Lymph drainage routes from the abdominal and pelvic cavities in beagle dogs were observed serially by following the time course of India ink administered intraperitoneally. Four systems of lymph drainage routes from the peritoneal cavity were observed in this study. The earliest drainage returned to the cranial mediastinal lymph nodes via the sternal lymph vessels; subsequently, the sternal lymph nodes located along the internal thoracic artery became involved. Then, a drainage route via the lymph vessel along the left vagus nerve was observed. The final drainage route flowed into the lateral lymph vessel through the thoracic duct located on the vertebra. These results show that India ink is absorbed from the peritoneal cavity, and that the lymph drainage first flows mainly towards the cranial mediastinal lymph nodes through the ventral lymphatic channels. Our serial observations suggest that, over time, the lymph drainage routes changed from the ventral abdominal to the dorsal thoracic lymphatic channels in the thorax.
The microvasculature of the eye of 10 pigs was investigated using scanning electron micrographs of corrosion casts. The ciliary body, iris and bulbar conjunctiva were supplied by the iridociliary ring artery via the long posterior ciliary artery. Capillaries of the ciliary process were of large diameter (23.2-27.5 μm) with an irregular bore, forming a thoroughfare channel draining blood in the ciliary arterioles into the pars plana venous vessels. Arterioles and venules in the iris exhibited a zigzag or spiral features. The third palpebra was supplied by the anterior ciliary artery. The capillary bed of the third palpebra was dense and was formed by many rows of fine hair-pin loops. Capillaries in the bulbar conjunctiva formed a sparse network disposing approximately parallel to the epithelium and formed a well-developed venous plexus, draining into the vortex veins. Retinal arterioles formed a slender and long course to capillaries. Retinal capillaries were extremely thin (3.0-4.0 μm in diameter). The choroid was supplied by the short posterior ciliary arteries. Choroidal arterioles exhibited a thick and short course to the choriocapillaris. The choriocapillaris was flat and sinusoid-like (8.9-13.9 μm in diameter), forming a dense sheet-like network. Blood from the choroid emptied into the episcleral vein via the vortex vein. Blood from the retina was drained by the posterior ciliary veins. The functional significance of this vascular architecture was discussed.
To clarify the strain differences in the morphology of the rat kidneys, we investigated the morphometrical characteristics of the kidneys of Slc:Wistar, Slc:SD, and F344/NSlc rats. The diameter of the renal corpuscles in female F344/N rats is smaller than that in female Wistar rats. Although sex differences (males > females) were shown in SD and F344/N rats, no effects of castration were detected in any of the groups. Strain-dependent differences in the percentage of renal corpuscles with a cuboidal parietal layer were found in both male and female groups. The highest percentage of them was noted in male Wistar rats. Effects of castration were observed in female Wistar and male F344/N rats, and the values after castration were significantly higher than those in the intact animals. As for the number of proximal convoluted tubular nuclei, no strain differences were detected in either the male or female groups. Although a sex difference was found in SD rats (female>male), no effects of castration were detected in any of the groups. In female F344/N rats, numerous numbers of PAS-positive granules, which were observed in the proximal convoluted and straight tubular epithelia, were noted. Orchiectomy induced an increase of these granules in male SD and F344/N rats, but ovariectomy showed no effects on them in any strains. This is the first study to clarify the strain differences in the morphological characteristics of the kidneys in ordinary rat strains.
The populations of retinal ganglion cell (RGC) groups (Groups I, II, III, IV) were similar each other between the central and intermediate zones, but the population in the peripheral zone were clearly different from those in the central and intermediate zones due to increase of Group III and IV cells and decrease of Group I cells. The dimensions of somal area and dendritic field of Group I cells increased very gradually toward the peripheral zone, but those of other three Groups grew steeply in the peripheral zone. The correlation index between somal area and dendritic field of RGCs showed high coefficient in the central (r=0.73) and intermediate (r=0.77) zones, but lowered clearly in the peripheral zone (r=0.64) due to increase of Group III cells, which showed nonlinear relation between somal area and dendritic field.
