Milk Protein Polymorphism in Kangayam Cattle

This paper reports the milk protein polymorphism, the allele frequencies of variants and the possible linkages among various combinations of milk protein phenotypes in the Kangayam cattle of south India. Milk samples from 156 Kangayam cows were typed by starch gel and polyacrylamide gel electrophoresis for caseins and whey proteins, respectively. All the four milk protein components studied, _s1-casein, -casein, -lactoglobulin and -lactalbumin, exhibited polymorphism with high allele frequencies of 0.9231±0.0151 for _s1-casein C, 0.9263±0.0148 for -casein A, 0.9135±0.0159 for -lactoglobulin B and a relatively high frequency of 0.6218±0.0275 for -lactalbumin A. The mean heterozygosity estimated over all the four milk protein loci was 0.2420. Genetic equilibrium was observed among all the loci studied, except -lactalbumin. Linkage analysis confirmed the non-independence between _s1- and -caseins and between caseins and -lactalbumin phenotypes.     

Plasma,Urinary and Biliary Residues in Cattle Following Intramuscular Injection of Nortestosterone Laurate

The synthetic androgen 19-nortestosterone (-NT) has been used illegally as a growth promoter in cattle production in the European Union. Elimination of -NT and its metabolites in plasma, urine and bile was studied in three cattle with cannulated gallbladders following intramuscular injection at a single site of 500 mg of the laurate ester (NTL) containing 300.5 mg -NT. Using enzyme immunoassay quantification, plasma Cmax of free -NT was 0.5±0.15 g/L (mean±SEM). Concentrations of free -NT in plasma were consistently greater than the assay limit of quantification (0.12 g/L) for 32.7±13.42 days. Mean residence time for free -NT in plasma was 68.5±20.75 days. Following sample preparation by immunoaffinity chromatography, high-resolution GC-MS was used to quantify -NT and -NT in urine and bile. -NT was detected irregularly in urine from two of the three animals post injection. The principal metabolite present in the urine, -NT, was detected for 160.3±22.67 days post injection. Cmax for -NT in urine was 13.7±5.14 g/L. Mean urinary AUC0–183 days for -NT was 845.7±400.90 (g h)/L.In bile, -NT was the only metabolite detected for 174.3±8.67 days post treatment. Cmax for -NT in bile was 40.8±12.70 g/L and mean biliary AUC0–183 days for -NT was 1982.6±373.81 (g h)/L. Concentrations of -NT in bile samples were greater than those in urine samples taken at the same time. The mean ratio of biliary:urinary AUC0–183 days was 3.0±0.72. It is concluded that bile is a superior fluid for detection of -NT following injection of NTL, owing to the longer period during which residues may be detected after administration.     

Leukocyte and Cytokine Accumulation in the Ovine Teat and Udder during Endotoxin-Induced Inflammation

Persson Waller, K., Colditz, I.G., Flapper, P. and Seow, H.-F., 1997. Leukocyte and cytokine accumulation in the ovine teat and udder during endotoxin-induced inflammation. Veterinary Research Communications, 21 (2), 101-115The accumulation of leukocytes, ovine serum albumin and the cytokines interleukin-1 (IL-1), tumour necrosis factor- (TNF-), interleukin-8 (IL-8), granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon- (IFN-) was studied during endotoxin-induced inflammation in lactating and dry ovine udders, and in the teat cisterns of dry ewes after surgical closure of the passage between the teat and udder cisterns. Samples were taken before infusion and hourly up to 10 h after infusion of 0.1, 1 or 10 µg of endotoxin, or infusion of pyrogen-free saline (PFS) as a control. Rectal temperatures were measured.A significant dose- and time-dependent accumulation of leukocytes, mainly neutrophils, was observed in the lactating udders and in the teat cisterns. In the dry udders, the leukocyte accumulation was significant for time but not for dose. Peak numbers of cells were reached at 3-4 h in the dry udders and in the teat cisterns, but not until 10 h after infusion in the lactating udders. The changes in the ovine serum albumin concentrations mostly paralleled changes in leukocyte numbers.A role was indicated for TNF-, IL-8 and GM-CSF, but not for IL-1 and IFN-, during endotoxin-induced inflammation in the ovine udder. Release of TNF-, IL-8 and GM-CSF was most prominent in lactating udders, peaking at 2 or 3 h after infusion, but was also detected in dry udders and teat cisterns. Detectable levels of IL-1 and IFN- were occasionally found in all three groups.     

