Preliminary Morphometrics of Spleen and Kidney Macrophage Aggregates in Clinically Normal Blue Gourami Trichogaster trichopterus and Freshwater Angelfish Pterophyllum scalare

Abstract

Macrophage aggregate (MA) morphometry and pigment composition are believed to be dependent on the species, age, and health status of the fish. The aim of this study was to characterize a “normal” morphometry baseline of spleen and kidney MAs in blue gourami Trichogaster trichopterus and freshwater angelfish Pterophyllum scalare. Three size-classes of clinically normal fish were analyzed. Blue gourami and freshwater angelfish were obtained from three local ornamental fish farms; for each size-class, 10 fish from each farm were analyzed. Hematoxylin- and eosin-stained tissue sections were analyzed by light microscopy at 100× magnification and an image analysis program. The percentage of tissue occupied by MAs, MA size, and MA number were calculated on three arbitrarily selected fields of view from each spleen and kidney. In clinically normal blue gourami, increases in the percentage of tissue occupied by MAs and in MA size were associated with an increase in fish size, but in clinically normal angelfish no correlation was observed. Furthermore, in angelfish, a high variability in MA morphometry was observed, even among fish from the same sample group. In both species, a significant difference in the value of the morphometric parameters was observed among farms. Because iridoviruses inhibit macrophage activity and (possibly) proliferation, MAs in 25 clinical cases of iridovirus-infected blue gourami were analyzed. Preliminary data indicate that in iridovirus-infected blue gourami, there is a decrease in MA size and MA number compared with those of healthy fish.     

Replacement of fish meal protein by surimi by‐product protein in the diet of blue gourami Trichogaster trichopterus fingerlings

Based on the nutrient requirement of Trichogaster trichopterus, a fish meal–based basal diet with 350 g/kg diet crude protein and 16.7 MJ/kg energy was formulated, in which the fish meal protein was replaced by surimi by‐product protein at 0.0 (control), 12.5, 25, 50, 75 and 100% levels. The formulated diets were fed ad libitum to T. trichopterus fingerlings (4.80 ± 0.03 g) in triplicate groups for 45 days in a closed water system. Eighteen fibre‐reinforced plastic tanks with 200 l of water were used for rearing the fish. Weight gain, specific growth rate, feed/gain ratio, protein efficiency ratio, nutrient retention and digestibility (protein and energy) of fish were not affected (p > 0.05) up to 50% fish meal protein replacement level by surimi by‐product protein. While whole‐body protein content of fish was marginally decreased, the lipid content was increased with increase in surumi by‐product incorporation level in the diet. The study results suggest that the fish meal protein, which is scarce and costly nowadays, could be replaced up to 50% by surimi by‐product protein in the diet of blue gourami without hampering the growth and nutrient utilization of fish.     

Effects of metomidate hydrochloride sedation on blood glucose and marketability of transported threespot gourami Trichogaster trichopterus

Our objectives were to determine whether sedation with metomidate hydrochloride (hereafter, "metomidate") during transportation of threespot gourami Trichogaster trichopterus would prevent an increase in blood glucose levels and improve fish marketability (i.e., based on appearance and behavior) in comparison with unsedated controls. Threespot gourami are obligate air-breathers that possess a labyrinth organ, enabling the fish to respire air above the water surface; these fish should be lightly sedated during transport. Fish were transported for approximately 24 h via truck and domestic airline. Blood was sampled at 0, 2, 6, and 12 h posttransport ation, and appearance and behavior were observed at 0, 2, 4, 6, and 12 h and 7 d posttransportation. Metomidate concentrations tested were 0.0 (control), 0.1, 0.2, 0.3, and 0.4 mg/L. At the concentrations tested, metomidate neither inhibited elevations in blood glucose nor improved marketability. Fish that were transported with 0.3-mg/L metomidate were less marketable based on behavioral indices, and fish that were transported with 0.4-mg/L metomidate had higher glucose levels than control fish. Use of metomidate as a transport sedative for threespot gourami should be considered with caution and may be problematic at the concentrations tested; however, further research examining additional indices of stress may clarify metomidate use for this species.     

