Measurement uncertainty could play an important role in the assessment of test results in laboratories and industries. We investigated measurement uncertainties possibly included in determination of flubendazole, a benzimidazole anthelmintic, in pork by HPLC. The concentration of flubendazole was 62.69 ng/g in a sample of pork. Uncertainty was estimated in the analytical procedure of flubendazole. A model equation was made for determination of flubendazole in pork. The four uncertainty components such as weight of sample, volume of sample, calibration curve, and recovery were selected to estimate measurement uncertainties. Standard uncertainty was calculated for each component and all the standard uncertainties were combined. The combined standard uncertainty was expanded to a sample population as an expanded uncertainty. The expanded uncertainty was calculated using k value on Student's t-table and effective degrees of freedom from Welch-Satterthwaite formula. The expanded uncertainty was calculated as 3.45 with the combined standard uncertainty, 1.584 6 and the k value, 2.18. The final expression can be ($62.69{\pm}3.45$) ng/g (confidence level 95%, k = 2.18). The uncertainty value might be estimated differently depending on the selection of the uncertainty components. It is difficult to estimate all the uncertainty factors. Therefore, it is better to take several big effecting components instead of many small effecting components.

We investigated the safety, immunogenicity and protectivity of mix-crude outer membrane protein (cOMP) vaccine against salmonellosis in animals. The mix-cOMP vaccine was extracted from Salmonella enterica serovar Typhimurium (ST) and Salmonella enterica serovar Enteritidis (SE) and Salmonella enterica serovar Braenderup (SB) isolated from pigs. The mix-cOMP vaccine gave significantly higher antibody response than ST-bacterin and ST-cOMP vaccine in guinea pigs. The survival rates of mix-cOMP vaccinated groups showed significantly higher (100%) than those (0-20%) of unvaccinated control group, challenged with 3 species of Salmonella (ST, SE and SB) in mice. Vaccinated groups in pigs showed reduction of clinical signs, increase of average weight gains, decrease of bacterial recovery rates, compared with unvaccinated groups. Especially, the survival rates (100%) of vaccinated groups in chickens showed higher than that (0%) of unvaccinated group. Based on these results, we suggest that the mix-cOMP Salmonella vaccine developed in this study will be effective for the protection against Salmonellosis caused by the various serotypes Salmonella species in animals.

Lactic acid bacteria (LAB) has been regarded as a useful microorganism and tried to manipulate plasmid DNA for increasing the usefulness. Although several methods have been developed to isolate plasmid DNA from Escherichia coli (E. coli), these methods were not sufficient to apply to LAB with exception of O'Sullivan's lysis method. So, we evaluated plasmid DNA extraction from LAB using general E. coli preparation methods and tried to improve the extraction yield and DNA purity by modifying O'Sullivan's alkaline lysis method. To improve the extraction yield, salt and carrier were added to precipitant and those were incubated at $-70{^{\circ}C}$. Only incubation at $-70{^{\circ}C}$ was the effective method of those modifications. Purity of plasmid DNA was improved by two times of each centrifugation and phenol/chloroform extraction. However, DNA was damaged by twice extraction with phenol/chloroform. Also, exclusion of ethidium bromide showed negative effect to purity. Additionally, it was recommended that improvement of the extraction yield may be due to centrifugation at high speed for more time and to dissolving complete DNA pellet before addition of 7.5 M ammonium acetate. Extraction using this modification produced higher quality of plasmid DNA.

Bovine viral diarrhea (BVD) is very important disease in cattle industry with a worldwide distribution. Detection and elimination of persistently infected calves with bovine viral diarrhea virus (BVDV) was valuable strategy for BVD eradication because those calves were main source for transmission. We surveyed persistent infection with BVDV by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC) using whole blood and skin. Five hundred thirty nine Korean native calves were tested. Four calves (0.7%) were positive for BVDV antigen for both tests. Those calves remained positive for follow-up by RT-PCR and IHC. Therefore they were identified as persistently infected with BVDV. We confirmed that immunohistochemistry using skin biopsy samples was very useful tool to detect persistently infected calves with BVDV. As far as we know, this would be first report on persistent infection with BVDV in Korea.

To provide information for the molecular pathogenesis and antigenic structures of Korean isolates of porcine epidemic diarrhea virus (PEDV), the small membrane (sM) protein gene of Chinju99 strain, which was previously isolated from piglets suffering from severe diarrhea was characterized and further analyzed with other PEDV strains. The sM gene of Chinju99 generated by reverse transcription and polymerase chain reaction had a single open reading frame with 231 bases consisting of 24.2% adenine, 18.6% cytosine, 18.1% guanine and 39.0% thymine nucleotides. Nucleotide sequence of the gene revealed 97.8% homology to those of Belgian strain CV777 and British strain Br1/87, and 97.0% to Chinese strain LZC. The gene encoded a protein with 76 amino acids, and putative amino acid sequence of the gene revealed 98.7% homology to those of CV777 and Br1/87, and 96.1% to LZC. The amino acids of Chinju99 sM gene consisted of mostly hydrophobic residues, and there were one potential N-myristylation site and one potential threonine (T)-linked phosphorylation site recognized. Also, there was a transmembrane region with 46 amino acids, and Chinju99 was more close to CV777 and Br1/87 than to LZC in phylogenetic analysis on the sM amino acid sequences.

