Euthanized Research Articles

OTX008 inhibits retinal neovascularization in oxygen-induced retinopathy mice

Objective To investigate the inhibitory effects and possible related mechanism of OTX008 [a selective inhibitor of galectin-1 (Galectin-1)] on retinal neovascularization (RNV) in mouse model of oxygen-induced retinopathy (OIR). Methods 7-day-old (P7) C57BL/6J mice were randomly (according to random number table) divided into 4 groups including normal group, OIR group, OIR-OTX008 group and OIR-phosphate buffered saline (PBS) group. To establish the OIR mouse model, mice from all groups except normal group were expose to (75±2)% oxygen for 5 days and then to room air. OIR-OTX008 group received an intravitreal injection of 1 μl (0.25 μg/μl) OTX008 at P12, OIR-PBS group received the equal volume (1 μl) of PBS injection. Mice from 4 groups were euthanized at P17, and retinas were collected for molecular biological analysis and morphological study. RNV was evaluated by counting the number of pre-retinal neovascular nuclei and the whole-mount immunofluorescent staining of mouse retina. Cyrosections of retinas were imaged via confocal microscopy to observe the enrichment of staining of Galectin-1. Protein levels of Galectin-1, Neuropilin-1 and phosphorylation of vascular endothelial growth factor receptor 2 (pVEGFR2) were determined with Western blot. Results At P17, Galectin-1 expressed higher in retinal ganglion cell layer, inner plexiform layer and inner nuclear layer from OIR group and OIR-PBS group than normal group. Galectin-1 expressed less in cryosection retinas from OIR-OTX008 group than OIR group and OIR-PBS group. The numbers of pre-retinal neovascular cell nuclei from OIR group and OIR-PBS group were obviously more than that from normal group (t=9.314,P<0.05). The number of pre-retinal neovascular cell nuclei from OIR-OTX008 group were obviously lower than those from OIR group and OIR-PBS group (t=8.038, 7.774;P<0.05). The RNV tufts area (t=13.250, 12.570), non-perfusion area (t=15.590, 12.430) and hypoxic area (t=9.542, 9.928) from OIR-OTX008 group were significantly smaller than those in OIR group and OIR-PBS group (P<0.05). Protein levels of Galectin-1 (t=24.800, 23.060), Neuropilin-1 (t=4.120, 3.530) and pVEGFR2 (t=25.880, 15.480) in the OIR-OTX008 group were significantly down-regulated than those from OIR group and OIR-PBS group (P<0.05). Conclusion Intravitreal injection of OTX008 inhibits RNV and ameliorates retinal hypoxia in mice model of OIR possibly through down-regulating Galectin-1, Neurolinpin-1 and pVEGFR2. Key words: Retinal neovascularization/prevention c Galectin 1/antagonists i Animal experimentation

Fertility and Embryo-Fetal Development Assessment in Rats and Rabbits with Evacetrapib: A Cholesteryl Ester Transfer Protein Inhibitor.

The purpose of these studies was to evaluate the effects of evacetrapib on male and female fertility and on embryo-fetal development (EFD). Evacetrapib, a potent and selective inhibitor of cholesteryl ester transfer protein (CETP), was administered daily by oral gavage starting 2 weeks (for female) or 4 weeks (for male) before mating, during cohabitation, and until necropsy in the male rat fertility study or through gestation day (GD) 17 in the female rat combined fertility/EFD study. For rabbit EFD studies, animals were dosed from GDs 7 to 19 or from 1 week before mating through GD 19. Dose levels of evacetrapib ranged from 60 to 600 mg/kg for rats and from 1 to 100 mg/kg/day for rabbits. Parental findings in rats included decreased body weight and food consumption and moribund euthanasia in animals given 600 mg/kg/day and decreased food consumption at 300 mg/kg/day. There were no adverse effects on estrus cycling, fertility indices, sperm parameters, maternal reproductive parameters, male reproductive tissue, or fetal viability, growth, or external/visceral morphology. An increase in the incidence of 14th rudimentary ribs, a minor, transient variation considered nonadverse, was the only significant developmental finding in rats given 600 mg/kg/day. Slight decreases in body weight and food consumption at 100 mg/kg/day were the only maternal effects observed in rabbits with no adverse developmental effects noted. No adverse effects on fertility or EFD were observed in rats at doses up to 600 mg/kg/day and no adverse effects on EFD were noted in rabbits at doses up to 100 mg/kg/day. Birth Defects Research 109:513-527, 2017. © 2017 Wiley Periodicals, Inc.

