A rostrocaudal gradient for neurotransmitter metabolites and a caudorostral gradient for protein in canine cerebrospinal fluid

Neurotransmitter metabolites (dihydroxyphenylacetic acid [DOPAC], homovanillic acid [HVA], and 5-hydroxyindoleacetic acid [5-HIAA]) in CSF of 10 healthy dogs were evaluated with reverse-phase high-pressure liquid chromatography and electrochemical detection. Neurotransmitter metabolite concentrations determined in CSF collected from the cisterna magna were compared with those values in CSF collected from the lumbar dorsal subarachnoid space. Amounts of DOPAC (P = 0.0444), HVA (P = 0.0001), and 5-HIAA (P = 0.0316) were significantly lower in lumbar spinal fluid compared with those values in the cervical spinal fluid. Metabolite concentrations in cervical and lumbar CSF were: DOPAC = 2.81 +/- 0.73 ng/ml of CSF and 1.28 +/- 0.57 ng/ml; HVA = 98.29 +/- 12.42 ng/ml and 4.68 +/- 1.61 ng/ml; and 5-HIAA = 46.29 +/- 8.17 ng/ml and 36.96 +/- 4.07 ng/ml, respectively. Cytologic evaluations of cervical and lumbar CSF revealed a similar concentration of 3 +/- 1 WBC/microliters in both fluids. A significant (P = 0.0002) difference in protein concentration between the 2 regions was observed, with 16.1 +/- 1.8 mg of protein/dl in the cervical CSF and 27.2 +/- 2.3 mg of protein/dl in the lumbar CSF. Between the cisterna magna and lumbar dorsal subarachnoid space of dogs, a rostrocaudal gradient existed for neurotransmitter metabolites, and a caudorostral gradient existed for protein.     

Analysis of equine cisterna magna cerebrospinal fluid for the presence of some monoamine neutrotransmitters and transmitter metabolites

Small volumes (0.05 ml) of cisterna magna cerebrospinal fluid (CSF) from 23 neurologically normal horses were analysed for the monoamine neurotransmitters dopamine, norepinephrine, epinephrine, serotonin and their metabolites using high pressure liquid chromatography and electrochemical detection. Two metabolites, homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA) were present in all CSF samples. The deaminated and methylated metabolite of dopamine, HVA, was present at a mean concentration of 42.33±3.14 ng/ml of CSF. The deaminated metabolite of serotonin, 5-HIAA, was present at a mean concentration of 45.52±3.65 ng/ml of CSF. A positive correlation was found between the CSF concentrations of HVA and 5-HIAA (r=0.72, p=0.0003). The mean ratio of HVA to 5-HIAA concentrations was 1.07±0.05. The ratios of HVA to 5-HIAA concentrations were found to be more indicative of the serotonergic metabolite 5-HIAA (r=–0.62, p=0.0016) than the dopaminergic metabolite HVA (r=0.11, p=0.60). There was a stronger relationship between the concentrations of 5-HIAA and the ratios of HVA to 5-HIAA in male CSF (r=–0.85, p=0.0006) than in female CSF (r=–0.54, p=0.01).Concentrations of CSF neurotransmitter metabolites were evaluated in horses grouped by age (0–4, 5–9 and 10–13 years). The youngest group of horses had a significantly (p=0.003) greater CSF concentration of HVA than of 5-HIAA. The 10–13-year-old horses had a significantly (p=0.02) lower mean CSF HVA concentration than the 0–4-year-old horses. No age-related differences in CSF 5-HIAA concentrations were detected.     