This study was carried out to investigate the pathogenesis and pathogenicity of the porcine circovirus type 2 (PCV2) Korean isolate from weaned pigs. Twenty four weaned pigs, PCV2, porcine reproductive and respiratory syndrome virus (PRRSV) and porcine parvovirus (PPV) antibodies free, were allocated to 4 groups (n = 6). Six pigs were inoculated intranasally with PCV2 alone, 6 with PCV2 and PRRSV, 6 with the combined PCV2/PRRSV/PPV inoculum, and 6 were remained as a uninoculated negative control. Pigs were killed 3 and 6 weeks after inoculation and tissue samples examined for gross and microscopic lesions and for the presence of PCV2 antigens and nucleic acids. Experimentally inoculated pigs were evaluated for 3 considerations: 1. development of postweaning multisystemic wasting syndrome (PMWS), 2. distribution of viral antigens by immunohistochemistry and polymerase chain reaction (PCR), and 3. cytokine mRNA levels in lymph nodes. Pigs inoculated with PCV2/PRRSV/PPV showed typical clinical signs, gross findings, and histopathologic characteristics of PMWS. In the PCV2/PRRSV/PPV inoculated group, the PCV2 antigen was widely distributed in various parenchymal organs such as brain, spinal cord, tonsil, lymph nodes, lung, heart, liver, kidney, spleen, and peyer's patch. Lymph node mRNA expression of IL-$1{\alpha}$, IL-2R and IL-8 was determined by real-time PCR. The pigs of PCV2/PRRSV and PCV2/PRRSV/PPV inoculation group, the mRNA expression was characterized by a decrease of IL-$1{\alpha}$, IL-2R and IL-8. The decrease of cytokine mRNA represent the state of T cell immuno-suppression in pig, and nicely support the evidence for the impairment of immune system in pigs with PMWS. In conclusion, PCV2 infection and some additional infectious causes such as PRRSV and/or PPV are warranted for the presence of PMWS in weaned pigs in Korea.

A 7-year-old Jeju native boar was requested to the Veterinary Pathology Laboratory of Cheju National University with a clinical signs of anorexia, melena, lethargy and sudden death. At necropsy, four coalescing firm masses were occupied in the abdominal cavity between ventral surface of stomach and pancreas. Individual mass was a yellowish white in color and up to 6 cm in diameter. These masses were not encapsulated and bulged from the cut surface. Liver and spleen were enlarged and pale nodules were presented in these tissues. Some yellowish white nodules up to 5 mm in diameter were scattered in kidneys. Histopathologically, lymphoblastic tumor cells were occupied in the abdominal masses, multifocal areas of liver, kidneys, and spleen. Morphologically lymphoblastic tumor cells were round to oval in shape, and medium to large in size. They had round to oval nuclei, moderate amount of eosinophilic cytoplasm, and many mitotic figures. Immunohistochemistry revealed that tumor cells were CD3-positive and $CD79{\alpha}$-negative, consistent with T-cell lineage. Based on gross, microscopic findings and immunohistochemistry, this case was diagnosed as porcine multi-centric T cell lymphosarcoma. In animals, as in human, the T-cell lymphomas are generally more aggressive than B cell types and respond less well to therapy. In our best knowledge, this is the first report for porcine T cell lymphosarcoma in Korea.

This study reveals the unusual case reported for the first time on the selective non digestion of yellow mealworm (Tenebrio molitor) larvae by arowana fish. In January 2005, an Asian arowana (Scleropages formosus) (red variety), from Daesang Tropical Fish Corporation, Seoul, Korea, mortality was observed due to unknown cause. No putative causal factors were suggested by bacteriological and parasitological examinations. Internal examination of the dissected stomach showed some undigested debris with mandible parts of mealworms attached to the mucosal lining of stomach wall. Feeding experiment of yellow mealworm (Tenebrio molitor) was conducted on the silver arowana (Osteoglossum bicirrhosum). Result showed that on the test group, fish released vomitous material containing undigested mandibles of mealworm was observed on the 24th day of the feeding experiment. Histopathological examination of the cross section of the stomach wall layers of the test groups, showed detached parts of the mucosal layer and gastric pits around the damaged area with intact mucularis tissues. While the control group fed with mealworm larvae without head part showed intact stomach wall layers consisting of gastric pit, mucosa and mucularis tissues. Fish on both treatments survived until the termination of experiment. The removal of mealworm larvae head before feeding probably help or aid in the fast digestion of these insect larvae.