First tick-borne encephalitis in a dog resident in Northern Germany

Tick-borne encephalitis (TBE) is a mainly tick-transmitted viral disease in humans and mammals. It is caused by several subtypes of tick-borne encephalitis virus (TBEV), genus flavivirus, endemic in Asia, southeastern and central Europe. In Germany most cases of tentatively infected dogs with neurologic signs were mainly investigated serologically. Intralesional demonstration of the causative agent and molecular characterisation has been described in dogs exclusively in other countries. A three and a half-year-old male-neutered Saint Bernard dog, resident in northern Germany, developed subsequent to routine vaccination paresis and dyspnoea and was euthanised due to poor prognosis. Pathomorphologically, a severe multifocal lympho-histiocytic meningo-encephalomyelitis with neuronophagia and intralesional TBEV antigen was found. Using PCR and sequencing, genome fragments of the European subtype of the virus were detected. Immunophenotyping of the inflammatory infiltrate revealed predominantly T-lymphocytes and macrophages/microglial cells with an extensive expression of MHC-II. TBE was confirmed in a dog resident in northern Germany for the first time. In most canine cases of TBEV infection, virus is eliminated rapidly without a central nervous inflammatory reaction. Whether routine vaccination in this case interacted with virus clearance, remains undetermined. TBE has to be regarded as differential diagnosis also in northern Germany in dogs with neurological signs. As prevention the application of effective repellents is recommended.

Effect of Concentrated Apple Extract on Experimental Colitis Induced by Acetic Acid.

Reactive oxygen and nitrogen species (ROS/RNS) play a crucial role in inflammatory bowel disease (IBD) exacerbating the chronic inflammatory process. Endogenous and diet antioxidants can neutralize these compounds. The apple is widely consumed, with several antioxidant activity compounds. The present study evaluated the effects of concentrated apple extract (CAE) in acetic acid induced colitis. 29 Wistar male rats were randomized into 5 groups. G1-Sham/saline solution, G2-CAE/control, G3-acetic acid/control, G4-curative- CAE treatment and G5-preventive-CAE treatment. Eight days later, the animals were euthanized and the colonic segment resected for macroscopic and histological analysis. Gene expression was evaluated for inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), catalase and copper and zinc superoxide dismutase (CuZnSOD) by quantitative real time PCR, while protein expression was assessed for iNOS, COX-2 and 8-hydroxy-20-deoxyguanosine (8-OHdG) via immunohistochemistry. The groups G3, G4 and G5 had weight loss, while G5 had weight increase at the end of the experiment. The treatment with CAE reduced the macroscopic and microscopic injury, decreased iNOS mRNA expression and increased CuZnSOD mRNA expression in animals with induced acetic acid-colitis. The findings of the present study suggest that CAE treatment exerts an antioxidant role by downregulating iNOS and upregulating CuZnSOD.

Mice aversion to sevoflurane, isoflurane and carbon dioxide using an approach-avoidance task

