Comparison of in vitro activities of biotransformation enzymes in pig, cattle, goat and sheep

In vitro activities of cytochromes P450 (7-alkyl/aryloxyresorufin dealkyl(aryl)ases, testosterone hydroxylase/oxidase, 6-chlorzoxazone hydroxylase, 7-methoxy-4-trifluoromethyl-coumarin demethylase, and lauric acid hydroxylases), reductases of carbonyl group (toward metyrapone, daunorubicin, glyceraldehyde, and 4-pyridine-carboxaldehyde) and conjugation enzymes (p-nitrophenol-UDP-glucuronosyl transferase, 1-chloro-2,4-dinitrobenzene glutathione-S-tranferase) in young adults, males, non-castrated (N=6) farm animals were studied and compared. Presence of proteins cross-reacting with anti-human CYP3A4, CYP2C9, and CYP2E1 IgG was detected in all farm species. Bovine microsomes differed from other microsomes of farm species in very high 7-ethoxyresorufin-O-deethylase activity (CYP1A1/2). Significantly higher 7-methoxy-4-trifluoromethyl-coumarin demethylase (2-3 times) and 12-lauric acid hydroxylases (4-10 times) activities (probably corresponding to CYP2C and CYP4A, respectively) were found in ovine microsomes. The highest 6beta-testosterone hydroxylase activity, which is usually considered to be a CYP3A activity marker, was found in pig. Reductases of all farm animals display considerable ability to reduce carbonyl group of xenobiotics. Significant differences in level and activity of many biotransformation enzymes tested suggest that extrapolation of pharmacokinetic data obtained in one species to another (even related) could be misleading.     

Mouflon (Ovis musimon) dicrocoeliosis: effects of parasitosis on the activities of biotransformation enzymes and albendazole metabolism in liver

Parasitic infections can modify the host's ability to metabolize drugs and other xenobiotics by altering the biotransformation enzymes; these changes may have various pharmacological, toxicological or physiological consequences. In our study, several activities of liver biotransformation enzymes and in vitro metabolism of albendazole (ABZ) were tested and compared in non-infected mouflons (Ovis musimon) and in mouflons infected by lancet fluke (Dicrocoelium dendriticum). Subcellular fractions of liver homogenates were isolated from 5+5 mouflon rams (1-year-old) parasitologically negative or naturally infected by fluke. From the eight enzyme activities that were assayed, only two activities significantly differ in the case of Dicrocoelium-infected versus non-infected animals. In infected mouflons, a significant increase (53%) of thiobenzamide-S-oxidase (TBSO) activity, corresponding mainly to the activity of flavine monooxygenase (FMO), and significant decrease (60%) of glutathione-S-transferase (GST) activity was observed. In addition, dicrocoeliosis caused the enhancement of ABZ hepatic biotransformation. The velocity of the formation of (+)-ABZ sulfoxide and ABZ sulfone was significantly increased. However, the shifts in ABZ biotransformation were very mild that undesirable alterations in ABZ pharmacokinetic are not expected. From this point of view, the use of ABZ in the therapy of mouflon dicrocoeliosis in young animals can be recommended. The treatment of the same mouflons by other drugs that are mainly conjugated with glutathione, seems to be more problematic; hence, all consequences of documented reduced GST activity should be accounted.     

Gross pathology and histopathology of haemonchosis in sheep and goats in Iraq

The response of Awassi sheep and a local breed of goats to H. contortus, the prevalent ovine and caprine strongyloid species in Iraq, was studied. A dose of 500 H. contortus (sheep origin) larvae kg-1 body wt. induced clinical and pathological changes of haemonchosis. Although the percentage recovery of worms was higher in sheep, the pathological (gross and microscopic) changes were more marked in infected goats. The extensive abomasal changes noted in goats were, in general, accompanied by a higher density or larvae in the gastric wall; the pathology did not necessarily depend upon parasite count alone and a definite relationship could not be established. The tissue changes in the goats are possibly of immunological origin, although a genetic resistance may operate, primarily at the level of worm establishment.     

A possible relationship between haemonchosis and haemoglobin polymorphism in Merino sheep in Kenya.

It is known that sheep show polymorphism for haemoglobin in that three phenotypes (HbA, HbB AND HbAB) are normally found. Since it has been suggested, on the basis of haematological values, that HbA type sheep are more able to survive in Haemonchus contortus endemic environments, a preliminary study was conducted on the performance of set-stocked Merino ewes and lambs in relation to their haemoglobin phenotypes. The results provided some evidence that, in Merino sheep at least, those with HbA type show 'self-cure' more frequently and effectively than those with HbB and HbAB types. Examination of the PCV values and total body weights of Merino eres showed a similar trend, ie, the HbA types had the highest haematological parameters and the heaviest body weights, while the HbB types were consistently lowest. It would appear that the practical significance of these results justifies investigation on a larger scale.     