Pharmacokinetics and distribution of sulphadimidine in plasma,milk and uterine fluid of female buffaloes

The pharmacokinetics of sulphadimidine after a single dose (200 mg/kg i.v.) was studied in five healthy lactating buffaloes. The study revealed that the drug attained its peak concentration of 314.0±13.0, 242.4±3.0 and 100.2±2.5 g/ml at 15 min, 30 min and 12 h in plasma, milk and uterine fluid, respectively. The pharmacokinetic parameters calculated by a 2-compartment open model gave values for t₁, t₁ and vd_area of 2.10±0.36 h, 12.36±0.57 h and 1.23±0.07 L/Kg, respectively. A high vd_area as well as a value of 0.74±0.08 for K₁₂:K₂₁- (tissue Plasma) indicates better penetration of the drug into the different body fluids and tissues, which is further supported by a high concentration obtained in milk and uterine fluid. The therapeutic concentration (50 g/ml) was maintained for around 24 h in plasma and milk and 12 h in uterine fluid. The results suggest that, apart from its use in systemic infections, the drug can be effectively used by the i.v. route in uterine and mammary gland infections. The dosages for maintaining concentration of 50 g/ml, 100 g/ml and 150 g/ml at convenient dosage intervals of 12 and 24 h were also determined.     

The pharmacokinetics of fenbendazole and oxfendazole in cattle

The pharmacokinetics of fenbendazole and oxfendazole in cattle are described. The pharmacokinetics of oxfendazole were not significantly different when administered orally and by intra-ruminal injection. At a dose rate of 4.5 mg/kg, administered orally, fenbendazole gave rise to mean peak concentrations in plasma of fenbendazole and oxfendazole of 0.11 and 0.13 g/ml respectively. Oral administration of oxfendazole, at 4.5 mg/kg body weight, gave rise to plasma peak concentrations of fenbendazole and oxfendazole of 0.10 and 0.20 g/ml respectively. Following intra-ruminal administration of oxfendazole, the peak concentrations were 0.11 and 0.18 g/ml respectively.     

Evaluation of Synchronization of Oestrus Based on Gonadotrophin-releasing Hormone and Its Potential Use for Fixed-Time Breeding in Tuli Beef Cows

The objective of the experiment was to compare the reproductive post-partum performance of beef cows synchronized for oestrus using prostaglandin F₂ (PGF₂) alone or with a gonadotrophin-releasing hormone (GnRH)-based drug. Fifty-five post-partum lactating Tuli cows were randomly allocated to three groups. Two groups were synchronized using either two injections of PGF₂ (500 g Prosolvin per injection) given 11 days apart (group 1), or GnRH (12.5 g Receptal per injection) followed 6 days later by an injection of 500 g PGF₂ (group 2). The cows were bred by artificial insemination 12 h after they were observed in oestrus. Group 3 was synchronized as for group 2, but a second injection of GnRH was given 54 h after the PGF₂ injection, at which time the cows were bred by artificial insemination (AI) without detection of oestrus. Blood samples were taken from the cows in group 3 and analysed for progesterone concentration to establish which cows were cycling and in oestrus before and at the time of breeding. Detection of oestrus and breeding by AI was done over 60 days. There were no significant differences (p>0.05) among the three groups in the first service and total conception rates. The percentage of cows in oestrus within 10 days of the synchronization treatment was not significantly different (p>0.05) between groups 1 and 2. The progesterone concentrations in the cows in group 3 showed that only those that were cycling at the start of the experiment responded to the synchronization treatment and conceived after fixed-time breeding. These results suggest that combinations of PGF₂ and GnRH may be of value in synchronizing oestrus and controlling breeding in Tuli cows. However, the benefit might be greater if only cows that are known to be cycling are bred in this way.     

