Nam Shik Shin

Cloning endangered gray wolves (Canis lupus) from somatic cells collected postmortem

The objective of the present study was to investigate whether nuclear transfer of postmortem wolf somatic cells into enucleated dog oocytes, is a feasible method to produce a cloned wolf. In vivo-matured oocytes (from domestic dogs) were enucleated and fused with somatic cells derived from culture of tissue obtained from a male gray wolf 6h after death. The reconstructed embryos were activated and transferred into the oviducts of naturally synchronous domestic bitches. Overall, 372 reconstructed embryos were transferred to 17 recipient dogs; four recipients (23.5%) were confirmed pregnant (ultrasonographically) 23-25 d after embryo transfer. One recipient spontaneously delivered two dead pups and three recipients delivered, by cesarean section, four cloned wolf pups, weighing 450, 190, 300, and 490g, respectively. The pup that weighed 190g died within 12h after birth. The six cloned wolf pups were genetically identical to the donor wolf, and their mitochondrial DNA originated from the oocyte donors. The three live wolf pups had a normal wolf karyotype (78, XY), and the amount of telomeric DNA, assessed by quantitative fluorescence in situ hybridization, was similar to, or lower than, that of the nuclear donor. In conclusion, the present study demonstrated the successful cloning of an endangered male gray wolf via interspecies transfer of somatic cells, isolated postmortem from a wolf, and transferred into enucleated dog oocytes. Therefore, somatic cell nuclear transfer has potential for preservation of canine species in extreme situations, including sudden death.

Detection of betanodaviruses in apparently healthy aquarium fishes and invertebrates.

Betanodaviruses are the causative agents of viral nervous necrosis (VNN) in cultured marine fish. A total of 237 apparently healthy aquarium fish, marine (65 species) and freshwater (12 species) fishes and marine invertebrates (4 species), which were stocked in a commercial aquarium in Seoul, South Korea, were collected from November 2005 to February 2006. The brains of the fish and other tissues of the invertebrates were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) and nested PCR to detect betanodavirus. Positive nested PCR results were obtained from the brains of 8 marine fish species (shrimp fish Aeoliscus strigatus, milkfish Chanos chanos, three spot damsel Dascyllus trimaculatus, Japanese anchovy Engraulis japonicus, pinecone fish Monocentris japonica, blue ribbon eel Rhinomuraena quaesita, look down fish Selene vomer, yellow tang Zebrasoma flavesenes), 1 marine invertebrate species (spiny lobster Pamulirus versicolor), and 2 freshwater fish species (South American leaf fish Monocirrhus polyacanthus and red piranha Pygocentrus nattereri). The detection rate in nested PCR was 11/237 (4.64%). These subclinically infected aquarium fish and invertebrates may constitute an inoculum source of betanodaviruses for cultured fishes in the Korean Peninsula.

Reproduction of Fowl Typhoid by Respiratory Challenge with Salmonella Gallinarum

Abstract Fowl typhoid is a disease of adult chickens and is caused by Salmonella Gallinarum infection via the alimentary tract. The experimental reproduction of fowl typhoid per os (PO) requires artificial conditions to minimize the effect of gastric acid, and several Salmonella serovars have been known to be transmitted via the respiratory route. Therefore, we have hypothesized the existence of a respiratory route for Salmonella Gallinarum infection and have attempted to reproduce fowl typhoid via intratracheal challenge. In accordance with our hypothesis, the intratracheal challenges of Salmonella Gallinarum reproduced exactly same lesions as fowl typhoid and induced higher mortality and morbidity than those of the PO challenge. Therefore, this study represents the first reproduction of fowl typhoid via respiratory route, and our findings may be useful for understanding the transmission of Salmonella Gallinarum in the field.

Hepatocellular adenoma in a Eurasian otter (Lutra lutra).

A 7-year-old female Eurasian otter (Lutra lutra) at the Seoul Grand Park, Korea, died after displaying depression, anorexia, weight loss and rough skin for several days. At necropsy, a solitary friable round mass, which was approximately 12 × 9 × 5 cm and mottled dark red and yellow, was found bulging from the right hepatic lobe. Microscopically, the nonencapsulated, poorly circumscribed mass was composed of solid sheets of neoplastic hepatocytes. In addition, numerous small tan foci, ranging from 0.5 to 1.0 cm in diameter, were evenly scattered throughout the pancreatic tissue. These foci were found to be nonencapsulated, well-demarcated hyperplastic nodules of the exocrine pancreatic gland. We observed neither intrahepatic nor extrahepatic metastases. Based on the gross and microscopic changes, we diagnosed the animal as having a hepatocellular adenoma accompanied by exocrine pancreatic nodular hyperplasia.