This experiment has been investigated in order to examine larval migration inhibition activity of ivermectin, doramectin and ethanol against Anisakis simplex (A. simplex) in vitro. A. simplex larvae were obtained from the mackerel acquired from the fish market of Cheongju. They were divided into many groups and placed in culture dishes (40 larvae each) containing RPMI-1640, in the absence or presence of different concentrations of ivermectin, doramectin and ethanol. Ivermectin had a complete inhibition of larval migration at 72 h in all groups ($10-300{\mu}g/ml$). Ethanol reduced the migration of the larvae, its maximum activity being an high doses (7.5%, 10% ethanol) when it was 100% efficacy at 4 h. Doramectin had no efficacy in vitro. Being needed that further studies with ivermectin and doramectin, it is recommended that in vivo test with laboratory animals should be carried.

Conventional processing of biological materials including nematode parasites for scanning electron microscopy includes fixation with glutaraldehyde and osmium, followed by dehydration in an ascending grade of ethanol, and finally freeze drying. This procedure takes about 8 to 12 h depending on the characteristics of samples. Microwave irradiation of 2,450 MHz enhance the action of cross-linking fixatives and can greatly accelerate various stages of tissue processing. In this study, samples of nematode parasites, Setaria digitata, were fixed by a combination of conventional chemical fixation and the microwave irradiation during the process. The microwave irradiation was also incorporated in the serial dehydration process with ethanol. The complete procedure from the initial fixation to the completion of dehydration with ethanol was reduced to 1 h with good preservation of the ultrastructural details of the specimens.

A 3-year-old female Cocker Spaniel dog acutely developed hepatopathy following ingestion of root of Cycad (Cycas revoluta). Seven hours after the ingestion, she showed acute continuous nausea, vomiting, and marked depression and was presented to the local veterinary clinic. At physical evaluation, the dog showed moderate weakness, pale mucous membrane, jaundice, and dehydration. Serum biochemical analysis revealed elevated alanine aminotransferase and aspartate aminotransferase, and mild azotemia. The dog was treated with intravenous fluid, antibiotics, and hepatic protectants for 10 days and recovered without any sequela. Although the ingestion of Cycad in dogs is rare, it can be treated successfully with appropriate supportive therapy.

A 14-month-old, neutered female Miniature Pinscher was referred due to severe renal failure. The dog was treated with hysterotomy and followed by ovariohysterectomy (OHE) 6 days before the presentation. Excretory urographic and ultrasonographic studies revealed bilateral hydroureter and hydronephrosis. Postmortem necropsy revealed that ligatures were placed around the caudal portion of bilateral ureters in front of entrance into the bladder. This case shows that identifying and protecting the ureters are important during hysterotomy with OHE as well as OHE.

Bone allograft had been increased the need because autogenous bone graft is associated with donor site morbidity and is restricted in quantity. The bone allograft implants have to prepare properly for increasing osteoinductive ability and decreasing immune responses before providing to graft. The purpose of this study was to investigate the efficacy on new bone formation in bone allografts by treatment of implants. Cortical bone allografts were transplanted to experimental defects on midshaft of diaphyseal fibulae in 15 rabbits, which were divided to 3 experimental groups according to the preparation methods-freezing, freeze-drying, defat-freezing. The grafted regions of fibulae of all groups had been radiographed biweekly for 16 weeks to observe new bone formation and union between implant and recipient bone. Bone Alkaline Phosphatase (BALP) in all groups was evaluated biweekly till the end of the experiment to determine osteoblast activities. Unions between implant and recipient bone were observed at 30% (3 of 10 cases) of freezing, 50% (5 of 10 cases) of freeze-drying and 80% (8 of 10 cases) of defat-freezing. BALP was increased over 100% from before graft at 2 weeks of graft procedures in all union cases of freezing and defat-freezing group, then gradually decreased till 16th week. In non-union cases, there is no significant variation in BALP value. Defat-freezing method for allograft implants might be more effective for osteoinductive efficacy of implants than freezing and freeze-drying method.

Vulvar agenesis in an eight-month-old toy poodle dog is described. Urinary incontinence and dysuria were the main clinical signs. The morphology of the urogenital system was assessed with contrast radiography. The constructers of lower urinary tract and reproductive system except for the vulva were normal features. Through episiostomy, a perineal stoma, resembling a vulva, was created, resulting in complete resolution of the clinical signs.

