = 11.03/group). There was no difference between methods in aggression (events/merger) (P = 0.771). However further analyses showed the new system produced 47% lower aggression (P = 0.048) among wild-caught males and 86% higher aggression (P = 0.031) in captive-bred males. In conclusion, success of the refined system depends on the animals’ origin highlights the importance of tailoring the management system to the characteristics of the animals. These findings have relevance for those working in a range of captive contexts including breeding facilities, laboratories, zoos, and sanctuaries with potential significant improvement in animal welfare during mergers.

Cutaneous Melanoma in a Cynomolgus Macaque (Macaca fascicularis) WM Burnside*, AP Garcia, KJ Hopper, JL Wagner Haman Ranch, The Mannheimer Foundation, LaBelle, FL An outdoor-housed 11-year-old female cynomolgus macaque (Macaca fascicularis) presented for a highly pigmented, discrete, firm, slightly raised, crusting cutaneous mass (ovoid, 3 × 2 × 1 cm) on the dorsal tail-base after routine physical examination. She weighed 4.59 kg and was in ideal body condition. Peripheral lymph nodes were within normal limits. Metastatic disease was not apparent on thoracic or abdominal radiographs, or abdominal ultrasound. After 10 days indoors, the mass had reduced in size (ovoid, 1 × 0.5 × 0.5 cm), was surgically excised and submitted for histopathology. Histopathological examination of the mass revealed epidermal hyperkeratosis and acanthosis. Diffuse, moderate infiltration of lymphocytes and plasma cells were present in the superficial dermis, while non-encapsulated proliferation of spindle to ovoid to polygonal cells that often contained a large amount of a brownblack pigment (melanin) within the cytoplasm and obscured the nuclei were present in the deep dermis. Adjacent to the mass, focal, subcorneal infiltration of neutrophils admixed with an eosinophilic material was present in the epidermis, while focal area of infiltration of multinucleated giant cells and macrophages surrounded a free hair shaft were apparent in the superficial dermis. The neoplastic cells did not extend to the surgical margins. Additional immunohistochemical stains including vimentin, cytokeratin S-100, and melan-A were performed to confirm the neoplasm origin. In addition, Ki67 stain was performed to determine the cell proliferation and growth fraction of the tumor. After 11 months, there were no signs of recurrence at the surgical site, new palpable cutaneous masses or peripheral lymph node enlargement, or abnormalities detected via repeated radiographic or ultrasonic imaging. Cutaneous melanoma is a malignant neoplasm originating from melanocytes; most commonly, oncogenesis is due to overexposure of poorly pigmented skin to ultraviolet light. This condition is uncommon and rarely described in nonhuman primates. Longevity of Passive Integrated Transponders in Nonhuman Primates: A 3-Brand Comparison WM Burnside*, AI Cameron, KJ Hopper, K Rivas-Wagner, JL Wagner Haman Ranch, The Mannheimer Foundation, LaBelle, FL Despite frequent use as an animal identification method, little peer-reviewed literature is available regarding the long-term survivability of commercially available passive integrated transponders (PITs). These subcutaneous implants are often used as an alternative or adjunct to external identification in research and companion animals. This 10-year retrospective study of data collected at 2 facilities examined the longevity of 3 common PIT brands in three species of nonhuman primate (n = 6003). Transponders were placed at 92.06 ± 66.91 (mean ± S.D.) days of age and scanned approximately every six months during routine physical examination. Survival analyses were used to compare the probability of transponder failure over time and included four factors: PIT brand, animal sex, species, and facility of origin. There was no significant difference between sexes (P = 0.142); however, highly significant differences among PIT brands (P < 0.0001) and species (P < 0.0001) were detected. The predicted median survival times were highest in cynomolgus macaques (Macaca fascicularis) at 12.21 ± 1.65 (median ± 95% CI) years, followed by hamadryas baboons (Papio hamadryas) at 7.26 ± 1.20 (median ± 95% CI) years and rhesus macaques (Macaca mulatta) at 6.50 ± 0.47 (median ± 95% CI) years. The highest predicted median survival time of the 3 PIT brands was 19.24 ± 2.85 (median ± 95% CI) years. In rhesus macaques, one PIT brand was significantly different between the two facilities (P = 0.0003) with a 1.26-year difference in predicted median survival time. Facility differences may be attributed to differences in genetics or management of rhesus macaques, or staff and training. Overall, the survivability of 1 PIT

