Recently, genetically engineering using recombinant DNA techniques has been applied to design new viral vaccines in order to reduce some problems which present viral vaccines have. Up to now, many viruses have been investigated for development of recombinant attenuated vaccines or live viral vectors for delivery of foreign immunogenic antigens. In this review, we introduced three kind of viruses; herpesviruses, vaccinia viruses, and adenoviruses, which have best widely been studied as recombinant vaccines or delivery vaccines for the veterinary use.
Approximately a half of swine examined had a fat storing area (F-area) in the quadrate lobe of the liver. This area was stained strongly by Sudan Black B (fat staining) and PAS (carbohydrate staining). The area was light brown to greyish yellow in appearance due to the deposition of lipid droplets. In cross section the F-area was wedge-shaped, and had a specific venous supply which directly connected it to the right gastric vein. The larger the connecting vein the wider the F-area, indicating that the connecting vein, not reported so far, is a key factor in the formation of the F-area. This would be a good model for the formation of partial fatty liver.
In order to elucidate the mechanism of germ cell degeneration in experimental cryptorchidism, we examined the testes of adult mice from a morphological standpoint. Adult ICR mice were made cryptorchid either unilaterally or bilaterally. In some mice, testes were surgically replaced back into the scrotum at 2 months after induction of cryptorchidism to observe the regenerative process. Morphological changes of cryptorchid and replaced testes have been studied by light and electron microscopy. Testes were also examined by the TUNEL (TdT-mediated dUTP-biotin nick end labelling) method to evaluate whether the degenerative cells, spermatocytes and spermatids, were dying by apoptosis or by any other process(es). At 8 weeks after the induction of cryptorchidism, the seminiferous epithelium consisted only of Sertoli cells, spermatogonia, and some spermatocytes of early meiotic stages. Soon after the replacement of testes to the scrotum, most of the seminiferous tubules resumed spermatogenic processes. Many degenerating cells, especially the spermatocytes, showed condensation of the nucleus and cytoplasm in cryptorchid testes. Although the cytoplasm was markedly eosinophilic under a light microscope to imply condensation of the cytoplasm, the extent of the condensation was not as pronounced under an electron microscope as reported in previous publications. The cytoplasm showed no expansion. By the TUNEL method, many of the degenerating cells, mainly the spermatocytes, have been shown as undergoing apoptosis. These data provide evidence that at least some of the cells die by apoptosis, or by a process similar to apoptosis.
Distributions of the cardiac plexuses and cardiac ganglia were gross-anatomically and histologically studied in eight Beijing ducks. The cardiac plexuses consisted of two components, the cardiac nerve arising from the sympathetic trunk and the cranial and caudal cardiac nerves arising from the vagus. Branches of these nerves made the cardiac plexuses on the epicardium. The cardiac plexuses could be divided into the six plexuses, that is, the right and left coronary plexuses, pericardiac transverse sinus plexus, caudal cardiac plexus, and right and left superior cardiac plexuses. There were small ganglia in the caudal cardiac plexus and the right and left coronary plexuses. These ganglia containing multipolar neurons were found like a linking chain in a single nerve.
The adherent chicken cell line, MDCC-MSB1-41C, was highly transplantable and metastatic in vivo, compared with the parental non-adherent cell line MDCC-MSB1 from Marek's disease (MD) lymphoblastoid tumor. For clarification of differences in extracellular matrix proteins in MSB1-41C and MSB1 cells, examination was made of various components of extracellular matrix proteins. A detachment experiment indicated the protein(s) recognizing the Arg-Gly-Asp (RGD) sequence, the minimum structure required for recognition by the cell-surface receptors, is essential for the adherent character. Immunoblot assay using antibodies showed increased expression of fibronectin, fibronectin receptors, and vinculin on MSB1-41C cell lines. RGD-directed integrins mediate important cell-cell adhesive interaction and these interactions with extracellular matrix proteins may thus possibly be requisite for migration, proliferation and metastatic dissemination of MDCC-MSB1-41C cells. The RGD-containing peptide in the culture medium could cause detachment of cultured adhesive lymphoid leukosis LSCC-1104X5 cells from the dish too.
