Artificial insemination (AI) with frozen or cooled semen is widely used in commercial fields of cattle and pig. Little is known about characteristics of canine sperm after freezing or cooling. For both practical and commercial goal, the canine semen treated with cooling and freezing should be carried out to exam the fundamentals, including sperm motility, survivability and fertilizing capacity. The aim of this study, thus, was to identify the effects of extended exposure to 4$0^{\circ}C$ on canine semen by motility, survivability, acrosomal changes following different duration. Fifteen ejaculates collected by digital manipulation twice per week from 3 dogs (Shih-Tzu) were divided to 16 aliquots after adding Tris-egg yolk (TE) buffer formulated by our laboratory, and cooled from 37 to 4$^{\circ}C$, by ramp rate of 0.6$^{\circ}C$/min. Each sample was evaluated by their motility, survivability and the acrosomal status at 0h (control), 2h, 12h and 1 d~10 d, respectively. The motility of spermatozoa was graded to 6 levels using the modified method of Seager. The survivability of sperm was assessed using an epifluorescence microscope after Fert/Light (Mole-cular Probes Inc.) staining. To estimate the proportion of the spermatozoa of intact acrosome, 200 spermatozoa were assessed in randomly selected fields, using epifluorescence microscope after FITC/PSA (Sigma) staining. At 2 h after cooling, the motility of most spermatozoa were assessed to be grade 0 and 1. At 12 h, high number of sperm were in grade 0 to 1, however, it was significantly (P<0.05) lower than that of 2 h. From 1 d to 4 d, ~50% of sperm was assessed to grade 0 to 1. On day 7, a little sperm were in grade 0 to 1. No sperm showed motility on day 10. Sperm motility was rapidly reduced by the percent of 10% of grade 0 to 1. From 2 h to 6 h, the number of live sperm was 90% and the sperm chilled for 10 days lived>50%. Acrosomal intact of spermatozoa exposed to 4$^{\circ}C$ for 2 h was 51%, supposed the sperm of control was 100%. Our results suggest that 1) this is easy to transfer and preservation for short periods 2) AI can be used by semen chilled for 6-Day.
Park, Seungjo;Kwon, Seong-young;Min, Jung-Joon;Choi, Jihye
Journal of Veterinary Clinics
/
v.33
no.1
/
pp.51-57
/
2016
In this study, the features of canine lymphoma on fluorine-18 fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT) were evaluated in three small breed dogs. In case 1, ultrasonography and CT indicated neoplastic involvement of the sternal, right axillary, submandibular, lower cervical, tracheobronchial, mesenteric, and sublumbar lymph nodes; spleen; and liver. However, intense FDG uptake on PET/CT images was detected only for the lymph nodes and spleen. No FDG uptake by the liver was detected for case 1 despite the confirmation of lymphoma by cytology. In case 2, ultrasonography and CT indicated neoplastic involvement of the axillary, mesenteric, and sublumbar lymph nodes and the spleen, while intense FDG uptake on PET/CT images was detected for the axillary and a few mesenteric lymph nodes, and the spleen. FDG uptake was additionally observed from popliteal lymph nodes, however there was no uptake by the sublumbar lymph nodes and some mesenteric lymph nodes. In case 3, neoplastic changes in the splenic, mesenteric, and sublumbar lymph nodes and spleen were suspected on ultrasonography, and lower cervical and popliteal lymph node involvements were additionally detected on PET/CT. Compared to ultrasonography, repeated PET/CT showed increased FDG uptake by the lymph nodes at an earlier stage after chemotherapy in case 3. This study illustrated the features of PET/CT in canine lymphomas and compared those to ultrasonography and CT findings. FDG uptakes were not detected from some lesions which were suspected to be neoplastic involvement in case 1 and 2. We could not clearly explain the reason of this result in the present study because cytological or histological examination was not performed for lesions that showed different results on ultrasonography, CT, and PET/CT. Further studies on the subclassification of canine lymphoma and the sensitivity and specificity of PET/CT for the detection of canine lymphoma are required. PET/CT data can provide useful information for predicting the therapeutic response at an early stage after treatment.
