Seventy-nine strains of enterotoxigenic Escherichia coli (ETEC) isolated from 35 diarrheic piglets (aged one to 43 days) reared on 22 farms in Okinawa Prefecture between 1989 and 1998 were submitted to PCR investigations for serotypes, biochemical characteristics, fimbrial adhesions, entrotoxin production, and the presence of virulence genes. The isolates belonged to 6 serogroups: 0149 (57), 0141 (8), 020 (7), 09 (3), 0141 (3), and 064 (1). All of the 0149 strains demonstrated β hemolytic activities, whereas the remaining 0 serogroups did not. On the basis of 20 biochemical features noted in all isolates, characteristic isolate patterns were divided into 10 distinct types: 40 isolates (50.6%) were classified as one type. The F4 (K88) ac fimbrial gene was detected in 64 strains (81.0%); the F5 (K99) gene in 3 strains (3.8%); and the F6 (987P) gene in 11 strains (13.9%). One strain (1.3%) was negative for all 4 of the adhesive genes. Thirty-six strains (45.6%) were shown to possess 4 enterotoxin genes (LT I, ST I, ST II, and EAST 1). LT I, ST II, and EAST 1 were present in 22 strains; ST I in 10 strains; ST I and ST II in 5 strains; ST I, ST II, and EAST 1 in 4 strains; and ST I and EAST 1 in 2 strains. These results indicate the dominance of the serogroup 0149 harboring 4 enterotoxin genes (LT I, ST 1, ST II, and EAST 1) and the F4ac fimbrial gene.
The following four methods for extracting viral DNA as templates for polymerase chain reaction (PCR) from formalin-fixed paraffin specimens (FFPS) were evaluated: phenol extraction and three commercial kits (Sepa-Gene, the DNA isolator PS kit, and DEXPAT). Extraction efficacies of these methods in materials from chickens infected with the chicken anemia virus (CAV) were compared by means of 2-step PCR. At 8, 14, and 22 days after inoculation, all methods detected PCR products of 279 bp. At 31 days after inoculation, however, products were detectable only in phenol-extracted FFPS. In the first step of PCR, when 480 bp was taken as the amplification region, products were detectable only in phenol-extracted FFPS collected at 8, 14, and 22 days after inoculation. When 279 bp was taken as the target sequence in the first step of PCR on the other hands, all the methods investigated detected CAV DNA. These results show phenol extraction to be most effective in extracting viral DNA from FFPS. But, it is time-consuming and requires the use of toxic reagents. In the light of these shortcomings, commercial kits too are useful since they successfully detect viral DNA from FFPS when 2-step PCR with a short target sequence is employed.
The angiotensin-converting enzyme inhibitor, benazepril was orally administered to dogs for a period 15days before and after renal ablation. Renal ablation by right-side nephrectomy and ligation of one branch of the left renal artery were performed in the animals to impair renal function. One group of dogs received a standard dose (0.5mg/kg) of benazepril; a second group received 20 times the standard dose (10mg/kg). During the period before surgery, neither the standard nor the increased doses produced remarkable changed in clinical signs and laboratory test results. In 1 1/2 nephrectomized dogs, renal plasma flow and glomerular filtration rate decreased to approximately 1/3 of what they had been before surgery, and BUN and plasma creatinine concentrations increased. These variables remained unchanged in both dose groups during the administration period also in 11/2 nephrectomized dogs.
A Shiba-breed dog (8 months old) referred to our hospital for progressive neurological symptoms exhibited ataxia, dysmetria, head tremor, corneal clouding, and visual defects. No severe abnormalities in hearing or olfactory functions were observed. Serum and cerebrospinal fluid analysis ruled out infection with the canine distemper virus. Radiographic evaluations were normal. Computed topography revealed no brain abnormalities. Although routine blood biochemistry values were within normal limits, in a blood smear, multiple large vacuoles were observed in many lymphocytes. On the basis of this finding, suspected lysosomal disease was diagnosed. Measurements of leukocytic lysosomal enzyme activities showed a deficiency of β-galactosidase (0.3% of the normal level). The dog was therefore diagnosed as having GM1 gangliosidosis.
Hematological examinations of a seven-year-old male mongrel dog with a one-month history of anorexia and depression revealed mild nonregenerative anemia and severe leukocytosis. Numerous blast cells resembling lymphoblasts were observed in smear samples from peripheral blood and bone marrow. A diagnosis of acute lymphoblastic leukemia was made. Although chemotherapy with cyclophosphamide, vincristine, and predonisolone was performed, the dog died 41 days after initial presentation. Histopathological examination revealed multifocal necrotic lesions with Candida albicans in the liver, spleen, heart, diaphragm, urinary bladder, and lymph nodes. Systemic candidiasis in dogs with acute leukemia has not been reported before but is presumed to become more frequent in the future.
From 1997 to 1999, epidemiological surveys of helminths in such wild mammals as foxes and rodents were conducted in Kanagawa, Yamanashi, Shizuoka, and Nagano Prefectures and in Tokyo. Each infected fox harbored two or more of the following helminth species: six species of nematodes (Taxocara canis, Trichuris vulpis, Anclystoma kushimaense, Arthrostoma miyazakiense, Strongyloides planiceps, and Molineus patens); two species of trematodes (Metagonimus takahashii and Concinnum ten); and one species of cestode (Taenia pisiformis). In total, nine different helminths were detected in 25 (78.1%) of 32 animals examined. The crysticerci T. taeniaeformis and T. crassiceps were found in Rattus rattus and in Apodemus speciosus. The pleroceroide of Spirometra erinaceieuropaei was detected for the first time in Urotrichus talpoides, heretofore never considered a host. No Echinococcus adults or larvae, however, were detected in the animals examined.
By the examination of Campylobacter species, 47 Campylobacter jejuni strains were isolated from 107 cattle cecal specimens obtained in a Saitama slaughterhouse brought from 20 farms in 5 prefectures. On the basis of the PCR-based randomly amplified polymorphic DNA (RAPD) method, the isolates were divided into 27 types. This method is recommended for discriminating strains of C. jejuni isolated from cattle for the purposes of epidemiological study.