Visual evoked potentials (VEPs) were recorded in 10 adult male guinea pigs with brain lesion. Lesions were produced in 5 animals by superficial suction of the occipital lobe. The other 5 animals were orally administered with hexachlorophene (about 35 mg/kg/day) for 28 days. In the VEP following the ablation of the occipital lobe, the peaks P₁₀, N₂₀, P₅₅, N₇₅, N₁₄₀ and P₂₀₀ disappeared in many cases. The amplitude of the peak N₄₀ decreased to approximately one half its control VEP. In the VEP obtained from the animals administered with hexachlorophene, the peak latencies of N₂₀, P₃₀, P₅₅, N₇₅ and P₁₀₀ were slightly prolonged after the 7th day following the first administration. On the other hand, there was no change in the latency of N₄₀ during the whole period of administration. The peak-to-peak amplitude showed some variability in different peaks. Histologically, diffuse status spongiosis were found in the white matter of the cerebrum, cerebellum, and brain stem. As described above, the ablation of the occipitallobe caused markedly depressed VEPs, however, the responses to the photic stimulation persisted after the injury. On the other hand, the VEPs of animals administered with hexachlorophene showed a high probability of peak appearance, and a decrease in amplitude was not marked.

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The fecal Escherichia coli isolated from wild Japanese serows living in mountainous areas away from humans and those from captive serows kept in human areas were examined for antimicrobial resistance and the possession of transferable R plasmids. Of 874 E. coli strains isolated from 283 wild serows in 1980-1981, only 11 (1.3%) were resistant to at least one of 6 antimicrobial drugs; ampicillin, streptomycin, tetracycline, chloramphenicol, kanamycin and sulfadimethoxin. Seven (2.5%) individuals were found to carry resistant E. coli. To heighten the isolation frequency of drug-resistant strains, fecal samples of 244 wild serows in 1983-1984 were cultured directly onto drug-supplemented media. Only 12 (4.9%) serows were shown to have drug-resistant E. coli. No transferable R plasmid was detected among a total of 87 resistant strains from wild serows. In contrast, all 33 captive serows except one which was kept only one day after capture, showed resistant E. coli and 20 (60.6%) serows were excreting R plasmid-carrying E. coli. Of 161 drug-resistant strains from captive serows, 50 (31.1%) were found to carry R plasmids. Wild serows seemed to readily change to harbor resistant E. coli almost as soon they were reared in human areas without direct exposure to drugs. These results lead to the conclusion that drug-resistant E. coli can probably be used as microbial indicator for natural environmental pollution.

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We have studied the effects of in vivo asbestos exposure on the surface immune-associated (Ia) antigen expression and distribution of alveolar macrophage subpopulations defined by continuous iso-osmotic Percoll gradients (density range: 1, 006 to 1.123 g/ml) using a rat model of asbestos inhalation. Two groups of rats were exposed by intermittent inhalation (6 hr/day for 5 days/week over 4 weeks) to either amphibole (crocidolite) or serpentine (chrysotile) asbestos. A group of control rats was sham-exposed to clean air only. Alveolar macrophages from rats of three groups were obtained by bronchoalveolar lavage. During exposure, distinct differences appeared within 7 days of asbestos exposure, and some of these findings persisted in the crocidolite-exposed group for as long as 2 to 5 months after the cessation of exposure. Furthermore, relatively greater proportions of Ia-antigen positive cells were detected in several density fractions obtained from both asbestos-exposed groups (especially the crocidolite-exposed group). Multinucleated alveolar macrophages were seen frequently in all Percoll fractions after both types of asbestos inhalation. A significant proportion of multinucleated alveolar macrophages in these fractions expressed surface Ia-antigen positivity. The finding of enriched numbers of higher-density phagocytes in bronchoalveolar lavage cell subpopulations from asbestos-exposed rats may reflect the presence of newly recruited-immature monocytes and/or macrophages at sites of intrapulmonary asbestos deposition. Also, increased proportions of Ia-antigen positive cells suggest that a part of them were functionally activated.

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A Japanese black calf with cyanosis, tachycardia, tachypnea and systolic murmur died of hypoxemia and cardiac insufficiency on the 38th day after birth. We could not establish the diagnosis during it's life. However, anatomically corrected malposition (ACM) with ventricular septal defect was confirmed at autopsy. There was situs solitus of the viscera and atria with atrio-ventricular discordance and ventriculo-arterial concordance. The ventricles demonstrated l-loop, i. e. on the right-sided ventricle there was a markedly enlarged morphologic left ventricle, and on the left-sided ventricle there was a hypoplastic morphologic right ventricle with a stenotic tricuspid valve and Ebstein-like deformity. The right posterior aorta originated from the left ventricle. The pulmonary artery arose from the left-sided right ventricle via infundibulum. There was a fibrous continuity between the aortic and mitral valve. We considered that this is the first reported case of bovine ACM.