Bacterial dermatitis is common disease that is necessary to treat with antibiotics. In recent, antibiotic-resistant bacteria is being increased in worldwide. The purpose of the present study was to evaluate the prevalence of resistant genes in Staphylococcus (S.) pseudintermedius isolated from dogs, and to compare the resistant gene profile with the result of antibiotic disc diffusion test. A total of seven S. pseudintermedius was included in the study. Bacterial identification was performed by 16S ribosomal RNA gene sequence analysis. S. pseudintermedius isolates had more than one antibiotic resistant gene (mecA, blaZ and aac(6')/aph(2"). While all isolates were PCR positive to blaZ gene, only two isolates were resistant to amoxicillin/clavulanate. Among five isolates harboring gentamicin resistance, one isolate was negative to aac(6')/aph(2")-targeted PCR. Taken together, the results suggest that resistant gene-targeted PCR and disc diffusion test are complementary to detect antibiotic resistance.
Recently, demographic studies of veterinary medical database have been conducted to understand patterns of disease occurrence. Understanding incidence of breed-related disease would provide appropriate guidance for future health care strategies and offer useful information for early diagnosis of disease. However, in veterinary medicine, theses research has not yet been investigated in the Republic of Korea. The purpose of this study was to investigate the prevalence of common medical disorders among dog breeds examined at primary-care veterinary clinics in Jeollabuk-Do, Republic of Korea. The data were analyzed based on World Health Organization's International Classification of Disease. A total 13,176 medical records of canine patients were analyzed from six primary veterinary clinics in Jeollabuk-Do from January to December 2016. Results showed that the most common health problems were 'disease of skin' (17.7%); followed by 'diseases of digestive system' (12.26%), 'preventive medicine' (10.08%), and 'diseases of ear and mastoid process' (10.4%). In seven out of ten breeds, the most common medical disorder was skin disease. For poodle such as Pomeranian and Chihuahua, digestive system disease was most prevalent. On the other hand, respiratory system disease was found to be higher in Pomeranian than other breeds; while ear and mastoid process disease was most common for Maltese and Poodle. This study can help owners, breeders, and veterinarians prevent and manage various diseases of popular breeds in Jeollabuk-Do in the future.
Swine whipworm(Trichuris suis) is cosmopolitan nematode which can cause serious pathology in immature stage(larva2~larva5) of infected pigs, such as anorexia, diarrhea, anemia, and death in heavy infections. In this larval stages, it is very difficult to diagnose the infection of whipworm and to differentiate from other common swine gastrointestinal disorders such as 21 day scours which are associated with TGE virus, rota virus, coccidium, and the stress of weaning. In this experiment, the isolated antigens of Trichuris spp. were carried out to examine the structure and specificity of antigens and to select the reasonable antigens which would be used in serological diagnosis by electrophoresis, Western blotting, ELISA. The results of this experiment were as follows; 1. The common fractions of each Trichuris suis antigen were identified 28,32,45, 80kDa by SDS-PAGE with silver stain and four major fractions could be detected in positive swine sera by Western blot analysis. 2. The OD(optical density) values of somatic and excretory-secretory antigens which were reacted against positive(negative) sera from pigs infected with Trichuris suis by ELISA reader were; 1) OD values($mean{\pm}SD$) of adult somatic antigen against positive(negative) sera were $0.30{\pm}0.12(0.09{\pm}0.006)$ and third-stage larva of somatic antigen were $0.28{\pm}0.038(0.10{\pm}0.005)$. And OD values of excretory-secretory antigens of adult and third-stage larva were $0.24{\pm}0.031(0.11{\pm}0.005)$ and $0.08{\pm}0.013(0.10{\pm}0.003)$, respectively. 2) OD values of adult somatic, larval somatic antigen and adult excretory-secretory antigen response to positive sera were significantly (p<0.01) associated with negative swine sera. And the Cut-off OD values(minimum positive value) were determined to be mean negative value plus 3 SD that would minimized the risk of false positives. 3. The OD values of somatic antigens of T suis and T vulpis against swine positive(negative) sera were $0.30{\pm}0.120(0.09{\pm}0.006)$ and $0.25{\pm}0.141(0.09{\pm}0.003)$. These data mean that the somatic antigens of T suis and T vulpis were able to diagnose T vulpis infection in dogs as well as T suis infection in pigs. These results suggest that somatic antigen of third-stage larva and excretory-secretory antigen of adult T suis could be used the diagnostic antigen by serological test(ELISA) in immature Trichuris spp. infection.