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brand of appeared to outperform the others. Transponders are more likely to fail earlier in rhesus macaques and hamadryas baboons than cynomolgus macaques. It is recommended that nonhuman primate facilities regularly monitor PIT function to evaluate failure rates and develop practices to improve transponder survivability. Evaluation of Indirect Blood Pressure Measurements in Anesthetized Rhesus Macaques (Macaca mulatta) S Kang*, K Jampachaisri, C Pacharinsak Stanford University, Stanford, CA The aim of the study was to evaluate indirect blood pressure (BP) measurement methods (Doppler ultrasound and oscillometry) compared to direct BP measurement in anesthetized rhesus macaques. A direct BP measurement for systolic arterial blood pressure (SAP) and mean arterial blood pressure (MAP) was performed via a saphenous arterial catheter connected to a pressure transducer. The cuff placement locations for Doppler ultrasound (Vet-Dop2; VD) and oscillometry (Surgivet; OS) were at the proximal tail and midantebrachium, respectively. All BP measurements were performed every 5 minutes for a total of 120 minutes. For VD, when MAP < 60 mmHg, VD-systolic was lower (8.8 ± 1.8 mmHg) than SAP, but higher (8.2 ± 1.2 mmHg) than MAP. When MAP > 60 mmHg, VDsystolic was lower (18.5 ± 4.2 mmHg) than SAP, but higher (17.2 ± 6.0 mmHg) than MAP. For OS, when MAP < 60 mmHg, OS-systolic was lower (1.0 ± 1.6 mmHg) than SAP; OS-mean was lower (12.2 ± 1.9 mmHg) than MAP. When MAP > 60 mmHg, OS-systolic was lower (15.6 ± 14.4 mmHg) than SAP and OS-mean was lower (14.0 ± 7.1 mmHg) than MAP. This study suggests that both Doppler and oscillometric BP measurements underestimate direct BP measurements in rhesus macaques.

bright, alert, and responsive. There was significant swelling of the scrotum and prepuce, with a visible mass in the right inguinal region. Physical examination revealed a soft mass that was unable to be manually reduced. Palpation of both testicles revealed no abnormalities. Radiographs indicated a soft tissue-density material in the right inguinal region and ultrasound demonstrated heterogenous tissue in the cranial scrotum with hypoechoic fluid accumulation. No gas loops were visualized, indicating a right-sided inguinal herniation of omentum. CBC and clinical chemistry showed no abnormalities. The patient was given 0.2mg/kg meloxicam SC for pain and inflammation, and a unilateral open herniorrhaphy with orchiectomy was performed the following day. For surgery, a 7cm longitudinal incision was made directly over the swelling in the right inguinal region, revealing peritoneum with omentum visible within. After the orchiectomy was complete, the herniated omentum was unable to be completely reduced; a 5cm incision was made through the skin and linea alba cranial to the pubic symphysis. The internal inguinal ring was located and the herniated omentum was pulled through from the abdominal side. Any remaining omentum was removed before the inguinal canal was closed with 2-0 PDS in a simple interrupted pattern. Both incisions were closed in 3 layers (2 simple interrupted patterns and skin apposition with intradermal 3-0 PDS reinforced with surgical glue). No complications occurred during surgery or recovery. Postsurgical analgesics (0.01mg/kg buprenorphine IM and 0.1mg/kg meloxicam PO) were administered for 3 days, along with 5mg/kg enrofloxacin IM for one week. The scrotal and prepucial swelling resolved over the following 3 weeks and herniation has not recurred. Periodontitis and Chronic Otitis Media in a Cynomolgus Macaque (Macaca fascicularis) KE Scott*, TJ Oura, V Bakthavatchalu, JG Fox, RP Marini

Characterization of Endometriosis-Associated Gross Pathologies in the Rhesus Macaque (Macaca mulatta)

Massachusetts Institute of Technology, Division of Comparative Medicine, Cambridge, MA

