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Feline immunodeficiency virus (FIV) proviral DNA was detected by the polymerase chain reaction method (PCR). PCR products were detected by gel electrophoresis and ethidium bromide staining. The P-10, P-15 and P-24 regions of the gag gene of FIV were chosen as the target sequences for amplification, and three primer pairs were prepared. The PCR products subjected to amplification with each primer pair were found to possess sites of digestion by a restriction enzyme, as hypothesized. They did not react with feline leukemia virus (FeLV)-infected or feline syncytium-forming virus (FeSFV)-infected cell-derived DNA, and specifically amplified FIV-infected cell-derived DNA. FIV proviral DNA was detected by the PCR method with either primer pair (one-step amplification: single PCR) in DNA derived from peripheral blood lymphocytes (PBL) from 7 of 12 FIV antibody-positive cats. When PCR products in each of the 12 cats were subjected to a second amplification using the same primer pair (two-step amplification: double PCR), FIV proviral DNA was detected in all of the cats. When PBL samples collected from three cats that were negative and three that were positive in the single PCR were cultured for a few weeks in the presence of interleukin 2, FIV proviral DNA was detected in all six cats by the single PCR method. The results suggest that either the use of cultured PBL as the sample or the performance of the double PCR method enables simple and specific detection of FIV proviral DNA in PBL.

ABSTRACT. The circadian variation of urinary N-acetyl-β-D-glucosaminidase (NAG, EC 3.2.1.30) and γ-glutamyl transpeptidase (γ-GTP, EC 2.3.2.2) was evaluated in cats. Urine and blood were collected at 4-hr intervals from adult cats (3 males, 9 females) weighing between 2.6 and 5.0 kg. There was no circadian variation in the urine volume, creatinine clearance, creatinine excretion, NAG excretion or γ-GTP excretion. The average NAG and γ-GTP indices in the 4-hr urine were similar to those for the 24-hr urine. However, the variance for the 4-hr urine samples was higher than that of 24-hr urine. In conclusion, although 4-hr urine samples can be used to estimate 24-hr urinary enzyme excretion, short-term spot urine samples may cause increased variation in the enzyme index. -KEY WORDS: creatinine clearance, kidney function, urine volume.J. Vet. Med. Sci. 60(9): 1033-1034, 1998 until analyzed. The activity of urinary NAG was determined colorimetrically, using the 580 nm wavelength of m-cresol purple released by NAG from the substrate sodio-mcresolsulfonphthaleinyl-N-acetyl-β -D-glucosaminide (MCP-NAG) (Shionogi Co., Ltd., Tokyo, Japan). Urinary γ-GTP activity was measured with a commercial testing kit using γ-glutamyl-p-nitroanilide as the substrate at 37°C and 405 nm using RaBA ∑ (Chugai Pharmaceutical Co., Ltd., Tokyo, Japan). Serum and urine creatinine levels were determined with a commercially available kit using the Jaffé reaction (Chugai Pharmaceutical Co., Ltd., Tokyo, Japan). The urine was diluted with distilled water (× 50) for creatinine determination. Creatinine clearance was calculated with the formula:where C cr is the creatinine clearance (ml/min/kg), U cr is the urine creatinine concentration (mg/dl), S cr is the serum creatinine concentration (mg/dl), V u is the urine volume (ml/ min), and W is the body weight (kg). Urinary NAG and γ-GTP excretion were calculated using the formula:NAG and γ-GTP excretion (U/kg)=U act × urine volume (liters) /W (kg), where U act is the urinary NAG or γ-GTP activity (U/l). The NAG and γ-GTP indices were determined from the ratio of urinary enzyme activity to creatinine: Index (U/g-Cr)=Urinary enzyme activity (U/l)/U cr (mg/ dl) × 100.Data were expressed as the mean ± SD. A repeatedmeasures analysis of variance procedure was used to determine the statistical significance of differences amongThe urinary enzymes N-acetyl-β-D-glucosaminidase (NAG, EC 3.2.1.30) and γ-glutamyl transpeptidase (γ-GTP, EC 2.3.2.2) are clinically useful for detecting early renal disorders in dogs [7,8]. Since renal diseases are also major diseases in cats, the early detection of renal disorders using urinary enzymes might be important for treating renal disease. A 24-hr urine sample is an accurate way of assessing urinary enzymes [6], but it is easy to lose or contaminate the sample. Spot urine sampling is a more feasible technique for performing urinary analysis in a veterinary clinic [1,4]. In dogs, there is no circadian variation of urinary enzyme excretion in spot urine samples, despite variatio...

We encountered 19 cases of canine babesiosis which were not previously reported in Aomori prefecture. The main clinical signs were depression, decrease of appetite, excretion of brownish urine and pallor of the visible mucous membrane. A remarkable regenerative anemia was peculiar in blood examinations. On Giemsa stained blood smears, parasitism of small protozoa was

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