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The pathogenesis associated with the 36 Megadalton (Md) plasmid of Salmonella choleraesuis subsp. choleraesuis serovar Enteritidis (S. Enteritidis) was assessed by using the plasmid-containing strain AL1190, plasmid-cured strain AL1192, and plasmid-reintroduced strain AL1193. After oral inoculation of strain AL1190 or AL1192 to C57BL/6 mice, mesenteric lymph nodes, spleens, and livers were examined for the numbers of viable bacteria and for the histopathological changes. The numbers of bacteria were greater and histopathological changes were severer in these organs of mice inoculated with strain AL1190, than in those with strain AL1192. Strains AL1190, AL1192, and AL1193 showed the equivalent survival rate to sera of guinea pig, calf, and pig, and grew equally well under the iron-limiting conditions. These results suggested that the 36 Md plasmid of S. Enteritidis contribute to the spread of the infection beyond small intestines to mesenteric lymph nodes, spleens, and livers, but not to serum resistance or acquisition of iron.

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Hemocytes from adult and juvenile specimens of a brackish-water clam, Corbicula japonica, were attracted chemotactically to live cells of Vibrio parahaemolyticus and Escherichia coli strains in a balanced salt solution, which was enhanced significantly in the presence of respective C. japonica plasma. Chemotactic attractions of adult's and juvenile's hemocytes were seen also in artificial seawater at a similar level to those in the balanced salt solution. Chemotactic attractions of juvenile's hemocytes to these strains were lower in level than those of adult's hemocytes. C. japonica plasma seems to facilitate for C. japonica hemocytes to recognize these organisms.

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Juxtaglomerular (JG) cells on the acute phase in two-kidney Goldblatt hypertensive (2KGH) rats and spontaneously hypertensive rats (SHR) were examined immunohistochemically. JG cells in 2KGH rats and SHR were positively stained with anti-renin serum and anti-angiotensin II (A II) serum. In 2KGH rats, the number of renin and A II immunoreactive JG cells in the clipped kidneys increased throughout the observation period. The number of renin and A II immunoreactive JG cells in the unclipped kidneys was almost the same as that in control rats, although immunoreactivity of these cells was weak and they were small in size. These changes in the unclipped kidneys became obvious with the time course after operation. We did not see any changes in these cells in SHR. In 2KGH rats treated with captopril, the number of renin immunoreactive JG cells in the clipped kidneys increased, whereas that of A II immunoreactive JG cells in the bilateral kidney decreased. When captopril was administered to SHR, the number of renin immunoreactive JG cells in the bilateral kidney increased, whereas that of A II immunoreactive JG cells in the bilateral kidney decreased. These results suggested that the JG cell in the bilateral kidney was closely related to the development of hypertension in 2KGH rats, but not in SHR. The increase of renin immunoreactive JG cells in 2KGH rats and SHR treated with captopril was probably due to the removal of negative feedback inhibition of A II on JG cells.

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By immunization of BALB/c mice with a feline T lymphoblastoid cell line, MYA-1 cells, two types of lymphocyte-specific monoclonal antibodies (mAbs) were obtained. The 220/205/190 kd protein defined by 2F11 mAb is highly expressed on the surface of MYA-1 cells and another feline T lymphoma cell line, FL74 cells. The protein is also expressed on normal feline thymocytes, splenocytes and feline peripheral blood mononuclear cells (PBMCs). Another mAb, 17B10, caused similar results as those of 2F11 except for its low reactivity with FL74 cells. The second type of mAb, 15B3, defined the 220 kd protein. The reactivities of this mAb with MYA-1 cells, FL74 cells, PBMCs and feline splenocytes were lower than the former two mAbs, and did not react to feline thymocytes. On the other hand, 17B10 and 15B3 defined partial populations of MYA-1 and FL74 cells recognized by 2F11. The cells defined by the 2F11 and 17B10 are all leukocytes in spleen and lymph node. In contrast, 15B3 defined most of the cells in B cell area and partially in T cell area. These results suggested that 2F11 and 17B10 recognized the specific antigen of 220/205/190 kd of the leukocyte-common antigen (L-CA) family, CD45R, with different epitopes, and that 15B3 defined the distinct antigen of 220 kd on CD45R.

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Three neonatal calves ranging in age from 4 to 14 days were examined pathologically and bacteriologically. The calves showed depression, anorexia, pyrexia, and difficulty or inability to stand followed by cloudiness of the ocular aqueous humor of cornea. Autopsy revealed congestion, petechiae, and cloudy areas in the meninges. Histologically, the central nervous system (CNS) lesions were prominent and limited to the meninges where fibrinous exudate and infiltrations of neutrophils, macrophages, and lymphocytes were present. There were mild or slight degrees of choroid plexitis and ependymitis. Endophthalmitis was seen as a concurrent lesion in all cases. Fibrinous or fibrinopurulent changes were found in the peritoneum and epicardium as well as in several other organs. Numerous Gram-positive cocci were detected in affected areas of the whole body. Bacteriologically, Streptococcus bovis was isolated from all examined materials consisting of the brain, cerebrospinal fluid, ocular aqueous humor, and several other organs. These results suggest that the lesions were associated with infection of the organism and that the present cases were in the process of septicemia.

