The distribution of motoneurons innervating the tail muscles, M. levator caudae, M. depressor caudae, M. pubocaudalis externus, M. pubocaudalis internus, M. lateralis caudae and M. flexor cruris, in the adult pigeon was examined by retrograde transport of horseradish peroxides. Labeled motoneurons innervating tail muscles were distributed in the longitudinal column of the spinal cord below the sacral spinal segment 5. The average diameter of cell bodies ranged from 23.7 to 62.5 μm. Each motoneuron pool was localized in a characteristic position in the ventral horn.
One hundred and thirty-eight strains of Staphylococcus hyicus and 21 strains of S. chromogenes isolated from animals were analyzed by pulsed-field gel electrophoresis (PFGE) after restriction endonuclease SmaI digestion of chromosomal DNA. Eighty-eight strains of S. hyicus from pigs with or without exudative epidermitis (EE) generated 16 to 26 fragments in the size range of <1 to 485 kb, and yielded 39 different patterns. With regard to the strains from pigs with EE, PFGE patterns differed according to the country of origin. Outbreaks of EE occurring on four separate pig farms in Japan involved S. hyicus with different PFGE patterns. The PFGE patterns shown by S. hyicus strains from 4 kinds of animals were compared. Strains from pigs differed from those isolated from chickens (n=45; 18 to 24 fragments of <1 to 425 kb), cows (n=3; 17 to 19 fragments of <1 to 475 kb), and goats (n=2; 16 or 17 fragments of <1 to 1,125 kb). Also, each of the chicken, cow and goat strains had a host-specific fragment. The results suggest that PFGE analysis might be a useful marker for distinguishing ecovars within S. hyicus. In contrast, strains of S. chromogenes from pigs and cows generated 17 to 24 fragments ranging from <1 to 545 kb. The PFGE patterns of S. chromogenes strains were more highly conserved than those of S. hyicus. S. chromogenes strains could be distinguished from S. hyicus strains by fragments within the range of 305 to 545 kb. The results indicate that PFGE analysis could be used to distinguish between S. hyicus and S. chromogenes. We conclude that PFGE analysis is a useful tool not only for species or strain identification but also for epidemiologic or ecologic studies of S. hyicus and S. chromogenes.
Isolation of mycoplasmas from the oropharynxes of 60 fantails reared under natural conditions at different zoological parks in Miyazaki prefecture was carried out. Mycoplasma columbinum, M. columborale and M. columbinasale were isolated from 28 (46.7%), 22 (36.7%), and 1 (1.7%) of 60 oropharynxes, respectively, but no Mycoplasma was isolated from 5 cloacas tested. Ureaplasma was not isolated from any of the 28 oropharynxes or 5 cloacas examined. We report that 41 (68.3%) of 60 fantails had one or two species of mycoplasmas in their oropharynxes, and make the first confirmation of M. columbinasale inhabiting a Japanese pigeon.
Although it is known that commercialized bovine serum is sometimes contaminated with mycoplasmas, it is not clear whether mycoplasmas can survive in horse serum. In this study, as a preliminary examination of the survival of mycoplasmas inoculated in horse sera, the survivability of 8 strains of 7 mycoplasmas was tested. The results obtained reveal that two strains of M. bovis and M. gallisepticum were found to survive in non-heated and inactivated sera for 94 to 330 days at 30 or 37°C. Three strains of M. bovirhinis, M. gateae and A. laidlawii lived for 7 to 330 days depending upon the temperature maintained or pH of the serum. Strains of M. bovigenitalium and U. diversum survived for a maximum of 8 days in all horse sera tested. Therefore, mycoplasmas are generally likely to survive for a long period in horse serum although the survival period depends on the species, strain and temperature.
The phenotype of high K (HK) red blood cells, which is an autosomal recessive, was found in dog groups from 10 of 13 breeds or populations in Japan. The incidence of HK was 26 to 38% in the San'in-Shiba, Shinshu-Shiba and Akita breeds, and the gene frequencies of HK ranged from 0.513 to 0.612. The highest incidence (42%) was found in the Jindo breed from Korea, and the gene frequency was 0.652. Two other groups from Korea also possessed this HK variation. However, although HK cells were not found in dogs from Taiwan, Indonesia, Mongolia and Sakhalin, Russia, the HK phenotype is clearly distributed now throughout Japan and Korea.
