Effect of bovine lactoferrin on the immune responses of captive bottlenosed dolphins (Tursiops truncatus) being transported over long distances.

Bovine lactoferrin was administered orally, in feed, to six bottlenosed dolphins (Tursiops truncatus) before they were transported for approximately six hours; their stress responses were compared with those of five untreated dolphins. During the journey the dolphins had an increased plasma concentration of cortisol, and lymphopenia, eosinopenia and mild neutrophilia, indicating a stress response. The administration of lactoferrin did not affect the function of the dolphins' polymorphonuclear leucocytes, but affected their leucogram by maintaining the number of circulating eosinophils.

The production of arthritis in beagles by an immunological reaction to bovine serum albumin.

Arthritis was produced in beagles by the immunological reaction to bovine serum albumin (BSA). Dogs immunized with BSA showed the development of delayed type-hypersensitivity response to BSA and the significant increase in the titer of serum anti-BSA antibodies. The development of arthritis and the increase in a number of nucleated cells in synovial fluid were observed by the injection with BSA into the knee joints of immunized dogs. The synovial membrane of BSA-injected joints revealed a remarkable villous hyperplasia of membrane, and an infiltration of lymphocytes and plasma cells around vessels, resulting in a lymphoid nodule-like formation. The depositions of IgG and C3 on the surface of the synovial membrane were also observed in BSA-injected joints. Histopathological and immunopathological findings indicated that the immune response to BSA in the knee joints could induce a rheumatoid arthritis like chronic synovitis in dogs.

Serum lipid and lipoprotein concentrations in obese dogs.

Serum lipid and lipoprotein concentrations in 10 obese and 16 control dogs were examined. The serum triglyceride (TG) concentration in obese dogs was significantly higher than in control dogs. The serum concentrations of TG and phospholipid (PL) in beta lipoprotein and PL in pre-beta lipoprotein were significantly higher in obese dogs, while the serum PL concentration in alpha 1 lipoprotein was significantly lower in obese animals. In the serum total cholesterol concentration in obese dogs, a higher tendency for beta and pre-beta lipoproteins and lower tendency for alpha 1 lipoprotein were observed. These abnormal lipoprotein profiles were similar to those in diabetes mellitus in men and acute pancreatitis in dogs.

Partial cloning of prohibitin cDNA from canine, feline, bovine, equine, and rabbit liver mRNA by RT-PCR.

Prohibitin is the protein which has an inhibitory function in cell growth, and its gene is suggested to be one of putative tumor suppressor genes. In this report, we described a partial cloning of prohibitin cDNAs from canine, feline, bovine, equine, and rabbit liver mRNAs by RT-PCR, and their homology analysis. The sequences of these RT-PCR products were compared with each other as well as those reported for human and rat. The homology in this region of prohibitin cDNA was approximately 90%, and the amino acid sequence of each RT-PCR product shared more than 95% identity. Therefore, it is concluded that all the RT-PCR products are a part of prohibitin cDNA of each animal.

Lipoprotein profile in canine pancreatitis induced with oleic acid.

Lipid and lipoprotein concentrations and apolipoprotein profile were investigated in canine pancreatitis induced by infusion with oleic acid (OA) into the accessory pancreatic duct. Pancreatitis was diagnosed by physical, hematological, biochemical and pathological examinations. In OA-treated dogs, serum triglyceride (TG) concentration was increased; however, there were no changes in serum total cholesterol (TC) and phospholipid (PL) concentrations. Serum concentrations of TG, TC, PL and total lipids (TL) in beta lipoprotein and those of TC, PL and TL in pre-beta lipoprotein were increased and those of TC, PL and TL in alpha, lipoprotein were decreased. In apolipoprotein profile, the proportions of apolipoprotein B100 in low density lipoprotein fraction and apolipoprotein A-IV in high density lipoprotein fraction were increased. In addition, decreased proportion of apolipoprotein A-I and the appearance of serum amyloid A protein in high density lipoprotein fraction were observed. These results suggest that lipoprotein profiles observed in canine acute pancreatitis are attributed to the alterations in apolipoprotein compositions.