Different gradients for neurotransmitter metabolites and protein in horse cerebrospinal fluid

The serotonin metabolite, 5-hydroxyindoleacetic acid (5-HIAA) and the dopamine metabolite, homovanillic acid (HVA) in the cerebrospinal fluid (CSF) of seven clinically normal horses were evaluated with reverse phase high pressure liquid chromatography and electrochemical detection. Comparisons of the neurotransmitter metabolite concentrations were made on CSF collected simultaneously from the atlanto-occipital and lumbosacral regions. There were significantly higher amounts of 5-HIAA and HVA in atlanto-occipital CSF than in lumbosacral CSF. Mean 5-HIAA concentrations in atlanto-occipital and lumbosacral CSF were 206.1±37.0 nmol/L and 68.5±12.6 nmol/L, respectively. The mean HVA concentration in atlanto-occipital CSF was 254.4±50.4 nmol/L. No HVA was detected in lumbosacral CSF. In contrast, significantly greater protein concentrations were present in lumbosacral CSF than in atlanto-occipital CSF. Lumbosacral and atlanto-occipital CSF contained 32.1±3.7 mg protein/dl and 25.0±1.3 mg protein/dl, respectively. Thus, between the atlanto-occipital and lumbosacral space of the horses, a rostrocaudal gradient existed for neurotransmitter metabolites and a caudorostral gradient existed for protein.     

The effect of acute restraint stress on regional brain neurotransmitter levels in stress-susceptible pietrain pigs

Concentrations of noradrenaline (NA), adrenaline (A), dopamine (DA), 5-hydroxytryptamine (5-HT), the DA metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) and the main 5-HT metabolite 5-hydroxyindole-3-acetic acid (5-HIAA) were measured using HPLC in 16 brain areas of control and immobilized Pietrain pigs. The animals were immobilized for 15, 30 and 60 min in the prone position. Control pigs showed patterns of regional distribution of brain monoamines similar to those described for rats, dogs and Duroc pigs. However, the absolute values of NA and A in the hypothalamus and preoptic area were much higher than those in rats and dogs, but similar to those in Duroc pigs. The concentrations of dopamine and its metabolites DOPAC and HVA were highest in the caudate nucleus, the nucleus accumbens, the olfactory tubercle and the ventral tegmental area. The distributions of serotonin and its metabolite 5-HIAA were similar in all examined structures. DOPAC/DA and HVA/DA ratios were higher in the cornu ammonis, the hippocampus posterior and the raphe nuclei than in other structures, which suggests brain structure-related differences in dopamine turnover. The greatest decreases in NA and A content were observed in the hypothalamus, the preoptic area and the olfactory tuberculum during the first 30 min of immobilization stress. 5-HT turnover was increased in the raphe nuclei, while DA turnover was affected in the raphe nuclei, the septum, the substantia nigra and the olfactory tubercle. We suggest that acute immobilization stress caused differences in regional patterns of brain biogenic amines, thereby maintaining adequate transmitter levels during stress in stress-susceptible pigs.     

Analysis of cat retina for dopamine,dihydroxyphenylacetic acid, 3-methoxytyramine and homovanillic acid

Twenty retinas from 10 cats were evaluted for dopamine (DA) and its metabolites dihydroxyphenylacetic acid (DOPAC), 3-methoxytyramine (3-MT) and homovanillic acid (HVA) by high pressure liquid chromatography and electrochemical detection. Dopamine was present in all 20 retinas at a mean concentration of 3.00±0.54 ng/mg protein. Dihydroxyphenylacetic acid, 3-MT and HVA were detected in 16, 14 and 9 retinas respectively.In retinas in which these metabolites were detectable, they were present in the following mean concentrations: DOPAC, 1.07±0.21 ng/mg protein; 3-MT, 3.44±0.97 ng/mg protein and HVA, 4.54±1.05 ng/mg protein. Significantly higher concentrations of 3-MT (p=0.0108, paired t test) and HVA (p=0.0166, paired t test) than DOPAC were present in cat retina. Linear correlation analysis between DA and its metabolites indicated that the 3-MT and DOPAC concentrations correlated well with each other and with the amount of DA in cat retina. The concentrations of the end product metabolite, HVA, had poor correlations with the concentrations of 3-MT, DOPAC or DA. These data indicated that once DA is released in cat retina it can be metabolized to 3-MT, DOPAC and HVA.     