A gram-positive, catalase-negative, facultatively anaerobic coccus was isolated from a lactating cow with hematuria and urodynia in Japan. The isolate was identified by 16S rRNA gene sequence analysis as Facklamia sourekii. The biochemical and culture characteristics of the isolate were well consistent with those of F. sourekii type strain. Since all F. sourekii strains reported so far were isolated from human clinical specimens, this is the first reported case of F. sourekii isolated from veterinary clinical specimen.
Ovary lipid of Skipjack tuna (OLS) (Katsuwonus pelamis) contains a high level of docosahexaenoic acid combined with phospholipids. In this study, we examined the effect of OLS in male Wistar rats given OLS mixed in feed (0.9%) for 42 days, using an animal model of anxiety, the elevated T-maze test. The avoidance latency at the 1st trial was significantly shorter in the OLS ingestion group than in the control group. Those at the 2nd and 3rd trials showed a similar tendency. There was almost no difference in escape latency at the 1st trial between the two groups but the escape latencies at the 2nd and 3rd trials tended to be longer in the OLS group. These results suggested that OLS inhibits anxious behavior in rats.
To clarify the relationship between cellular immune status and nutritive condition in periparturient dairy cows, feeding content, blood profiles, and immune condition were observed in cows from two dairy herds with different types of feed content. Immunological analyses such as leukocyte population and peripheral blood mononuclear cell (PBMC) mRNA of IFN-γ, TNF-α, IL-4, and IL-10, quantified by real-time RT-PCR were performed. With regard to feed content during dry periods, there were six cows in the herd with insufficient non-structural carbohydrate (NFC) intake (group I) and six cows in the herd with sufficient NFC intake (group II). Significantly lower levels of blood glucose were observed in group I between weeks -12 and 16 compared with group II. Serum cholesterol level was significantly lower in group I between weeks 2 and 10 than in group II. The numbers of CD3+ and CD4+ T cells in group I were significantly lower than those in group II in weeks 6 and 14. The numbers of CD21+ B cells were significantly lower in group I than in group II in weeks -16, -12, 2, and 10. On the other hand, the CD4+/CD8+ ratio in group II was significantly higher than group I between weeks 2 and 14. The IFNγ/IL-4 mRNA rate in group I was significantly lower than group II in week 6. We concluded that cellular immune depression occurrs after calving in dairy cows with low nutritional status in the periparturient period.
Slit, a secreted protein, functions as a chemorepellent factor in axon guidance and neuronal migration and as an inhibitor in leukocyte chemotaxis. In humans, slit2 protein attracts endothelial cells and promotes tube formation in the tumor angiogenic mechanism. In this study, we cloned a part of the canine slit subfamily and examined the expression of slit subfamily mRNAs in 3 normal canine mammary glands and 11 mammary tumor samples by RT-PCR. The cloned part of the slit gene sequences showed high similarity to those of the human, mouse, and rat. The mRNAs were expressed at low levels in the normal mammary gland. The expression levels of slit1 mRNA were low in both the normal and tumor tissues. In contrast, the expression of slit2 mRNA increased in most of the malignant mammary tumors, and an increase in slit3 mRNA expression was observed in 2 of the malignant mixed tumors. These results suggest that the expression of slit2 plays an important role in tumor angiogenesis in canine mammary gland tumors and that slit2 can be a putative marker for malignancy diagnosis of these tumors.
The inhibition of Bcl-xL mRNA expression and the acceleration of apoptotic cell rates in canine mammary tumor cell line (CF33) by the small interfering RNA (siRNA) were analyzed. The level of Bcl-xL transcripts in CF33 was decreased when cultured with siRNA, suggesting that siRNA might inhibit the expression of Bcl-xL mRNA in the CF33. Apoptotic cell rates in CF33 cultured with siRNA in Oligofectamine medium, with double strand RNA in Oligofectamine medium, without siRNA in Oligofectamine medium and in DMEM alone were 60.9%, 30%, 28.7% and 11.6% at 48-hr incubation, respectively, when evaluated by TUNEL assay. From these results, it was suggested that canine Bcl-xL might be an anticancer target of canine tumors.