C Cullin*, B Zwerling, L Colgin

A 15-year-old, intact, singly housed male cynomolgus macaque (Macaca fascicularis) with a head-post implant presented with an acute focal swelling infraorbital, overlying the maxillary bone (1 cm diameter), which ulcerated and had serous discharge within 48 hours post presentation. Two and a half years prior to presentation, he had been treated for purulent otitis externa and an oral abscess from which Staphylococcus aureus and Streptococcus pyogenes were cultured. Multiple radiographic studies revealed no oral or otic pathology. Biopsies of the facial swelling were nondiagnostic and aerobic, anaerobic, and fungal cultures were negative. An MRI was performed and during the imaging session, the right ear bar was difficult to place due to hyperplastic tissue in the ear canal. Blood drained from the canal after the procedure. MRI revealed that the rostral aspect of the facial swelling had a focal hyperintense linear tract that extended 5mm into the underlying tissues and a less intense soft tissue material extending through a small focal defect in the maxillary bone to the level of the root of the right maxillary canine tooth. MRI also revealed heterogenous T2W hyperintensity filling the right tympanic bulla with extension of strongly T2W hyperintense material (fluid) into the mastoid air cells of the right temporal bone. The bulla wall remained intact and no abnormalities were detected in the cochlea and adjacent brain parenchyma. Based on MRI-findings, a diagnosis of otitis media and periodontitis was confirmed. Myringotomy and tympanocentesis was attempted but unsuccessful due to the depth and narrow diameter of the external ear canal. The right maxillary canine was extracted, and histology of associated soft-tissue showed an infiltration of a low number of neutrophils, macrophages, lymphocytes, and plasma cells. Due to the small size of the tympanic bulla of cynomolgus macaques, radiographic examination for bulla-associated pathology is not as sensitive as in other species. In this animal, the otitis media remained subclinical, without signs of discomfort or neurologic signs. Otitis media can remain subclinical chronically and is difficult to confirm and culture in cynomolgus primates.

Oregon National Primate Research Center, Beaverton, OR Endometriosis is a chronic disease characterized by the presence of ectopic endometrial glandular and stromal tissue in menstruating primates. It has been well documented in rhesus macaques, and we report here the associated gross morphologic sequelae of endometriosis in a large breeding population. Retrospective analysis of health records for females born on-site over a 15-year period from 2000-2014 identified 152 cases of endometriosis which were confirmed via necropsy and subsequent microscopic examination. Age at diagnosis ranged from 4 to 29 years; the calculated incidence among all sexually mature females (>4 years) during this period was 7.9%. Of those animals diagnosed with endometriosis, 58 (38.2%) had complications associated with the condition. Gross pathologies included abdominal hemorrhage in 20/152 (13.2%), fibrous adhesions resulting in intestinal stricture formation in 14/152 (9.2%), and unilateral or bilateral hydroureter and hydronephrosis in 26/152 (17.1%). Four animals had both intestinal stricture and hydroureter with hydronephrosis. In addition, 6 females had concurrent reproductive pathology, including uterine leiomyoma (n = 5) and uterine leiomyoma with endometrial polyp (n = 1). Because of the relatively high prevalence of pathologies and the severe health consequences associated with spontaneous endometriosis, affected animals should be closely monitored in order to provide appropriate clinical care. Inguinal Hernia Repair in a Rhesus Macaque (Macaca mulatta) C Tansey*, B Gottstein, A Peregrine, PV Turner University of Guelph, Guelph, Ontario, Canada A 9-year-old, 15.3kg, intact, male rhesus macaque was reported for reddened, swollen testicles. On cage-side examination, he was

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Mineralized Trichobezoars in a Rhesus Macaque (Macaca mulatta)

