The mandibles of the Japanese wild pigs (Sus scrofa leucomystax) from various geographical locations, which have been stored in the University of Tokyo and National Science Museum, Tokyo were examined. The mandibles from northern localities were larger in size than those from southern localities. The Oita population was significantly smaller than the Honshu populations. The Mie population was smaller than the Hyogo population in length from the angle of the mandible. In the multivariate analysis, the Kyushu populations could be obviously distinguished from the Honshu populations in the principal component charts. However, each Honshu population did not show the locality-specific distribution of the principal component plots.
Substance P-immunoreactive neurons projecting from the midbrain to the spinal cord of the chicken were examined by the use of the retrograde tract-tracing method combined with immunohistochemical techniques. Many small neurons were densely clustered in the rostral midline area of the midbrain (RMA), and showed substance P-immunoreactivity. These substance P-immunoreactive neurons sent axons to the intermediomedial cell column (avian autonomic preganglion) and its vicinity in the lumbar spinal segments. On the basis of the strong neuroanatomical analogy in the cytoarchitectural features, immunoreactivity, and fiber connections, the RMA was assumed to be the avian homologue of the anteromedian nucleus in the mammalian midbrain.
Water-in-oil emulsions containing bovine serum albumin (BSA) as a model antigen were prepared using aliphatic saturated hydrocarbons with carbon number from 12 to 18, and were tested in chickens. Straight-chain hydrocarbons induced higher antibody titers against BSA after administration than branched-chain hydrocarbons. n-C 16H34 and n-C18H38 maintained high antibody titers even at 32 weeks after administration, compared with n-C12H26, n-C14H30 and n-C15H32. n-C12H26 and n-C14H30 raised concentrations of sialic acid and creatine kinase in plasma, both of which are important markers of inflammatory responses, immediately after administration. n-C 16H34 and n-C18H38 did not raise the values of these markers. These results indicated that n-C16H34 and n-C18H 38 induced elevated and sustained immune responses without severe adverse reactions in chickens.
Since the turbidity of nuclear suspensions is known to be correlated with the nuclear morphology, the effects of long-wave ultraviolet (UVA) radiation on Mg2+-dependent structural transition of chromatin in isolated chicken liver nuclei were monitored by measuring the relative turbidity of nuclear suspensions. UVA radiation of the nuclei inhibited the Mg2+-dependent change in relative turbidity of nuclear suspensions in a UVA dose-dependent manner under aerobic conditions but not under N2 conditions. No inhibitrory effect of UVA radiation on the change in relative turbidity was observed in the presence of 50 mM NaN3, which scavenges singlet oxygen (1O2) and hydroxyl radicals (·OH). In contrast, 100 mM dimethyl sulfoxide, which primarily scavenges ·OH, did not show the inhibitory effect of UVA radiation. The amounts of DNA-protein crosslinks increased with UVA dose under aerobic conditions but not under N2 conditions. The present study showed that UVA radiation of isolated nuclei inhibited the Mg2+-dependent unfolding of condensed chromatin and that 1O2 is likely to be involved in this process. Furthermore, the formation of DNA-protein crosslinks may contribute to the inhibition.
In this study, bovine β2-m was purified from urine by ion-exchange chromatography and gel chromatography, and the characteristics were compared with those of colostral β2-m by the immunological reactivity, isoelectric points, peptide map, and amino acid sequence. The characteristics of purified urinary β2-m were consistent with those of the colostral β2-m. The urinary and colostral β2-m possessed the same polypeptide chain consisting of 98 amino acids, and its molecular weight is 11.8 kDa. Furthermore, four isoforms of β2-m were found. The isoelectric points were different from each other.
The regional variation in the intraepithelial lymphocytes (IELs) in the small intestine was examined in BALB/c male and female mice and C3H/He and C57BL/6 male mice. The small intestines were taken from 11 to 12-week-old mice and divided equally into 3 parts (the proximal, middle and distal parts). IELs were isolated from each part of the intestine and analyzed with flow cytometer. The number of IELs was highest in the proximal part and lowest in the distal part. The distribution of IEL subsets was markedly different between the proximal and the distal parts, and that in the middle part showed the intermediate pattern. The percentage of αβ T cells were higher in the distal part. In αβ T cell subset, the percentage of CD8αα T cells was higher in the proximal part, whereas those of CD4 and CD4CD8αα double positive T cells were higher in the distal part. In γδ T cell subset, no regional variations were found. The regional variations in the number and subsets of IELs showed almost the same patterns between male and female BALB/c mice and similar patterns among three strains of mice. This strongly suggests that the regional variations in the small intestinal IELs are common to mouse species.