Uterine motility of the sow during the oestrous cycle and early pregnancy

With electromyography and strain gauges the uterine motility of the sow during the oestrous cycle and early pregnancy was studied. Special attention was paid to characterization of myometrial activity at the time of intrauterine migration of blastocysts.From recordings of 4 animals (3 became pregnant) 3 types of electrical spiking activity (each could be correlated with an elevation of the strain gauge tension) were discerned. Two of them appeared regularly and were suitable for analysis: High Voltage Slow Acticity (with relative high amplitude and low frequency of spikes) and Low Voltage Fast Activity (with relative low aplitude and high frequency of spikes).The sexual status markedly influenced myometrial activity: during prooestrus and oestrus only one type of electrical activity was present whereas the myoelectrical complex (an episode of this activity and the subsequent interval of inactivity) was shorter than 10 min. During dioestrus the myoelectrical complex was longer than 10 min and High Voltage Slow Activity (solely on the cervix and bifurcation) and Low Voltage Fast Activity occurred simultaneously in episodes which mainly appeared to originate on the bifurcation.The characteristics of uterine activity during pregnancy were similar to those of a cyclic sow until day 12. It was only on day 12 that gestation appeared from an increased frequency of myoelectrical complexes.It is concluded that Low Voltage Fast Activity as it was found on the uterine horn at days 8–9 might be involved in the process of intra-uterine migration of blastocysts. In cyclic and in pregnant animals the patterns of Low Voltage Fast Activity were similar. Therefore, the occurrence of Low Voltage Fast Activity is independent of the actual presence of blastocysts. It seems to be exhibited in dependence on the ovarian hormones.     

Cholinergic reactivity of the bovine fetal ruminal wall in vitro

Strips of rumen wall from bovine fetuses were incubated in an organ bath with acetylcholine only (0.16 to 5.12 g/ml) or in the presence of neostigmine (0.20 g/ml) or atropine (0.05g/ml). The highest reactivity was observed in the period of 4.0–5.9 months of fetal age. This reactivity could be associated with the starting point of rumen papillary development.     

Kinetics of Interferon-Gamma Production and its Comparison with Anti-Listeriolysin O Detection in Experimental Bovine Listeriosis

The kinetics of the production of interferon gamma (IFN-) in whole blood culture and its comparison with anti-listeriolysin O (ALLO) detection by ELISA were studied during oral infection of calves with Listeria monocytogenes. Culture filtrate antigen (CFA), listeriolysin O (LLO), and sonicated antigen (SA) were used to prime the peripheral blood mononuclear cells (PBMCs) and the plasma from orally infected calves. IFN- and ALLO appeared as early as day 7 of an oral infection. IFN- was detected earlier with LLO than with SA. The Max50 interleukin (IL-2) activity and IFN- estimated in the culture supernatant from PBMCs primed in vitro with different antigens of L. monocytogenes revealed high induction of IL-2 and IFN- by CFA, LLO and live antigen. IFN- assay and ALLO detection were used for testing cases of repeat breeding in dairy cattle. It appeared that detection of IFN- employing LLO can be used to diagnose listerial infections.     

Intestinal atresia and stenosis: A review comparing its morphology

A review of the literature on intestinal atresia of domestic animal species and humans was done. The 5 types of intestinal occlusions described in human infants are atresia type 1, atresia type 2, atresia type 3, stenosis, and the apple peel or Christmas tree deformity. The intestinal defects described in domestic animal species such as the bovine, equine and porcine are similar to those of human infants. The T-formation, an intestinal defect of the bovine resembling atresia type 3, and rectal stricture, an acquired intestinal defect of the porcine resembling stenosis, were described recently. Intestinal atresia is similar in several species and these similarities raise the questions as to whether the pathogenic mechanism and possibly etiologies of intestinal atresia are similar in these species.     