University of Guelph, Guelph, Ontario, Canada

C Wallace*, J Wilson, A Brice, L Makaron

A 1-year-old, male ring-tailed lemur from an exotic animal sanctuary presented to a small animal clinic with a distended abdomen. Radiographs demonstrated moderate ascites; larval cestodes were noted on cytology of the abdominal fluid. Antihelminthics were administered intermittently over 4 months until the animal developed liver failure and was hospitalized for supportive care. The lemur was subsequently discharged and died at the sanctuary. At necropsy, the animal was in poor body condition. Occupying ~75% of the abdominal cavity and replacing ~90% of the liver was a multiloculated mass, composed of thick-walled, tan nodules with central cavitations containing cloudy tan fluid. Similar nodules were present adjacent to the kidneys, lumbar vertebrae, and thoracic inlet. Microscopically, nodules were composed of multiple cysts lined by trilaminar, hyalinized membranes, and contained numerous protoscoleces each with a single, circular arrangement of hooklets. Echinococcus multilocularis was suspected and subsequently confirmed by PCR. A second lemur died 1 week later of unrelated causes and was subsequently found to have similar gross and microscopic findings. These animals were cohoused with other primates, as well as foxes, dogs, and lynx, with indoor-outdoor access on a large rural property. The private facility was open to the public and animals were frequently taken to local schools. Daily care and husbandry of the animals was provided by volunteers. Echinococcus multilocularis is enzootic in wild canids living in northern Canada and Alaska. Primates are an aberrant host and infection occurs following ingestion of eggs shed by canids or ingestion of intermediate hosts. Once infected, treatment is aimed at controlling larval replication. Although lemurs can’t directly transmit the parasite to humans, there is zoonotic risk from the source of infection, as well as welfare concerns for the primates. These represent the first documented cases of E. multilocularis in lemurs in North America.

University of Pennsylvania, Philadelphia, PA A naïve 5-year-old female Rhesus macaque (Macaca mulatta) was evaluated upon entry into quarantine. Physical examination revealed thin body condition and a large, round, hard structure palpated mid abdomen. Serology was negative for Herpes B virus, Simian T-cell Leukemia Virus, Simian Immunodeficiency Virus and positive for Simian Retrovirus (SRV) and Measles Virus. The animal was later sedated for additional diagnostics. Weight loss was evident and the original mass was still palpable in addition to a smaller, more caudal mass of similar composition. Radiographs revealed 3 radioopaque masses: one in the upper right abdomen and 2 located mid abdomen. Differentials included parasitic cysts, SRV with fibromatosis, fat necrosis, foreign body, and granulomas due to tuberculosis. Abdominal tuberculin skin test was negative and thoracic radiographs were unremarkable. An abdominal exploratory surgery revealed 3 freely movable masses in the cecum which were surgically removed via typhlotomy. The masses were 2-3 cm in diameter with hard, dark, mottled outer shells and firm fibrous cores, diagnosed as cecal trichobezoars covered by a mineralized shell. Microscopically the masses were composed of abundant hair shafts, mineral, bacteria, food particles, and small bone fragments. While trichobezoars have been reported in Rhesus macaques, baboons, and spider monkeys, this is the first report to our knowledge of mineralized trichobezoars in a Rhesus macaque. Echinococcus Multilocularis in 2 Lemurs from an Exotic Animal Sanctuary N Compo*, B Gottstein, A Peregrine, PV Turner

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May 2017

March 19–21: IACUC Conference, New Orleans, LA, http:// www.primr.org/iacuc17/

May 1–6: Assisted Reproductive Technologies (ARTs) in the Laboratory Mouse, Jackson Laboratory, Bar Harbor ME, www. jax.org/course-schedule

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June 2017

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September 2017 September 18–23: Surgical Techniques in the Laboratory Mouse, Jackson Laboratory, Bar Harbor, ME, www.jax.org/courseschedule September 21–24: Southwest Veterinary Symposium, Henry B. Gonzalez Convention Center, San Antonio, Texas, http://www. swvs.org/about_the_show.cfm September 24–29: Advanced Surgical Techniques in the Laboratory Mouse, Jackson Laboratory, Bar Harbor, ME, www.jax. org/course-schedule

October 2017 October 1–4: The 14th Transgenic Technology Meeting, Snowbird Resort, Salt Lake City, Utah, www.tt2017org October 9–14: Workshop on Phenotyping Mouse Models of Human Lung Disease, Jackson Laboratory, Bar Harbor, ME, www. jax.org/course-schedule October 15–19: American Association for Laboratory Animal Science National Meeting , Austin, TX, (901)754 -8620 AALAS or [email protected]. https://www.aalas.org/national-meeting/ general-information/future-meetings

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Information for Authors General

The American Association for Laboratory Animal Science (AALAS) currently publishes two journals containing datadriven, peer-reviewed articles. The mission of Comparative Medicine (CM) is to disseminate high-quality, peer-reviewed information that expands biomedical knowledge and promotes human and animal health through the study of laboratory animal disease, animal models of disease, and basic biologic mechanisms related to disease in people and animals.