A two-year old male Welsh Corgi was referred for persistent thrombocytosis and occasional seizure. Hematological findings indicated marked thrombocytosis, eosinophilia, basophilia and moderate anemia. Bone marrow examination revealed marked megakaryocytic hyperplasia with morphologic abnormality. A diagnosis of essential thrombocythemia was made and the treatment was initiated with combination chemotherapy and maintained by prednisolone and busulfan. The dog successfully achieved complete remission on 100 days after initial presentation and has been good in health without chemotherapy since then.
To compare the changes in the insulin reaction of Holstein dairy cows and Japanese Black cows (JB) during the periparturient period, the insulin resistance test in vivo and lymphocytes proliferation with insulin in vitro were performed. Ten healthy Holstein dairy cows (Holstein group) and 10 healthy JB cows (JB group) used in this study were observed on days 60, 40, and 20 before calving and days 7 and 20 after calving. In insulin resistance reaction in vivo and in vitro, a low insulin-stimulated glucose disposal rate and lymphocyte proliferation with insulin were observed in the Holstein group compared with the JB group during the experimental period. An analysis of the lymphocytes cultured with insulin showed that the percentage of CD4+CD45R- T cells in the Holstein group was significantly lower than that of the JB group before day 20. These findings indicate that T cells reaction to insulin in healthy periparturient Holstein cows is lower than that in Japanese Black.
We evaluated hepatic T lymphocyte phenotypes in a dog with chronic hepatitis. Before treatment, numerous CD3+ lymphocytes were demonstrated in the liver, and the ratio of CD4+/CD8+ was remarkably high (2.96; reference range, 0.33 ± 0.12). After treatment, CD3+ lymphocyte infiltration in the liver was reduced, and the ratio of CD4+/CD8+ decreased to 0.31. Therefore, hepatic T lymphocytes, especially CD4+ lymphocytes, might play a central role in the pathogenesis of this dog with chronic hepatitis.
A crossbred Maltese dog, 6-year-old, male, was presented to us for examination due to coagulopathy. On examination of blood coagulation screening tests, activated partial thromboplastin time (APTT) was markedly prolonged (63.6 sec). Therefore, a defect in the intrinsic pathway of coagulation was suspected. An additional serum test was also examined and APTT was returned to within the normal range. Furthermore, factor IX coagulation activity was markedly low (2.3%). On the basis of these results, the dog was diagnosed with hemophilia B. The dog has since been presented to us because of hemorrhage problems again after 5, 10, and 16 months, but blood transfusions have maintained good control of its coagulopathy for more than two years.
A 15-year-old female maltese was referred to us because of a 3-month history of ataxia, circling, and acute blindness. A mass was noted in the brainstem on brain magnetic resonance images. A cerebellar herniation was also detected on T1-weighted sagittal images. The lateral, third and fourth ventricles and central canal of the cervical spinal cord were enlarged. Based on diagnostic imaging findings, cervical syringomyelia secondary to a brainstem tumor was suspected. The clinical signs were controlled well by lomustine and the dog survived for 8 months after the initial diagnosis. The mass was diagnosed as a meningioma based on histopathological findings. This report describes the clinical findings and imaging characteristics of an acquired syringomyelia resulting from a brainstem meningioma.
Chondrocytes isolated from proximal femoral articular cartilage from 3 adult cat cadavers were expanded in monolayer culture and subsequently cultured in alginate microspheres for 24 days. Cell proliferation and production of proteoglycans in alginate microspheres were observed during day 18 and 24. Quantification of chondroitin sulfates (CS) by capillary electrophoresis revealed that cultured chondrocytes synthesized CS6 but not CS4. Three-dimensional culture using alginate microspheres is a useful in vitro technique to study proliferation and metabolism of chondrocytes; however, further modifications are needed to apply the technique to feline articular chondrocytes.