The trivalent vaccine of A. pleuropneumoniae serotype 1, 2 and 5 (AP3V) was prepared in the oil-in-water type adjuvanten form. At an SPF farm, the vaccinated pigs were observed for their antibody response, finishing rate, and lung lesions at the time of slaughter and for injection scars. The CF titers against serotype 1, 2 and 5 started to rise after the second injection, showed the highest titer at 30 days after injection and then gradually decreased in vaccinated pigs. The finishing rate in the vaccinated group was 91.6% and that in the control group immunized with commercial vaccine was 60%. The lungs in the control pigs showed severe pneumonia with hyperemia, pleural adhesion and abscess. In contrast, vaccinated pigs showed slight pneumonia. Injection scars were not observed in vaccinated pigs 100 days after the second injection. In conclusion, the pigs immunized with AP3V were sufficiently protected against A. pleuropneumoniae infection and the trial proved to be satisfactory in the safety of the vaccine under field conditions.
We established a versatile method for the measurement of indocyanine green maximal removal rate (ICG Rmax) to detect hepatic functional mass in conscious rats using a repeated blood sampling procedure. On investigation of the optimal technical conditions, the appropriate intravenous administered doses of ICG were 2.5, 5, 10 or 20 mg/kg, and the best blood collection times for calculating plasma half-life at these doses were immediately before, and 4, 7 and 10 min after ICG injection. The interval among the respective ICG injections was more than 4 hr. In hepatectomized rats, the ICG Rmax value was reduced to about 50% and 20% of sham-operated rats in mean 2/3 and 4/5 liver resections, respectively, suggesting that it would almost extrapolate to hepatic surviving reserves under these experimental conditions. In rats treated subcutaneously with carbon tetrachloride (CCl4, 0.1 and 0.25 ml/kg) thrice weekly during a 17-week period (120 days), a decrease in ICG Rmax value did not correlate with increases in serum alanine transaminase (ALT), alkaline phosphatase (ALP) and total bilirubin values throughout the experimental periods. However, the reduced ICG Rmax well correlated with decreases in serum albumin and cholinesterase (CHE) values from day 50. Histological examinations in the liver revealed that nodules of hepatocytes were separated by thick fibrous bands, defining the typical aspect of cirrhosis on day 30 to 90. These results suggest that the measurement of ICG Rmax is a valuable tool for the estimation of hepatic functional integrity in rats.
This study was undertaken to characterize 20 cases of dominance aggression seen at Tufts University School of Veterinary Medicine and to investigate the efficacy of our non-confrontational behavior modification program for 8 weeks. The 20 cases included 18 pure breed and 2 mixed breed dogs. Thirteen of the dogs were male. The dogs' ages ranged from 7 to 84 months (mean 32.1 ± 22.64 SE). There was no correlation between the severity of dominance aggression and the signalment of the dogs. At the conclusion of the eight week follow up period, 14 dogs (70%) were reported to have responded to the treatment to some degree. Six dogs did not demonstrate any noticeable reduction in aggressive behavior or became more aggressive. The results of the study is powerful evidence of the efficacy of the non-confrontational behavior modification program.
Certain macromolecules of human and canine cutaneous basement membrane zone (BMZ) have shown to have responsibilities for pathogenesis of mechanobullous skin diseases. Salt-split skin by 1 M NaCl have been used for diagnosis of human mechanobullous diseases. However, there have been no studies to characterize canine salt-split skin. Electron microscopy of canine salt-split skin showed the separation within lamina lucida. Indirect immunofluorescence revealed the roof of the cleft was labeled by human patient serum with bullous pemphigoid, whereas laminin, laminin 5, type IV and type VII collagen were labeled at the bottom of the cleft. It is suggested that immunomapping of salt-split skin may be useful for the differential diagnosis of canine mechanobullous diseases.