Determination of homovanillic acid, 5-hydroxyindoleacetic acid and pressure in the cerebrospinal fluid of Collie dogs following administration of ivermectin

Twelve adult Collie dogs were studied to determine the effects of ivermectin on neurotransmitter metabolites released from the brain into the cerebrospinal fluid (CSF) and on CSF pressure. Ten of the 12 Collies were given ivermectin orally at a concentration of 200 g/kg body weight. Three of these 10 Collies showed clinical signs of ivermectin-induced toxicosis which progressed into a state of unresponsive recumbency in 2 dogs.Cerebrospinal fluid pressures and neurotransmitter metabolite concentrations in cisterna magna CSF were analysed 49 to 50 hours after administration of ivermectin in 6 of the 10 treated dogs, and in the 2 untreated control Collies. Cerebrospinal fluid pressures were within normal limits in all dogs. However, compared to the CSF concentrations in the 2 untreated and 3 non-reactive Collies, the 2 ivermectin-reactive Collies still in recumbency had elevated CSF concentrations of homovanillic acid (HVA), a metabolite of dopamine, and 5-hydroxyindoleacetic acid (5-HIAA), a metabolite of serotonin. These findings suggest an association between altered neurotransmission and severe ivermectin-induced neurological abnormalities. No evidence of elevated intracranial pressure was found.     

Cerebrospinal fluid S-100B concentrations in normal and diseased cattle

We measured the concentrations of S-100B, a marker protein used in humans to detect brain damage, in the cerebrospinal fluid (CSF) of clinically normal cattle (n=15, mean age +/- SD: 31.8 +/- 37.5 months) and of cattle with various inflammatory disorders (n=43, 70.6 +/- 31.9 months). The mean +/- SD CSF S-100B level was 2.9 +/- 1.6 ng/ml in the normal group and 7.0 +/- 7.4 ng/ml in the diseased group. Thirteen diseased cattle that had developed no obvious neurological signs showed abnormally high S-100B concentrations (> 8.0 ng/ml), whereas the two cattle with neurological disorders did not. No particular disease could be related to the S-100B rise. Therefore, it remains inconclusive whether measurement of CSF S-100B concentration is useful in veterinary neurological diagnosis.     

Effects of endophyte-infected fescue on concentrations of prolactin in blood sera and the anterior pituitary and concentrations of dopamine and dopamine metabolites in brains of steers

An experiment was conducted to determine if the decrease in circulating concentrations of prolactin in cattle consuming endophyte (Acremonium coenophialum) -infected tall fescue (Festuca arundinacea) was associated with changes in prolactin concentrations in the anterior pituitary and concentrations of dopamine (DA) and its metabolites 3,4-dihydroxyphenyl-acetic acid (DOPAC) and homovanillic acid (HVA) in the stalk median eminence (SME), preoptic area (POA) and hypothalamus (HP). Six crossbred steers that grazed high-endophyte (greater than 90% infected) fescue and four steers that grazed low-endophyte (less than 1% infected) fescue from April to September were slaughtered. Brains and pituitaries were removed and dissected. Extracts from neural tissue were analyzed for DA, DOPAC and HVA using high performance liquid chromatography/electrochemical detection. Pituitary extracts and sera from blood samples taken 5 d prior to slaughter were subjected to prolactin radioimmunoassay. Consumption of high-endophyte fescue was associated with decreased concentrations of prolactin in serum (P less than .01) and in the anterior pituitary (P = .08), decreased (P less than .05) concentrations of DA in the SME and decreased (P less than .01) concentrations of HVA in the POA and HP, but it did not influence levels of DOPAC. These results suggest that endophyte toxins may reduce prolactin synthesis and release and may alter activity of dopaminergic neurons.     