Euthanasia of laboratory animals is one of the main concerns for animal welfare, especially the use of carbon dioxide (CO2) which has been described to induce aversive-like behaviours. In order to refine mass euthanasia in rodents, this study aims to determine which gas, sevoflurane, isoflurane or CO2 induce less aversion in mice. Contrarily to carbon dioxide, the other gases do not induce formation of carbonic acid in the mucosa and are often used to anaesthetized mammals.A new approach-avoidance test was performed using 26 male NMRI mice. Animals had the possibility to choose between two gas environments delivered in different chambers at a 0.2L/min:1) 8% sevoflurane or 4% isoflurane (sevo vs iso), 2) 8% sevoflurane or 100% CO2 (sevo vs CO2) and 3) 4% isoflurane or 100% CO2 (iso vs CO2). Mice were placed in a central elevated compartment called the air chamber, and moved freely between the rewarded gas chambers until ataxia was observed or when 300s elapsed. Time spent in each chamber, the number of visits per minute, and eaten rewards were measured to evaluate aversion.Mice spent significantly more time in the sevoflurane chamber than in the air chamber in the sevo vs CO2 treatment. In the iso vs CO2 treatment, mice spent less time in the isoflurane chamber than in the air chamber. Also, the introduction of isoflurane and CO2 in the testing trial induced mice to spend less time in these gas chambers compared with the time spent when no gas was present during the habituation period. Moreover, animals also ate fewer pellets in the isoflurane chamber compared with what was eaten when no gas was present.These observations indicate that sevoflurane may induce lower levels of aversion, contrarily to CO2, and isoflurane. Thereby the use of sevoflurane may contribute to the refinement of mice euthanasia. Sevoflurane may be used to induce unconsciousness before a potential more aversive gas is used to finish the euthanasia process.

Expression change of microRNA-124 and its correlation with axon regeneration after traumatic brain injury in mice

Objective To observe the expression changes of microRNA-124(miRNA-124) following traumatic brain injury (TBI) in mice and investigate the correlation of miRNA-124 with neural axon regeneration. Methods Ninety-one C57BL/6 mice were assigned into TBI group (n=63) and control group (n=28) according to the random number table. Mice in TBI group were subjected to controlled cortical impact and euthanized at 12 hours and 1, 3, 7, 14, 21, 28 days postinjury for the collection of brain tissue in the trauma zone. Mice in control group underwent craniectomy only. Trauma zone observation was done using the HE staining. Expression of miRNA-124 was detected using the real-time PCR. Levels of Nrp-1, Gap-43 and Tau were detected using the Western blot and immunohistochemical staining. Results After injury, study of mice behavior and HE staining indicated the establishment of experimental model was successful. Expression of miRNA-124 reached the peak at 3 days postinjury (3.80±0.22), expression of Nrp-1 reached the peak at 7 days postinjury (2.006±0.179), expression of Tau reached the peak at 14 days postinjury (2.063±0.172), and expression of Gap-43 sustained high level since 12 hours after injury(1.355±0.093) (P 0.05), but the morphology in TBI group changed obviously. Although the positive cells of axon marker decreased at 1 day postinjury, expressions of miRNA-124, Nrp-1, Tau and Gap-43 in TBI group were significantly increased compared to the detections in control group (P<0.05). Conclusion Increased expression of miRNA-124 in trauma zone may closely related to axon regeneration after TBI in mice. Key words: Brain injuries; Axons; microRNA-124

Even with rehydration, preservation in ethanol influences the mechanical properties of bone and how bone responds to experimental manipulation

Typically, bones are harvested at the time of animal euthanasia and stored until mechanical testing. However, storage methods are not standardized, and differential effects on mechanical properties are possible between methods. The goal of this study was to investigate the effects that two common preservation methods (freezing wrapped in saline-soaked gauze and refrigerating ethanol fixed samples) have on bone mechanical properties in the context of an in vitro ribosylation treatment designed to modify mechanical integrity. It was hypothesized that there would be an interactive effect between ribose treatment and preservation method. Tibiae from twenty five 11week old female C57BL/6 mice were separated into 2 preservation groups. Micro-CT scans of contralateral pairs assessed differences in geometry prior to storage. After 7weeks of storage, bones in each pair of tibiae were soaked in a solution containing either 0M or 0.6M ribose for 1week prior to 4 point bending tests. There were no differences in any cortical geometric parameters between contralateral tibiae. There was a significant main effect of ethanol fixation on displacement to yield (−16.3%), stiffness (+24.5%), strain to yield (−13.9%), and elastic modulus (+18.5%) relative to frozen specimens. There was a significant main effect of ribose treatment for yield force (+13.9%), ultimate force (+9.2%), work to yield (+22.2%), yield stress (+14.1%), and resilience (+21.9%) relative to control-soaked bones. Postyield displacement, total displacement, postyield work, total work, total strain, and toughness were analyzed separately within each preservation method due to significant interactions. For samples stored frozen, all six properties were lower in the ribose-soaked group (49%–68%) while no significant effects of ribose were observed in ethanol fixed bones. Storage in ethanol likely caused changes to the collagen matrix which prevented or masked the embrittling effects of ribosylation that were seen in samples stored frozen wrapped in saline-soaked gauze. These data illustrate the clear importance of maintaining hydration if the eventual goal is to use bones for mechanical assessments and further show that storage in ethanol can alter potential to detect effects of experimental manipulation (in this case ribosylation).