Circulating immune complexes (CIC) in the sera from 45 clinically healthy cats and 23 feline immunodeficiency virus (FIV) infected cats were measured by an immune adherence hemagglutination (IAHA) method. The level of CIC in the sera from FIV sera-negative healthy cats was 47.2 ± 47.3 μg/ml when expressed as heat aggregated feline IgG equivalent value in IAHA reactivity. On the other hand, the level of CIC in the sera from FIV infected cats was 757.4 ± 910.5 μg/m l, which was significantly higher than that of healthy ones. CIC levels of 11 symptomatic cats and 12 asymptomatic ones were 837.8 ± 1138.2 μg/ml and 683.0 ± 684.2 μg/ml, respectively. These results showed that IAHA method was reliable to detect CIC levels of cat sera and that CIC levels in the sera of cats infected with FIV were higher than those of healthy ones.
The gene encoding the envelope glycoprotein (E2) of bovine viral diarrhea virus (BVDV) was expressed in a baculovirus. The expressed protein was detected on the surface of infected cells by immunofluorescence. Western blotting analysis showed the presence of the expressed protein of a similar molecular size to the E2 protein. The antigenicity of expressed protein were tested in guinea pigs and cattle. The immunized animals developed neutralizing antibodies against BVDV.
Wild Japanese serows (Capricornis crispus) were found to have parasitic lesions in tendons that attached the musculus tricepus brachii to the olecranon. Histopathological study of the lesions showed chronic tendinitis with multiple granulation nodules around the worms. The lesions were found in 138 of the 353 serows examined and were more frequent in aged animals than young ones. Transverse ridges on the cuticle of the female midbody, the sizes and morphological features of the spicules, and the arrangement of the caudal papillae of the males showed the parasite to be Onchocerca skrjabini. Therefore, O. skrjabini causes olecranon lesions in addition to fibrous bursa formation in carpal and tarsal regions of the Japanese serows.
A malignant aortic body tumor was observed in a 5-year-old female Holstein cow. The neoplastic mass, of 22 × 17 × 15 cm in size, was located at the base of the left atrium, having irregular lobular structures. The tumor cells had slightly eosinophilic cytoplasm, and a round or oval nucleus. Metastasis was only present in the premediastinal lymph node. The tumor cells exhibited intense immunoreactivity for neuron-specific enolase (NSE) and synaptophysin, and were moderately positive for chromogranin A. Electron-microscopy revealed membrane-limited granules in the cytoplasm. The cultured cells were spindle in shape, and having projectional cytoplasm. They were intensely positive for NSE, synaptophysin, chromogranin A, and neurofilament (200 kD). Consequently, this case was diagnosed as a malignant aortic body tumor from the neuroecrodermal origin.
A comparative study on the adhesion of attaching and effacing Escherichia coli (AEEC) to the enterocytes between the colon of a calf and the jejunum of a piglet showed differences in the developmental process of attaching and effacing (AE) lesions. In the calf, pedestals consisted of fused microvilli, while in the piglet they developed from the apical epithelial cell membranes after effacing microvilli. Microvilli adjacent to the AEEC attachment site were atrophic in the calf, whereas they were elongated in the piglet. The production of AE lesions in the calf may be indicative of a novel developmental process with AEEC infection.
Effects of soft X-ray irradiation on the natural killer (NK) cell activity and the percentage of asialo GM1-positive cells of spleen cells in mice were investigated by using a soft X-ray generator intended for non-destructive radiological examination. Soft X-ray irradiation in graded doses of more than 25.8 mC/kg indicated dose dependent reductions in the NK cell activity in the spleen of irradiated mice. Significant reductions in the population of asialo GM1-positive cells in spleen cells were also observed. These results suggest that a soft X-ray generator could also be useful in immuno-irradiation studies.