Effect of dietary protein and lipid levels on growth,nutrient utilization and whole‐body composition of blue gourami,Trichogaster trichopterus fingerlings

Nine semi‐purified diets were prepared with three levels each of protein (300, 350 and 400 g/kg) and lipid (60, 80 and 100 g/kg) and fed ad libitum to Trichogaster trichopterus fingerlings (0.61 ± 0.03 g) in triplicate groups (10 fish/replicate) for 90 days to determine optimum dietary protein and lipid levels. Twenty‐seven flow‐through fibre‐reinforced plastic tanks (200 l capacity each with 100 l of water) were used for rearing the fish. The dietary protein, lipid and their interactions had significant effects (p < 0.05) on weight gain, feed conversion ratio, specific growth rate, nutrient retention and digestibility, but not on hepato‐ and viscerosomatic indexes (p > 0.05). Dietary protein and the interaction of protein with lipid had significant effect (p < 0.05) on whole‐body dry matter, lipid and energy contents, but not on protein and ash contents (p > 0.05). But, the dietary lipid had significant (p < 0.05) effect on whole‐body dry matter, protein, lipid and energy contents except the ash contents (p > 0.05). For each level of dietary protein, the increase in dietary lipid resulted significant increase (p < 0.05) in whole‐body lipid contents without affecting the protein and ash contents (p > 0.05). Based on better growth and dietary performances, the optimum dietary protein and lipid levels of blue gourami fingerling are 350 and 80 g/kg diet respectively.     

Differentiation and growth of myotomal muscles in a non-model tropical fish Pterophyllum scalare (Teleostei: Cichlidae)

Somite differentiation, muscle fibres formation and growth were analysed in a non-model tropical fish Pterophyllum scalare. In this study, it was found that during somite differentiation, a primary myotome appears. The primary myotome is filled with multinucleated myotubes that constitute the major part of the somite. Subsequently, Pax-3 (paired-box protein)-positive cells, located externally to the myotomes, appear. In post-hatching stages, mononucleated proliferating cell nuclear antigen-positive cells are observed in the inter-myotomal spaces and within the myotomes. The mononucleated cells, situated in the myotomes, first express desmin in their cytoplasm and then Pax-7 (paired-box protein) in their nuclei. Expression of desmin indicates that they will enter myogenic pathway, whereas expression of Pax-7 suggests their role of satellite cells. We assume that mononucleated intramyotomal cells are myogenic precursors involved in muscle growth. In advanced (post-hatching) stages of myogenesis, myotomes contain both primary and new muscle fibres. Morphometric analyses show that in Pterophyllum scalare, growth of muscle fibres is mainly a result of hypertrophy.     

Pharmacokinetics and pharmacodynamics of pantoprazole in clinically normal neonatal foals

REASONS FOR PERFORMING STUDY: Proton pump inhibitors (PPIs) are a mainstay of treatment for acid-related ulceration in man and horses. Currently, only an oral preparation of omeprazole is approved for use in horses in the USA. Intravenous administration of a PPI would provide a useful therapeutic alternative for those foals in which oral medication is not feasible. OBJECTIVE: To investigate the pharmacokinetics and pharmacodynamics of pantoprazole following i.v. or intragastric administration in healthy neonatal foals. METHODS: Seven healthy foals age 6-12 days at the start of the study were evaluated. Treatments included no drug administration, i.v. pantoprazole (1.5 mg/kg bwt) and intragastric pantoprazole (1.5 mg/kg bwt). Intragastric pH was recorded for 24 h after drug administration for pharmacodynamic evaluation. Plasma pantoprazole concentrations were measured using high-performance liquid chromatography. RESULTS: Plasma concentrations of pantoprazole were detectable at the 5 min sampling point following i.v. or intragastric administration. Bioavailability of intragastric-administered pantoprazole was 41%. Baseline mean hourly pH was 1.5-6.1. There was a statistically significant increase in mean hourly pH relative to untreated foals 2-24 h after i.v. or intragastric pantoprazole administration. CONCLUSIONS: Based on these data, i.v. or intragastric administration of pantoprazole results in a significant, prolonged increase in intragastric pH. POTENTIAL RELEVANCE: The i.v. formulation of pantoprazole may provide a clinically useful alternative means of acid suppression in foals unable to tolerate enteral administration of a PPI, such as those with pyloric outflow obstruction.     

