Lipopolysaccharides (LPSs) were purified from Actinobacillus pleuropneumoniae serotype 2, Bordetella bronchiseptica and Haemophilus parasuis serotype 5, which were used for vaccine production in Japan, by the phenol-water procedure. In SDS-PAGE analysis, A. pleuropneumoniae LPS, as well as Escherichia coli LPS, demonstrated a typical ladder profile of a smooth-type LPS. On the other hand, B. bronchiseptica and H. parasuis LPSs lacked the ladder profiles. It was found that the biological activity of these LPSs was comparable to those of E. coli LPS in terms of activation of the clotting enzyme of Limulus amoebocyte lysate, mitogenic activity of mouse spleen cells, stimulation of TNF-α and nitric oxide production, but IL-6 production could hardly be observed in any LPS.
In order to establish a rapid diagnostic method for contagious equine metritis (CEM), we developed and evaluated a polymerase chain reaction (PCR) test. Species-specific PCR primer sets were derived from the DNA sequence of a cloned DNA fragment of Taylorella equigenitalis that did not hybridize with the genome of a taxomonically related species, Oligella urethralis. Single step PCR with primer set P1-N2 and two-step semi-nested PCR with primer sets P1-N2 and P2-N2 detected as low as 100 and 10 CFU of the bacteria, respectively. Single-step PCR detected T. equigenitalis from genital swabs of experimentally infected mares with sensitivity comparable to that of bacterial isolation. Furthermore, two-step PCR was more sensitive than the culture method. Upon examination of field samples, 12 out of 3, 123 samples were positive by single-step PCR while only 2 were positive by bacterial culture. The 12 PCR-positive samples originated from 5 mares, of which 3 animals were considered to be carriers based on previous bacteriologic and serologic diagnoses for CEM. The PCR test described in this study would provide a specific and highly sensitive tool for the rapid diagnosis of CEM.
The D4 dopamine receptor (D4DR) polymorphic region, which is possibly related to the personality trait known as novelty seeking in humans, was examined in 34 dogs from two breeds (Golden retriever and the Japanese indigenous breed, Shiba) by the polymerase chain reaction (PCR), and the DNA sequences of each allele were determined. The polymorphic region of the dog D4DR gene was composed of 39- and 12- base pair (bp) units, and four alleles (A-D) were identified based on the number and/or order of these units. Intra- and inter-breed allele variations were observed. The frequency of the short A allele was dominant (78.9%) in the Golden retriever, while the long D allele was most common (46.7%) in the Shiba. These findings suggested that the allele frequency varied significantly between different breeds, and that analysis of the polymorphism in D4DR might be of use for understanding the behavioral traits of dogs.
The bottle-nosed dolphin (Tursiops truncatus) IL-1α and IL-1β cDNA were cloned from mitogen stimulated peripheral blood mononuclear cells (PBMC) RNA utilizing the reverse transcription-polymerase chain reaction (RT-PCR). The sequences of these cDNAs showed that dolphin IL-1α and IL-1β clones contained open reading frames encoding 265 and 266 amino acids, respectively. Comparison of the deduced amino acid showed that dolphin IL-1α sequence shared 77, 77, 77, 74, 71, 65 and 57% similarity with the bovine, ovine, porcine, equine, feline, human and mouse IL-1α sequences, respectively. Similarly, the amino acid sequence showed that dolphin IL-1β shared 77, 77, 74, 69, 65, 64 and 63% similarity with the bovine, ovine, porcine, equine, feline, human and mouse IL-1β sequences, respectively. The relatedness of dolphin IL-1α and IL-1β were relatively distant with 21% amino acid homology.