A cDNA expression library from the salivary glands of hard tick, Haemaphysalis longicornis, was constructed. Immunoscreening was performed using sera of the rabbit repeatedly infested with ticks and seventeen positive clones were obtained. A BLASTP search suggested that 8 sequences matched with that of hypothetical H. longicornis sequence and one clone encoded HL35 antigen U from the same tick species. Eight of 17 gave no match to any sequence reported in the database. The proteins expected from these novel sequences possess common characteristics with cement proteins which assist ticks in their attachment to the host during blood feeding. The expression of these genes in salivary glands was confirmed by RT-PCR. Four of the 8 sequences showed to be upregulated upon blood feeding. These immunodominant antigens are of particular interest as candidates for future cement protein based-tick vaccine.
Detection rates from the samples including a small number of Cryptosporidium parvum oocysts were compared between the sugar floatation and the sugar centrifugal floatation methods. As the results, the oocysts were detected from 70 and 80 of 100 samples including 6.0 × 102 and 1.0 × 103 oocysts per 1 ml by the floatation method, respectively, whereas from 52 and 53 of the same samples by the centrifugal floatation method. Therefore, it was considered that the floatation method is the most suitable method for the detection from samples including a small number of Cryptosporidium oocysts. It is also suggested that results of the sugar floatation method were reliable for samples including more than 1.0 × 103 oocysts /ml.
To define the characteristics of malignancy we performed routine histology and an immunohistochemical study on seventeen aortic body tumors in dogs. We essayed tumors using a panel of immunohistochemical markers: neuron specific enolase (NSE), chromogranin A (CrA) and S-100. Among 17 cases, the neoplastic cells were positive for NSE (17 cases, 100%), S-100 (9 cases, 53%), and CrA (8 cases, 47%), respectively. The sustentacular cells density and chief cell staining intensity were both inversely related to tumor grade. The most relevant data was consistent with a negative staining of S-100 correlated with absence or decreased number of sustentacular cells in tumors grade III. This report indicates that the immunohistochemical panel has utility for the diagnosis of chemodectoma and the negative staining to CrA and S-100 markers in tumors grade III expresses an indication of malignant behaviour of the tumor.
ParalluxTM, a solid-phase fluorescence immunoassay (SPFIA) developed for antibotics residue detection in milk, was applied for analysis of fish tissue. The recommended therapeutic doses of oxytetracycline (OTC, 100 g/ton water, withdrawal period 30 days) and tetracycline (TC, 150 g/ton water, withdrawal period 30 days) were treated to a group of 35 olive flounders (Paralichthys olivaceus) using dipping administration. Muscle was sampled before and after drug treatment 1st, 2nd, 3rd, 5th, 7th, and 14th day. The concentration of oxytetracycline in muscle, determined by SPFIA, was compared with that of internal standard (100 ppb as oxytetracycline). The S/C ratio of sample inhibition value to cutoff inhibition value was employed as an index to determine the muscle residue in olive flounder. To investigate the recovery rate, and standard solutions were added to muscle samples to give final concentrations in muscle of 0.1 and 0.5 μg/ml. The recovery rates of all spiked samples were >89% of the spiked value. OTC and TC were detected in muscle of fishes treated until the 3rd day of withdrawal period. The present study showed that the SPFIA can be easily adopted in predicting tissue residues for OTC and TC in farmed fishes.