The aim of this study was to examine renal hemodynamics at the hypovolemic and recovery phases in two different hypovolemic shock models using Doppler ultrasonography, and to compare this with systemic hemodynamics. In experiment 1, the hypovolemic phase was induced in 6 mongrel dogs by removing arterial blood at 30 ml/kg for 60 min. In the recovery phase, this blood was reinfused at 30 ml/kg over 60 min. In experiment 2, hypovolemia was induced in 12 beagle dogs by rapid blood removal until blood pressure decreased to 40 mmHg and was maintained at this pressure for 30 min. Six of the dogs were then infused with 20 ml/kg hydroxyethyl starch over 5 min, and the other 6 were infused with 60 ml/kg lactated Ringer's solution also over 5 min. Parameters for systemic and renal hemodynamics were measured by using a polygraph and the Doppler method, respectively. The decrease of diastolic blood flow, resulted in an increase of vessel resistance, and was detected in the hypovolemic kidney by the Doppler method. The rapid and large volume infusion of resuscitation fluids was effective for the recovery of both systemic circulation and renal blood flow, however this induced an increase of kidney vessel resistance, a result of the autoregulation mechanism of the kidney. The changes in these parameters at the main renal artery and interlobar artery were similar.
Effects of ether stress on the hypothalamo-hypophysial-gonadal axis in adult male rats were examined. To clarify the role of adrenal glucocorticoids in gonadal function, the effects of adrenalectomy and Dexamethasone treatment were also investigated. Ether stress increased the plasma concentrations of ACTH and corticosterone, but decreased the plasma concentrations of LH, FSH, inhibin and testosterone. The pituitary responsiveness to LH-RH for LH release and testicular responsiveness to the endogenous LH for testosterone release were maintained in stressed rats. Adrenalectomy caused an increase in the plasma concentrations of ACTH, but decreased the plasma concentrations of LH, FSH and testosterone. Dexamethasone treatment in adrenalectomized rats recovered the levels of plasma gonadotropins to control levels. The concentration of plasma inhibin did not change in adrenalectomized rats, but it was decreased compared to control rats by Dexamethasone treatment. Treatments of Dexamethasone in intact male rats resulted in a decline in plasma levels of testosterone and inhibin without a decrease in the levels of LH and FSH, indicating the direct effect of Dexamethasone on the testes. These results indicate that increased ACTH secretion in stressed rats is probably due to hypersecretion of CRH from the hypothalamus, which suppresses gonadotropin secretion via the inhibition of LH-RH. The decreased levels of testosterone may be caused by a stress-induced decrease in plasma LH concentrations and increased secretion of corticosterone in the ether stressed rats. The low levels of plasma inhibin in stressed rats was also probably due to the direct effect of corticosterone on the Sertoli cells.
Recently, transvaginal ultrasound-guided follicle aspiration technology has been found to be of great value for in vitro fertilization (IVF) programs, even though the oocyte recovery rate and cleavage rate of transferrable embryos were low. In this study, we investigated the effect of the removal of the dominant follicle at different stages of the estrous cycle on the ovarian response of donor cows. Four experiments (EXPs) were devised. In EXP 1, 3 cows received 20 mg FSH on Day 1, ovulation occurred on Day 0, and on Day 3 follicles were aspirated. In EXP 2, the dominant follicle of the first wave was removed on Day 6 from 3 cows which received 20 mg FSH on Day 7 and on Day 9 follicles were aspirated. In EXP 3, 2 pregnant cows received 20 mg FSH on 70 d of pregnancy and 48 hr later follicles were aspirated a total of 5 times at 5-day intervals. In EXP 4, after ovulation on Day 0, 9 cows received 20 mg FSH on Days 8 to 14 of the estrous cycle and 48 hr after the last injection, follicles were aspirated once. The respective mean ± SD numbers of aspirated follicles and recovered oocytes were higher (p<0.01) in EXP 1 (13.4 ± 1.7 and 8.7 ± 2.3), EXP 2 (12.1 ± 1.4 and 7.7 ± 1.7) and EXP 3 (10.7 ± 2.1 and 7.0 ± 2.2) than in EXP 4 (5.8 ± 2.3 and 3.1 ± 1.6). The oocyte recovery rates were higher (p<0.05) in EXP 1, EXP 2 and EXP 3 than in EXP 4. Similarly, the respective numbers of viable oocytes and cleavage rates were higher in EXP 1, EXP 2 and EXP 3 (6.0 ± 1.3, 5.0 ± 1.1 and 4.6 ± 1.5 viable oocytes (p<0.01); 66, 73 and 65% cleavage rates (p<0.05)) than in EXP 4 (2.4 ± 1.1; 46%). The numbers of morulae and blastocysts were higher (p<0.05) in EXP 1, EXP 2 and EXP 3 than in EXP 4. In conclusion 1) removal of the dominant follicles from lactating and pregnant cows enabled viable oocytes to be recovered constantly and repeatedly by aspiration at different reproductive stages, and that viable blastocysts can be produced after IVF. 2) The presence or absence of a dominant follicle significantly affects the ovarian responses to FSH treatment. 3) This ultrasound-guided procedure proved to be an effective, repeatable and safe method for viable oocyte recovery from valuable pregnant donors.