Pharmacokinetics of Selamectin in Dogs after Topical Application

Some pharmacokinetic parameters of selamectin were determined in male (n = 5) and female (n = 5) Beagle dogs following a topical application at a dose rate of 6 mg/kg. The plasma concentration versus time data for the drug were analysed using a one-compartment model. The maximum plasma concentrations of 12.72 +/- 5.13 ng/ml for males and 22.65 +/- 11.95 ng/ml for females occurred around 5 days after administration. The area under the concentration-time curve (AUC) was 192.08 +/- 63.85 ng.day/ml for males and 370.97 +/- 146.87 ng.day/ml for females. The mean residence time was the same in males and females (12.55 days). This study reveals a sex-influence on the disposition of selamectin in the plasma of dogs, which implies that further information will be needed for correlation with efficacy studies in dogs.     

Differences in catecholamine metabolism and behaviour in neonatal broiler and layer chicks

1. To clarify the difference in behavioural activities and catecholamine metabolism between layer and broiler-type chicks two experiments were conducted. 2. In experiment 1, 1-d-old male layer and broiler chicks were placed in an open-field area and their responses were investigated for 10 min. The responses of the two strains were remarkably different, with broilers being less active than layers. Vocalisations rapidly decreased in broilers whereas those of layers remained elevated during the 10 min. 3. In experiment 2, 1-d-old chicks of both strains were killed and brain catecholamine concentrations were determined in three parts of the brain: telencephalon, optic lobe and brain stem. 4. In the whole brain, dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) were significantly higher in broilers. However, the values for norepinephrine (NE), epinephrine (E) and 4-hydroxy-3-methoxyphenylacetic acid (HVA) were similar between strains. The ratios of metabolite/precursor were also calculated: HVA/DOPAC was higher in layers, while NE/DA, E/NE and DOPAC/DA were not significantly different between strains. 5. These results suggest that behavioural activities differ greatly, while there are some differences in catecholamine metabolism between the two strains.     

Cerebrospinal fluid glutamine,tryptophan, and tryptophan metabolite concentrations in dogs with portosystemic shunts

OBJECTIVE: To determine whether glutamine (GLN), tryptophan (TRP), and tryptophan metabolite concentrations are higher in cerebralspinal fluid (CSF) dogs with naturally occurring portosystemic shunts (PSS), compared with control dogs. ANIMALS: 11 dogs with confirmed PSS and 12 control dogs fed low- and high-protein diets. PROCEDURE: Cerebrospinal fluid and blood samples were collected from all dogs. Serum and CSF concentrations of GLN, alanine, serine, TRP, 5-hydroxyindoleacetic acid (5-HIAA), and quinolinic acid (QUIN) were measured. RESULTS: Cerebrospinal fluid concentrations of GLN, TRP, and 5-HIAA were significantly higher in PSS dogs, compared with control dogs fed high- or low-protein diets. Cerebrospinal fluid QUIN concentration was significantly higher in PSS dogs, compared with control dogs fed the low-protein diet. Serum QUIN concentration was significantly lower in PSS dogs, compared with control dogs fed either high- or low-protein diets. CONCLUSIONS AND CLINICAL RELEVANCE: An increase in CNS GLN concentration is associated with high CSF concentrations of TRP and TRP metabolites in dogs with PSS. High CSF 5-HIAA concentrations indicate an increased flux of TRP through the CNS serotonin metabolic pathway, whereas high CSF QUIN concentrations indicate an increased metabolism of TRP through the indolamine-2,3-dioxygenase pathway. The high CSF QUIN concentrations in the face of low serum QUIN concentrations in dogs with PSS indicates that QUIN production from TRP is occurring in the CNS. High concentrations of QUIN and other TRP metabolites in the CNS may contribute to neurologic abnormalities found in dogs with PSS and hepatic encephalopathy.     

