• Journal of Veterinary Clinics
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• v.31 no.1
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• pp.19-24
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• 2014

The purpose of the present study was to determine the priming effects of progesterone that affect the emergence of LH surge mode secretion by three different progesterone levels. In previous studies, we have shown that LH surge occurred in follicular levels of progesterone, whereas there was no surge mode secretion of LH and FSH in either the subluteal or luteal levels of progesterone. In this study, the hypothesis was that the priming effects of progesterone on the timing of the LH surge induced by exogenous estradiol are same between subluteal and luteal levels of progesterone. Long-term ovariectomized Shiba goats that had received implants of estradiol capsules (Day 0) and three different progesterone silastic packet inducing follicular, subluteal and luteal levels of progesterone were divided into three groups such as non-P, low-P and high-P group. Blood samples were collected daily throughout the experiment for the analysis of gonadal steroid hormone levels. On Day 7, all devices of progesterone packets were removed but estradiol capsules were maintained during the experiment, and blood samples were collected at 1 hr interval for 12 h from the time of progesterone removals to determine peripheral changes of estradiol and progesterone concentration. Then all animals were infused estradiol on the Day 7 after 13 h from the removals of progesterone devices with a peristaltic pump into jugular vein at a rate of 3 ${\mu}g/h$ for 36 h. For analysis of peripheral LH and estradiol concentration, blood samples were collected via another jugular vein at 2 h intervals for 52 h (from 4 h before the start of estradiol infusion to 48 h after the start of estradiol infusion). In all animals of the three groups treated with estradiol infusion, an LH surge was expressed but the peak time of LH surge was different. This time interval from estradiol infusion until the peak of LH surge was gradually and significantly extended by the different levels of progesterone treated before estradiol infusions in the three groups.

• Journal of Veterinary Clinics
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• v.31 no.1
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• pp.25-30
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• 2014

This study tested the hypothesis that the priming effects of progesterone on the timing of the LH surge induced by exogenous estradiol are more potentiated the negative feedback actions of progesterone on LH secretion by the existence of estradiol. In previous studies, the time interval from estradiol infusion until the peak of LH surge was gradually and significantly extended by the different levels of progesterone treated before estradiol infusions. Longterm ovariectomized Shiba goats that had received implants of estradiol capsules (Day 0) and three different progesterone silastic packet inducing follicular, subluteal and luteal levels of progesterone were divided into three groups such as non-P, low-P and high-P group. Blood samples were collected daily throughout the experiment for the analysis of gonadal steroid hormone levels. On Day 7, all devices of progesterone and estradiol packets were removed but estradiol capsules were maintained during the experiment, and blood samples were collected at 1 hr interval for 12 h from the time of progesterone removals to determine peripheral changes of estradiol and progesterone concentration. Then all animals were infused estradiol on the Day 7 after 13 h from the removals of progesterone devices with a peristaltic pump into jugular vein at a rate of 3-6 ${\mu}g/h$ for 36 h. For analysis of peripheral LH and estradiol concentration, blood samples were collected via another jugular vein at 2 h intervals for 52 h (from 4 h before the start of estradiol infusion to 48 h after the start of estradiol infusion). In all animals of the three groups treated with estradiol infusion, an LH surge was expressed but the peak time of LH surge was different. This time interval was not extended by the different levels of progesterone treated before estradiol infusions and the difference was not significant during this interval between the Low P and the High P groups. Progesterone pretreatment may contribute to regulating the neural system that is responded by estradiol, and estradiol existence potentiates the negative feedback effect of progesterone on GnRH/LH surge-generating system.

• Journal of Veterinary Clinics
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• v.29 no.5
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• pp.377-383
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• 2012