Exploring the Grief Experience Among Callers to a Pet Loss Support Hotline

ABSTRACTThe present study explored the grief experience among callers to the Ontario Veterinary College Pet Loss Support Hotline (PLSH). Summary notes (n = 75) of the conversations between callers and PLSH volunteers were qualitatively analyzed using content analysis. Major content categories emerging from the analysis include personal pet narrative; perceived support sources; grief experiences; relationship with pet; memorialization; grief coping strategies; grief triggers; decision to euthanize; and veterinary interaction. Findings support that pet loss grief is a broad, complex construct, which can be complicated by a pet owner’s perceptions that they have minimal social support sources and negative veterinary interactions. Pet owners involved in the current study appeared comfortable sharing personal stories about their pet’s death when asked. In addition, memorialization appeared to be widely used by PLSH callers and may have helped them cope with their loss. Importantly, the interaction with veterinary staff was identified to potentially aggravate or alleviate a pet owner’s grief experience. Implications for mental health counselors and veterinary staff are that asking pet owners to tell their pet’s story and explore their support network may have value following the loss of a pet. Also, effectively communicating about memorialization and grief coping strategies may be beneficial when talking to owners experiencing grief. Finally, there continues to be a need for curricula and continuing professional training courses for veterinarians and mental health counselors to address topics such as the human–animal bond, grief, euthanasia, and memorialization practices.

Anatomy, histology and histochemistry of accessory sex glands in male Persian squirrel (Sciurus anomalus)

Persian squirrel (Sciurus anomalus) is a species of squirrels in the Middle East. There is little information about the anatomy and histology of various organs of this species. Moreover, there are no practical data about the accessory male sex glands of this species of squirrel. In this study the anatomical, histological and histochemical properties of the male reproductive system accessory glands of Persian Squirrel was evaluated. Eight adult male squirrels were anesthetized and euthanized. The pelvic area was dissected and the male reproductive system was separated. The accessory sex glands were investigated for gross anatomical aspect. Samples were fixed in formaldehyde for histological and histochemical studies. The coagulating glands, prostate and bulbourethral glands were observed at gross anatomical level. A single heart-shape and compact prostate gland was situated on the dorsal side of pelvic urethra. Two small coagulating glands were observed on the cranio-dorsal side of prostate. Two large spiral shape bulbourethral glands were situated out of the pelvic cavity near the root of penis on both sides of anal area. Histologic studies revealed that all accessory sex glands were alveolar glands with cuboidal to columnar epithelium. The most positive reaction for PAS stain was observed in the trabeculae of glands. There was no positive reaction for lipids in glands stained with Oil red O and Sudan Black. Some anatomical differences were observed in the accessory sex glands of Persian Squirrel in comparison to other rodents, however the histology of glands was almost similar to other rodents.

Anesthesia and Euthanasia of Amphibians and Reptiles Used in Scientific Research: Should Hypothermia and Freezing Be Prohibited?