Experimental and clinical studies on plasma leptin in obese dogs

Leptin is a protein synthesized and secreted primarily by adipocytes, and the circulating leptin concentration is elevated in obese humans and rodents. Recently, we have established a sandwich enzyme-linked immunosorbent assay for canine leptin. In the present study, plasma leptin concentrations were measured in experimentally developed obese beagles and in clinically obese dogs. When 5 male beagles were given a high-energy diet for 3 months, all of them became obese and the plasma leptin concentration significantly increased from 2.4+/-1.2 to 4.9+/-0.9 ng/ml, positively correlating with body fat content estimated by the deuterium oxide dilution method (r=0.87). The leptin concentrations of plasma samples collected from 59 dogs in veterinary practices were compared with their body condition scores (BCS). The plasma leptin concentrations of obese dogs were 9.7+/-0.7 and 12.3+/-1.5 ng/ml at BCS=4 and BCS=5, respectively, which were significantly higher than those of optimal (BCS=3) dogs (2.7+/-0.3 ng/ml). There was no significant effect of sex and breed. A weak positive correlation (r=0.37) was found between the plasma leptin concentration and age, probably due to the lesser content of visceral fat in puppies younger than 1 year old. These results indicate that plasma leptin is a good index of adiposity in dogs regardless of breed, age and sex, and may be useful for quantitative assessment of obesity in small animal practice.     

Prepartum changes of plasma concentrations of prostaglandin F and 13,14-dihydro-15-ketoprostaglandin metabolites in pregnant animals exposed to Sarcocystis cruzi or Campylobacter fetus

Pregnant cows at 4- to 5-months' of gestation were exposed to Sarcocystis cruzi or Campylobacter fetus. Plasma prostaglandin F (PGF) and 13,14-dihydro-15-ketoprostaglandin metabolite (PGM) concentrations were determined at intervals from before exposure until abortion or parturition. The plasma PGF concentration of pregnant infected cattle remained at 0.02 +/- 0.04 ng/ml until 24 to 48 hours before abortion or parturition when it increased 5-fold to 0.11 +/- 0.12 ng/ml. The plasma PGM concentration of these cattle remained at 0.10 +/- 0.07 ng/ml until 24 to 48 hours before abortion or parturition when it increased over 10-fold to 1.36 +/- 0.60 ng/ml. This change in PGF and PGM was similar to that of cattle exposed to each of the infective agents and to that of normal cows at parturition. Thus, changes in PGF and PGM concentrations in bovine plasma cannot be used as a diagnostic tool to determine fetal distress or fetal death for these infections.     

Thyroid hormones in the cerebrospinal fluid of the third ventricle of adult female sheep during different periods of reproductive activity

Thyroid hormones (THs) are obligatory for transition from breeding season to anestrus in sheep. In this process, THs act during a very limited time of the year and primarily within the brain. In ewes chronically equipped for sampling cerebrospinal fluid (CSF) from the third ventricle, we have characterized the concentrations of total and free thyroxine (T4), triiodothyronine (T3), and total reverse T3 (rT3) in the CSF during breeding season, anestrus and during a critical period required for transition to anestrus (December-March). The total T4, T3, rT3 and free T3 average concentrations (+/- SEM) in CSF were 1.5 +/- 0.07 ng/ml, 14.5 +/- 1.2 pg/ml, 43 +/- 7.4 pg/ml, and 0.6 +/- 0.05 pg/ml, respectively, and all were significantly lower (p < 0.001) than in blood plasma except free T4 (12.6 +/- 1.1 pg/ml), which was similar to that in plasma. There was a seasonal trend (p < 0.05) in the concentration of total T3 (highest in December) and free T4 (highest in November) in the CSF that does not follow that in blood plasma. During the period of transition to anestrus the CSF total T3/TT4 molar ratio and free T3/T4 ratio were significantly lower (p < 0.05 and p < 0.01, respectively) than in blood plasma, while the total rT3/T4 ratio was significantly higher (p < 0.01) at the end of this period (March). Additionally, the CSF total rT3 concentrations were also significantly correlated with the CSF total T4 levels (r = 0.57; p < 0.05). In conclusion, the CSF in sheep may serve as a considerable source of thyroid hormones for neuroendocrine events. The lack of significant changes in THs concentrations in the CSF during the period of transition to anestrus indicate that neither seasonal changes of THs circulating in the blood plasma nor THs circulating in the CSF actively drive the transition to anestrus.     