In order to obtain basic information about bovine interleukin-1 (IL-1β), levels of IL-1β in sera and milk of clinically normal mature Holstein cattle before and after parturition and in sera of newborn calves were examined by ELISA. The level of IL-1β was undetectable in sera of mature cattle around the time of artificial insemination, but the concentration gradually increased and reached a peak at parturition and then decreased again to an undetectable level. IL-1 β in milk was detected on the day of parturition but not thereafter. IL-1β mRNA was detected by reverse transcription-polymerase chain reaction in the cells from milk collected during 20 days before and 2 to 3 days after parturition, but was not detected thereafter. Although IL-1β was not detected in all the sera of newborn calves, the concentration transiently increased with peak titers on day 3 and became undetectable by day 14 after birth. Newborns that showed serum IL-1β on day 3 had been fed on colostrum in which the IL-1β concentration was significantly higher than that in colostrum that had been fed to newborns having no detectable IL-1β on day 3. These results indicate that IL-1β is induced in association with pregnancy in healthy dairy cattle and that the cytokine might be transferred to neonates via colostrum.
Changes of endotoxin in plasma, and the response of the coagulation system and blood cells in septicemia of Haemophilus parasuis infection were examined by inoculation with H. parasuis in specific pathogen-free (SPF) pigs. Eight pigs were inoculated intratracheally with 105, 106 and 107 colony formation units (CFU) of the strain Nagasaki (serovar 5). All pigs died 28 to 42 hr after inoculation. Haematologically, severe leukopenia occurred 24 hr post inoculation (hpi) until death. Glucose concentration decreased from 24 hpi to death. In the coagulation system, decrease of platelet counts, prolongation of prothrombin time, activated partial thromboplastin time, and increase of fibrinogen-fibrin degradation products were observed in all inoculated pigs. Endotoxin was detected in the plasma of all the inoculated pigs from 16 hpi to death, and its concentration rose dramatically just before death. H. parasuis was re-isolated from the blood of all inoculated pigs from 16 hpi to death, and also from almost all organs and body fluids of the pigs. The pigs had microthrombi in the kidney, liver and lungs, and many also had pneumonia, meningitis and serositis. H. parasuis antigen was detected in the lesions by the immunoperoxidase technique. The results indicated that disseminated intravascular coagulation (DIC) and endotoxin shock involved aggravation of clinical signs and death on the pigs induced to septicemia of H. parasuis.
Values of blood gas, serum chloride, and potassium were tabulated for 21 dairy cows with coliform mastitis. Severe cases showed marked clinical signs such as loss of appetite and depression of digestive tract motility, and metabolic alkalosis such as an increase in blood pH, hypochloremia and hypokalemia compared with normal and mild cases (p<0.01). The results showed that metabolic alkalosis can be detected more easily than acidosis in cases of severe coliform mastitis.
For clarifying the relation between the amount of daily urinary creatinine (Cr) and the body weight (BW), fifty-eight healthy castrated male Landrace pigs ranging from 28.6 kg to 93.5 kg in body weight (BW) were examined. There was a significant positive correlation between the BW (X=kg) and Cr (Y=mg/day), and a linear regression formula of Y=40.7X-224.9 (R2=0.985, P<0.001) was obtained. The coefficient of variation of the daily urinary Cr was small (5.1 ± 2.4%). On the other hand, in the relation between the BW and quantity of urine in 24 hr, and in the relation between the daily urinary creatinine and daily urinary volume, no significant correlation was recognized (P>0.2), respectively.
Esherichia coli endotoxin was administered intravenously to 7 Holstein adult cows, to evaluate the effect of endotoxin on acid-base balance. Endotoxin shock was observed immediately after the administration of endotoxin. A loss of appetite and depression of digestive tract motility continued for about 120 hr after the challenge. Metabolic alkalosis following hypochloremia and hypokalemia were particularly pronounced at 12 to 72 hr after the administration of endotoxin.
Cosmid clone containing swine endothelin-1 (EDN1) gene, cosEDN1, was isolated from swine cosmid library using swine EDN1 cDNA as a probe. The sequence analysis of cosEDN1 DNA revealed that the swine EDN1 gene consists of 5 exons, spanning approximately 6.5 kb. In the 5'-upstream region of the EDN1 gene, AP-1 and NF-1 elements were found, suggesting the possibility that the expression of swine EDN1 gene is controlled by protooncogene products Fos and Jun, and TGF-β. Fluorescence in situ hybridization (FISH) using cosEDN1 DNA as a probe demonstrated that EDN1 gene resides on swine chromosome 7p13->pter.
We attempted to establish a system to generate monoclonal antibodies (mAbs) recognizing CD antigens on lymphocytes of domestic animals using the expressed CD antigens by a baculovirus expression system. For this purpose, we selected feline CD4 (fCD4) antigen and expressed it in an insect cell line (Sf9 cells). To obtain mAbs, BALB/c mice were immunized with feline peripheral blood mononuclear cells (fPBMCs) or Sf9 cells expressing the fCD4 (Sf-fCD4 cells), and then hybridomas secreting antibodies were screened by indirect immunofluorescence assay against the opposite antigens of Sf-fCD4 cells or fPBMCs, respectively. Five mAbs recognizing the fCD4 were obtained in total. The system established here might be useful to obtain mAbs recognizing CD antigens of domestic animals.