To evaluate clinical effects of autogenous toxoid-bacterin treatment for Staphylococcus aureus subclinical mastitis in lactating cows, 22 cows which had at least one S. aureus infected quarter were selected from among cows at a S. aureus prevalent dairy farm. Eleven cows were injected with their own autogenous toxoid-bacterin and the others were maintained as non-injected control. In the toxoid-bacterin injected group, 270f infected quarters were cured during the 12-week trial, compared to 5 0n the control group. New intramammary infections with S. aureus were only detected in 3 quarters of the control group. Mean IgG antibody titer against S. aureus somatic antigens and α-toxin in serum and milk were significantly increased in the toxoid-bacterin injected group (p<0.05) and remained higher than those of the control group which showed no significant changes (p<0.05). In contrast to the control group, from 3 weeks after the second injection of the toxoid-bacterin injected group, mean S. aureus cfu/ml in milk samples from injected quarters with S. aureus was significantly decreased until the end of the study (p<0.05). In the toxoid-bacterin injected group, significant decreases of mean SCC were detected from milk samples from infected quarters with S. aureus from week 7 to week 10 (p<0.05). These data show that autogenous toxoid-bacterin treatment against S. aureus subclinical mastitis in lactating cows may increase the cure rate of the infections, reduce the severity of the infections and also prevent occurrence of the new infections.
A new chemolytic therapy with amino acid preparation was tried for use on struvite renal stones in two dogs, which were admitted to Azabu University Veterinary Teaching Hospital. No other special diet, for the purpose of dissolving the stones, was fed during the treatment. Dissolution of the stones began at week 1, and no stone was detected on radiography at week 4 in both cases. The amino acid preparation thereafter has been administered every 2-3 days to prevent formation of the stones. After 6 months from treatment, complete dissolution was confirmed, and no relapse has been observed. Therefore, it is concluded that chemolytic therapy with amino acid preparation might be useful for the treatment of nephrolithiasis with struvite renal stones in dogs.
With radiography and ultrasound, reversed positioning of the fundus ventriculi and pylorus, a duodenum running on the left side, transposition of the kidneys, and normal thoracic organs were found in a 5-month-old miniature dachshund that presented with anorexia and weight loss. The case was diagnosed as partial heterotaxia. Gross observation revealed partial heterotaxia, polysplenia, abnormal lobulation of the liver, and absence of the greater omentum. These findings were consistent with those observed in asplenia-polysplenia syndrome in humans.
A fungus was isolated from the skin of an Otaria byronia and from the water of the pool in which the animal was kept. It formed creamy colonies with soft texture on Dixon agar and grew well without supplements of long-chain fatty acids. Cells were ovoid to cylindrical in shape, budded from a broad base, and budded and divided at the same location. Thus, the isolate was identified as M. pachydermatis. We compared this very rare isolate from a marine mammal with four strains of M. pachydermatis using the freeze-etching electron-microscopy technique. The cells showed the same characteristic ring-swellings on the protoplasmic membrane on the neck site between the mother and the daughter parts, and the same accumulation of circumvallate bulgings in a small area near the straight sections of spiral grooves as four reference strains. Thus, in terms of morphology and ultrastructure, the isolate could be regarded as a typical M. pachydermatis.
Porcine genomic DNA encoding a 55 kDa subunit of interleukin-2 receptor (IL-2R), which is termed α chain (IL-2R α), was cloned by repeated plaque hybridization using IL-2Rα cDNA as a probe. Two different lambda phage clones, one of which encoded exon 1 and the 5'-upstream flanking region of IL-2Rα gene and another encoded the sequence from exon 2 to exon 8, were isolated. By analysis of the 5'-upstream region of the gene, putative binding motifs for transcription factors such as GATA family proteins, Ikaros, NF-κB, NF-IL2R α and SRF, were found as described in human, murine and bovine genes. Two additional motifs for STAT4 binding were also found in this region. Moreover, using the FISH technique, we assigned the porcine IL-2Rα locus to the distal end of the long arm of chromosome 10 (10q6-qter) where the vimentin gene had been assigned nearby.