Comparative pharmacokinetics of meloxicam in clinically normal horses and donkeys

OBJECTIVE: To determine the disposition of a bolus of meloxicam (administered IV) in horses and donkeys (Equus asinus) and compare the relative pharmacokinetic variables between the species. ANIMALS: 5 clinically normal horses and 5 clinically normal donkeys. PROCEDURES: Blood samples were collected before and after IV administration of a bolus of meloxicam (0.6 mg/kg). Serum meloxicam concentrations were determined in triplicate via high-performance liquid chromatography. The serum concentration-time curve for each horse and donkey was analyzed separately to estimate standard noncompartmental pharmacokinetic variables. RESULTS: In horses and donkeys, mean +/- SD area under the curve was 18.8 +/- 7.31 microg/mL/h and 4.6 +/- 2.55 microg/mL/h, respectively; mean residence time (MRT) was 9.6 +/- 9.24 hours and 0.6 +/- 0.36 hours, respectively. Total body clearance (CL(T)) was 34.7 +/- 9.21 mL/kg/h in horses and 187.9 +/- 147.26 mL/kg/h in donkeys. Volume of distribution at steady state (VD(SS)) was 270 +/- 160.5 mL/kg in horses and 93.2 +/- 33.74 mL/kg in donkeys. All values, except VD(SS), were significantly different between donkeys and horses. CONCLUSIONS AND CLINICAL RELEVANCE: The small VD(SS) of meloxicam in horses and donkeys (attributed to high protein binding) was similar to values determined for other nonsteroidal anti-inflammatory drugs. Compared with other species, horses had a much shorter MRT and greater CL(T) for meloxicam, indicating a rapid elimination of the drug from plasma; the even shorter MRT and greater CL(T) of meloxicam in donkeys, compared with horses, may make the use of the drug in this species impractical.     

Mycoplasmas and mites in the ears of clinically normal goats

SUMMARY Mycoplasmas were detected in the external ear canal of goats by swabbing and culture. Up to 10⁸ colony forming units were recovered from single swabs. The resulting cultures were usually mixtures of mycoplasmas containing up to 5 species. The species present in sequential swabs varied. Pathogenic species (M.agalactiae, M.capricolum, M.mycoides subsp. capri, M.mycoides subsp. mycoides of the large colony (LC) type, M.putrefaciens) were isolated from the ears and in addition 3 untyped mycoplasmas G, U and V were often present. The same mycoplasmas were found in large numbers in the mites Psoroptes cuniculi and Raillietla caprae which were sometimes present in the external ear canal. The role of the mycoplasmas in the external ear canal as a source of infection and disease and of the mites in the spread of infection requires further elucidation.     

Quantification of hepatic and renal cortical echogenicity in clinically normal cats

OBJECTIVE: To quantitatively determine echogenicity of the liver and renal cortex in clinically normal cats. ANIMALS: 17 clinically normal adult cats. PROCEDURE: 3 ultrasonographic images of the liver and the right kidney were digitized from video output from each cat. Without changing the ultrasound machine settings, an image of a tissue-equivalent phantom was digitized. Biopsy specimens of the right renal cortex and liver were obtained for histologic examination. Mean pixel intensities within the region of interest (ROI) on hepatic, renal cortical, and tissue-equivalent phantom ultrasonographic images were determined by histogram analysis. From ultrasonographic images, mean pixel intensities for hepatic and renal cortical ROI were standardized by dividing each mean value by the mean pixel intensity from the tissue-equivalent phantom. RESULTS: The mean (+/- SD) standardized hepatic echogenicity value was 1.06 +/- 0.02 (95% confidence interval, 1.02 to 1.10). The mean standardized right renal cortical echogenicity value was 1.04 +/- 0.02 (95% confidence interval, 1.01 to 1.08). The mean combined standardized hepatic and renal cortical echogenicity value was 1.02 +/- 0.05 (95% confidence interval, 0.99 to 1.04). CONCLUSIONS AND CLINICAL RELEVANCE: Quantitative determination of hepatic and renal cortical echogenicity in cats is feasible, using histogram analysis, and may be useful for early detection of diffuse parenchymal disease and for serially evaluating disease progression.     