Bovine diarrhea is a major economical burden to the bovine industry in Korea. Since multiple infectious agents can be involved in bovine diarrhea, differential diagnosis is essential for effective treatment. Therefore, a panel of two multiplex real-time PCR assays which can simultaneously detect six major bovine enteric pathogens [i.e., bovine viral diarrhea virus (BVDV), bovine coronavirus (BCoV), group A bovine rotavirus (BRV), Salmonella spp., Escherichia coli (E. coli) $K99^+$, and Cryptosporidium parvum] was developed and applied to test 97 fecal samples collected from cattle farms in Korea. In addition, microscopic examination was also preformed on the samples to detect Coccidium oocyst. The estimated sensitivity of the multiplex PCR was 0.1 $TCID_{50}$ for BVDV, BCoV and group A BRV, 5 and 0.5 CFU for E. coli $K99^+$ and Salmonella, respectively, and 50 oocysts for Cryptosporidium. The amplification efficiency of the multiplex PCR ranged between 0.97 and 0.99 for each pathogen. Among 97 samples, 36 samples were positive for at least one of the 6 major pathogens and 6 samples were simultaneously positive for 2 pathogens by the multiplex PCR assay. Coccidium oocysts were also detected in 48 samples, which were all collected from over 1 month old calves. In conclusion, the multiplex real-time PCR panel can be a useful tool for fast and accurate diagnosis of calf diarrhea associated with BVDV, BCoV, group A BRV, E. coli $K99^+$, Salmonella, and/or Cryptosporidium and Coccidium may be an important target which needs to be included in the multiplex PCR panel in the future.

• Journal of Veterinary Clinics
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• v.29 no.2
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• pp.141-147
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• 2012

This work describes the characteristics of $Malassezia$ $pachydermatis$ isolated from dog ear canals and the effect of essential oils on the growth of this organism. Sterile cotton swabs were used to collect specimens from the external ear canal and culture tests were performed to detect the population size of $Malassezia$ yeast. Using three different isolation media, included Sabouraud dextrose agar (SDA) to isolate common $M.$ $pachydermatis$, and SDA supplemented with olive oil (SDAO) and Leeming's medium (LM) to detect lipophilic yeast, $Malassezia$ spp were isolated from 14 of 18 dogs (77.8%); isolation rates were 33.3% in SDA, 72.2% in SDAO and 66.7% in LM media. All $Malassezia$ spp isolates were identified as $M.$ $pachydermatis$ according to results of PCR amplification, but gross colony morphology and SDA growth rates suggested four different subtypes. Large (LC) and medium colony (MC) types respectively describe large colony (diameter > 3 mm) and medium colony (around 2 mm) after 72 hour incubation, and small (SC) type refers to smaller colony (< 1 mm) even after 5 days incubation; lipid dependent colonies did not grow onto SDA. Large Colony type strains were isolated from 4, 11, and 11 samples, MC type strains from 2, 3 and 1 and SC type strains from 1, 2 and 1 in SDA, SDAO and LM, respectively. Lipid-dependent $M.$ $pachydermatis$ (Lipo) were isolated from 3 samples each in SDAO and LM. Anti-$M.$ $pachydermatis$ activity testing was done using disc-diffusion assays and well diffusion tests. Most essential oils inhibited the growth of $M.$ $pachydermatis$ in a range from 0.5% to 1.0% of essential oils. MIC90 and MIC50 were variable depending upon the nature of essential oils. Thyme oil was found to be highly effective in inhibiting the growth of $M.$ $pachydermatis$ in a range from 0.125% to 0.0625% while marjoram and then tea tree oil exhibited lower inhibitory capacity.

• Journal of Veterinary Clinics
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• v.27 no.5
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• pp.533-539
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• 2010

Stem cell-based therapy is under intensive investigation to treat acute renal failure (ARF). The purpose of this study was to evaluate available ARF models, and suggest a model appropriate to therapeutic evaluation of the stem cells in preclinical approach by determining the optimum concentration of nephrotoxic agents and duration of ischemia induction. Three different types of available acute kidney injury (AKI) animal models were analyzed using rats: Cisplatin (saline, 5 and 7.5 mg/kg, IP) or glycerol (saline, 8 and 10 ml/kg, IM)-induced nephrotoxicity as toxic models and ischemia-induced (sham, 35 and 45 minutes) nephropathy as an ischemic model. The relevance and applicability to investigate especially the regenerative ability of stem cells were evaluated regarding morphology, renal function and survival at this time point. In the point of renal function, 10 ml glycerol/kg and 7.5 mg cisplatin/kg model in toxic models and 45 min model in ischemia models showed significant decrease for the longer observation time compared to 8 ml glycerol/kg, 5 mg cisplatin/kg and the 35 min ischemia models, respectively. All groups were observed no mortality except 45 min-ischemia model with 50% survival. Histological significant alterations including cast formation in the tubular lumen, tubular necrosis and apoptosis were revealed on the second day in either ischemiaor glycerol-induced models, and on day 5 in cisplatin-induced models. The results indicate that ischemia 35 min-, cisplatin 7.5 mg/kg- and glycerol 10 ml/kg-induced AKI would be ideal animal models to monitor a outcome parameter related to the therapeutic effects on renal function with noninvasive techniques in the same animal at multiple time points. Our findings also suggest that the best time points for the functional or histological interpretation of renal will be on day 2 in both glycerol- and ischemia-induced AKI models and on day 5 in cisplatin-induced AKI.