Facts and Statistics CM • Average submission to final decision time: 56 days • 512 pages printed in 2016 • 55% acceptance rate JAALAS • Average submission to final decision time: 63 days • 844 pages printed in 2016 • 57% acceptance rate • • • •

No submission fee. No page charges. No color charges. Complementary uploading to PubMed Central.

The mission of The Journal of the American Association for Laboratory Animal Science (JAALAS) is to disseminate high-quality, peer-reviewed information on animal biology, technology, facility operations, management, and compliance as relevant to the AALAS membership. The types of articles accepted are Case Report, Case Study, Research Report, Overview, and Letter to the Editor. Definitions of and distinctions between article types are given in the online version of the Information for Authors. The editorial style of AALAS journals is based on that described in Scientific Style and Format: The CSE Manual for Authors, Editors, and Publishers (seventh edition). Consult previous issues of the relevant journal for information not addressed in Scientific Style and Format or the following material. An annotated version of the Information for Authors is found at https://www.aalas.org/publications/information-for-authors/ cm-and-jaalas.

Title page

Manuscript Preparation

On the first page of the manuscript, include the • Full Title—a concise informative description of the presented work including the common name of the animals used (add genus and species in parentheses for nonhuman primates and species whose common name may be unfamiliar to readers). • List of authors—the first name, middle initial (or first initial and middle name), and last name of each author • Institutional affiliation of each author—the Department (or Program), Institution (or Company), City, State (or province), and Country (if not USA) at which the described work was done.

Use superscripted Arabic numerals to indicate respective institutions ■ Spell out all locations in full • Corresponding author—the person who readers can contact regarding information or reagents ■ Indicate with * in the list of authors and provide that author’s email address • Running title—a descriptive phrase of no more than 72 characters (including spaces) to be used as a running head on each printed page • Abbreviations and acronyms—a list of all nonstandard acronyms and abbreviations used throughout the manuscript and their definitions ■ Standard Abbreviations need not be included on the title page. ■ Do not abbreviate terms used fewer than five times in a manuscript. ■

Abstract

Provide a single paragraph (no subheadings) of 250 words or fewer (preferably no more than 200 words) that sequentially summarizes the background, rationale, methods, results, and conclusions of the work.

Body

Use coded or nonproprietary language throughout the manuscript. Cite the proprietary, brand, or vendor name associated with an assay, instrument, machine, service, or compound only in Materials and Methods. Please do not include website links within the body of the manuscript. Online resources should be cited and added to the reference section. For specific details, refer to previous issues of JAALAS or CM for examples of citation formats. Define all nonstandard abbreviations and acronyms at first use. Limit the number of novel abbreviations used. A list of Standard Abbreviations that can used without definition is found in the Manuscript Preparation section in the online version of the Information for Authors. The length of and subsections comprising the body of the manuscript will vary depending on the type of article submitted. The manuscript may include some or all of the following sections: • Introduction ■ Provide the rationale and supporting background for the presented work and its importance and relevance. ■ Extensive reviews of the existing literature are inappropriate for research reports and case studies/ reports. • Materials and Methods ■ Describe the animals, husbandry, tests, equipment, procedures, reagents, and services used in sufficient detail to permit replication of the work, with citation of published references as consistent with brevity and clarity. ◆ Clearly define use of the term ‘specific pathogenfree’ by including specific criteria (for example, tests, organisms surveilled, housing, husbandry conditions) or citing publications providing that information. ◆ Include statistical methods where relevant and attribute (name of software program used and name and location of vendor) or reference them appropriately. In addition, provide the P value used 227