Serial ultrasonographic examinations were performed on 9 pregnant Korea Jin-do dog from days 15 to 60 to determine the size of gestational structures throughout pregnancy. Gestational age was timed from the day of ovulation (Day 0), which was estimated to occur when plasma progesterone concentration was first increased above 4.0 ng/ml. Extra-fetal structures were measureable from days 17 to 49. Outer uterine diameter increased from $7.0{\pm}0.7$ ($mean{\pm}SD$)mm at day 17 to $54.0{\pm}2.2mm$ at day 49 and inner chorionic cavity diameter increased from $3.0{\pm}0.7mm$ at day 17 to $37.5{\pm}0.6mm$ at day 49. Uterine wall thickness increased from $2.8{\pm}0.4mm$ at day 17 to $8.3{\pm}0.5mm$ at day 49, placental thickness increased from $1.0{\pm}0.1mm$ at day 22 to $5.7{\pm}0.2mm$ at day 49 and length of chorionic cavity or zonary placenta increased from $5.5{\pm}1.3mm$ at day 20 to $52.3{\pm}2.2mm$ at day 49. Inner chorionic cavity diameter, outer uterine diameter and placental length each increased at a linear rate through day 37, after which time, each had a marked plateau in growth. Of the extra-fetal structures, inner chorionic cavity diameter was the most accurate for estimation of gestational age until day 37. Fetal structures were measureable from days l7 to 60. Crown-rump length, increased from $3.0{\pm}0.7mm$ at day 22 to $118.7{\pm}3.1mm$ at day 49, fetal body diameter increased from $4.0{\pm}0.7mm$ at day 25 to $55.8{\pm}1.7mm$ at day 60 and fetal head diameter increased from $4.3{\pm}0.6mm$ at day 26 to $29.8{\pm}0.8mm$ at day 60. Of the fetal structures, fetal head diameter was the most accurate for estimation of gestational age from day 37 until day 60.
Central nervous system of two dogs with natural canine distemper was investigated histopathologically and immunocytochemically with antisera to MBP, MAG and GFAP. Histopathologically, there were neuronal degeneration and diffuse gliosis in the cerebrum, vacuolar degeneration, hypertrophy of astrocytes and demyelination in cerebellar white matter adjacent to the 4th ventricle and optic tracts showing non-inflammatory demyelinating encephalomyelitis (Summers and Appel, 1987). Immunohistochemically, there was a concurrent disappearance of MBP and MAG in the well developed demyelinating lesion in the cerebellar white matter. At the margin of demyelination, Loss of both MBP and MAG varied on the stage of demyelinating process. GFAP-positive astrocytes were hypertrophied and contained canine distemper virus intranuclear inclusions. GFAP-positive fibers were increased at the early stage of demyelination, and then were not immunoreaeted at the well developed demyelination. Hypertrophic astrocytes with intranuclear inclusions were commonly identified in the interfascular layer without myelin vacuolation and demyelination. This is the first study of primary demyelination and astroglial reactions in natural CDE investigated using immunocytochemistry of two myelin proteins and GFAP. Concurrent loss of MBP and MAG suggest that the myelin sheath is the target in the demyelinating process in CDE.
Pulmonary function is the determinant of blood gas tension. However, Acid-Base disturbances can also alter partial pressures of oxygen and carbon dioxide in arterial blood. During respiratory acidosis $PO_2$ will be lowered and reverse changes will be produced during respiratory alkalosis. On the other hand, in metabolic acidosis $PO_2$ will be elevated and $PCO_2$ will be lowered by the respiratory compensation, and reverse response will be induced in metabolic alkalosis. Urinary gas tension has many influencing factors than arterial blood and difficult to estimate the tendency of its alterations. Urinary $PO_2$ and $PCO_2$ are not always identical level as venous blood. It is to be altered by blood gas tension, flow rate of urine, metabolic rate of kidney, and Acid-Base status of blood. Particularly countercurrent exchange of oxygen and carbon dioxide in the renal medulla will make larger alteration of gas tension than venous blood. After induction of Acid-Base disturbances [disturbances] arterial and urinary $PCO_2$, $PO_2$, urinary volume, and osmolarity were determined in dogs, and the relationships between arterial and urinary $PCO_2$ , $PO_2$ Acid-Base disturbances, urinary volume, and osmolarity were investigated. 1. During the acute Metabolic and Respiratory disturbances urinary pH did not respond on respiratory origin. However, there were immediate urinary response in pH on metabolic origin. 2. Urinary $PO_2$, $PCO_2$, did not always follow arterial or venous gas tension and Acid-Base disturbance. Urinary $PCO_2$, correlate well with the urinary volume. The larger the urinary volume, $PCO_2$ lowered to the venous level. The smaller the urinary volume, urinary $PCO_2$ tends to be higher. However urinary $PO_2$ did not have any particular correlation with urinary volume. 3. Correlation between urinary $PCO_2$ and $PO_2$ were inversely proportional to arterial blood. Differences of $PCO_2$ between arterial blood and urine also did not have any particular correlation with urinary volume. This may suggest that changes on blood gas tensions can influence on urinary $PCO_2$. 4. There were eminent clear inverse correlation between urinary $PCO_2$ and osmolar concentrations of urine. Above results strongly suggest that partial pressure of gas in urine primarily depend upon counter-current exchanges in renal medullary tissues.