This study was carried out to evaluate the effects of Dochetang for the treatment of chronic diarrhea in a 4 months-old-female Korean native calf. The calf was presented to the Wow Animal Clinic, Iksan with the history of chronic diarrhea for several weeks. The feces test did not reveal the presence of the parasites or microbes causing diarrhea in calf. The blood test was also negative to the virus that causes of diarrhea in calf. Adminstration of parenteral antibiotics resulted in improvement of the condition temporarily but diarrhea was recurred again after 2-3 weeks. Then the calf was treated with Dochetang administered orally once a day in an empty stomach for 15 days. Feces was significantly reduced in moisture on 7 days after initial treatment. On 9 days after initial treatment, the calf had normal appetite and defecation in physiological conditions. Blood samples were collected before administration and on 1, 2 and 3 weeks after initial administration of Dochetang for hematology and biochemistry. A significantly differences were observed in the white blood cell (WBC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), albumin (ALB), glutamic pyrubic transaminase (GPT), inorganic phosphorus (IP) and magnesium (Mg), while no significant differences were seen in the red blood cell (RBC), hemoglobin (Hb), hematocrit (HCT), platelet (PLT), glucose (Glu), total protein (T-pro), glutamic oxaloacetic transaminase (GOT), blood urea nitrogen (BUN) and creatine (CRE). This study suggests that Dochetang administration can be a successful alternative therapeutic agent in instead of antibiotics for the treatment of chronic diarrhea in calves.

Parallux (IDEXX Laboratories, USA), a solid-phase fluorescence immunoassay (SPFIA) developed for antibiotics residue detection in milk, was applied for analysis of antibiotics in muscle tissue of olive flounder (Paralichthys olivaceus), rockfish (Sebastes schlegeli), and red sea bream (Pagrus major). Fishes were dipped in streptomycin 20 g/ton water, the recommended therapeutic dose, for 3 consecutive days. Muscle was sampled after 1st, 2nd, 3rd, 4th and 5th day drug treatment. The concentration of streptomycin in muscle, determined by SPFIA, was compared with that of internal standard (200 ppb as streptomycin). The absorbance ratio of sample to internal standard (S/C) was employed as an index to determine the muscle residues in fishes. The standard solutions were added to muscle samples to give final concentrations in muscle of 0.2 and 0.5 mg/ml to investigate the recovery rate. The recovery rates of all spiked samples were > 84% of the spiked value. Streptomycin was detected in muscles of fishes treated after the 1st day of withdrawal period. The present study showed that the SPFIA can be easily adopted in predicting muscle tissue residues for streptomycin in farmed fishes.

This study was performed to identify the normal anatomic orientation of pulmonary arteries and to obtain the normal baseline parameters and the optimal contrast material delivery methods of computed tomographic pulmonary angiography (CTPA) on normal beagle dogs. Based on the contrast injection flow rate, the contrast volume, and the administration methods, the experimental groups were divided into 4 groups such as group 1 : 2 ml/s, 3 ml/kg, and monophasic administration; group 2 : 5 ml/s, 3 ml/kg, and monophasic administration; group 3 : 5 ml/s, 4 ml/kg, and monophasic administration; group 4 : 5 ml/s and 2 ml/kg in first phase, 0.3 ml/s and 2 ml/kg in second phase, as biphasic administration. Normal anatomic orientation of pulmonary arteries in CTPA was evaluated through reformatted and 3D images after retro-reconstruction. Normal parameters for great arteries and peripheral pulmonary arteries were obtained on the factor of basement hounsfield unit (HU) values, contrast enhanced HU values, delay time, and peak time. And the optimal contrast delivery methods were evaluated on the factor of contrast enhanced HU values, image quality, and artifact. The monophasic administration with 5 ml/s contrast injection flow rate and 3 ml/kg contrast volume was optimal in canine CTPA.

Familial renal amyloidosis was found in a four-year-old male Shar Pei dog. The dog had intermittent fever with signs of renal failure. Another sibling of this dog also showed subclinical signs of renal amyloidosis. Despite aggressive therapy with peritoneal dialysis, the dog died after 10 days of the first presentation. With special staining for amyloid, the renal amylodosis was confirmed.

The choice of input distribution in quantitative risk assessments modeling is of great importance to get unbiased overall estimates, although it is difficult to characterize them in situations where data available are too sparse or small. The present study is particularly concerned with accommodation of uncertainties commonly encountered in the practice of modeling. The authors applied parametric and non-parametric bootstrap simulation methods which consist of re-sampling with replacement, in together with the classical Student-t statistics based on the normal distribution. The implications of these methods were demonstrated through an empirical analysis of trade volume from the amount of chicken and pork meat imported to Korea during the period of 1998-2005. The results of bootstrap method were comparable to the classical techniques, indicating that bootstrap can be an alternative approach in a specific context of trade volume. We also illustrated on what extent the bias corrected and accelerated non-parametric bootstrap method produces different estimate of interest, as compared by non-parametric bootstrap method.