Current research guidelines for ectothermic tetrapod vertebrates prohibit the use of cold as an adjunct to gaseous anesthesia, and they prohibit freezing as a means of euthanasia of these same animals. Here, we argue that those guidelines merit re-evaluation. Under natural conditions, numerous amphibians and reptiles experience large variations of body temperature, and life at low temperatures is natural. In tropical species less tolerant of cold, nociception is likely to be extinguished at low temperatures because of the anesthetizing actions of cold on membranes and cold block of nerve conduction. Physical principles and physiological data suggest that smaller ectothermic vertebrates do not experience pain attributable to ice crystals that form during freezing. Therefore, whole-body cooling, followed by freezing, should be a humane form of euthanasia for numerous smaller ectothermic species. In addition, we believe that cooling offers a humane and useful means of supplementing currently acceptable methods of anesthesia.

Performing good death at the veterinary clinic: experiences of pet euthanasia in Finland

In contemporary pet‐keeping culture, the death of an animal is managed by the veterinary profession. The situation of euthanising the pet at the clinic is not an easy one for the owner of the animal, who has to manage the emotions involved in the death of a pet, while at the same time worrying about animal welfare in euthanasia. In this paper I explore the performances of good death in pet euthanasia. Drawing on pet owners’ experiences, I scrutinise the practice of euthanasia in the space of the veterinary clinic, emotions felt by owners about pet loss, the role of animal agency and the expertise of the veterinarian in providing the animal with an ending to its life. Theoretically, the paper draws on recent discussions about human–animal relationships as performances, as productive processes in which the relationship comes into being. The data consists of written narratives from a nationwide writing collection organised in Finland in 2014–2015. According to the analysis, the veterinary clinic as a site of pet euthanasia makes the human–pet relationship vulnerable by shifting it away from the home, the space in which the relationship is otherwise experienced and lived. Pet euthanasia nevertheless has the potential to become a relational achievement between the agency and bodies of the owner, the veterinarian and the pet. As such, it is a situated practice in which the animal can be killed at the same time that its relationship with humans is celebrated – an act of responsible killing and of care, with a possibility to provide the animal a good ending to its life.

Effects of Anadara granosa shell combined with Sardinella longiceps oil on oesteoblast proliferation in bone defect healing process

Background: Alveolar bone damage is the most common case in dentistry. One way to fix the bone damage is by using bone graft. Anadara granosa shell is a potential bone substitute since it is rich in calcium which can be processed into hydroxyapatite. The addition of Sardinella longiceps oil rich in omega-3 can modulate inflammation, thus accelerating the healing process. Purpose: This study aimed to determine effects of application of Anadara granosa shell combined with Sardinella longiceps oil on osteoblast proliferation in the healing process of bone defects. Method: The subjects were 32 male rats type Wistar divided into 4 groups (n = 8). Making defect was performed on the right bone of the femurs with a half of the diameter of round Mcisinger® Germany bur sized 18. The first group (K) is a negative control group that was not given anything. The second group (AG) was given Anadara granosa pasta. The third group (AM10) was given Anadara granosa pasta combined with 10% Sardinella longiceps oil. And, the fourth group (AM30) was given Anadara granosa pasta combined with 30% Sardinella longiceps oil. Next, preparations and animal euthanasia were performed on the 7th day after the treatment. The number of osteoblasts then was measured after making preparations for HPA with Hematoxylin eosin staining (HE). Afterward, tabulation of data followed by statistical analysis of Anova and HSD Tukey was carried out. Result: The average number of osteoblasts in Groups K, AG, AM10, and AM30 was 19.00, 34.63, 33.50, and 38.50. The results of Anova test showed a significant difference (p&lt;0.05). Similarly, the results of Tukey-HSD test also showed significant differences (p&lt;0.05) between Group K and all other groups (AG, AM10, and AM30). Nevertheless, there were no significant differences between Group AG and Groups AM10 and AM30, as well as between Group AM10 and Group AM30. Conclusion: The application of the combination of Anadara granosa shell and Sardinella longiceps oil can not increase the proliferation of osteoblasts in the healing process of bone defects.