The influence of clenbuterol on prostaglandin F2α-induced dyspnoea in calves

Pulmonary function tests were performed in six healthy calves. Prostaglandin F₂ causes severe narrowing of both upper and lower airways (total lung resistance increased, dynamic compliance decreased). Clenbuterol administered intravenously fifteen minutes prior to prostaglandin F₂ aerosol, and in increasing doses (0, 0.4, 0.8, 1.2 g/kg), on days 1, 2, 4 and 6 of the experiment, effectively but not entirely suppressed these responses.These data indicate that -adrenergic receptors are present in the bovine airways and that the use of clenbuterol (0.8 g/kg) may be effective in treating clinical respiratory disease such as bronchopneumonia in calves.     

β-Adrenoceptors in Equine Trachea and Heart

The density and subtype pattern of -adrenoceptors in equine tracheal epithelium, tracheal smooth muscle and heart from 6–9 horses were investigated by radioligand binding studies using the non-selective -adrenoceptor antagonist 125I-cyanopindolol (ICYP). The specific binding of ICYP was 341±162 fmol/mg protein (mean±SD) for epithelium, 42±13 fmol/mg for smooth muscle and 124±39 and 101±19 fmol/mg for the cardiac atrium and ventricle, respectively. The Kd value of ICYP was 6.7–10.2 pmol/L.In competition studies, different concentrations of either the 2-selective drug ICI 118551 or the 1-selective CGP 20712A competed with 25 pmol/L ICYP for the binding sites. The competition curves for tracheal smooth muscle and epithelium were monophasic with an approximate Kd value for ICI 118551 of 1 nmol/L and for CGP 20712A of 10 000 nmol/L. This corresponds to known Kd values for these substances binding to 2-adrenoceptors. 2-Adrenoceptors were also found in the heart, most pronounced in the atrium, where the density was 29%±6% (mean±SD) of the total receptor density. CGP 20712A and ICI 118551 bound to the dominating binding site of 1-adrenoceptors in the heart with Kd values of approximately 1 nmol/L and 100 nmol/L, respectively.     

Clinical Value of Fructosamine Measurements in Non-healthy Dogs

Ninety-three unhealthy dogs (including some with diabetes mellitus or insulinoma) of different ages, sex and breeds were divided into 10 groups according to their pathology. Serum fructosamine concentrations were determined using a commercial colorimetric nitroblue tetrazolium method. Diabetic dogs had the highest fructosamine concentrations (454.85±149.34 mol/L). Dogs with insulinoma had significantly lower fructosamine concentrations (202.80±31.22 mol/L), similar to those with leishmaniosis (202.83±99.83 mol/L). Fructosamine concentrations in non-healthy dogs, except those with diabetes mellitus, insulinoma or leishmaniosis, were within the reference limits previously reported.     

Feed intake and rumen motility in dwarf goats. Effects of some α-adrenergic agonists,prostaglandins and posterior pituitary hormones

The purpose of the present investigation was to test the hypothesis that drug-induced changes in rumen contractions influence feed intake in dwarf goats. Intravenous (i.v.) administration of clonidine (1 g kg^-1 min^-1 for 10 min), xylazine (1 g kg^-1 min^-1 for 10 min), and PGF-2 (10 g kg^-1 min^-1 for 15 min) caused bradycardia and inhibition of rumen contractions. However, no appetite-stimulating effect of these drugs was observed. Other clinical changes induced by the ₂-adrenergic agonists included slight sedation and a decrease in body temperature; all clinical effects of clonidine and xylazine were partly antagonized by tolazoline pretreatment (10 g kg^-1 min^-1 for 30 min). These results suggest that the CNS control of feeding differs in ruminants and monogastric species.In dwarf goats fasted for 2 h, i.v. administration of oxytocin (0.01 IU kg^-1 min^-1 for 15 min), vasopressin (0.01 IU kg^-1 min^-1 for 15 min), octapressin (0.003 IU kg^-1 min^-1 for 15 min) or PGE (0.8 g kg^-1 min^-1 for 15 min) did not change feeding behaviour during the two observation periods (0–30 min and 180–210 min after drug infusion, respectively). In previous studies, similar doses of these drugs induced changes in heart rate and inhibition of rumen contraction in goats. These findings demonstrate that drug-induced changes in forestomach contractions do not simply cause changes in feeding behaviour. The i.v. infusion of the PGF₂ analogues etiproston (10 g kg^-1 min^-1 for 15 min), luprostiol (30 g kg^-1 min^-1 for 15 min), cloprostenol (1 g kg^-1 min^-1 for 15 min) and tiaprost (1 g kg^-1 min^-1 for 15 min) induced hypophagic effects and stimulated intestinal propulsion.     