In order to identify the alternative effective chemotherapeutic agents for murine babesiosis, some selected drugs were examined for their efficacy against protozoan infection in the mouse-Babesia rodhaini (B. rodhaini) model. Clindamycin was not completely effective for elimination of parasites in a dose of 50 mg or 100 mg/kg BW/day b.i.d. but effective to prolong the life span of hosts, while it completely cured B. rodhaini infections in a dose of 200 mg. On the other hand, a double therapy consisting of 2 treatments with 100 mg clindamycin and 100 mg clindamycin and with 100 mg clindamycin and 100 mg tetracycline; respectively, and a single therapy with 100 mg tetracycline or 200 mg clindamycin, had a possibility to clear away B. rodhaini organisms from hosts. However, almost all the treatment groups, had a relapse of the infection within 10 days post treatment or re-treatment. Cured mice by treatment with clindamycin and clindamycin, or clindamycin and tetracycline showed complete resistance against challenge with B. rodhaini, while mice cured by administration of clindamycin at 200 mg or tetracycline at 100 mg showed incomplete resistance to challenge infection. The present data suggest that the two former chemotherapies can induce effective protective immunity (premunization), but the latter two chemotherapies induce incomplete premunization.
Autumn crocus poisoning of cattle is characterized by severe diarrhea caused by alkaloid colchicine. Previously, we examined pathologically this poisoning in cattle and reported that enterotoxic lesions were closely associated with apoptosis. To examine enterotoxicity of autumn crocus more precisely, a reproductive study was performed using guinea pigs and mice, and pathological findings associated with autumn crocus poisoning were compared with those of colchicine. Each group of guinea pigs given the bulb of autumn crocus or colchicine exhibited severe diarrhea. Histopathological findings in intoxicated guinea pigs were entirely consistent with those in the autumn crocus-poisoned cattle. In contrast, each group of mice administered with the bulb or colchicine did not develop diarrhea. Our results confirmed that the toxicity of autumn crocus bulb is attributable to the toxicity of ingredient colchicine, and revealed that the guinea pig has high reproducibility of autumn crocus poisoning in cattle and colchicine poisoning in humans. It has been reported that the physiological mechanism of the apoptotic process for eliminating the enterocytes in the mouse and rat differs from that of the guinea pig, monkey, cattle and horse. Taking the observation that the former animals do not develop diarrhea, whereas the latter animals do so in the autumn crocus or colchicine poisoning into consideration, it would seem that the species-difference in enterotoxicity of autumn crocus may be closely associated with the physiological mechanism of eliminating the effete enterocytes.
A polymerase chain reaction (PCR)-based method using a herpesvirus consensus primer was assessed for the identification of herpesviral infections in tortoises. A single band of about 230 bp was detected in PCR products from two out of twenty swabs taken from the oral cavity, three out of three paraffin-embedded tissue sections from the liver (two cases) and oral mucosa (one case), and one out of two fresh tissue samples from the oral mucosa. Nucleotide sequencing of these PCR products indicated that the herpesvirus present in these tortoises might belong to the alphaherpesvirinae. PCR using swabs and biopsy tissues was a sensitive and highly specific method for the diagnosis of herpesviral infections in tortoises.
An intrathoracic sarcoma from a 7-month-old female pig was studied by light and electron microscopy and immunohistochemistry. The tumor tissue of varied cell density consisted chiefly of spindle-shaped cells, some of which grew around blood vessels in a concentric fashion. Many tumor cells were positive for alpha smooth muscle actin and not for desmin, though some cells were reactive for both antigens. A majority of tumor cells had ultrastructural features characteristic of myofibroblasts, but a few cells resembled vascular smooth muscle cells in the synthetic state. This neoplasm may have arisen from vascular smooth muscle with features indicating a transition to myofibroblasts.