• Journal of Veterinary Clinics
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• v.28 no.5
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• pp.486-496
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• 2011

A special mesenchymal tissue layer called perichondrium has a chondrogenic capacity and is a candidate tissue for engineering of cartilage. To overcome limited potential for chondrocyte proliferation and re-absorption, we studied a method of cartilage tissue engineering comprising chondrocyte-hydrogel pluronic complex (CPC) and cultured perichondrial cell sheet (cPCs) which entirely cover CPC. For effective cartilage regeneration, cell-sheet engineering technique of high-density culture was used for fabrication of cPCs. Hydrogel pluronic as a biomimetic cell carrier used for stable and maintains the chondrocytes. The human cPCs was cultured as a single layer and entirely covered CPC. The tissue engineered constructs were implanted into the dorsal subcutaneous tissue pocket on nude mice (n = 6). CPC without cPCs were used as a controls (N = 6). Engineered cartilage specimens were harvested at 12 weeks after implantation and evaluated with gross morphology and histological examination. Biological analysis was also performed for glycosaminoglycan (GAG) and type II collagen. Indeed, we performed additional in vivo studies of cartilage regeneration using canine large fullthickness chondrial defect model. The dogs were allocated to the experimental groups as treated chondrocyte sheets with perichondrial cell sheet group (n = 4), and chondrocyte sheets only group (n = 4). The histological and biochemical studies performed 12 weeks later as same manners as nude mouse but additional immunofluorescence study. Grossly, the size of cartilage specimen of cPCs covered group was larger than that of the control. On histological examination, the specimen of cPCs covered group showed typical characteristics of cartilage tissue. The contents of GAG and type II collagen were higher in cPCs covered group than that of the control. These studies demonstrated the potential of such CPC/cPCs constructs to support chondrogenesis in vivo. In conclusion, the method of cartilage tissue engineering using cPCs supposed to be an effective method with higher cartilage tissue gain. We suggest a new method of cartilage tissue engineering using cultured perichondrial cell sheet as a promising strategy for cartilage tissue reconstruction.

• Journal of Veterinary Clinics
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• v.30 no.6
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• pp.442-448
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• 2013

The aim of the present study was to develop glycerol-free TRIS extender using glucose for dog sperm cryopreservation. We determined the appropriate concentration of glucose in glycerol-free TRIS and the exposure time in glycerol-free TRIS containing 0.3 M glucose at $4^{\circ}C$. Ejaculates of six dog sperm were cooled in glycerol-free TRIS through $4^{\circ}C$ for 100 min, cooled at $4^{\circ}C$ in TRIS with different glucose concentrations 0 M, 0.04 M, 0.1 M, 0.2 M and 0.3 M, respectively for 30 min followed by cryopreservation. After thawing at $37^{\circ}C$ for 25 sec, membrane and acrosome integrities of dog sperm were evaluated. In addition, the effect of exposure time (10, 30, 50 and 70 min) of sperm to glycerol-free TRIS containing 0.3 M glucose at $4^{\circ}C$ on progressive motility, viability, and DNA integrity following sperm cryopreservation was studied. Membrane integrity and acrosome integrity were assessed by 6-carboxyfluoresceindiacetate (6-CFDA)/propidium iodide (PI) fluorescent staining and Pisum sativum agglutinin conjugated to fluorescein isothiocyanate, respectively. DNA integrity was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling, using flow cytometry. Sperm frozen in glycerol-free TRIS supplemented with 0.2 M or 0.3 M glucose have an intact plasma membrane (CFDA+/PI-) after cryopreservation than sperm frozen in the extenders with lower glucose concentrations (p<0.05). Acrosome integrity was significantly higher in the 0.3 M group than less than 0.1 M groups (p<0.05). The sperm DNA fragmentation index did not differ according to exposure time, although progressive motility was significantly higher in the 50 min exposure group than the other groups (p<0.05). These results indicate that cryopreservation of dog sperm is feasible and yields more motile sperm following freezing and thawing in glycerol-free TRIS containing 0.3 M glucose with the exposure time for 50 min at $4^{\circ}C$.