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to define statistical significance. Include a statement regarding Institutional Animal Care and Use Committee approval (or equivalent) for procedures and protocols involving animals. ■ Provide assurances regarding humane care and use of animals, citing AALAS Position Statements and national standards as appropriate to the country in which the work was performed. ■ For research involving human subjects, identify the committee that approved the experiments and include a statement that informed consent was obtained from all subjects, that measures are in place to protect the identity of all subjects, and that no coercion was used to solicit subjects. ■ Provide the vendor’s name and location for any solesource item or service. ■ Insert callouts (in parentheses) for all Figures and Tables, which are numbered in order of their mention in the text. ■ Follow correct nomenclature for laboratory animals, genes, genetic markers, alleles, mutations, and microbes. ■ Wherever possible, use International System of Units base and derived units for numerical data. • Results ■ Use headings as needed to guide readers. ■ Accompany statements of differences between groups with appropriate statistics. ■ Summarize selected data from Figures and Tables in the Results section; do not merely repeat all information presented in graphics. ■ Save interpretation of data for the Discussion section. • Discussion ■ Begin the Discussion with a brief summary of the key findings. ■ Limit discussion of study findings to those that have been presented in the Results. ■ Address any limitations of the study and directions for potential future research. ■

Acknowledgments

Recognize (with their permission) people and institutions whose contributions of funding, technical assistance, reagents, data collection and analysis, and other services do not meet the criteria for authorship (http://www.icmje.org/ethical_1author.html).

References

Provide complete and accurate bibliographic information for all cited materials. Only information that is published or is already accepted for publication (that is, “in press”) can be used as references. We will not accept citations of unpublished information or materials (for example, personal communications, unpublished data, manuscripts still undergoing review) in a reference list or parenthetically in the text. Journals published by AALAS follow a modified version of the citation style found in Scientific Style and Format. Refer to previous issues of JAALAS or CM or the online Information for Authors for examples of citation formats for specific types of referenced material. Organize references numerically in strict (‘letter-by-letter’) alphabetical order. List references by the same author(s) chronologically. Templates for RefMan and EndNote are available in the online version of the Information for Authors.

Figure Legends

Provide complete, concise descriptions of all figures in order of their mention in the text. Define additional abbreviations or

acronyms. Indicate the original magnification of images by citing the magnification factor in the legend or by using scale bars within images themselves. Use of previously published material (in whole or part) must be cited in the legend and accompanied by a signed Permission for Use form. Appropriate manipulation of digital images must be made explicit in the accompanying legend.

Supplementary Materials

Supplementary materials are not permitted. Manuscripts submitted with supplementary figures (or tables) will be returned to the author to have the information incorporated into the manuscript, which could potentially delay publication.

Tables

Tables must be generated by using the Table function of Microsoft Word. Tables generated by using other programs (for example, Microsoft Excel) will be returned to the authors for reformatting, potentially delaying publication of the article. Number tables in order of their mention in the text and provide a brief title describing the information presented. Footnotes to tables are indicated by using superscript lowercase letters. Tables are used to present numerical data and are not wholly narrative. According to AALAS journal style, narrative information in a tabular format (for example, descriptions of scoring systems, experimental protocols) is submitted as a figure.

Preparation of Figures

Figures are submitted electronically, separately from the manuscript. Do not embed any images within the manuscript file. Also, all submitted micrographs must be in color; black and white micrographs will not be accepted for publication. File formats accepted are TIFF (preferred), EPS, high-resolution JPG, and high-quality PDF (no image compression). PowerPoint slides, Excel graphs, and images embedded in Word are not acceptable. Minimal resolution: 600 dpi for line art (for example, graphs in black and white); 300 dpi for color (save as CMYK; not RGB or indexed) or grayscale images (save black and white images as grayscale); 1200 dpi for scanned line art (save as TIFF). Photos taken with a digital camera must have a resolution of at least 4 megapixels. Create figures with a width of 89 mm (single column) or 187 mm (double column); do not enlarge created figure to meet these dimensions. Designate panels of figures by using uppercase letters (no periods) in the upper left corner of the image; keep size of lettering and other labels (at least 3 mm in height as submitted) consistent between panels of a figure and between figures. Embed fonts within digital images. Indicate the magnification factor of an image by including an appropriate size bar in its lower right corner. Minimize the use of color in charts, graphs, and drawings to that necessary for clarity of communication and ease of understanding. Use solid fill or percentage screens (not pattern or textured fills) and a minimum line weight of 0.5 pt throughout. Additional information regarding generating and formatting figures is available by emailing Amy Tippett (tgstudio@ carolina.rr.com).

Manuscript Submission

Letters to the Editor are submitted by email (journals@aalas. org; subject line: Letter to the Editor, CM or JAALAS) or through the USPS (Letter to the Editor, CM or JAALAS; AALAS; 9190 Crestwyn Hills Drive, Memphis, TN, 38125).