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Natural cytotoxicity against bovine leukemia cells (PC-3 cells) was found in bovine peripheral blood lymphocytes (PBL), and in non-adherent cells but not in adherent cells to nylon-wool column. Natural cytotoxic cells (NCC), which have natural cytotoxic activity, are found in T cell-rich fraction. When NCC were cocultured with PC-3 cells, natural cytotoxic factor (NCF) was released rapidly from NCC, and dose-response curve for NCF was almost linear induction. Cytotoxicity against PC-3 cells by NCC or NCF was increased with an increment of incubation period. Cytotoxicity against K562 cells, CL-1 cells, M1 cells or EL-4 cells by NCF was almost the same level as that against PC-3 cells, but that against those cell lines by NCC was not found. NCF activity in culture fluid from NCC cocultured with K562 cells or CL-1 cells was lower than that from NCC cocultured with PC-3 cells.

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Effects of dehydration on vasopressin-secreting cells (VP cells) of the supraoptic nucleus (SON) and the paraventricular nucleus (PVN) in the Mongolian gerbils were studied immunocytochemically and morphometrically. The plasma osmolality was measured at the time of sacrifice of individual animals and the body weight was measured every day during dehydration. The plasma osmolality increased significantly on day 3 of dehydration, followed by a gradual increase to reach nearly its equibilium state on day 10. The body weight decreased rapidly until day 10, followed by a gradual decrease thereafter. The area of VP cells increased significantly in both the SON and PVN on day 1 of dehydration, the level being nearly the same until days 3 to 5 and going up on day 7 to reach the plateau after day 15. These findings seem to reflect a compensation mechanism between the volume of body fluid and the plasma osmolality and to reflect responses of VP cells to the osmotic stimuli. Electron microscopic observation revealed that, at the beginning and late stages of dehydration, the increase in the area of VP cells was in parallel with the expansion of the Golgi area and with the distension of cisternae of the rough endoplasmic reticulum.

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Rats and guinea pigs were experimentally infected by intradermal and intraperitoneal inoculation of T. suis. These animals were observed for pathologic changes at various stages. Macroscopic observation disclosed visible grayish white soft nodules at the injection site in those inoculated intradermally and many grayish white nodules in the peritoneum of those inoculated intraperitoneally. Histopathological examination revealed the presence of characteristic microbial elements in the centre of the lesions. Various stages of the lesions were recognized; the early stage being abscess, the advanced stage being pus-forming granuloma and the final stage being residual granuloma. The microbial elements were composed of spores, thalli and multilocular tubers. Clubs were formed around microbial elements in rats inoculated intradermally and in guinea pigs inoculated intraperitoneally. The characteristic T. suis lesions in pigs were reproduced by experimental infection of this organisms in rats and guinea pigs, and thus the pathogenicity of T. suis was confirmed.

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To examine effects of thromboemboli due to dead worms on pulmonary arterial pressure (PAP), 20 to 50 dead heartworms were inserted into the pulmonary arteries of 4 heartworm uninfected dogs (uninfected group) and 11 dogs infected with heartworms (infected group). In the uninfected group, the mean PAP rose 1 week after worm insertion (10.9 to 166. mmHg), but it recovered by the 4th week. Clinical signs, hemodynamics and blood gas findings also deteriorated at the 1st week, but recovered at the 4th week. Angiographic and pathological findings indicated that blood flow recovered through the spaces between thromboemboli and vessel walls at the 4th week. The infected dogs were divided into three groups. In the infected-I group (5 dogs), the intimal lesions of the pulmonary arteries were slight, and clinical and laboratory findings showed changes similar to those of the uninfected group. In the infected-II group (4 dogs), the pulmonary arteriallesions were severe and the mean PAP was higher (25.7 mmHg) than in the uninfected group before worm insertion. An increase in PAP (34.1 mmHg) and worsening of clinical and laboratory findings were noticed till the 4th week. Thromboemboli adhered extensively to the vessel walls. Two dogs in the infected-III group died of severe dyspnea on the 9th and 10th day, and the mean PAP rose remarkably at the 1st week (from 19.4 to 28.2 mmHg). Severe pulmonary parenchymal lesions with edema or perforation were observed. From the above results, it was clarified that effects of dead worms on PAP and clinical signs depended on the severity of pulmonary arterial lesions before worm insertion. These results may explain the clinical diversity in spontaneous canine heartworm disease.