Plasma methylguanidine and creatinine concentrations in cats with experimentally induced acute renal failure.

Plasma methylguanidine (MG) and creatinine (CRN) concentrations were measured in 11 cats with experimentally induced acute renal failure by a two-stage surgical procedure. According to the progression of renal failure, both plasma MG and CRN levels increased. A significant positive correlation (y = 0.187X - 0.379, lambda = 0.9176, P < 0.001) was found between plasma MG and CRN levels. These results suggested that the increase in plasma MG level was an available indicator for uremic status in cats.

Evaluation of plasma erythropoietin levels in normal adult dogs by in vivo bioassay using concentrated plasma.

Measurement of plasma erythropoietin level in normal dogs by in vivo bioassay has been considered to be impossible so far. In the present study, we successfully determined it by using concentrated plasma 60 times which allowed the lower limit to 2.7 mU/ml. This normal plasma erythropoietin level was the first to be determined as an in vivo bioactivity and was 9.14 +/- 7.81 mU/ml in 75 normal adult dogs. This value was sufficiently reliable in terms of accuracy of determination and considered to be meaningful as the low level in vivo bioactivity that hasn't been known to date. Furthermore, erythropoietin levels in normal plasma were within a certain lower range and showed neither difference in plasma erythropoietin level between males and females or among breeds nor correlation between erythropoietin and hemoglobin level.

The iothalamate clearance in cats with experimentally induced renal failure.

Plasma iothalamate (IOT) disappearance rates were measured after a single-injection of IOT (113.8 mg/kg, IV) in cats with experimentally induced renal failure. The disappearance rates especially fitted into the one compartment model. The mean value of plasma disappearance rates of IOT in these cats with induced renal failure (2.16 +/- 0.240 x 10(-3) micrograms/ml/min) was markedly lower than that of clinically healthy cats (4.10 +/- 1.00 x 10(-3) micrograms/ml/min). These results demonstrate that IOT clearance is available for evaluation of renal function in cats.

Plasma concentrations of substances suspected as uremic toxins in experimentally induced and spontaneous uremic dogs.

Plasma concentrations of four substances, a pyridine derivative (S7a), uric acid (UA), hippuric acid (HA) and kynurenic acid (KA), suspected as uremic toxins in dogs were determined in dogs with experimentally induced uremia by the ligations of renal arteries, spontaneous uremic dog patients and normal dogs. In experimentally induced uremic dogs, plasma concentrations of S7a, HA and KA showed continuous increase after the ligation of renal arteries together with a significant correlation to plasma creatinine concentration (Cre). Plasma UA concentration increased rapidly, but it showed a varying fluctuation without showing any correlation to Cre. Plasma concentrations of S7a, UA, HA and KA in spontaneous uremic dogs were almost within the ranges of those of experimentally induced uremic dogs.

Evaluation of erythropoietin production in dogs with reduced functional renal tissue.

This study was designed to determine the quantity of functional renal tissue needed to maintain homeostasis of red blood cell production by evaluating the erythropoietin (EPO) production response to phlebotomic stimulation in the 1/2- and 1/4-kidney dogs surgically prepared. The results showed that the reduction in functional renal tissue caused a decrease in EPO production, which led to the delay in recovery from anemia. In the anemic progress stage, the plasma EPO level showed a transition proportional to the quantity of functional renal tissue immediately after the operation for tissue reduction. The 1/2-kidney dog group still kept such proportional relation even in the recovery stage. Thus, the half of the normal renal tissue was considered sufficiently contributory to EPO production needed to maintain homeostasis of red blood cell production. However, the 1/4-kidney dog group precipitously decreased in plasma EPO level in the recovery stage and fell into an extremely unfavorable anemia. This indicated that homeostatic maintenance in erythropoiesis would be impossible more below a quarter of normal renal tissue. These findings disclosed that reduction in functional renal tissue quantity would sensitively influence homeostatic maintenance of red blood cell production through the decrease in EPO production, even if it does not affect renal function concerned with urine production.

Evaluation of phlebotomy-induced erythropoietin production in the dog.