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Information for authors

Research Reports, Case Reports, Case Studies, and Overviews are submitted electronically through the Manuscript Central system. To avoid publication delays, ensure that all portions of the manuscript conform to the specifications in the Information for Authors. To submit a manuscript for possible publication, you need • A Microsoft Word file of the manuscript itself. • All associated image files. • A list of MeSH terms (maximum, 5; http://www.nlm.nih. gov/mesh/authors.html) for use as key words appropriate for indexing of the article. • Names, institutional affiliations, and email addresses (maximum, 4) of suggested persons to include or exclude as potential reviewers. Submit all necessary files for CM manuscripts at http:// mc.manuscriptcentral.com/aalas-cm. Submit files for JAALAS manuscripts at http://mc.manuscriptcentral.com/aalas-jaalas. As part of the submission process, return signed copies of the Copyright Transfer and Declaration of Potential Competing Interests by email ([email protected]) or fax (901-334-5152). The corresponding author must retain editable electronic copies of all text and illustrations used in the submitted manuscript.

CONTAINMENT

ANIMAL

TRANSFER

STATION

Manuscript Review and Status

The Editor-in-Chief reviews all submissions and makes an initial determination regarding suitability for publication. If an Associate Editor transfers a submitted manuscript from CM to JAALAS (or vice versa), the contact author will be notified by email and may opt to withdraw the manuscript from consideration. Before being sent for peer review, newly submitted manuscripts are screened to ensure that the text, figures (charts, graphs, images), and tables comply with the criteria described in the Information for Authors. All manuscripts undergo thorough peer review (including digital assessment for plagiarism prior to acceptance), typically by three reviewers with relevant experience. Selection of the panel of reviewers ultimately is the prerogative of the Associate Editor. AALAS gives timely review the highest priority. Once all reviewers for a manuscript have been assigned, authors can check the status of their submission through Manuscript Central. Whether a manuscript is accepted, requires revisions, or is rejected for publication typically is decided within 4 weeks of being sent for review. To avoid publication delays, regularly verify and update authors’ contact information (http:// mc.manuscriptcentral.com/aalas-cm for manuscripts submitted to CM; http://mc.manuscriptcentral.com/aalas-jaalas for manuscripts submitted to JAALAS). Changes requested by reviewers must be completed within 2 months or an extension requested (email [email protected] or call the Journals Office at 901-754-8620). Without timely return of revisions or a request for extension, the manuscript will be withdrawn from the publication process. Approximately 5 weeks before the slated publication date, the contact author receives a copyedited version of the manuscript, which will have undergone a final review by the Editor-in-Chief. After any additional queries that arise during copyediting and final review are addressed satisfactorily, the contact author receives a PDF of the page proofs of the article. At this late stage in the publication process, only minor revisions can be accommodated. Approved page proofs must be emailed to journals@aalas. org within 48 hours of receipt. After publication, the manuscript will be submitted to PubMed Central for indexing. Articles will become available to the general public 6 months after publication. Updated 2/17

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ORDERING INFORMATION No returns accepted. Please allow up to 2 weeks for processing. • Member ID must match the shipping or billing information to receive the member price. If you are a third party purchasing service ordering on behalf of an AALAS member, member information must be included with your order to receive the member discount or to use a purchase order for an AALAS institutional or commercial member. Please contact [email protected] for the authorization form. • All prices are subject to change without notice. • We will accept credit card orders by fax, mail, or online. • Orders are not accepted by phone. • Order fulfillment is subject to credit approval. • Purchase orders will be accepted from commercial and institutional members only with an official purchase order. For all other orders, payment must accompany order. • Checks and money orders from outside the USA must be issued on a USA bank. Non-USA bank drafts are not acceptable. • Receipt for purchase will be mailed to the shipping address on this form. • Duty charges on international orders are the responsibility of the purchaser. • All orders are shipped within the US, Canada, or Mexico via UPS Ground unless other arrangements are made and paid for accordingly. • Orders shipped to other countries are shipped via USPS. • Delivery cannot be guaranteed on international shipments through USPS and no refunds or credits will be issued. • Orders cannot be shipped to PO boxes.