Measurement of thyroid hormones (thyroxine, T4; triiodothyronine, T3) in captive nondomestic felids.

The aim of this research was to obtain basic values for the evaluation of thyroid function in nondomestic felids. Serum thyroid hormone concentrations (thyroxine, T4; triiodothyronine, T3) were measured by radioimmunoassay in 145 cats, representing nine species of captive nondomestic felids: jaguar (Panthera onca), n = 49; puma (Puma concolor), n = 10; ocelot (Leopardus pardalis), n = 22; oncilla (Leopardus tigrinus), n = 12; geoffroy (Oncifelis geoffroyi), n = 4; jaguarundi (Herpailurus yaguarondi), n = 8; margay (Leopardus wiedii), n = 7; lion (Panthera leo), n = 26; and tiger (Panthera tigris), n = 7. For each species, mean +/- SEM of T3 and T4, respectively, were as follows: jaguar, 0.56 +/- 0.03 and 9.7 +/- 0.8 ng/ml; puma, 0.67 +/- 0.04 and 11.2 +/- 1.2 ng/ml; ocelot, 0.48 +/- 0.03 and 13.8 +/- 1.5 ng/ml; oncilla, 0.43 +/- 0.03 and 10.0 +/- 1.6 ng/ml; geoffroy, 0.44 +/- 0.04 and 8.0 +/- 0.16 ng/ml; jaguarundi, 0.7 +/- 0.03 and 5.0 +/- 1.0 ng/ml; margay, 0.48 +/- 0.04 and 12.2 +/- 2.3 ng/ml; lion, 0.43 +/- 0.02 and 5.7 +/- 2.6 ng/ml; and tiger, 0.66 +/- 0.03 and 12.6 +/- 0.9 ng/ml. Within species, T3 and T4 concentrations did not differ (P > 0.05) between males and females.     

Endocrine patterns in two strains of Japanese black cattle with growth retardation

Endocrine patterns were compared in 2 strains of Japanese black cattle with growth retardation; MHO- and HSK-paternal strains (MHO and HSK cattle, respectively). MHO cattle (n=8) displayed lower serum concentrations of insulin-like growth factor-1 (IGF-1), triiodothyronine (T3), thyroxine (T4), and cortisol (31.1+/-20.7 ng/ml, 73.9+/-51.9 ng/dl, and 2.9+/-2.9 microg/dl, 1.3+/-0.7 microg/dl, respectively) than those in both HSK cattle (n=5) (64.9+/-47.6 ng/ml, 97.8+/-40.7 ng/dl, 4.1+/-2.1 microg/dl and 1.8+/-1.1 microg/dl, respectively), and the controls (n=6) (314.7+/-197.2 ng/ml, 140.2+/-21.3 ng/dl, 5.8+/-1.7 microg/dl, and 3.0+/-1.4 microg/dl, respectively). The area under the concentration curve of growth hormone (GH-AUC 0-600 min) in MHO cattle (22210+/-18951 ng.min/ml) tended to be greater than those in HSK (7887+/-6340 ng.min/ml) and the controls (2811+/-1275 ng.min/ml). MHO cattle showed a high GH-AUC0-600 min in contrast to a low serum IGF-1 concentration, as well as lower serum T3, T4, and cortisol concentrations. HSK cattle exhibited the same secretory patterns, but much more moderately. Growth retardation in Japanese black cattle exhibits some variations based on pedigree.     