Erythropoietin (EPO) production in dogs was studied by reducing red blood cells with phlebotomy. In this study, the hemoglobin reduction rate (delta %Hb) was newly taken into account as the regulating factor for EPO production, and its usefulness to estimate the stimulating intensity to EPO production was examined. As the result, plasma EPO was highly correlated with delta %Hb showing the importance related to regulation of EPO production, in the increasing plasma EPO by different degrees of phlebotomy, in the change of plasma EPO through the anemia progress and recovery period after severe phlebotomy, and in the initial variation of plasma EPO induced by chronic mild phlebotomy. On the other hand, increasing EPO production appeared at least within 6 hr after acute severe phlebotomy, which revealed significantly higher plasma level compared with the mild chronic phlebotomy, suggesting the effect of time leading to red blood cell reduction on EPO production response. Simultaneously, considering an in vivo EPO half life of 8.4 hr calculated from plasma EPO disappearance after bilateral nephrectomy, endogenous plasma EPO accumulation should be taken into consideration in rapidly increasing of delta %Hb.

Detection of uremic peaks in dogs by anion exchange high performance liquid chromatography.

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.

Expression patterns of the erbB subfamily mRNA in canine benign and malignant mammary tumors.

ErbB subfamily genes, known as proto-oncogenes, encode receptor tyrosine kinases, and are expressed in relation to tumorigenesis of the mammary gland in humans. In this study, we examined the expression of erbB subfamily mRNAs in two canine normal mammary glands and 12 mammary tumor samples by reverse transcriptase-coupled polymerase chain reaction (RT-PCR). Each primer set was designed from the nucleotide sequence of the region conserved in erbB subfamily cDNA among other species. No erbB subfamily mRNAs were expressed in the normal mammary gland. In contrast, all of the subfamily mRNAs were expressed in a benign mammary tumor, and more than one type of the subfamily mRNA were observed in 11 malignant mammary tumors. The length of RT-PCR products were 380 bp for erbB1, 500 bp for erbB2, 644 bp for erbB3, and 416 bp for erbB4. These sequences were highly homologous to the cDNA sequences of other species. Therefore, these results suggest that the expression of erbB subfamily mRNAs in canine mammary tumors plays an important role in tumorigenesis of the mammary gland.

A control of a golden retriever with renal dysplasia.

A six-month-old male Golden Retriever with a three-month history of polyuria and polydipsia was examined. Hematological examinations revealed nonregenerative anemia, azotemia, high serum creatinine level, hypercalcemia, hyperphosphatemia, hypercholesterolemia, hyperamylasemia, and low level of total serum protein. Urinalysis indicated mild proteinuria, and low specific gravity. Radiographic and ultrasonographic examinations revealed bilateral small sized kidneys. Histological examination by renal biopsy confirmed the diagnosis of renal dysplasia. Treatment with a dietary protein restriction, oral adsorbents, and dried aluminum hydroxide gel have been performed in this dog, and then, azotemia, high serum creatinine level, hypercalcemia, and hyperphosphatemia were improved. During 10 months after the initiation of treatments, no significant clinical change except polydipsia and polyuria has been observed.

In vitro cytotoxicity of recombinant human-TNF-alpha and actinomycin D on canine normal and tumor cells.

The in vitro cytotoxicity of recombinant human tumor necrosis factor-alpha (rh-TNF-alpha) and actinomycin D (ACT-D) on canine normal and tumor cells was investigated. rh-TNF-alpha showed dose-dependent cytotoxic and cell-growth inhibitory effects on cultured canine kidney carcinoma cells (CKCa-1). rh-TNF-alpha alone produced little cytotoxic effect on canine normal cells. However, combined with ACT-D, it showed moderate cytotoxicity on normal canine cells from the kidney medulla, spleen, heart muscle and lung. When the effects on the spontaneous tumor cells were examined, the combination of rh-TNF-alpha and ACT-D produced substantial cytotoxic effect on 60% of the tumor cells. All mammary mixed tumors and perianal gland tumors tested were susceptible to this combination. The data indicated the combination of rh-TNF-alpha and ACT-D have in vitro cytotoxicity on certain canine tumor cells.