The use of cultured hepatocytes from goats and cattle to investigate xenobiotic oxidative metabolism

The oxidative metabolism of aldicarb (ALD), a carbamate pesticide, and fenbendazole (FBZ), an anthelmintic, was studied using cultured hepatocytes obtained from 4 goats and a bullock and incubated with ALD (50 mol/L) and FBZ (10 mol/L). The parent compounds and the metabolites were measured by HPLC. Both compounds are metabolized at the sulphur atom via two sequential oxidations, first to the sulphoxide (aldicarb sulphoxide and oxfendazole, respectively) and then to the sulphone. Oxfendazole and fenbendazole sulphone from FBZ, and aldicarb sulphoxide from ALD were found in both species. Aldicarb sulphone was not produced by the hepatocyte preparations from the bullock. The good correlation obtained comparing the in vitro results of FBZ metabolism with published in vivo dat obtained on FBZ kinetics in the same species confirmed the usefulness of in vitro models for predictive analysis of in vivo xenobiotic biotransformations.Abbreviations ALD aldicarb - ALDSON aldicarb sulphone - ALDSOX aldicarb sulphoxide - BSA bovine serum albumin - ID internal diameter - EGTA ethylene glycol bis(-aminoethyl ether) N,N,N,N-tetraacetic acid - FBZ fenbendazole - FBZSON fenbendazole sulphone - HBSS Hanks' balanced saline solution - HPLC high-pressure liquid chromatography - LDH lactate dehydrogenase - MFO mixed function oxidase - NCS newborn calf serum - OXF oxfendazole - WME Williams' Medium E     

Disposition kinetics and distribution of demeclocycline in various biological fluids in female goats

A pharmacokinetic study of demeclocycline was carried out following intravenous administration at 5 mg/kg body weight in lactating goats. Demeclocycline appeared within 5 min in plasma, interstitial fluid (isf) and urine, while it appeared at 1 h in milk. Peak concentrations of 21.70±4.06, 2.67±0.23, 5.65±0.45 and 82.23±10.06 g/ml were attained at 5 min and at 6, 8 and 8 h in plasma, isf, milk and urine respectively. A potentially therapeutic concentration of 0.5 g/ml was maintained from 5 min–36 h, 30 min–30 h, 1–36 h and 5 min–48 h in plasma, isf, milk and urine respectively. The drug was detectable in all the above biological fluids for at least 48 h. A low distribution half life (t_1/2) of 0.44±0.04 h and a high elimination half life (t_1/2) of 19.24±1.22 h denote rapid distribution but very slow elimination of the drug in goats. A high tissue plasma concentration ratio [K₁₂:(K_2I–)] of 5.12±0.97 during the elimination phase and a Vd_area of 1.59±0.18 L/kg indicate uniform distribution of demeclocycline in the tissues and body fluids of goats. The dosage regimen for maintaining minimum plasma concentration (C min = MIC) of 0.5, 1.0 and 1.5 g/ml at selected dosage intervals of 12 and 24 h was also calculated.     

The effect of induced fever on the biokinetics of norfloxacin and its interaction with probenecid in goats

The kinetic profiles of norfloxacin were evaluated in afebrile, febrile and probenecid pre-treated (70 mg/kg orally) febrile goats after a single intravenous (i.v) dose (5 mg/kg). Fever was induced and maintained for 12 h by injecting Escherichia coli endotoxin (0.2 g/kg, i.v.) and repeating it in half the dose (0.1 g/kg) 5 h later. The plasma pharmacokinetic values for norfloxacin were best represented using a two-compartment open model. The peak norfloxacin plasma level of 90.52±3.18 g/ml attained in the probenecid pre-treated febrile goats was higher than that in the febrile (75.46±0.72 g/ml) or afebrile goats (62.25±1.23 g/ml). Cl_B and K _el values were significantly (p<0.01) decreased in febrile compared with afebrile goats. These values were further reduced in febrile goats after probenecid pre-treatment. However, t _1/2 was not affected by the fever-probenecid interaction. Norfloxacin may be used as an infusion with probenecid in caprine diseases where very high plasma levels are required to combat resistant organisms such as Bacteroides.Abbreviations MIC minimum inhibitory concentration - LPS lipopolysaccharide - i.v. intravenous(ly)     