PAYMENT & SHIPPING INFORMATION Subtotal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . $_____________________ Discount or Coupons (if applicable) . . . . . . . . . $_____________________ Shipping Charge . . . . . . . . . . . . . . . . . . . . . . . . $_____________________ TOTAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . $_____________________

SHIP TO: Member ID #___________________________________________________ q Check if not a current AALAS national member (If no member ID is indicated, or you are not the member, you will be charged the non-member price.) Name_________________________________________________________ Street Address (no PO box)_______________________________________ ______________________________________________________________ ______________________________________________________________ City___________________________________________________________

DISCOUNTS AALAS offers discounts on select products based on the quantities ordered. • The discount is applied to the price of the actual product, not the total order. • Items on the order form marked with an * are eligible for discount.

Phone ________________________________________________________ Email__________________________________________________________

PAYMENT METHOD:

20% discount: Training manual: quantities of 15 or more

q Check q Money Order

SHIPPING CHARGES Shipping charges are based on your total order amount and the country to which the order will be shipped.

Month Year

Expiration Date

United States

Canada/ Mexico

$0* to $200

$11

$30

$45

$201 to $600

$17

$55

$100

$601 to $1000

$40

$110

$275

$1001 to $2000

$60

$160

$325

$2001 to $4999

$100

$220

$400

free

free

free

$5000 and up

State__________ Zip______________ Country________________________

International

*There is a minimum shipping charge when you order only free items, except for public outreach products (those with product numbers beginning with 08-). AALAS covers the shipping costs for public outreach materials when they are ordered with purchased items; you will still be responsible for shipping charges on the purchased items. For orders of public outreach materials only, shipping costs are supported by the AALAS Foundation.

q Master Card q Visa

q American Express q Discover

Account Number—please include all digits For your identity protection, please do NOT e-mail your credit card information in the body of an e-mail or in an attachment. in any form.

Print name exactly as it appears on card

Signature

CREDIT CARD BILLING ADDRESS:

(If different from above. Must be exact for card approval.) Name_________________________________________________________ Street Address (no PO box)_______________________________________ ______________________________________________________________ ______________________________________________________________ City___________________________________________________________

CANCELLATIONS & REFUNDS No cancellations or refunds.

State__________ Zip______________ Country________________________ Phone ________________________________________________________

Fax orders to (901) 334-5157. Order online at www.aalas.org in the Bookstore. Prices subject to change. Form revised 03/04/11. page 3 of 3 Code:


3/9/2017 1:31:48 PM

2016 Trauma Association of Canada Annual Scientific Meeting Abstracts.

Association of Primate Veterinarians 2014 Nonhuman Primate Housing Survey.

2016 Canadian Society of Surgical Oncology Annual Scientific Meeting Abstracts.

Abstracts of scientific papers 19th world congress on disaster and emergency medicine.

Abstracts of the 52nd Workshop for Pediatric Research : Frankfurt, Germany. 27-28 October 2016.

The Association of Clinical Pathologists: 102nd scientific meeting. Abstracts.

2017 Trauma Association of Canada Annual Scientific Meeting Abstracts.

Pre-course workshop abstracts.

Abstracts from the 2016 American College of Medical Toxicology (ACMT) Annual Scientific Meeting.

ESGAR 2016 Book of Abstracts.

ISMB 2016 Proceedings Papers Committee.

Abstracts from Hydrocephalus 2016.

British Association of Clinical Anatomists. Abstracts of papers presented at Summer Meeting, 1978.

British Association of Clinical Anatomists. Abstracts of papers presented at the Annual General Meeting, 1979.

2016 Canadian Surgery Forum Abstracts.

2016 Bethune Round Table Abstracts.

2016 Canadian Spine Society Abstracts.

Scientific papers should not be boring.

How many scientific papers are not original?

Editorial: "Writing scientific papers in English".

Abstracts of the Seventh Scientific Meeting of The TMJ Association, 7-9 September 2014, Bethesda, Maryland.

Completion publication of abstracts presented at the scientific meetings of the Pan-African Pediatric Surgical Association.

Abstracts of the Hong Kong Urological Association Annual Scientific Meeting, November 9, 2014, Hong Kong.

Sixth Scientific Meeting of the British Oncological Association. 30th June-2nd July 1991, Bath. Abstracts.