The effects of quaternary ammonium compounds (QACs) with sodium hydroxide on swine vesicular disease virus (SVDV), an enterovirus were studied. Didecyldimethylammonium chloride (DDAC) with 0.1% NaOH showed a stronger effect against SVDV than other QACs with 0.1% NaOH. The effect of DDAC with 0.1% NaOH was strong at 40°C. DDAC was effective against SVDV at pH values around 11.0, but not in the distilled water control. The effect of DDAC with 0.1% NaOH was already observed at 1 min after mixing of the DDAC with SVDV. Observation under an electron microscopy revealed that the probable mechanism of inactivation of DDAC with 0.1% NaOH is as follows: The virus particles were partially destroyed by 0.1% NaOH. DDAC gathered these affected particles and formed a micelle, then SVDV lost its infectivity. From these results, QACs with 0.1% NaOH are considered to be very effective against SVDV representing enteroviruses.
Non-cytopathogenic (NCP) viruses of bovine viral diarrhea (BVD) virus were detected at a low ratio by the reverse plaque formation method from virus samples after several plaque clonings of cytopathogenic (CP) BVD viruses; NADL and Osloss strains. This phenomenon suggests that the NCP BVD viruses are produced at a low ratio during the propagation of CP BVD viruses in vitro. To investigate the differences between the parent CP BVD virus and the NCP BVD virus as a real progeny, the regions flanking the insertion of cellular mRNA in the p125 domain of NADL and Osloss strains were amplified by RT-PCR and cloned into pGEM 3Z plasmid vector, and then sequenced. Consequently, it was confirmed that sequences of cellular mRNA insertion of CP BVD viruses, NADL and Osloss strains, were completely and exactly deleted from the NCP BVD viruses which were real progeny of CP BVD viruses, NADL and Osloss strains. These results suggest that NCP BVD viruses may revert from CP biotype to NCP biotype by the deletion of cellular mRNA insertion in the viral genome of CP BVD viruses (NADL and Osloss strains).
Six strains of feline immunodeficiency virus (FIV) classified into subtypes A, B and D were examined by cross-neutralization test using Kumi-1 cells (CD4+, CD8-, and CD9+), an interleukin-2 dependent feline T-lymphocyte cell line. Neutralizing activities against these six FIV strains were also investigated in 50 FIV-antibody-positive serum samples collected from different geographical regions in Japan. The cross-neutralization test revealed that antisera against the six strains tended to possess high neutralizing activity against the homologous strain. These antisera were also capable of neutralizing viral strains of the same subtype. However, some of the antisera were broadly crossreactive with all six FIV strains. Serum samples collected from naturally infected cats in the field showed various neutralization patterns for the six FIV strains. These observations reflect the antigenic diversity in FIV strains prevailing in the field. There were also broadly crossreactive serum samples, and 36% (18/50 samples) showed neutralization for all six FIV strains.