Induction of fragile sites by fluorodeoxyuridine and caffeine accompanies with misincorpolation of endogenous uridine nucleotide into DNA of feline fibroblasts.

Fluorodeoxyuridine, an inhibitor of thymidylate synthetase, is known to induce chromosomal fragile sites. The drug treatment may cause deprivation of intracellular thymidine nucleotide pool followed by a serious imbalance of deoxynucleotide pool. Though the stress is probably related to the induction of folate-sensitive fragile sites, the exact mechanism is still to be investigated. The present study has been carried out to test the possibility that the fragile sites are originated, at least in part, from incorpolated uracil residues. The incorpolated uracil residue can be detected by a novel assay for abasic sites after treatment with uracil-DNA N-glycosylase (UDG). About 2.7 abasic sites per 10(4) nucleotides were detected in the DNA extracted from feline fibroblasts after the treatment with FUdR and caffeine. By digesting the DNA with UDG prior to the assay, significant increase in the number of abasic sites were observed. These results indicate that the large amount of uracil residues are present in the feline fibroblast DNA under the condition which induces chromosomal fragile sites.

Computed tomography on renal masses in dogs and cats.

Computed tomography (CT) was performed on renal tumors (Wilms' tumor and renal cell carcinoma) and renal cysts in dogs and cats. CT images in renal tumors were well correlated with macroscopic findings, and contrast CT images were quite useful in differentiating tumoral regions from non-tumoral ones. On renal cysts, intravenous pyelography and ultrasonography were as effective as CT images in morphological diagnosis, but CT was considered to be superior for evaluating three-dimensional (3-D) relationships in complicated lesions.

C-cell adenoma containing variously sized thyroid follicles in a horse.

A thyroid gland tumor, showing unusual histology, was identified in a 13-year-old male Andalusian horse. Microscopically, the tumor consisted of neoplastic proliferation of C-cell (parafollicular cell) with cytoplasmic fine granules, containing diffusely distributed, variously sized colloid-containing follicles. Immunohistochemically, the neoplastic C-cell were positive for calcitonin and follicleforming epithelial cells showed a positive reaction for thyroglobulin. Ultrastructurally, membrane-bound secretory granules up to 250 nm in diameter were found in the cytoplasm of the parafollicular cells, whereas the follicular epithelial cells had microvilli, junctional complex, and well-developed endoplasmic reticulum.

Urinary bladder rhabdomyosarcoma (sarcoma botryoides) in a young Newfoundland dog.

A 13-month-old female Newfoundland dog suffered from urinary bladder tumor. Histologically the tumor consisted of round or fusiform cells, occasionally having eosinophilic cytoplasms. Apparent mature rhabdomyoblasts possessing elongated eosinophilic cytoplasm and cross striations were infrequently observed. The tumor cells exhibited immuno-positive for anti-myoglobin, desmin and vimentin antibodies. Ultrastructurally, tumor cells have abundant myofibrils in their cytoplasm and Z bands were also detected. The present tumor was diagnosed as a urinary bladder rhabdomyosarcoma in a Newfoundland dog, which has not been frequently reported in dogs.

Novel putative fragile sites observed in feline fibroblasts treated with aphidicolin and fluorodeoxyuridine.

Fragile sites are non-randomly distributed chromosomal breaks and gaps observed in the cells cultivated under certain conditions. Feline fragile sites were analyzed using skin fibroblast strains after the treatments with aphidicolin and fluorodeoxyuridine in combination with caffeine. Three aphidicolin-induced fragile sites (A1q21, C2q13 and E1p21) as well as a folate-sensitive site (B1q14) were observed in all the 3 fibroblast strains tested for each treatment group. The loci in A1q21 and B1q14 are very close to that reported previously for peripheral blood lymphocytes and lung cells. Two chromosomal break points in C2q13 and E1p21 seem to be new fragile sites. Fifteen candidates for feline fragile sites were also assigned their locations in feline chromosomes. Both the incidence and distribution of feline fragile sites in skin fibroblasts seem to be different at least in part from those in lymphocytes.