Central and local actions of opioids upon reticulo-ruminal motility in sheep

The effects of opioids and naloxone on cyclical forestomach motility were determined in anaesthetized and conscious sheep. To assess central or peripheral opioid actions, differential routes of administration were used. Possible dynamic effects along the innervating vagovagal reflex arc were investigated electrophysiologically at the cervical level of the vagus nerve. Further, direct influences on the smooth muscle were evaluatedin vitro on isolated longitudinal reticular strips. Additionally, the effects of some spasmogenic agents were studied for comparative purposes. In anaesthetized sheep, opioids depressed in an identical manner both the amplitude of spontaneous cyclical contractions and contractions evoked by electrical stimulation of the distal end of the cut cervical vagus. In conscious sheep, low doses of normorphine and loperamide inhibited frequency and amplitude centrally (20 g/kg and 4 g/kg via carotid artery respectively), whereas locally higher dose levels (200 g/kg and 10 g/kg via coeliac artery respectively) affected only the amplitude of cyclical contractions. Furthermore the opioid peptides Leu-, Met-enkephalin and [D-Ala²-Met⁵]-enkephalinamide preferentially depressed the amplitude of cyclical motility most efficiently if administrated via the coeliac artery. These results indicate the presence both of a central opioid action depressing frequency and amplitude and of a local opioid action depressing only the amplitude of cyclical reticulo-ruminal motility. Opioids did not alter the resting discharge of afferent tension units and similarly failed to modulate tone of reticular stripsin vitro, suggesting that the opioids act locally on the intramural neuronal plexus, possibly by diminishing the output of excitatory transmitter. Whether substance P could play a role as a vagal excitatory transmitter besides the classically implicated acetylcholine has been discussed. The central opioid mechanism is probably not situated within the gastric centres but elsewhere in the brain. Naloxone ( 100 g/kg, jugular vein) stimulated the frequency of cyclical ruminal motility only in well-defined experimental conditions, probably via a central mechanism.     

Pharmacokinetics and Dosage Regimen of Enrofloxacin in Buffalo Bulls after Intramuscular Administration

The disposition kinetics and dosage regimen of enrofloxacin were investigated in breeding buffalo bulls following a single intramuscular administration of 5 mg/kg. The absorption half-life, half-life of the terminal phase, apparent volume of distribution and total body clearance were 0.262±0.099 h, 1.97±0.23 h, 0.61±0.13 L/kg and 210.2±18.6 ml/(kg.h), respectively. Therapeutic plasma levels (1 g/ml) were maintained for up to 6 h. A satisfactory intramuscular dosage regimen for enrofloxacin in buffalo bulls would be 8.5 mg/kg followed by 8.0 mg/kg at 8 h intervals.     

Failure of 3-methylindole to contract the bovine pulmonary vein or to release mediators of anaphylaxis from the bovine lung in vitro

Strips of smooth muscle from the pulmonary vein, pulmonary artery, trachea and bronchus of calves were incubated in an organ bath with 3-methylindole (3MI) and 3-methyloxindole (3MOI). 3MI and 3MOI (5–640 g/ml) did not cause contraction of any of the isolated smooth muscle preparations. No evidence for the release of mediators of anaphylaxis was obtained when chopped bovine lung preparations were incubated with 3MI (20 g/ml) and 3MOI (25 g/ml). Results of the present work diminish the possibility that the pneumotoxic effect of 3MI is due to a primary hydrodynamic imbalance across the alveolocapillary membrane resulting in excess filtration over reabsorption.