To evaluate whether hemodialysis with a dialysate containing no calcium (Ca-free HD) can induce hypocalcemia and restore the clinical signs and blood biochemical changes in naturally occurred hypocalcemic disorder in ruminants, the clinical signs and the changes in plasma electrolytes and minerals concentrations were observed in goats during 6-hr hemodialysis. The four goats received hemodialysis with the dialysate containing calcium (Ca HD), and 10 days later they had Ca-free HD. The plasma ionized Ca (Ca++) and total Ca (TCa) concentrations were not affected by Ca HD, whereas the levels significantly decreased during whole period of Ca-free HD. The Ca++ and TCa concentrations were 0.69 ± 0.06 mmol/l and 5.9 ± 0.3 mg/dl at 6 hr of Ca-free HD, respectively. The clinical signs observed during Ca-free HD seemed to resemble to those in naturally occurred hypocalcemic cases that were reported previously. Therefore, Ca-free HD was suggested to be one of the possible methods to induce experimental hypocalcemia in ruminants.
Transcranial Doppler ultrasonography (TCD) may be useful for determining alterations in cerebral blood flow (CBF) during excessive hemodynamic changes by non-invasive measurement of the CBF velocity. The purpose of this study was to measure the correlation between CBF and the middle cerebral artery (MCA) and basilar artery (BA) flow velocities, as measured by TCD during excessive hemodynamic changes produced by hypertension and hypotension in adult dogs. The peak, diastolic, and mean flow velocities were measured by TCD. Arterial hypertension was induced by administration of dopamine at 5 and 15 μg/kg/min, and hypotension was induced by hemorrhage. During the hemodynamic changes, the BA velocity correlated more closely with the alteration in the CBF than the MCA velocity. In terms of percentages of the values during anesthesia, there was good correlation between CBF and the MCA and BA velocities. In conclusion, our findings indicate that MCA and BA velocity measurements, as a percentage of the values during anesthesia, both give an equally accurate indication of alterations in CBF during excessive hemodynamic changes.
DNA polymorphism in twelve starains of Eimeria tenella isolated from various places in Japan was examined using 1.1 kb small subunits ribosomal RNA amplified by PCR. Genetic variation was evaluated by random amplification of polymorphic DNA (RAPD) analysis. DNA fingerprint patterns were grouped into two, indicating that at least two DNA polymorphisms exist in Japanese E. tenella strains.
In the winter of 1998-1999, we collected parasitological data from 54 wild carnivores in the north-western part of Tohoku region, Japan. These consisted of 38 martens (Martes melampus melampus), 14 raccoon dogs (Nyctereutes procyonoides viverrinus) and 2 foxes (Vulpes vulpes japonica). Collected helminth parasites were 11 nematode, 10 trematode, 3 cestode, and a single acanthocephalan species, including 5 hitherto unknown species for this research area or the mainland of Japan (Honshu). Mesocestoides paucitesticulus was for the first time recorded from martens as well as from carnivores distributed in Honshu. Brachylaima tokudai originally recorded from Urotrichus talpoides in the central part of Honshu was for the first time found from a raccoon dog.
A gelatinous focus with cystic spaces, was found in the posterior funiculus of the 2nd to 3rd lumbar levels of the spinal cord of a Japanese Black heifer, 2 years old, with clinical signs of severe dysstasia. Histopathological examination revealed that the spinal lesion consisted of multifocal and diffuse proliferation of round cells with abundant vacuolar cytoplasm and hyperchromatic nuclei. In the lesions there was a number of cystic spaces containing aggregates of small round cells. The neoplastic foci showed a honeycomb structure divided by thin blood vessels, representing typical lesions of oligodendroglioma. Diffuse and multifocal proliferation of these round cells were also recognized in the subarachnoidal space in the sacral spinal cord. Immunohistochemically, the proliferating round cells were negative for glial fibrillary acidic protein. Based on these morphological features, the case was diagnosed as lumbar spinal oligodendroglioma with diffuse arachnoidal dissemination.
Lymphoma was observed in a 4-month-old female Japanese White rabbit. Grossly, the markedly enlarged mesenteric lymph nodes, prominent Peyer’s patches of jejunum, splenomegaly, and enlargement of tracheobronchial lymph nodes, adrenal glands and ovaries were observed. Histologically, neoplastic lymphoid cells proliferated diffusely showing frequent mitotic figures and a characteristic ‘starry sky’ appearance. Their basophilic cytoplasm contained a few lipid droplets. The mesenteric lymph nodes, Peyer’s patches of jejunum, and tracheobronchial lymph nodes were largely replaced by the tumor tissues. The stomach, small intestines, especially the jejunum, liver, spleen, ovaries, and adrenal glands were heavily infiltrated with neoplastic cells. These results suggest that the present lymphoma may have originated from the gastrointestinal lymphoid tissue.
