Growth suppression and branchitis in trout exposed to hydrogen peroxide.

This study was designed to investigate the effect of single short-duration bath-treatment with hydrogen peroxide (1000, 1250 or 1500 mg/l for 20 min), as currently used in treatment for sea lice, on the growth rate and gill morphology of rainbow trout. All three dose levels significantly reduced the specific growth rate of fish (22-36% reduction) during the first 3 weeks after treatment. A significant effect on growth rate (10-11% reduction) persisted over an additional 3 weeks for the two highest doses. Gill lesions occurred at all three dose levels and also at an additional peroxide concentration of 750 mg/l; these lesions were characterized by large foci of epithelial hyperplasia in which lamellar fusion, pillar cell necrosis, and pillar channel aneurysms had developed. The proportion of damaged filaments showed a significant positive linear relationship with dose during the first 2 weeks after treatment. A significant decline in number of lesions occurred at 1000-1500 mg/l over the 3-week sampling period. During healing, necrotic lamellae were replaced by means of a pillar channel regenerative process.

Effects of formalin, chloramine-T, and low salinity dip on the behavior and hemolymph biochemistry of the American lobster.

The purpose of this research was to investigate the salinity and formalin sensitivity of a ciliate parasite (Anophryoides haemophila) of the American lobster (Homarus americanus), and to examine the target-animal (lobster) safety of chemical-bath treatments involving low salinity, formalin, or chloramine-T that could be used to control this parasite in lobster pounds. "Bumper car" disease, caused by An. haemophila, is an important concern to lobster pound operators in eastern North America, because of the implicated lobster mortality rate and the general lack of preventive and therapeutic intervention regimes. We determined, using an in vitro method, that formalin at 50 mg/L, or low salinity at 8.0 parts per thousand (ppt) for 1 hour killed 100% of the parasites. When healthy lobsters were exposed to formalin at 200 mg/L, there were no negative behavioral responses and no significant differences in a panel of hemolymph biochemical indices. Similar results occurred when lobsters were exposed to chloramine-T, a common finfish therapeutic agent for topical bacteria and protozoa, at 10 mg/L for 1 hour. The low salinity treatment (8.0 ppt) resulted in significant adverse changes in lobster behavior and biochemical indices; however, these changes did not persist for more than 1 week after treatment ended. Although these treatments are unlikely to kill parasites that have already invaded the lobster carapace, they should be effective in reducing parasite loads on the gill and carapace surface of the lobster and in the environment of the impoundment housing.

Evaluation of an automated colorimetric assay for the measurement of lipase activity in canine sera.

An automated colorimetric method for determining lipase activity in canine sera was evaluated for precision, linearity and correlation to existing assay methods. The colorimetric method was a commercial reagent that used a series of enzymatic reactions based on the hydrolysis of 1,2 diglyceride by pancreatic lipase. Within-run and between-run coefficients of variation were < 6.8% and < 8.3%, respectively. Linearity was determined to be at least 1366 U/L. Canine serum lipase concentrations attained using the colorimetric method were compared to both titrimetric and dry-film methods for measuring serum lipase activity, resulting in significant (P < or = 0.05) correlation coefficients of 0.92 and 0.77, respectively. Canine serum lipase concentrations measured using the colorimetric assay on 2 different automated analyzers had a significant (P < or = 0.05) correlation coefficient of 0.92. A laboratory reference range using serum samples from 56 healthy dogs (0-561 U/L) was established. There were no significant (P < or = 0.05) differences in mean serum lipase concentrations comparing male and female dogs or comparing young dogs (< or = 3 y) to mature (4-7 y) and older (> 7 y) dogs using this assay. It was concluded that the automated colorimetric assay was a reliable indicator of canine serum lipase activity and offered several advantages, including small sample volume and short analysis time.

Use of genomic DNA probes for the diagnosis of acute sarcocystosis in experimentally infected cattle.

Two clones of 1.4 and 4.33 kilobase pairs (kbp) DNA inserts, were selected from a Sarcocystis cruzi sporozoite genomic library constructed in bacteriophage lambda gt10. These clones strongly hybridized with sporozoite and merozoite DNA and were evaluated as probes for detection of merozoite DNA in clinical samples. Of five calves in the experiment, four were each orally dosed with approximately 200,000 S. cruzi sporocysts; one calf served as non-infected control. Subsequently, blood was collected from the calves twice weekly for 3.5 months and fractionated into buffy coats, polymorphonuclear cells, and plasma. Total cellular DNA extracted from these fractions was dot blotted on nylon membranes and hybridized with the probes radiolabeled with [alpha-32P]dATP. The probes detected merozoites on Day 22 post infection in the buffy coats and intermittently from Day 25-39 in the granulocyte fraction. Parasitemia (i.e. merozoites in blood) was also detected by indirect fluorescent antibody technique (IFAT) and direct microscopy, Diagnosis of sarcocystosis in cattle using genomic DNA probes by dot blot hybridization provides an alternative method of detecting parasitemia that is more rigorous than the other two tests (IFAT, direct microscopy) which rely on morphology of the merozoite and visualization by the examiner. As probes detected merozoite DNA in the granulocyte fraction, polymorphonuclear cells may be involved in the pathogenesis of S.cruzi; however this hypothesis requires further study.