10 Effects of Eupatorium adenophorum (flavonoids) in the rat model of benign prostatic hyperplasia

Objectives This study investigated the effect of total flavonoids on benign prostatic hyperplasia (BPH) rats though the administration of Lycopus decoction and Eupatorium adenophorum. Methods Ninety male Wistar rats, weighing 280–300 g, received an intraperitoneal injection of hydrazine hydrate, and the testes were removed bilaterally. Another 10 rats underwent a sham operation. Intramuscular injections of penicillin 200 000 units/kg were given. The rats were randomly and evenly divided into a control group, model group, Longbishu group, large, medium and small doses of Lycopus decoction groups, and large, medium and small doses of E. adenophorum flavonoids groups. On the 8th day after castration, rats were injected with testosterone propionate 4 mg/kg/day and irrigated with large, medium and small water decoctions of E. adenophorum (4 g/kg, 2 g/kg and 1 g/kg, 0.2 mL/100 g), large, medium and small doses of total E. adenophorum flavonoids (400 mg/kg, 200 mg/kg and 100 mg/kg, 0.2 mL/100 g) and Longbishu capsule suspension (300 mg/kg, 0.2 mL/100 g). The model group and blank control group were given the same volume of normal saline, continuing for 30 days. Then 2 hours after the last administration, the rats were weighed and euthanized. Prostate tissue was removed, the wet weight was measured, and the prostate index was calculated. Testosterone and oestradiol levels in prostate homogenate, and the levels of bFGF, TGF-β1, IGF-I and EGF were determined. Other samples of prostate tissue were fixed in formalin solution, and cytological morphology was examined under a light microscope. Results E. adenophorum water decoction and total flavonoids significantly reduced the rat prostate index, decreased serum testosterone levels, increased serum oestradiol levels, decreased bFGF, EGF and IGF levels in the rat prostate, elevated TGF-β1 levels, reduced rat prostate density, and increased the specific surface area. Conclusions The results of this study showed that treatment of benign prostatic hyperplasia (BPH) had a good effect and suggest a new therapy for BPH. Acknowledgments This research was financially supported by the Henan University of Chinese Medicine Graduate Student Innovation Fund (Grant No. 2014YCX011) and Research Topics of Traditional Chinese Medicine in Henan Province (Grant No. 2014ZY02007).

Novel Hemi-Staple for the Fusionless Correction of Pediatric Scoliosis: Influence on Intervertebral Disks and Growth Plates in a Porcine Model.

In vivo porcine model utilized to evaluate the influence of an intravertebral fusionless growth modulating device (hemi-staple) on intervertebral disks and growth plates. To evaluate the radiographic and histologic changes in disks and growth plates with the purpose of measuring influence of the explored hemi-staple. Fusionless growth modulation for the early treatment of scoliosis should insure the long-term viability of the intervertebral disk and successfully reduce or arrest local growth. A novel hemi-staple that proved effective in the control of coronal spinal alignment warranted further analyses of its influence on the disk health and growth-plate morphology. A hemi-staple that inhibited local vertebral growth exclusive of the disk was introduced over T5-T8 in 4 immature pigs (16 vertebrae; experimental), whereas 3 underwent surgery without instrumentation (sham) and 2 had no intervention (control). Three-month follow-up before animal euthanasia provided radiographic (disk height and health) and histologic (growth plate morphology, disk health, and type X collagen distribution) analyses. No postoperative complications were experienced. Radiographic data returned inverse disk wedging (greater disk height adjacent to device, 2.6±0.7 mm compared with the noninstrumented side, 1.8±0.5 mm) in experimental segments and suggested disk viability. Histologic data confirmed device growth modulation through significant local reduction of growth plate hypertrophic zone (125.64±16.61 μm and 61.16±8.25 μm in noninstrumented and instrumented sections, respectively) and cell height (16.14±1.87 μm and 9.22±1.57 μm in noninstrumented and instrumented sections, respectively). A variability of disk health, dependant of device insertion location, was observed. Type X collagen was consistently identified in experimental growth plates and absent from intervertebral disks. Hemi-staples decreased growth plate hypertrophic zone and cell height, and, depending on device insertion site, showed positive signs of disk health sustainability. Spinal growth modulation achieved exclusive of disk compression, as practiced by this method, offers unique advantages over other fusionless techniques. This technique may provide a suitable and attractive alternative for the early treatment of idiopathic scoliosis.