The activity of enzymes in serum and tissues of clinically normal sheep

Extract

Feline infectious peritonitis was first described as a distinct disease entity in 1966 in the United States (Wolfe and Griesemer, 1966), although it had been observed prior to that date (Holzworth, 1963). The disease is widespread in that country (Disque et al., 1968: Hardy and O'Reilly, 1969; Ward and Pederson, 1969; Colby and Low, 1970; Colgrove and Parker, 1971) and has been recorded in Canada (Stephenson et al., 1971), England (Ingram, 1970), Ireland (Hartigan and Wilson, 1972), Japan (Konishi et at., 1971), the Netherlands (Mieog and Richter, 1971), Switzerland (Stunzi and Grevel, 1973) and most recently in Australia (Watson et al., 1974; Jones and Hogg, 1974). Two cases of feline infectious .peritonitis have, been seen in New Zealand (R. C. Gumbrell, pers. comm.). One experimental cat inoculated with peritoneal fluid from this case developed clinical signs and lesions said to be consistent with feline infectious peritonitis.     

Evaluation of intraocular pressure in eyes of clinically normal llamas and alpacas

OBJECTIVE: To estimate intraocular pressure (IOP) in eyes of healthy camelids, using applanation tonometry. ANIMALS: The eyes of 34 camelids (16 llamas [Lama glamal and 18 alpacas [L. pacos]) that did not have major abnormalities of the ocular surface or intraocular abnormalities. PROCEDURE: Tonometry measurements were obtained from each eye 3 times during a 24-hour period. Each measurement was the mean of several corneal applanations obtained by use of an applanation tonometer. Data were analyzed, using an ANOVA for a repeated-measures design. RESULTS: Mean (+/- SEM) IOP of llamas and alpacas was 13.10+/-0.35 and 14.85+/-0.45 mm Hg, respectively. Range of IOP was 7 to 18 mm Hg for llamas and 11 to 21 mm Hg for alpacas. Mean IOP of llamas was significantly less than the mean IOP of alpacas. Significant differences in IOP were not detected between the right and left eye of animals. Significant differences in IOP were not attributed to sex, age, or time of measurement within llamas or alpacas. CONCLUSIONS AND CLINICAL RELEVANCE: Establishing the mean and range of IOP of clinically normal llamas and alpacas provides a frame of reference that is important for use in a complete ophthalmic examination of camelids, which can assist clinicians in the diagnosis of glaucoma and uveitis. Reasons for the difference in mean IOP between llamas and alpacas are unknown. Although the difference may be unimportant clinically, this finding reiterates the fact that caution must be used when extrapolating IOP among species.     

Comparison of the disposition of carbimazole and methimazole in clinically normal cats

The oral disposition of the antithyroid drugs methimazole and carbimazole were compared in nine clinically normal cats. After the administration of 5 mg of methimazole, serum concentrations of methimazole increased in all the cats, with mean drug concentrations reaching peak values (1·37 μg ml⁻¹) at 30 minutes. After administration of 5 mg carbimazole, serum concentrations of carbimazole remained low, but serum methnnazole became readily measurable, with mean drug concentrations reaching peak values (0·79 μg ml⁻¹) at 120 minutes. When serum concentrations of methimazole attained after administration of the two antithyroid drugs were compared, the mean maximum serum methimazole concentration achieved after administration of methimazole was approximately two-fold higher than peak concentrations measured after administration of carbimazole. In addition, the mean area under the serum concentration curve (AUC) after administration of methimazole was approximately two-fold higher than the mean AUC determined after administration of carbimazole. When the differences in molecular weight between the two drugs was taken into consideration, however, these methimazole:carbimazole ratios of 2:1 were nearly equivalent to the molar ratio of the 5 mg doses of the drugs given (1·63). Results of this study indicate that carbimazole is nearly totally converted to methimazole after oral administration to cats, similarly to the findings in man. The finding of less available serum methimazole after administration of a 5 mg tablet of carbimazole than after methimazole is also consistent with published antithyroid drug dosages needed to control hyperthyroidism in cats.     

Composition and analysis of cerebrospinal fluid in clinically normal adult cattle.