Cerebellum progesterone concentration decreased in canine distemper virus infection

Progesterone has neuroprotective effects including augmentation of myelination in the central and peripheral nervous system. This study was designed to determine if demyelinating lesions in the cerebellum resulting from canine distemper virus (CDV) infection are associated with progesterone levels. Progesterone was measured using radioimmunoassay in samples of the cerebellum, corpus callosum, medulla oblongata, parietal, frontal, temporal, and occipital cortices as well as cerebrospinal fluid (CSF) and plasma collected from ten CDV infected and six non-infected dogs. The cerebellum progesterone level was significantly different between CDV infected (0.66+/-0.09 ng/g) and control dogs (1.14+/-0.09 ng/g) (p<0.001); however, no difference was observed for the other CNS regions, plasma and CSF (p>0.05). The cerebellum progesterone level was also significantly different between acute (0.71+/-0.0 5 ng/g) and chronic cases (0.61+/-0.09 ng/g) (p<0.05). The CDV infected cerebella were also categorized histopathologically according to the severity of demyelinating lesions as mild (n=5), moderate (n=2), or severe (n=3) among which the cerebellum progesterone level was significantly different (p<0.05). Progesterone concentration was 0.71+/-0.05 ng/g in mild, 0.65+/-0.10 ng/g in moderate, and 0.56+/-0.07 ng/g in severe cases. In conclusion, progesterone concentration decreases in the cerebellum in CDV infection and the severity of demyelinating lesions is the greatest in cerebella with the lowest progesterone concentrations. The results suggest that local impairment of progesterone metabolism may be associated with the initiation and progression of cerebellar lesions in CDV infection.     

Relationships between force-feeding and some physiological parameters in geese bred for fatty liver

The susceptibility of geese of different genotypes and sexes to force-feeding, some plasma biochemical parameters (thyroid hormones, cholesterol, retinoids, total protein and albumin) of force-fed geese, and the relationship between force-feeding, fat storage and the above-mentioned parameters were studied. Sixty (30 male and 30 female) geese of three genotypes (Hungarian, Landes and their crossbred called Babat Hybrid) were divided in two groups at 12 weeks of age. Geese in one group (5 males and 5 females from each genotype) received mixed feeding ad libitum. Birds in the other group were force-fed with maize. After 3 weeks all birds were bled, blood samples were taken, and the above-mentioned plasma parameters were determined. Thyroxine (T4) levels were significantly lower in force-fed (11.6 +/- 3.5 ng/ml) than in control geese (22.7 +/- 4.09 ng/ml). Plasma triiodothyronine (T3) level was also lower in the force-fed than in the control group, but the difference was not significant (1.87 +/- 0.23 ng/ml and 2.11 +/- 0.28 ng/ml, respectively). Plasma total protein (TP, 45.2 +/- 4.5 g/l), albumin (ALB, 16.51 +/- 2.8 g/l), beta-carotene (BC, 3504 +/- 3107 micrograms/l), retinol (ROL, 1160 +/- 505 micrograms/l), retinyl palmitate (RP, 1745 +/- 405 micrograms/l) and total cholesterol (TCh, 4.32 +/- 0.55 mmol/l) levels were elevated in the force-fed group as compared to the control (TP = 36.4 +/- 5.1 g/l, ALB = 15.6 +/- 0.9 g/l, BC = 1657 +/- 1681 micrograms/l, ROL = 687 +/- 375 micrograms/l, RP = 1398 +/- 607 micrograms/l, and TCh = 2.83 +/- 1.98 mmol/l). All differences were significant except those found for albumin and beta-carotene. No significant sex- or genotype-related effects were observed for these parameters.     