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The complete nucleotide sequence of Clostridium botulinum type D strain CB16 neurotoxin was determined and the deduced amino acid sequence is reported here for the first time. The structure and function of botulinum type D neurotoxin is discussed from a molecular biological viewpoint. DNA was extracted from toxin-converting phage d-16φ of C. botulinum type D strain CB16; and a fragment (about 10 kbp) coding for the neurotoxin was cloned into Escherichia coli using λgt11. A 21-mer oligonudleotide which corresponds to Phe⁷ to Val¹³ of the partial amino acid sequence near the N-terminus of the type D neurotoxin was synthesized and used as a probe to identify the gene encoding type D neurotoxin. The nucleotide sequence contained a single open reading frame coding for 1, 275 amino acids (molecular weight of 146, 785) and the deduced amino acid sequence corresponded exactly to the partial amino acid sequences determined by direct microsequencing of the neurotoxin fragments. In the dichain molecule of the neurotoxin, Thr² and Asn⁴⁴³ formed the N-termini of the light chain (M. W. 50, 410) and heavy chain (M. W. 96, 394) respectively, and these two chains were linked with a disulfide bond between Cys⁴³⁷ on the light chain and Cys⁴⁵⁰ on the heavy chain. The nucleotide sequence of the D-CB16 neurotoxin differed from that previously reported for type D neurotoxin by three nucleotides.

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Physiological significance of body temperature responses to ambient heat (BTRAH) with high overshoot or a high equilibrium phase was studied in relation to thermoregulatory ability. Subsequently, an attempt was made to predict survival time (ST) by measuring the period required to attain a colonic temperature (Tco) of 42.0°C(t42.0), whereas ST hitherto having been calculated on the basis of a period until Tco attains 42.5°C(t42.5). Tco of rat was monitored continuously with a Cu-Co thermocouple during exposure to an ambient temperature (Ta) of 42.5°C. The lower the overshoot temperature (Tos) or the equilibrium temperature (Teq), the longer the rats survived in a hot environment. The present findings further suggest that these two types of BTRAH are immature variations of the typical triphasic BTRAH which is characteristic of heat resistant individuals. A new regression line of ST (Y) as a function of the t42.0 (X) was obtained for most rats as follows: Y=0.963X+43.85 (male); and Y=0.973X+39.10 (female). This equation enabled to calculate ST without thermal death. However, the former approach based on t42.5 must be applied yet in small number of rats which showed the irregular BTRAHs with Tos or Teq higher than 42.0°C. Delayed influence on survival and fecundity at a Ta of 24°C were not found during one year following this hyperthermia up to 42.5°C.

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From the end of September to November 1988, a compact scale of bovine ephemeral fever (BEF) outbreaks occurred suddenly in Tanegashima island of Kagoshima Prefecture, southern part of Kyusyu island of Japan. The BEF outbreak pattern showed epidemical characteristics as follows; (1) outbreak spread from few foci to zone during one month, and (2) the disease might be transmitted in farms with a fixed probability of adequate contact. By using the above aspects, we attempted to analyze the disease theoretically with the application of Poisson distribution and Reed-Frost model. The BEF incidence in farms was in well accord with the Poisson distribution. As the very rare event occurred in unit time or in unit area in this epidemic, the cattle population at risk were equivalently susceptible to BEF virus in this island, due to the influence of no vaccination to BEF control before the first outbreak. Similarly, the epidemic curve of the Reed-Frost model was proved to fit well the incidence observed in a farm, and the probability of adequate contact was induced as p=0.226. If the cattle population is less than 5 in this farm, the outbreak would not occur in the first instance.

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The lysate of Theileria sergenti piroplasms was tested for proteinases using sodium dodecyl sulfate-polyacrylamide gel electrophoresis in which substrate was included in gel matrix. Six proteinases of molecular weight 330, 125, 98, 94, 67 and 58 kilodalton (kDa) were detected. From the results of the Triton X-114 phase partition, 330, 125 and 58 kDa proteinases were partitioned into aqueous phase, which indicated that they were not associated with parasite membranes. All these three enzymes were classified into metalloproteinase family because of their sensitivities to metal-ion chelating compounds, ethylenediaminetetraacetic acid (EDTA) and 1, 10-phenanthroline. On the other hand, 98 and 94 kDa proteinases were membrane-associated metalloproteinases which were preferentially inhibited by 1, 10-phenanthroline. Another metalloproteinase of 67 kDa which was inhibited by EDTA and 1, 10-phenanthroline was not associated with parasite membranes. Proteinases of 98 and 94 kDa degraded heat-denatured hemoglobin.