Cerebrospinal fluid and serum were obtained from 16 clinically normal adult cows (11 dairy, 5 beef). Sodium, potassium, magnesium, total protein, and albumin concentrations, osmolality, and lactate dehydrogenase and creatine kinase activities, were quantified in CSF and serum. Total and differential cell counting, protein electrophoresis, and IgG quantification were performed on CSF. Statistical analyses of these variables, including mean, SEM, range, and 95% confidence intervals, were performed. Effects of blood contamination were evaluated, and were found to be negligible for all measured constituents. Correction factors for CSF creatine kinase and lactate dehydrogenase activities accounting for cellular contamination were developed. Total nucleated cell count was similar to counts in CSF of other species, but higher than values in healthy people. Differential leukocyte count in CSF was similar to that reported in CSF of other domestic animals: mostly lymphocytes, fewer monocytoid cells, and scant neutrophils. Cerebrospinal fluid protein concentration was higher than concentration reported for dogs, goats, and people, but was similar to values reported for horses. Beef cows had higher CSF total protein concentration than did dairy cows; also, beef cows had higher CSF gamma-globulin concentration. The concentration of sodium in CSF was slightly higher than the value in serum, and potassium concentration was lower than the value in serum. In contrast to studies of human beings, CSF osmolality was generally less than serum osmolality in the cows studied. Reference values for CSF electrolyte concentrations and osmolality are useful for diagnosis of salt poisoning and for assessment of the effects of fluid therapy. Magnesium concentration was lower in CSF, compared with serum.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cytological and bacteriological findings in guttural pouch lavages of clinically normal horses

Percutaneous washes of the guttural pouches were obtained from two groups of 15 clinically normal horses, one lightly exercised and the other heavily exercised. Microbiological and cytological studies showed a wide variation in the differential cell counts. The cytological pattern of the normal lavages (< 5 per cent neutrophils) was characterised by a large proportion of ciliated columnar epithelial cells, a few non-ciliated cuboidal epithelial cells, and less than 1 per cent monocytes, lymphocytes, and eosinophils. Abnormal lavages (with more than 5 per cent neutrophils) had higher levels of bacterial growth than normal lavages. There were significant differences between the bacterial growth and total cell count, and also between the neutrophil contents of the lavages from the two groups of horses.     

Evaluation of plasma clearance of inulin in clinically normal and partially nephrectomized cats

OBJECTIVE: To evaluate accuracy of measuring plasma clearance of inulin as an alternative renal function test for estimation of glomerular filtration rate (GFR) in cats. ANIMALS: 10 cats, first studied with intact kidneys and subsequently studied following partial nephrectomy. PROCEDURE: Clearance studies were performed in 10 clinically normal cats; those same cats then underwent partial nephrectomy, and clearance studies were performed again. Plasma concentration of inulin was determined after administration at 50 mg/kg of body weight to cats while renally intact and 45 mg/kg after the partial nephrectomy. Plasma clearance of inulin (PCin) was determined by dividing the dose by the area under the plasma inulin concentration versus time curve. Results for PCin were compared with values obtained simultaneously for urinary clearance of exogenously administered creatinine (Ccr), a widely accepted method for measurement of GFR in cats with intact kidneys and cats with reduced renal mass. RESULTS: Results of PCin were strongly correlated (r2 = 0.912, P < 0.001) with Ccr. Repeatability of determination of PCin was similar to that of Ccr. Sensitivity and specificity of PCin were superior and equivalent to that of Ccr, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Determination of PCin provides a reliable estimate of GFR in cats and is a promising alternative to determining Ccr in cats.     

Osmotic fragility of erythrocytes in clinically normal dogs and dogs infected with parasites

The osmotic fragility of the erythrocytes was measured in blood samples collected from randomly selected healthy and infected dogs at a dogs' rescue shelter. The dogs were classified into six groups on the basis of the final diagnoses from clinical, post mortem and laboratory findings. The minimum (less than 5 per cent) and maximum (more than 90 per cent) haemolysis of the erythrocytes of the clinically normal dogs (group 1), occurred in 0·60 per cent and 0·30 per cent solutions of sodium chloride (NaCI). For the non-anaemic hookworm-infected dogs (group 2a) the respective values were 0·8 per cent and 0·4 per cent NaCl, and for the anaemic hookworm-infected dogs (group 2b) they were 0·85 per cent and 0·5 per cent NaCl, respectively. The erythrocytes from dogs with Babesia canis (group 3), concurrent hookworm and B canis (group 4) and Ehrlichia canis infections (group 5) had minimum haemolysis in 0·75 per cent NaCl and maximum haemolysis at between 0·20 per cent and 0·35 per cent NaCI solutions. The derivative fragiligrams for groups 2a, 2b, 3 and 4 were shifted to the left, whereas the fragiligram for group 5 was similar to that for the clinically normal dogs (group 1). The left shift for the hookworm-infected dogs was due to the increased osmotic fragility of a minor sub-population of the erythrocytes, but for the dogs, infected with B canis major sub-populations of the erythrocytes had an increased osmotic fragility.