Plasma concentrations of folic acid, vitamin B12 and progesterone of cyclic bitches, bitches during pregnancy and induced abortion and bitches with pyometra

The aim of this study was to investigate the plasma concentrations of folic acid, vitamin B12 and progesterone at different stages of the sexual cycle and pregnancy, during induced abortion and in bitches with pyometra. Bitches (n = 97) were assigned to groups as follows: a) oestrous cycle (n = 42) b) pregnancy (n = 25) c) induction of abortion (n = 10 and d) pyometra (n = 20). Oestrous cycle stages were determined by vaginal inspection and cytology. Pregnancies were estimated by ultrasound (5.0 Mhz; linear transducer; Schimadzu) at days 15-25, 35-45 and 46-63 of pregnancy. Treatments for the induction of abortion were started between days 25 and 35 after mating (5 microg/kg cabergoline daily, Galastop; 5-10 microg/kg Alfaprostol every other day, Gabbrostim). Diagnosis of pyometra was confirmed by ultrasound and vaginoscopy. Folic acid and vitamin B12 concentrations did not differ among different stages of the oestrous cycle. The mean concentration of folic acid during early pregnancy (days 15-25) exceeded levels of later stages (days 46-63): 9.4 +/- 3.7 microg/ml and 4.7 +/- 1.8 microg/ml, respectively (p < 0.01). A positive correlation between folic acid and vitamin B12 was determined in pregnant dogs ( r = 0.925; p < 0.02). Before the induction of abortion, the concentration of folic acid was 9.6 +/- 5.2 microg/ml; during abortion it decreased to 5.0 +/- 3.2 microg/ml (p < 0.01). A significant correlation (r = 0.925; p < 0.02) between progesterone and folic acid was obtained in bitches with abortion. The mean concentration of folic acid in bitches with pyometra significantly differed from that of bitches at different stages of the oestrous cycle (p < 0.05). The mean concentration of folic acid was significantly lower in metoestrous bitches when compared to bitches with pyometra (p < 0.05). The decrease of serum concentrations of folic acid during pregnancy and induced abortion show that fetal growth and abortion caused higher consumption of folic acid. Concerning bitches did not show any deficiency symptoms, which is why it can be concluded that this decrease is physiological.     

Pharmacokinetics of flunixin in mature heifers following multiple intravenous administration

The pharmacokinetics of flunixin meglumine was determined after its multiple (altogether 4 doses at 24-hours intervals) intravenous administration at a dose of 2.2 mg/kg body weight in six mature clinically healthy heifers. Plasma flunixin and its metabolite 5-hydroxyflunixin concentrations were analyzed with high-pressure liquid chromatography using an assay with a lower limit detection of 0.03 microg/ml for both substances. Plasma concentrations versus time curves were described by a two compartment open model. Mean plasma flunixin concentrations were similar on day 1 and 4, and than rapidly decreased (within 2 hours) from initial concentrations higher than 10 microg/ml to the concentrations lower than 1 microg/ml. The distribution phase of flunixin was short (t0.5 alpha = 0.29 +/- 0.16 and 0.18 +/- 0.04 on day 1 and 4, respectively) and the elimination phase was more prolonged (t0.5 beta = 3.30 +/- 0.60 and 3.26 +/- 0.22 on day 1 and 4, respectively). The mean residence time of flunixin was similar on day 1 (1.83 +/- 0.83) and 4 (1.88 +/- 0.46), and for 5-hydroxyflunixin this parameter was insignificantly (P > 0.05) higher on day 1 (5.49 +/- 2.22) as compared to that found on day 4 (3.99 +/- 2.17). The clearance of flunixin was similar on both examined days (0.23 +/- 0.12 on day 1 and 0.31 +/- 0.15 on day 4), and for 5-hydroxyflunixin was insignificantly (P > 0.05) lower on day 1 (2.37 +/- 1.21) as compared to that determined on day 4 (3.23 +/- 1.06). Our data indicate that multiple administration of flunixin did not alter significantly the parent drug and its metabolite concentrations in plasma, however may cause some small changes in pharmacokinetic parameters.