Participation of nitric oxide, vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase activating peptide (PACAP) in nonadrenergic, noncholinergic (NANC) relaxation of longitudinal muscle of various intestinal regions in Sprague Dawley rats (8-week-old) was studied in vitro. Nitric oxide was suggested to participate in NANC relaxation of every intestinal region studied. But the participation was partial and its extent varied among the regions: significant in the proximal colon and rectum, and moderate in the jejunum, ileum and distal colon. Participation of PACAP in NANC relaxation was suggested only in the distal colon, while that of VIP was not detected in any of regions. Results obtained in the present study indicate that extent of participation of nitric oxide in NANC relaxation in Sprague Dawley rat intestine is more significant than those of other strains, Wistar and Wistar-ST.
Our previous report demonstrated that high concentration of taurine is present in rat milk for the first few days of lactation and plays an important role in the body growth of rat pups. In the present study, gene expression of rate-limiting enzyme for taurine biosynthesis, cysteine sulfinate decarboxylase (CSD) were examined in rat mammary gland. By RT-PCR, CSD mRNA was found to be expressed in rat mammary gland like that in the liver. The expression level of CSD mRNA in the mammary gland was higher in the earlier lactational stage (days 1 and 6 of lactation) than that in the later lactational stage (day 14). CSD mRNA expression in the mammary gland of non-pregnant rats was only a trace level. By in situ hybridization analysis, CSD mRNA was demonstrated in the epithelial cells of the mammary gland. These results suggest that high concentrations of taurine in the milk are at least partially resulted from de novo synthesis of taurine in mammary gland epithelial cells and that the expression pattern of CSD mRNA may be responsible for the changes in taurine levels in the milk during a lactational period.
Since enterohemorrhagic Escherichia coli O157, Salmonella, etc., sometimes contaminate animal feces and may cause infectious diseases to humans, it is important to remove pathogenic bacteria from domestic animal waste. For the purpose, we examined the antibacterial activity of chaff vinegar. We found that the chaff vinegar inhibited the growth of pathogenic bacteria immediately in vitro but not efficiently spores and lactic acid bacteria. Further, it removes bacteria, especially Enterobacteriaceae, from animal feces and the surface of the concrete-floor in the cattle barn. Chaff vinegar is advertised as a natural chemical substance for a soil conditioner, to promote the composting and to deodorize their smell. Chaff vinegar may be useful for organic agriculture without enteric pathogenic bacteria.
A recombinant bovine herpesvirus type 1 (BHV-1), designated BHV-1/ TF17-1, which expresses pseudorabies virus (PrV) glycoproteins gB, gC, gD, gE and gI in combination was constructed. To test the protective immunity, 10 mice were inoculated with BHV-1/TF17-1 and three weeks later 10 mice were intraperitoneally (i.p.) challenged with 20 LD50 virulent PrV (YS-81). BHV-1/TF17-1 protected all the mice from the PrV lethal challenge while all the control mice died in around 3 days. Mice vaccinated with BHV-1/TF17-1 acquired high PrV-neutralizing antibody titers and demonstrated strong delayed type hypersensitivity responses and moderate in vitro lymphocyte proliferative responses to PrV antigen. Since the major PrV glycoproteins were integrated into virions (probably into viral envelope), BHV-1/17-1 was neutralized with anti-PrV antiserum. However, the susceptibility of BHV-1/TF17-1 to anti-PrV antiserum is 2- to 4-fold lower than that of PrV vaccine lines. Our results demonstrated the possibility of BHV-1/17-1 as a vaccine to protect piglets from Audjesky's disease where maternal antibodies against PrV interfere attenuated live PrV vaccines.
The prevalence of infections with three feline retroviruses; feline leukemia virus (FeLV), feline immunodeficiency virus (FIV) and feline foamy virus (FeFV), was examined in domestic cats (Felis catus) and leopard cats (Felis bengalensis) in southern Vietnam in 1998. We then compared this data with our previous study in northern Vietnam in 1997. None of the cats had FeLV antigens in both the northen and southern areas. In contrast, there is a great distinction in the seropositivity of FIV. Twenty-two percent of domestic cats had FIV antibodies whereas no FIV positive cats were detected in northern area. FIV may have entered southern Vietnam recently and spread rapidly. FeFV infections were found in both areas, suggesting that FeFV might be present in the cat populations in Vietnam from the earliest time.