C-reactive protein concentration in dogs with inflammatory leukograms.

Serum C-reactive protein (CRP) concentration was measured, using an automated immunoturbidimetric assay, in 44 clinically normal dogs and 67 dogs with band neutrophil count > or = 10(9) cells/L, and values were found to be significantly (P < or = 0.05) different. Correlation of serum CRP concentration and band neutrophil count in the 67 dogs with > or = 10(9) band neutrophils/L resulted in a statistically significant (P < or = 0.05), but low correlation coefficient of 0.34. Serum CRP concentration and CBC values were determined for 6 clinically normal dogs undergoing anesthesia (controls) and 6 clinically normal dogs undergoing anesthesia and ovariohysterectomy. Significant alterations in CBC results and serum CRP concentration, compared with baseline values, were lacking in dogs of the control group. Serum CRP concentration was significantly (P < or = 0.05) increased above baseline values in dogs undergoing surgery and was significantly (P < or = 0.05) increased, compared with values in control dogs by 12 hours after surgery. In dogs undergoing surgery, serum CRP concentration was also significantly (P < or = 0.05) different from values in control dogs at 28 and 36 hours, but not at the 76- and 124-hour sample collection times. Alterations in CBC values compatible with possible or convincing inflammation were detected in 83% of the dogs undergoing surgery at the 8- and 12-hour postsurgery sample collection times, 100% of dogs at 16, 22, 28, and 36 hours after surgery, 83% of dogs at 52 and 76 hours after surgery, 67% of dogs at 100 hours after surgery, and 0% of dogs at 124 hours after surgery.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes in alkaline phosphatase isoenzyme activity in tissues and plasma of Atlantic salmon (Salmo salar) before and during smoltification and gonadal maturation.

Changes in tissue and plasma isoenzymes of alkaline phosphatase (ALP) were qualitatively and quantitatively determined for male and female Atlantic salmon parr, silvery parr, smolt, immature grilse, prespawning grilse and postspawning grilse using cellulose acetate electrophoresis, densitometry and spectrophotometry. Tissue ALP isoenzymes were isolated from intestine, kidney, bone, liver, and gonad and compared to plasma isoenzymes. Parr plasma displayed three isoenzymes from bone and liver (slow and fast). During smoltification, ALP activity increased in tissue extracts from liver, gonad, and kidney of males and females. Total plasma ALP activity also increased and was due to slow and fast liver isoenzymes. During ovarian development, total ALP plasma activity increased in females and was due mostly to liver isoenzymes and an incompletely identified isoenzyme or isoenzyme mixture (band 2). However, in males total ALP plasma activity did not increase during maturation and no band 2 was evident. In male and female maturing adult grilse, bone ALP activity declined and the isoenzyme band evident in parr plasma could not be detected. ALP activity declined in the plasma of postspawning males and females. In females this was due partly to the total clearance of band 2 from the plasma, together with lowered levels of liver isoenzymes. Treatment of postspawned grilse in February and March with triiodothyronine and thyroxine elevated plasma thyroid hormone levels and increased plasma ALP levels. In conclusion, plasma ALP isoenzyme activities change with physiological state, and knowledge of the conditions governing these changes is important when using these enzymes as a diagnostic tool.

Production of monoclonal antibodies specific for antigens derived from tissue of chinook salmon (Oncorhynchus tshawytscha) affected with plasmacytoid leukemia.

Two distinct monoclonal antibodies (MAB) were prepared for testing with kidney, spleen, and retrobulbar tissue imprints made from chinook salmon (Oncorhynchus tshawytscha) affected with plasmacytoid leukemia. (PL). Hybridomas were prepared from mice immunized with whole and lysed cells purified from renal or retrobulbar PL-positive tissues, which had been obtained from naturally and experimentally infected fish from British Columbia, Canada. The MAB reacted with at least 4 morphologically different cell types; fluorescence was associated with the plasma membrane and cytoplasm. The MAB also reacted with kidney imprints made from chinook salmon affected with a PL-like lymphoproliferative disease in California, indicating that these 2 diseases might be caused by a similar agent. The MAB did not react with any of the kidney or spleen imprints made from wild chinook salmon collected from a river in Ontario, Canada.

Alkaline phosphatase isoenzymes in feline serum using an agarose gel alkaline phosphatase kit method.

Total serum alkaline phosphatase (ALP) activity is the product of the combined activity of isoenzymes from a number of tissue sources. In this study, a commercially available kit for electrophoretic separation of ALP isoenzymes in an agarose gel was used to separate ALP isoenzymes in feline tissue extracts and serum. Five separate bands of ALP activity were identified. These bands were numbered 1 to 5 with band 1 having the most anodal migration. The tissue of origin corresponding to the migration position of the isoenzymes are as follows: Band 3 was the liver isoenzyme, band 4 was the bone isoenzyme and ALP isoenzymes of both intestine and kidney migrate in the position labelled band 5. Band 1 appears to be related to albumin and does not represent true ALP activity. The tissue source of band 2 (a and b) was not identified. Serum ALP activity of mature, healthy cats is primarily of liver origin. Immature cats (< 1 year of age) have a greater proportion of the bone isoenzyme in the serum.