Renal excretion of creatinine, electrolytes, protein, and enzymes in healthy sheep

Urinary indices of renal function and damage were measured in 6 healthy, mature ewes over a 48-hour period. Endogenous creatinine clearance, total and fractional electrolyte excretion rates, protein excretion, urine volume, and urine gamma-glutamyltransferase and beta-glucuronidase activities were measured. Significant variations in the excretion rates of creatinine, electrolytes, and protein were not found between intervals within the 48-hour urine collection period. Total urinary electrolyte excretion rates were significantly (P less than 0.001) correlated with fractional electrolyte excretion rates normalized for creatinine concentration; however, coefficient of determination was low.     

Benzimidazole drugs and modulation of biotransformation enzymes

Benzimidazole drugs (e.g., anthelmintics albendazole, fenbendazole, oxfenbendazole, thiabendazole, mebendazole; inhibitors of proton pump omeprazole, lansoprasole, pantoprasole) represent substances used in both human and veterinary medicine; however, from the point of view of induction and inhibition of biotransformation enzymes, research has been carried out mainly due to the initiative of human pharmacologists. The purpose of the present review is to inform about inductive and inhibitive effects of benzimidazole drugs in man, animals and cell cultures. Pharmacological and toxicological consequences of modulation of biotransformation enzymes are discussed and the significance of studies in the field of modulation of biotransformation enzymes in food-producing animals is explained. Since the modulating effect of benzimidazoles strongly varies depending on structure of the individual substances, the particular attention is paid to structure-modulation relationships.     

Blood gas values in healthy sheep and in sheep with lung diseases]

In 24 healthy and 101 sheep with different lung diseases like catarrhalic pneumonic bronchopneumonia, lung abscesses, lung worm infections, Maedi and Sheep Pulmonary Adenomatosis (SPA) in arterial and venous samples blood gas analysis was performed. The alveolo-arterial-oxygen-difference was the most sensitive parameter to detect impaired ventilation, it was 11.40 +/- 6.39 mm Hg in healthy sheep. Global insufficiency occurred mainly in sheep with SPA and in mixed infections of SPA with Maedi.     

Serodiagnosis of haemonchosis with a somatic antigen (Hc26) in several breeds of sheep.

Sera from 53 sheep belonging to Castellano, Churro, Manchego, and Merino breeds were analyzed to test the diagnostic value of a 26-kD antigen from adult Haemonchus contortus at prepatency and early and late patency of experimental haemonchosis. Animals that received zero, 1, or 2 infections with the parasite were tested. In addition, sera from 20 experimentally infected and 10 noninfected Texel sheep were used to test the antigen. Sera from 37 infected animals at prepatency as well as at patency in primary and secondary infection were found positive with the 26-kD antigen. However, sera from 10 animals with the lowest worm burdens (second infection) did not recognize the antigen during early patency (day 28 postinfection). IgG1 was the only isotype implicated in antigen recognition because IgG2, IgA, and IgM, in the same sera, showed no reactivity with the peptide. Antigen specificity was confirmed because hyperimmune sera against infective larvae and adult stages of the most common gastrointestinal nematodes found in natural infections in sheep (Trichostrongylus colubriformis and Teladorsagia circumcincta) did not recognize this peptide. The antigen was recognized only by anti-adult H. contortus hyperimmune sera and appeared to be absent in the L3 parasite stage. In addition, the partial N-terminal amino acid sequence of the diagnostic peptide is reported.     

Phenolsulfonphthalein test in healthy sheep and in sheep with reductions in functional renal mass

The phenolsulfonphthalein (PSP) plasma clearance and urinary excretion tests were applied to sheep before and after 50% and 75% reductions in functional renal mass. The PSP determinants found most useful as indicators of renal mass reduction were the 15-minute urinary excretion percentage and the 60-minute (PSP60) plasma concentration. Although both of these determinants could be used to detect renal mass reduction, the 15-minute PSP excretion percentage was the more sensitive. The PSP60 value was influenced by factors other than reduced nephron numbers; the contraction of the PSP volume of distribution that occurred after renal mass reduction was one important influencing factor. Overall, the PSP tests more accurately reflected the volume of blood delivered to the kidney than the proximal tubular secretory capacity.     