• Journal of Veterinary Clinics
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• v.10 no.2
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• pp.171-184
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• 1993

In Seoul area, there are so many kennel clubs, veterinary hospital, pet establishment and breeding confers that various problems have occurred. They are crowed pet houses, poor sanitation, stress to puppies, sudden environmental changes to puppies and unvaccination against parvovirus, canine distemper virus, parainfluenza virus, infectious hepatitis caused by Adenovirus type I and Leptospira. Several studies were made to survey the infectious agents involved in acute diarrhea of poppies in Seoul are, such as history taking, physical examination, complete blood count and serum chemistry, histopathological finding, bacterial isolation and identification, and hemagglutination test in feces and hemagglutination inhibition test in serum against parvovirus, respectively. The results obtained are summarized as followed. 1. The percent of PCV (30.5$\pm$5.6) and concentration of Total protein(5.0$\pm$0.8) resulted from statistical analysis are significantly lower than normal values (p<0.05), respectively. In addition, fibrinogen (505$\pm$326) was significantly higher an normal value (p<0.001), Band neutrophil (22.9$\pm$12.7) showed signifiant difference (P<0.01). decreased monocyte (3.2$\pm$2.1) and eosinophil (0.7$\pm$0.8) values appeared statistically significant (p<0.001), lymphocyte (16.7$\pm$9), as well. 2. The concentrations of calcium .(8.0$\pm$2.8), glucose (40.1$\pm$31.4), and albumin (2.0$\pm$0.39) were lower than normal values (p<0.01, p<0.01, p<0.001), respectively. Also Inorganic phosphate (7.1$\pm$2.4), pH (p<7.9f:0.2), and Blood Urea Nitrogen (40.41=37.1) were significantly higher than normal values (p<0.001, p(0.001, p<0.05), respectively. 3. Simple and mixed infections occupied 18% and 82% in the distribution of causes in puppies with acute diarrhea, respectively. 4. As puppies got older, incidence of acute diarrhea caused by Staphylococcus aureus was decreased to 13% and infection of canine distemper virus was increased to 53%, but E coli and canine parvovirus always showed high frequency of outbreak in the body weight ranged from 35g to 7.8Kg. 5. As showed in table 5, infections of E coli and Canine Parvovirus showed high outbreak regardless of the age which is classified into three stages, 35~50 day, 60 day and 75~day to 1 year, canine distemper virus appeared increased, but in case of Staphylococcus aureus, visa versa. 6. In comparison wi methods for the laboratory diagnosis diagnosis parvovirus, Hemagglutination test showed positive reaction in 25% and mean serum antibody titer measured by Hemagglutination inhibition test showed 2779 (n=20). In addition, positive reaction was 90% (18/20). 7. In histopathological studies, enteritic and pneumonic lesions were indicated in 53.7% and 39.5% of samples, respectively. 8. Etiologic diagnosis based on the history taking, clinical signs and histopathological findings in puppies with acute diarrhea and vomiting indicated that 50% of puppies were infected by canine parvovirus and distemper virus. 9. In the parasitological examination made by simple flotation with saturated zinc sulfate tour parasites found were Isospora canis, Toxocara canis, Ancylostoma spp and Toxocara spp. Isospora canis and Toxocara canis were more frequently found among those parasites of diarrhoeic causes in puppies ranged from 35 days to 1 year. Their infestation rates were 15% and 13% respectively.