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A dog suspected to be affected with rabies attacked 31 animals in Kesarahatti village of Gangavathi taluk, Koppal District in Karnataka on 14 th June 2015. Of these, 26 animals (14 cattle, 5 buffaloes and 7 dogs) were given the first aid followed by post exposure anti rabies vaccine at 1 (15 th June 2015), 3, 7, 14 and 28 days interval whereas the remaining 5 animals (2 cattle and 3 buffaloes) reportedly died of suspected (based on clinical manifestation) rabies after 8 days of dog bite episode. These 5 animals were not provided first aid and post exposure prophylactic (PEP) vaccination as their exposure to the rabid dog bite went unnoticed. Further, the serum samples could be collected from only 21 out of 26 vaccinated animals on 28 th and 90 th day post vaccination. These serum samples were tested by Rapid Fluorescent Focus Inhibition Test (RFFIT) and the titre of neutralizing antibodies ranged from 1-8 IU/ml from the serum collected on 28 th day post vaccination indicating protection in all the vaccinated animals. But the titre ranged from 0.5-4 IU/ml in 18 animals and less than 0.5 IU/ml in three animals (2 cows and a dog) on 90 th day post vaccination. The protective level of neutralizing antibody titre on day 28 post vaccination and non-development of disease suggests that the PEP vaccination could be effective in livestock avoiding euthanasia and culling of animals. Sunilkumar KM 2 , Isloor S 1,* , Ansar Kamran C 2 , Santosh AK 1 , Ramesh PT 2 , Sharada R 3 , Yathiraj S 2 , Rathnamma D 1 , Suryanarayana T 2 , Patil SS 4 and Abhinandan Patil 4

Bariatric Embolization: Pilot Study on the Impact of Gastroprotective Agents and Arterial Distribution on Ulceration Risk and Efficacy in a Porcine Model

PurposeTo assess whether the number of fundal arteries embolized and use of gastroprotective agents have an impact on ghrelin suppression and gastric ulceration rates. Materials and MethodsTwenty-two healthy, growing swine (mean, 38.4 kg; range, 30.3–47.0 kg) were evaluated. Six control swine underwent a sham procedure. Gastric embolization was performed by the infusion of 40-µm microspheres selectively into some or all gastric arteries supplying the gastric fundus. In group 1, 6 swine underwent embolization of all 4 arteries to the gastric fundus. In group 2, 5 swine underwent embolization of 2 gastric fundal arteries. In group 3, 5 swine underwent embolization of 1 gastric fundal artery. Animals in groups 2 and 3 were treated with gastroprotective agents (sucralfate and omeprazole). Weight and fasting plasma ghrelin levels were analyzed at baseline and at week 4. Upon animal euthanasia, gross analysis was performed for identification of ulcers. ResultsOnly group 1 animals exhibited changes in serum ghrelin levels that rendered them significantly lower than those in control animals (P = .049). Group 3 animals exhibited marked elevations in serum ghrelin levels compared with control animals (P = .001). Gross pathologic evaluation revealed 0 ulcers in the control animals, 3 ulcers (50%) in group 1, 2 ulcers (40%) in group 2, and 2 ulcers (40%) in group 3. ConclusionsAdministration of gastroprotective agents and embolization of fewer arteries to the gastric fundus did not prevent gastric ulceration in treated animals. Only animals that underwent embolization of all gastric arteries exhibited significant decreases in serum ghrelin levels.

SHEEP AS AN EXPERIMENTAL MODEL FOR BIOMATERIAL IMPLANT EVALUATION.