Hemangiosarcoma in a South American Sea Lion (Otaria byronia)

Abstract A 10-yr-old male South American sea lion (Otaria byronia) died after several weeks of depression, anorexia, weight loss, and progressive respiratory distress. At necropsy, three confluent, lobulated, dark-red masses were noted in the mesentery. Similar masses were also observed in the lung and both kidneys. Hemangiosarcoma was diagnosed based on gross findings, histopathology, and immunohistochemistry. This is the first case of hemangiosarcoma reported in pinnipeds.

Evaluation of effects of olfactory and auditory stimulation on separation anxiety by salivary cortisol measurement in dogs

Separation anxiety (SA) is a serious behavioral problem in dogs. In this study, salivary cortisol was studied to determine if the owners odor or voice could reduce SA in dogs. Twenty-eight dogs with SA were divided into three groups: group 1 (control), group 2 (with owners clothes during the separation period; SP) and group 3 (a recording of the owners voice was played during SP). The dogs saliva was collected after the owner and their dog were in the experimental room for 5 min (PRE). The dog was then separated from the owner for 20 min and saliva collected four times at intervals of 5 min (SP1–4). Finally, the owner was allowed back into the room to calm the dog for 5 min, after which saliva was collected (POST). Evaluation of salivary cortisol concentrations by ELISA revealed that the ratios of SP1 concentration to PRE or POST concentrations were significantly higher in group 1 than in group 2 or 3. Additionally, the concentrations of SP1–PRE and SP1–POST among groups differed significantly. These findings indicate that the owners odor or voice may be helpful to managing stress in dogs with SA.

Comparison of Stress Levels Induced by Two Types of Pet Dryers Using Salivary Cortisol Measurement in Dogs

Comparison of Stress Levels Induced by Two Types of Pet Dryers Using Salivary Cortisol Measurement in Dogs The common pet dryer (CD) is typically used to dry dogs after bathing, but the excessive heat and noise can induce stress. Dog owners and facilities housing many animals, such as research facilities, have begun to widely adopt the pet dry room (PDR) as a more convenient drying alternative. In the present study, the stress induced by CD or PDR was assessed by measuring the salivary cortisol. Ten healthy beagles at a research laboratory were included. The mean cortisol level before drying (S1) with CD was 0.25 μg/dl and significantly increased to 0.38 μg/dl 10 minutes after drying (S2) and 0.56 μg/dl 20 minutes after drying (S3). The cortisol level at S1 with PDR was 0.33 μg/dl, and increased to 0.38 μg/dl at S2 and 0.40 μg/dl at S3, but this difference was not statistically significant. Comparing the values from S1, S2 and S3, use of CD or PDR were not significantly different each other. However, the difference between concentrations S1 and S3 and the S3-to-S1 ratio differed significantly between the groups. These results indicate that PDR may induce less stress in dogs compared with CD. In conclusion, this facility may be convenient for owners or managers, especially those of large-scale facilities such as for laboratory dogs or centers for abandoned dogs, and could be more comfortable for dogs in that they induce less stress. The equipment also has the potential to be widely used in other animal welfare programs.

Effects of alprazolam on capture stress-related serum cortisol responses in Korean raccoon dogs (Nyctereutes procyonoides koreensis).

The purpose of this study was to evaluate the effect of alprazolam on the stress that Korean raccoon dogs (Nyctereutes procyonoides koreensis) may experience while caught in a live trap by measuring their serum cortisol response. The animals were placed in a live trap with or without being pretreated with oral doses of alprazolam. In both groups, pre-trap blood samples were initially collected without anesthesia before the animals were positioned in the live trap; then post-trap blood samples were collected after the animals had remained in the live trap for 2 h. Changes in cortisol levels were observed using a chemiluminescent immunoassay. The level of cortisol increased in the control group and decreased in the alprazolam-pretreatment group (p < 0.05). In this study, we demonstrated that alprazolam pretreatment reduced stress during live trap capture.