Peripheral nerve sheath tumor was found in a 7-year-old male mongrel dog. The tumors were located in the right cheek subcutis and oral submucosa. Histologically, neoplastic cells were arranged in streaming bundles, occasionally interlacing bundles or whorls of elongated and spindle cells. Cellular atypia was poor and mitotic figures were rarely observed. Ultrastructurally, neoplastic cells had basement membrane, typical of Schwann cells. One bundle of normal peripheral nerve fibers and some myelinated axons were seen within the tumor tissues. Immunohistochemically, neoplastic cells reacted to vimentin, glial fibrillary acidic protein, S-100 protein and neuron specific enolase. In addition to the above immunoreactions, the included nerve fibers were positive for myelin basic protein and neurofilament protein. This paper also discusses immunohistochemical findings on differential diagnosis in comparison with those of canine hemangiopericytomas reported hitherto.
The purpose of this study was to examine the role of the nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) system in the regulation of the ductus arteriosus (DA) patency in fetal rats. Pregnant rats were administered NG-nitro-L-arginine methyl ester (L-NAME, 50 mg/kg, ip), an NO synthase (NOS) inhibitor; methylene blue (30, 50 and 100 mg/kg, ip), a soluble guanylate cyclase inhibitor; or indomethacin (3 mg/kg, po), a cyclooxygenase inhibitor, at various times before cesarean section. Dams were decapitated to obtain the fetuses by cesarean section, and fetuses were rapidly frozen in an acetone-dry ice mixture. Using rapid freezing and shaving methods, the calibers of the DA, pulmonary artery (PA) and descending aorta (Ao) were measured to evaluate the effects of treatment. L-NAME reduced the DA calibers to 86% of the initial values, but recovery to the control levels occurred 6 hr after the injection. Indomethacin decreased the DA calibers to 34% of the control values and sustained the DA constriction until 24 hr after the treatment. Methylene blue caused DA constriction to almost the same degree as indomethacin, but the levels normalized within 24 hr after the treatment. We conclude that L-NAME caused a slight constriction of the DA, whereas methylene blue and indomethacin caused marked constriction of the vessels, suggesting that the NO-cGMP system as well as prostaglandins contribute to the DA patency.
Avian tuberculosis accompanied with many tubercular lesions in the liver and spleen was found in a painted quail at a zoological garden in Japan. Mycobacterium avium complex (MAC) serovar 9 without insertion sequence of IS901 was isolated from the liver (1.3 × 108 CFU/g), oviduct (9.4 × 107 CFU/g), and intestine (1.5 × 105 CFU/g). The isolates were inoculated intravenously to chickens. The inoculated chickens showed clinical symptoms of avian tuberculosis. Birds are susceptible to MAC serovar 9 without IS901.
Apically and basally released Sendai viruses (SeV) were obtained after infection of polarized Madin-Darby canine kidney (MDCK) cells grown on permeable membrane culture inserts. After 20 passages of adaptation in MDCK cells, we compared their in vivo and in vitro pathogenicity with the parental Mol-strain of SeV. These viruses had comparable in vitro pathogenicity, but the in vivo pathogenicities were varied. The apically released MDCK-adapted virus showed comparable pathogenicity with the parental virus, in contrast with the basally released MDCK-adapted virus, which showed in vivo attenuation.
A serosurvey of feline herpesvirus type 1 (FHV-1), feline calicivirus (FCV), and feline parvovirus (FPV) in cats from Ho Chi Minh City area in southern Vietnam was conducted in December 1998, and we compared the results with our previous results in northern Vietnam (Hanoi area). The positive rate of FHV and FCV in domestic cats were 44% and 74%, respectively. They were rather higher than those in Hanoi area, while the seropositivity of FPV (44%) was similar to that in Hanoi area. In leopard cats, the positive rate of FPV was high (3/4) and it indicated that FPV was prevailing in leopard cats in Vietnam.