ABSTRACTObjective:Based on a literature review and on our own experience, this study proposes sheep as an experimental model to evaluate the bioactive capacity of bone substitute biomaterials, dental implant systems and orthopedics devices. The literature review covered relevant databases available on the Internet from 1990 until to date, and was supplemented by our own experience. Methods:For its resemblance in size and weight to humans, sheep are quite suitable for use as an experimental model. However, information about their utility as an experimental model is limited. The different stages involving sheep experiments were discussed, including the care during breeding and maintenance of the animals obtaining specimens for laboratory processing, and highlighting the unnecessary euthanasia of animals at the end of study, in accordance to the guidelines of the 3Rs Program. Results:All experiments have been completed without any complications regarding the animals and allowed us to evaluate hypotheses and explain their mechanisms. Conclusion:The sheep is an excellent animal model for evaluation of biomaterial for bone regeneration and dental implant osseointegration. From an ethical point of view, one sheep allows for up to 12 implants per animal, permitting to keep them alive at the end of the experiments. Level of Evidence II, Retrospective Study.

A Simple Experiment Demonstrating Hormonal Control of Cutaneous Drinking in Toads

Simple teaching experiments demonstrating endocrinological concepts are difficult to come by. I discuss a simple experiment demonstrating control of cutaneous drinking by the hormone angiotensin II in terrestrial toads, designed after methods published in the primary literature and presented herein to make this exercise more widely accessible to ABT readers. This experiment is notable in that it (1) permits students to quantify both a physiological and a behavioral response to exogenous hormone administration; (2) can be implemented in relatively small class sizes; (3) does not require animal euthanasia; and (4) can be accomplished in one class period. Furthermore, data collection can be easily carried out by students using common laboratory supplies, and analysis and interpretation of the collected data are straightforward. Finally, in demonstrating the function of the highly conserved renin-angiotensin-aldosterone system, this hands-on experiment has obvious clinical connections relevant to human medicine (e.g., blood-pressure regulation and treatment of hypertension).

The difference between dead and away: An exploratory study of behavior change during companion animal euthanasia

Abstract Anecdotal reports suggest that animals might have a sense of dying and death, although little sustained scientific research has been conducted on death awareness in animals. We set out to systematically gather data from veterinarians regarding their own personal observations of animal behavior when another animal is euthanized. In our survey of 153 veterinarians in South Carolina, 54% reported that they rarely, sometimes, often, or always observe a change in nearby animals' behavior at the time of euthanasia. Behavior change was most often mentioned as occurring among dogs (sudden silence, agitation, and barking), yet horses were also noted as exhibiting behavior change. The reasons offered by veterinarians for these behavior changes were detecting scent of chemical changes in the dying body, awareness that the animal is ill and/or deceased, empathy, responding to changes in the physical and emotional states of the animal, release of the euthanized animal's spirit, similar grief to that of humans, and reaction to the emotions of humans.

A Good Death? Report of the Second Newcastle Meeting on Laboratory Animal Euthanasia.

Simple SummaryMillions of laboratory animals are killed each year worldwide. However, there is a lack of consensus regarding what methods of killing are humane for many species and stages of development. This report summarises research findings and discussions from an international meeting of experts and stakeholders, with recommendations to inform good practice for humane killing of mice, rats and zebrafish. It provides additional guidance and perspectives for researchers designing projects that involve euthanasing animals, researchers studying aspects of humane killing, euthanasia device manufacturers, regulators, and institutional ethics or animal care and use committees that wish to review local practice.Millions of laboratory animals are killed each year worldwide. There is an ethical, and in many countries also a legal, imperative to ensure those deaths cause minimal suffering. However, there is a lack of consensus regarding what methods of killing are humane for many species and stages of development. In 2013, an international group of researchers and stakeholders met at Newcastle University, United Kingdom to discuss the latest research and which methods could currently be considered most humane for the most commonly used laboratory species (mice, rats and zebrafish). They also discussed factors to consider when making decisions about appropriate techniques for particular species and projects, and priorities for further research. This report summarises the research findings and discussions, with recommendations to help inform good practice for humane killing.