• Journal of Veterinary Clinics
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• v.10 no.1
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• pp.65-94
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• 1993

This study examined metabolic profiles of 1349 Holstein cows from 91 commercial herds. Thirteen parameters which are consisted of twelve blood components and body condition score were examined and their mean values. standard deviations and standard limits, which are 80% confidential limits, in each lactational stage were reported. The variations of each parameter affected by season, individual milk yield, adjusted corrected milk yield of herd. and lactation number were also reported. A model of metabolic profile test applicable to this country where the average number of cows in a herd is small as to be fifteen is designed. Metabolic profiles as reflected in each parameter were discussed in relation to adequacy of dietary intake for production, milk production, reproductive performance, and diseases, and the possible measure to improve productivity of dairy cows were proposed. Much of the variation in parameters was due to differences between herds, and less to differences between seasons, differences between individual milk yield, and differences between lactational stages. As the average herd size in this country is small, it is believed that all the cows in a herd must be sampled, and the individual result of each parameter was compared with the standard limit for each lactational stage, and the percentage of cows which are outside the standard limits in a herd was calculated to use as a criteria for evaluation of the herd. Data outside the 99% confidential limits were to be deleted at first, but when the trends of the data outside the 99% confidential limits are same as the trends of the data within 99% confidential limits, the deleted data must be reviewed again, otherwise some important informations would be missed. The mean concentration of blood urea nitrogen in this study was much higher than that was reported in England, U.S.A. and Japan, and it was similar to the upper limits reported in England, U.S.A. and Japan. So it was thought that the concentration of blood urea nitrogen is improper as a criteria for protein intake. The increase of serum total protein cocentration beyond standard limits was due to increase of serum globulin concentration in most of the cows. The correlation coefficient between serum and protein and serum globulin concentration was 0.83. Serum globulin concentration was negatively related to adjusted corrected milk of herd. Serum albumin, calcium and magnessium concentrations were negatively related to adjusted corrected milk of herd, which indicate that high-producing individual or high-producing herd have not taken sufficient protein/amino acids, calcium and magnessium. Packed cell volume was negatively related to adjusted corrected milk of the herd, and the trend was same In each lactational stage. The correlation coefficient between serum and packed cell volume was 0.16 and the correlation was very weak. Blood glucose concentration was lowest in early lactational stage, which indicates negative energy balance in early lactational stage. Blood glucose concentration was negatively related to adjusted corrected milk of herd from peak to late lactational stage, which indicates negative energy balance during the period in high-producing individuals or high-producing herds. Correlation coefficient between serum aspartate aminotransferase activity and serum ${\gamma}$-glutamyltransferase activity was 0.41, and this indicates that serum ${\gamma}$-glutamyltransferase should be included as a parameter of metabolic profile test to evaluate liver function. Body condition score of dairy cows in this country was lower than that of Japan in every lactational stages, and the magnitude of increase in body condition score during middle and late lactational stages was small. Metabolic profile can not be evaluated with solely nutritional intake. When an individual or large percentage of cows in a herd have adnormal values In parameters of metabolic profile test, veterinary clinician and nutritionist should cooperate so as to diagnose diseases and to calculate the e of no운ents simultaneously.

• Journal of Veterinary Clinics
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• v.31 no.6
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• pp.495-501
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• 2014

The objective of this study was to evaluate the differences and reproducibility of Hounsfield unit (HU) value and volume measurements on different computed tomography (CT) scanner types and different collimations by using a gelatin phantom. The phantom consisting of five synthetic simulated calculus spanning diameters from 3.0 mm to 12.0 mm with 100 HU was scanned using a two-channel multi-detector row CT (MDCT) scanner, a four-channel MDCT scanner, and two 64-channel MDCT scanners. For all different scanner types, the thinnest possible collimation and the second thinnest collimation was used. The HU values and volumes of the synthetic simulated calculus were independently measured three times with minimum intervals of 2 weeks and by three experienced veterinary radiologists. ANOVA and Scheff$\acute{e}$ test for the multiple comparison were performed for statistical comparison of the HU values and volumes of the synthetic simulated calculus according to different CT scanner types and different collimations. The reproducibility of the HU value and volume measurements was determined by calculating Cohen's k. The reproducibility of HU value and volume measurements was very good. HU value varied between different CT scanner types, among different beam collimations. However, there was not statistically significant difference. The percent error (PE) decreased as the collimation thickness decreased, but the decrease was statistically insignificant. In addition, no statistically significant difference in the PEs of the different CT scanner types was found. It can be concluded that the CT scanner type insignificantly affects HU value and the volumetric measurement, but that a thinner collimation tends to be more useful for accurate volumetric measurement.