Babesia caballi infected erythrocytes were collected from the blood of an experimentally infected horse and could be continuously cultivated in vitro with parasitemia ranging from 2-4% in RPMI 1640 medium supplemented with 2 mM L-glutamine, 20 mM HEPES and 40% adult horse serum in a low oxygen atmosphere (2% O2, 5% CO2 and 93% N2). All attempts to increase parasitemia failed using other culture media, serum concentrations and culture vessels. However, parasite growth was enhanced by transfer of cultures from a low oxygen to 5% CO2 in air, with parasitemia ranging from 8-10%.
Immunoglobulin G (IgG) of Indian soft-furred rat, Millardia meltada, was purified by an immunoaffinity chromatography and antibodies against it was raised in rabbit. Using this rabbit anti-M. meltada IgG antibody, sensitivity of enzyme-linked immunosorbent assay (ELISA) to measure parasite-specific antibodies in the sera of M. meltada was markedly enhanced than the previous method using rabbit anti-mouse IgG and rabbit anti-rat IgG antibodies, which could cross-react to M. meltada IgG. Since M. meltada could effectively produce circulating antibodies against two intestinal helminths, Strongyloides venezuelensis and Nippostrongylus brasiliensis, the high susceptibility of this animal to an array of parasites seems to be not due to general immunological deficiency.
The fiber composition of the superior laryngeal nerve (SLN) which is served as the laryngeal afferent pathway was clarified in rats and guinea pigs. The proportions of the number of myelinated and unmyelinated fibers in the SLN were almost the same in both rats and guinea pigs. The unmyelinated fibers show the peak distribution of axon diameter ranging from 0.79 to 1.00 μm in both species, whereas the peak distribution of myelinated fibers ranged from 2.51 to 3.16 μm in rats and from 3.98 to 5.01 μm in guinea pigs. The mean axon diameter of unmyelinated fibers was significantly larger in rats (mean: 1.12 μm) than in guinea pigs (0.96 μm), whereas that of myelinated fibers was significantly larger in guinea pigs (4.04 μm) than in rats (3.30 μm). Such findings would reflect the cardiopulmonary reflexes elicited from the larynx in these animal species.
Canine rabies remains a serious public health problem in Thailand. The Queen Saovabha Memorial Institute (QSMI) is the principal rabies diagnostic center in central Thailand. The retrospective study of canine rabies cases submitted in 1992-1995 revealed that: (1) The prevalence of rabid dogs decreased, which was associated with an overall decrease in the number of animals examined. However, the proportion of FA positive dogs examined remains the same at approximately 50%. (2) The majority of rabies cases occurred in domestic dogs rather than stray dogs but the ratio of positive cases between domestic and stray animals remained the same. (3) More than 60% of domestic rabid dogs were below age one. Dogs at this age are thought to be more active and also most likely not adequately vaccinated. It should be noted that approximately 70% of rabid dogs were never vaccinated against rabies.
LH release in response to pulsatile administration of small amounts of GnRH analogue in cows with follicular cysts was examined. The pulsatile administration of GnRH analogue induced a LH-surge like peak over 10 hr in both normal cows and cows with follicular cysts. The mean peak value of LH in follicular cystic cows did not differ significantly from that of normal cows. All the cows with cysts resumed normal estrous cycles with ovulations within 3 weeks of this treatment. These results suggest that the function of the anterior pituitary for LH release in response to GnRH analogue is not abnormal in cows with follicular cysts, and that cystic cows recover to normal conditions after the pulsatile administration of GnRH analogue.
The method of the poly A-linked colorimetric reverse transcriptase assay (PAC-RTA) was developed and evaluated for the measurement of Mg2+-dependent reverse transcriptase (RT) activity of feline immunodeficiency virus (FIV). PAC-RTA was first evaluated for the detection of RT activity in the culture supernatant of FIV Petaluma strain. The detection limit of RT activity by PAC-RTA was about 10-fold better than that by the conventional non-radioisotopic RT assay kit. Then, PAC-RTA was evaluated for the indication of FIV isolation from cats naturally infected with FIV. FIV was isolated from peripheral blood mononuclear cells of 9 FIV-seropositive cats. The time course appearance of RT activity measured by PAC-RTA corresponded with the analysis of FIV antigen expression by indirect immunofluorescence. Finally, PAC-RTA evaluated the drug susceptibility of FIV. MYA-1 cells (feline T-lymphoblastoid cells) were infected with FIV and were cultured in the presence of various concentrations of anti-human immunodeficiency virus agents such as azidothymidine (AZT) or dextran sulfate. An inverse relationship between the RT activities and the concentrations of these agents in the culture supernatant was confirmed by PAC-RTA. PAC-RTA is easy to perform without using radioactive materials, and one plate can handle 96 samples at one time. By monitoring the RT activity, this assay is a useful method for FIV studies such as viral replication and drug susceptibility.