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Microvascular changes in the mammary gland in mice during pregnancy and lactation were investigated by scanning electron microscopy (SEM) with a corrosion cast method, transmission electron microscopy (TEM) and morphometry. By SEM, duct-associated capillary plexuses were sparsely distributed to branch into adipocytes during virgin period. With advance in pregnancy, both branches from the capillary plexuses and branches from the vessels surrounding adipocytes extended further to form capillary networks. The basket-like architecture was completed by day 18 of pregnancy. These findings may indicate that angiogenesis occurs frequently during this period. During lactation, the basket-like architecture still remained and the capillaries surrounding alveoli meandered. After weaning, the regression of microvasculature followed the degeneration of alveoli. By TEM and morphometry, the density of binocytotic vesicles (PVs) (number of PVs per μm² of endothelium cytoplasm) increased twofold from day 18 of pregnancy to day 5 of lactation, furthermore increased threefold from days 10 to 20 of lactation, and subsequently decreased after weaning. Marginal folds and microvillous processes gradually increased in length with advance in pregnancy, reached the maximum from days 5 to 15 of lactation, and thereafter decreased. In addition, the capillaries with thinner walls were in close contact with alveoli during the late stage of pregnancy and during lactation. Furthermore, the alveolar epithelial cells had well-developed basal infoldings during lactation. These findings suggest that the capillaries play an important role in transporting materials necessary for milk production.

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To clarify the immunologic effects of vaccination in dogs, we monitored total leukocyte and lymphocyte counts, humoral antibody responses, blastogenic responses of lymphocyte, and delayed type hypersensitivity (DTH) responses after vaccination. Mixed vaccines were administered on day 0 except for canine parvovirus (CPV) vaccine which was readministered on day 21. The puppy and adult dogs had a significant decrease in leukocyte and lymphocyte counts on day 7. The puppies showed a significant increase in the blastogenesis of lymphocytes after each vaccination, whereas the adult dogs had no significant changes. However, the adult dogs were divided into two groups, high responders and low responders in blastogenesis of lymphocytes. The dogs with higher or lower response in SI values on day 0 tended to show decrease or increase after the first vaccination, respectively. Since almost all dogs developed high titers of humoral antibody, it is considered that vaccination acts in an immunomodulative fashion. DTH responses to phytohemagglutinin (PHA) and CPV vaccine monitored at 0, 3, and 8 weeks after the first vaccination produced strong reactions, in particular those to CPV vaccine rose significantly after vaccination and maintained the higher responses for at least 2 months. These results suggest that DTH responses to PHA and CPV vaccine are helpful to monitoring non-specific and specific immune functions in vivo, therefore, DTH could be used as simple and rapid immunologic tests in canine practice.

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Sera of dogs with gentamicin-induced uremia were analyzed by high performance liquid chromatography system with strongly basic macroreticular anion exchange resin. Satisfactory separation of peaks was achieved with good reproducibility after deproteinization of sera with trichloroacetic acid at a final concentration of 3%, confirming that the system was suitable for qualitative analysis of uremic serum. The chromatograms showed that the number of peaks and the peak area had relation to concentrations of serum urea nitrogen or creatinine and severity of uremia. Four peaks were selected as suspected canine uremic peaks with high correlation to serum creatinine concentrations which were hardly influenced by extrarenal factors. The results suggested that these four fractions might contain uremic toxins.

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Field trials were carried out in calves using a live bovine respiratory syncytial (BRS) virus vaccine prepared from the attenuated BRS virus, strain rs-52. Two hundred seventy-five and 353 calves were vaccinated intranasally and intramuscularly, respectively. No undesirable postvaccinal reactions were observed in the vaccinated calves. Of the serum neutralizing (SN) antibody negative calves 89.7% (26/29) and 92.8% (90/97) developed SN antibody 1 month after intranasal and intramuscular vaccination, respectively. Most of the calves having SN antibody titers of 1:1 or 1:2 at the time of vaccination showed a significant increase in SN antibody titer. About 70% and 90% of the calves vaccinated intranasally and intramuscularly, respectively, maintained SN antibody for 6 months after vaccination. In a field trial, a natural BRS virus infection occurred about 5 months after the start of the trial. Ten of the 16 unvaccinated control calves showed respiratory symptoms due to BRS virus infection. On the contrary, all of the 68 vaccinated calves exhibited no symptoms at all, indicating efficacy of the vaccine.

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During a virus survey carried out in the period 1989-90 with 148 fecal samples collected from cats in Japan, three reovirus strains were isolated in feline cell cultures. Two strains (Nos. 114 and 140) were from 48 diarrheal fecal samples and another strain (No. 32/41) was from 100 normal fecal samples. The strains grew in feline and simian cell cultures with producing typical intracytoplasmic inclusion bodies in which virus particles were densely packed. All strains, especially Nos. 32/41 and 140 strains, showed trypsin-dependent growth in vitro. Their ultrastructural and genomic properties were characteristic of genus reovirus in the Reoviridae. All strains agglutinated erythrocytes of human type O but not of bovine. Although they were identified as serotype 2 by hemagglutination-inhibition test with the hyperimmune sera against human reovirus prototype strains, No. 114 strain was typical and the other two strains were atypical serotype 2 reoviruses. Furthermore, from the reason that Nos. 32/41 and 140 strains possessed some common properties though derived from cats in distant locations, they were considered to be reoviruses having been maintained in the cat population. Seroepizootiologic survey revealed that the prevalence of serotype 3 infection was most widespread and serotype 2 was least among three serotypes of reovirus in a cat population.