Interspecies cloning may be a useful method to help conserve endangered species and to study nuclear-cytoplasm interaction. The present study investigated in vitro development of goral (Naemorhedus goral) intergeneric nuclear transfer embryos produced by fusing goral fibroblasts with enucleated metaphase II (MII) bovine oocytes. After two to five passages, serum-starved or non-starved goral skin fibroblast cells were transferred into enucleated MII bovine oocytes. Couplets were electrically fused and chemically activated, and then cultured in either modified synthetic oviduct fluid (mSOF) or tissue culture medium-199 (TCM-199) supplemented with 10% FBS. Serum starvation of donor cells did not affect the fusion rate and or development to of cells to the two-cell stage, to more than 9-cells, or to morulae, regardless of culture medium. Three blastocysts from 202 fused embryos were obtained when embryos reconstructed with non- serum- starved donor cells were cultured in mSOF. However, no blastocysts were obtained when the embryos reconstructed with serum-starved donor cells were cultured in mSOF. The total cell number of goral intergeneric embryos averaged 130.3 (range 105-180). In conclusion, this study demonstrated that bovine oocytes can support blastocyst development after intergeneric SCNT with goral fibroblasts.
Ultrasonography-guided transabdominal needle aspiration was carried out to remove 3 to 14 ml of purulent matter from the cavities of prostatic abscesses in 6 dogs, and the same volume of tea tree oil was injected into the cavities to treat the abscesses. The same treatment was repeated 3 weeks later in 4 dogs, and subsequent disappearance of the purulent matter in the cavities and a marked reduction in the volume of the cavities were observed. These findings indicate that the treatment of prostatic abscesses by aspiration of the purulent matter and injection of tea tree oil into the cavities is very effective in dogs.
Previously we reported that immunization with pseudorabies virus (PRV), harboring chimeric Fc on the surface of the virus particles (PRV/Fc), induced higher immune responses than normal PRV particles. The chimeric Fc was fused with mouse transferrin receptor of transmembrane domain (mTR) and the Fc region of immunoglobulin G1. Since it has been reported that some chimeric protein of Fc and self-antigen induce auto-reactive antibodies, in this present study, we examined whether PRV/Fc induces auto-reactive antibodies that react with mTR. PRV/Fc immunized mice produced higher levels of anti-PRV antibodies and antibodies that reacted with mouse-derived 3T3/A31 cells (A31 cell), compared to normal PRV immunized mice. However, antibodies that reacted with mTR in A31 cells were not detected in both Western blot analyses and indirect immunofluorescence assay. The antibodies reacted with an antigen of approximately 16 kDa in A31 cells, but this antigen has a different molecular mass from that of mTR. The antibody also reacted with the antigen of approximately 16 kDa in RK13 cells in which the virus had been propagated. In addition, antibodies induced by immunization with normal PRV also reacted with the same antigen in A31 and RK13 cells. Moreover, neither kidney disorders, in which high levels of mTR were expressed, nor clinical symptoms of autoimmune diseases were observed in mice immunized with either PRV or PRV/Fc. These results indicated that the antibodies were not induced by mTR-Fc, but were instead induced by trace amounts of RK13 derived antigens contained in PRV or PRV/Fc preparations, and cross-reacted with equivalent molecules in mouse derived A31 cells. Therefore, this study confirmed that immunization with PRV/Fc did not induce harmful auto-reactive antibodies.
The virus neutralization (VN) antibody titers of serum samples from 18 individuals representing 8 carnivore species vaccinated with commercial polyvalent vaccines optimized for domestic cats containing inactivated feline panleukopenia virus (FPLV) were evaluated against canine parvovirus type 2 (CPV2). In addition, the titers among 5 individuals from 4 carnivore were evaluated against antigenic variants of feline parvoviruses; FPLV, CPV2, CPV2a, CPV2b, CPV2c, mink enteritis virus type 1 (MEV1) and MEV2. The polyvalent vaccines induced cross-reactive VN titers against antigenic variants of feline parvoviruses in nondomestic felids. However, we observed very low cross-reactive VN antibody in lions and Siberian tigers, therefore we should pay attention to CPV infections in these animals even if they were vaccinated with inactivated FPLV vaccines.