Heparin would have been used for preventing clotting of blood during extracorporeal circulation and subsequent use of protamine sulfate and made possible the neutralization of heparin. This procedure has been adopted for eliminating one of the great causes of bleeding, especially in cardiac surgery. In this experiment, the hypocoagulability of blood induced by heparin followed by neutralization with treatment of protamine sulfate were estimated by the Lee-White clotting time [CT], partial thromboplastin time [PTT] and protamine titration test. The results were as follows: 1] Comparison of clotting time between the heparinized [2.0 mg/kg] and non-heparinized dogs was done using CT and PT`I` of the blood. In heparinized group [Group I], the CT lasted infinitively and prolongation of PTT [4 times than normal] until 60 minutes. The CT [2 times] and PTT [3 times] has been shortened after 90 minutes, however they returned to normal limit level within 180 minutes. 2] The determination of appropriate ratio of heparin and protamine In vivo were performed. The group II [heparin 2.0 mg/kg, protamine 1.0 mg/kg] revealed rapid decrease of CT and PTT, but returned to normal after 120 minutes. The group III [heparin 2.0 mg/kg, protamine 2.0 mg/kg] returned rapidly to normal within 15 minutes. The group IV [heparin 2.0 mg/kg, protamine 3.0 mg/kg] recovered its normal level after 60 minutes. The group V [heparin 2.0 mg/kg, protamine 4.0 mg/kg] recovered its normal level after 90 minutes. 3] In the combined experimental study In vivo and vitro, the protamine titration test was done using the dog which were given 2.0 mg/kg and 3.0 mg/kg of heparin, respectively and coagulation time were checked after 15, 30, 60 and 120 minutes. The complete neutralization was showed to be heparin-protamine ratio of 1:1 to 1.5. 4] In vitro study, fresh blood was drawn into known amount of heparin content [20, 40, 60 and 100/ug per 1 ml of blood] syringe, thereafter protamine titration test was done. In all cases, the complete neutralization was found in heparin-protamine ratio of 1:0.85 to 1.5. 5] It was found by the present experiment that the ideal heparin-protamine ratio was 1:1 within 60 minutes and 1:0.5 after 60 minutes for avoiding the serious side effect due to overadministration of protamine sulfate.
A variety of experiments concerning the development of ideal prosthetic grafts for correcting circumferential tracheal defects have been performed. The requirements for an ideal tracheal prosthesis are impermeability to air, consistency to prevent collapse, and acceptance by the host tissue causing a minimum inflammatory reaction, allowing fibroblastic infiltration and epithelialization. The synthetic material, polyurethane(PU), is known as a biocompatible polymer with an inert component. In this study, the tracheal prosthesis was made from microporous PU(30 micrometer in diameter) coated with gelatin and reinforced with isoplastic rings. This procedure provides the prosthesis with a compression strength. The out side diame er of the prosthesis was 20 mm with a length of 30 mm. The gelatin used in the study was obtained from pig skin and immobilized and cross-linked by irradiation(60 Co gamma ray) to promote host tissue incorporation and render the prosthesis epithelization after implantation. Animal experiments using 10 mongrel dogs were performed to compare three kinds of prosthesis; gelatin coated polyurethane graft, uncoated polyurethane graft, and prosthesisf pericadium complex graft. After 6 weeks of implantation, the epithelialization of implants was seen on the gelatin-coated and prosthesisfpericadium complex grafts. Implanted prosthesis were complicated by airway obstruction due to anastomosis granuloma. Early tracheal stenosis was found in the uncoated graft group. Two kind of anastomosis techniques were tested on the gelatin-coated prosthesis. Everted anastomosis resulted severe granuloma than the inverted anastomosis. In the prosthesislpericadium complex graft, bacteria and inflammation at a anastomotic site was found. Based on these results, gelatin coated porous polyurethane trachea prosthesis is biocompatible and may be useful in clinical application with further investigation.
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