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In 100 dogs autopsied, glomerular IgA deposition was examined by the immunofluorescence technique and the histopathological features of glomeruli with IgA deposition were examined by light and electron microscopy. The incidence of the IgA deposition was age-related but there were no sex and breed predisposition. Deposition of IgA was observed mainly in mesangial areas in approximately a half (47%) of dogs examined. IgG, IgM and C3 often co-deposited. Histopathology of the glomeruli with IgA deposition indicated increase of mesangial cells, crescent formation, hemispherical deposits in paramesangial areas and glomerular sclerosis. Ultrastructurally electron dense substances positive for IgA deposited in mesangial and paramesangial areas. The examination to know the relation between the severity of IgA deposition and the number of mesangial cells or percent of the cells to total glomerular cells indicated that mesangial cells increased at the early stage of the disease and subsequently epithelial and endothelial cells proliferated as the increasing amount of IgA. Dogs suffering from enteritis or liver diseases showed high incidence of glomerular IgA deposition.

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To study the roles of conglutinin (Kg), mannan-binding protein (MBP), and serum amyloid P component (SAP) in the protection of cattle against infections, the concentrations of these proteins in the sera from cows with mastitis were determined by sandwich enzyme-linked immunosorbent assays. The Kg and MBP concentrations in the sera from cows with mastitis were much lower than those from uninfected (or clinically healthy) ones, and increased after recovery. No significant difference in the SAP concentrations was found between healthy and infected cows. With the sera from cows experimentally infected with bovine leukemia virus, the Kg concentrations were also lower than those from uninfected ones, whereas the MBP concentrations were not. From these findings, the Kg concentration is suggested to be serologically a possible indicator for clinical diagnosis of treatment for mastitis although both Kg and MBP concentrations were found to decrease in cows with mastitis and to increase after recovery.

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The present study was designed to investigate baroreflex control capacity of arterial pressure (AP) in the conscious Watanabe heritable hyperlipidemic (WHHL) rabbit. The control capacity of the baroreflex system was assessed with overall open-loop gain (G). Seven WHHL and 14 normal Japanese white rabbits were chronically implanted two catheters in the aortic arch through the left subclavian and common carotid arteries. A small amount of blood (2 ml/kg, body weight) was rapidly extracted into a syringe via the left common carotid artery in the conscious state. Mean arterial pressure (MAP) was monitored with a catheter-transducer system through the left subclavian artery. The MAP responses to the rapid hemorrhage were averaged 8 times by a computer. G was calculated as G=ΔAP_I/ΔAP_S-1, where ΔAP_I was an immediate MAP fall after the hemorrhage and ΔAP_s was a steady-state error 1-2 min after the hemorrhage. The values of G in the conscious normal and WHHL rabbits were 7.35±0.24 and 1.91±0.29 (mean ± SE, p<0.01), respectively. To investigate effects of pentobarbital anesthesia on baroreflex system, the hemorrhage experiment was repeated several times under pentobarbital anesthesia (20 mg/kg, i.v.). The values of G in the anesthetized normal and WHHL rabbits were 6.69±0.23 and 1.68±0.34 (mean ± SE, p<0.01), respectively. G in the normal and WHHL rabbits did not show any significant change in the presence and absence of pentobarbital anesthesia (p>0.05). It is concluded that the reduced baroreflex control capacity of AP was partly caused by decreased sensitivity of the baroreceptors due to spread of sclerotic lesions and that pentobarbital anesthesia did not affect overall AP control capacity of the baroreflex system.

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Vagal afferent activities and respiratory reflexes during drug-induced bronchoconstriction were studied in 31 anesthetized, spontaneously breathing or artificially ventilated guinea pigs. Histamine (5, 10, 20 μg/kg), ACh (10, 20, 40 μg/kg) and endothelin-1 (2 μg/kg) were intravenously injected to the animals in order to induce the bronchoconstriction. In spontaneously breathing and vagi intact animals, a considerable respiratory change characterized by rapid-shallow breathing was elicited by histamine. Such respiratory change was abolished by bilateral vagotomy, indicating that the vagal pathway fairly participated in the respiratory change during bronchoconstriction. Indeed, recordings of electrical activities of single vagal afferent nerve fibers from pulmonary stretch and irritant receptors elucidated that the bronchoconstriction by the three drugs markedly influenced these receptor activities. The response of stretch receptors to bronchoconstriction was grouped into four types: two of those types showed a marked increase in their activities and the other two a decrease or no change. Such uneven response was assumed to be derived from heterogenous contraction and aeration among the intrapulmonary small airway. On the other hand, irritant receptors were invariably stimulated by increased transmural pressure during bronchoconstriction. Administration of isoprotereno1 (20 μg/kg) which inhibited the smooth muscle contraction abolished stimulatory effect of the drugs to irritant receptors, suggesting that the effect was due to indirect action through the muscle contraction rather than their direct action to the nerve endings.

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A unique morphological cell type, "microfold or membranous (M) cell-like cell", was detected electron-microscopically in the cecal tonsil epithelium of the chicken. M cell-like cells possessed a few short microvilli of irregular arrangement and a large number of lymphocytes and macrophages wedged into their basal surfaces. Triticum vulgaris was found to bind to M cell-like cells. With horseradish peroxidase (HRP) treatment, M cell-like cells showed an active HRP uptake just as did the neighbouring usual absorptive epithelial cells. No uptake of colloidal carbon particles from the intestinal lumen was recognized in any part of the intestinal epithelium. These results suggest that M cell-like cells of the chicken possess some M cell-characteristic morphological and histochemical features, but that their active uptake of foreign materials is not so developed as in mammalian M cells.

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Histochemical activities of several enzymes were investigated in the olfactory epithelium (OE) and vomeronasal organ (VNO) of the golden hamster. Activities of adenosine triphosphatase, lactate dehydrogenase and succinate dehydrogenase were intense in the OE, and the sensory (VSE) and respiratory epithelium (VRE) of the VNO. The activity of acid phosphatase was intense in both the OE and the VSE, while that of non-specific esterase was intense in the VSE alone. The activity of alkaline phosphatase was detectable only in the VRE. Activities of monoamine oxidase and acetylcholine esterase were negative in all of the OE, VSE and VRE. These similarities and differences in the histochemical distribution of enzymes between OE and VSE may reflect the common olfactory function and/or functional specialization in these epithelia. On the other hand, the VRE was considerably different from the OE and VSE in the enzymatic distribution. This may reflect the non-olfactory function of this epithelium.

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Effects of pulsing electromagnetic fields (PEMFs) on ligament healing were investigated using 80 rabbits. All animals received square resection (4 mm×4 mm) of both patellar ligaments in full thickness at their center. They were divided into 4 groups of 20 rabbits each and stimulated with different electromagnetic intensity of 0 (control), 2, 10, and 50 gauss (G) for 6 hr daily. Pulse frequency and pulse width were 10 Hz and 25 μsec, respectively. After PEMFs stimulations for 1, 2, 3 and 4 weeks, 5 animals of each group were euthanized and the regenerated tissue at the defective portion was investigated histologically and biomechanically. Histologically, the tissue stimulated by PEMFs showed an earlier increase in capillaries and fibroblasts and more matured, prominent longitudinal orientation of collagen fibers than those of controlgroups. Among the rabbits stimulated electromagnetitally, those stimulated at 50 G revealed the earliest ligament healing. Tensile strength of regenerated ligament tissues of any PEMFs groups increased significantly at 1 and 2 weeks after operation, however, at 3 and 4 weeks after operation, there were no significant differences between groups. Among these values, those of 50 G group were the highest consistently during most of the experimental period. From the above results, PEMFs enhanced the earlier stage of ligament healings and 50 G gauss seemed to be the most effective among the 3 field intensities used. This promoting effect may potentiate the earlier recovery of the function after the ligament injury.

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Immunohistochemical analysis of constituents of senile plaques and cerebro-vascular amyloid in the brain of aged dogs was performed using antisera against beta protein, cystatin C, ubiquitin, tau, and neurofilament (NF). All types of senile plaques and cerebro-vascular amyloid in aged dogs were labeled by anti-beta protein serum. Cystatin C immunoreactivity was detected in neuronal cell bodies, primitive or classical plaques, and amyloid deposited around cerebral capillaries, but not in diffuse plaques and amyloid deposited in the media tunica of cerebro-meningeal arterioles. Ubiquitin-positive granules distributed widely in both gray and white matter of aged dogs, while they were very small in number in young dogs. Swollen neurites-like materials in primitive plaques or classical plaques were immunoreactive for anti-ubiquitin serum. Tau immunostaining labeled commonly axons and several neuronal or glial cells after hydrate autoclave pretreatment. Tau-positive components were observed very rarely in the corona of classical plaques. Most of swollen neurites-like structures of primitive or classical plaques were not reactive for anti-NF serum, and only a few plaques contained small numbers of NF-positive elements.

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Cytotoxin production by Campylobacter species isolated from proliferative enteropathy in swine was examined. Twenty-one of 29 strains of C. hyointestinalis, 10 of 27 strains of C. mucosalis and 10 of 10 strains of C. coli were cytotoxin positive. By the gelfiltration chromatography of C. hyointestinalis culture filtrate, cytotoxin activities were observed in two peaks (fraction I and fraction II). Most of the cytotoxic activities lay in fraction I, which is heat-labile, trypsin-sensitive and the molecular weight was estimated at 40, 000. On the other hand, fraction II cytotoxin was heat-stable, trypsin-insusceptible and molecular weight was approximately 1, 000.

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The dependency of the secretory function of the feline major vestibular gland (Bartholin's gland) on the ovarian steroid hormones was studied histochemically. After estrogen (estradiol-17β or diethylstilbestrol), progesterone or a combination of these was administered to cats which were previously ovariohysterectomized, the major vestibular glands were removed, embedded in paraffin; sectioned, and stained by alcian blue, periodic acid-Schiff or peroxidase-labeled lectin (peanut agglutinin and wheat germ agglutinin). The vividly positive reactions to the stainings applied were observed in the secretory epithelial cells of the major vestibular glands in the estrogen treated animals. The dependency of the secretory function of the major vestibular glands on the estrogen was demonstrated in this study.

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The plate-in-bottle method of Mitsuoka et al. (1969) for the counting of fecal bacteria in beagle dogs was superior to an anaerobic jar method. Comparisons of three nonselective media, such as medium 10 supplemented with 10% cecal extract of dogs (designated as M10C), M10 with 10% fecal extract of dogs (M10F), and M10, by the plate-in-bottle method indicated that the visible bacterial counts for M10C were higher than those for M10 and M10F. The high percentage (18.4%) of numbers of the extremely oxygen-sensitive anaerobes to the fecal total counts by using the plate-in-bottle method with M10C was also observed.

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The alteration of trypsin resistance in rat sperm head was investigated after the sperm were incubated in the culture media (199-Earele, 1% BSA) containing the inhibitors of protein synthesis. The head of non-iticubated testicular sperm was easily digested by trypsin (1 mg/ml) while the head of the incubated sperm showed resistance to trypsin digestion after 3 hr-incubation. Trypsin resistance in epididymal sperm head did not change after the sperm was incubated. Neither cybloheximide (100 μg/ml) nor chloramphenicol (100μg/ml) affected the trypsin resistance in the testicular sperm head.

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Activities of certain cytoplasmic enzymes were measured in bovine T lymphoma (BTL-PC3 cells). The activities of hexokinase, pyruvate kinase and glucose-6-phosphate dehydrogenase in PC3 cells were elevated as much as 2 or 3-fold of those in bovine thymocyte. The high activities of these enzymes derived from activation of glucose metabolism may reflect the high growth potential of PC3 cells.

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The effect of cobra venom factor (CoVF) treatment was examined to clarify the mechanism of resistance of mice to Clostridium chauvoei infection. In CoVF-treated mice inoculated with spores of C. chauvoei, no death occurred and the organisms in the infected muscle progressively decreased, similar to that of non-treated control mice. These results indicated that C3 did not play a significant role in the resistance of mice against C. chauvoei infection.

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A 2-year-old female Japanese domestic cat weighing 3.6 kg was presented due to marked abdominal distention and was diagnosed as hepatic cyst. The bile acid concentration of the hepatic cyst was as low as the serum bile acid concentration. The membrane of the cysts was excised from the edge of the compressed hepatic tissue and no other surgical manipulation was performed. This surgical procedure is simple and quick compared to hepatic lobectomy, however, it is necessary to evaluate the fluid of the cyst before performing this procedure.

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We found that the oral administration of active egg white product (AEWP) enhances host resistance to infections. The prophylactic administration of AEWP enhanced nonspecific resistance in mice to systemic infection with E. coli. Moreover, the therapeutic administration of AEWP was effective against S. aureus infection in mice.

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The inhibitory effect of sulfamonomethoxine and other sulfonamides on the capsule formation of sulfonamide-resistant Bordetella bronchiseptica was investigated. All the sulfonamides having MeO(-OCH₃) groups inhibited the capsule formation of B. bronchiseptica. Strong inhibition was obtained with sulfamonomethoxine. Inhibition was not seen with sulfonamides having no MeO groups.

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The possibility of estrus prevention in the queen by the oral administration of chlormadinone acetate was examined. The animals used were 29 mature ar1d 15 immature queens. For 16 mature females, 4-12.5 mg was given daily by mouth for 7 days every 3 months. Ten of the 16 queens given this treatment came into estrus within 4 months of the first treatment. For 28 females including the immature, 2-12.5 mg was given once a week throughout the experiment. This treatment prevented estrous activity for at least 1 year. In the queens in this study, the side effects were not observed excepting an increase in body weight during treatment. Our results showed that oral administration of this drug weekly is safe and reliable for long-range prevention of estrus in queens.

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