Changes in activities of enzymes related to energy metabolism in canine lymphoma cells

Alterations in the activities of enzymes related to energy metabolism in canine lymphoma cells were investigated. Cytosolic pyruvate kinase (PK) and mitochondrial malate dehydrogenase (MDH) activities in lymphoma cells were significantly higher than those in lymphocytes obtained from lymph nodes of healthy dogs, whereas cytosolic lactate dehydrogenase (LDH) activity was significantly lower in lymphoma cells. The cytosolic M/L ratio (MDH activity/LDH activity), which is considered to be a good indicator of energy metabolism related to glucose utilization in animal tissues, was significantly higher in lymphoma cells than in the normal lymphocytes.     

Modulation of porcine biotransformation enzymes by anthelmintic therapy with fenbendazole and flubendazole

Fenbendazole (FEN) and flubendazole (FLU) are benzimidazole anthelmintics often used in pig management for the control of nematodoses. The in vivo study presented here was designed to test the influence of FLU and FEN on cytochrome P4501A and other cytochrome P450 (CYP) isoforms, UDP-glucuronosyl transferase and several carbonyl reducing enzymes. The results indicated that FEN (in a single therapeutic dose as well as in repeated therapeutic doses) caused significant induction of pig CYP1A, while FLU did not show an inductive effect towards this isoform. Some of the other hepatic and intestinal biotransformation enzymes that were assayed were moderately influenced by FEN or FLU. Strong CYP1A induction following FEN therapy in pigs may negatively affect the efficacy and pharmacokinetics of FEN itself or other simultaneously or consecutively administered drugs. From the perspective of biotransformation enzyme modulation, FLU would appear to be a more convenient anthelmintic therapy of pigs than FEN.     

The FAMACHA system for managing haemonchosis in sheep and goats by clinically identifying individual animals for treatment

Escalating anthelmintic resistance has made it essential to develop alternative ways of worm management for reducing selection for worm resistance, and one of the most promising approaches is to treat only those animals unable to cope with worm challenge, thus favouring unselected worms originating from untreated animals. Only clinical evaluation of anaemia (FAMACHA system) and the body condition score are regarded as being of practical value or having potential, respectively, for repeatedly examining flocks or herds and identifying individuals for treatment. Only the FAMACHA system has been tested well enough for use under practical farming conditions. However, further investigation is needed on its effect on animal production, and methods to reduce labour. Trials over several seasons showed that most sheep under severe Haemonchus contortus challenge required no, or only one treatment over a full summer season. A small minority (usually < 5%) needed more than two treatments. Most sheep could cope without regular treatment. With sufficient training, clinical evaluation of anaemia was found reliable for practical use. The overwhelming majority of trainees (some poorly literate) were able to implement the FAMACHA system successfully. The dynamics of haemonchosis in a flock can easily be monitored. Farmers, farm workers and veterinarians all rated the system very highly (> 80%) and treatment costs dropped by approximately 58%. The heritability of FAMACHA values obtained by clinical evaluation was high at 0.55 +/- 0.17% in a Merino stud with +/- 550 young rams and ewes which were the progeny of 21 sires. The FAMACHA system may not be as applicable to goats as to sheep, but further work is necessary. The main benefits of the system are the reduction in treatments, its use for discriminating between animals of varying ability to cope with infection (thus allowing genetic selection), and its lowering of selection pressure on H. contortus for anthelmintic resistance.     

The pathogenicity of haemonchosis in sheep and goats in Iraq: 1. Clinical, parasitological and haematological findings

The response of Awassi sheep and the local breed of Iraqi goats to primary infection with a prevalent trichostrongylid nematode, Haemonchus contortus, was compared by examining their worm burden and the pathogenic effects of the parasite on these hosts following experimental infection. The results showed that the goats had lower worm burdens and had suffered less severe clinical and haematological disturbances than the Awassi sheep. The worm counts in sheep were higher than in goats but the count of the mucosal phase of the larvae was higher in goats. These differences are possibly immunologic in origin though a genetic resistance may operate, primarily at the level of worm establishment. The pathophysiological changes are correlated with parasitological and clinical observations in the two host species.     

The effects of fascioliasis on the activities of some drug-metabolizing enzymes in desert sheep liver.

Desert sheep experimentally or naturally infected with Fasciola gigantica were used to study the influence of infection on the activities of some drug-metabolizing enzymes found in the liver. The enzymes investigated were aminopyrine N-demethylase, aniline 4-hydroxylase and UDP-glucuronyltransferase. The experimental infection was confirmed histologically by detection of Fasciola eggs in faeces and by measuring the activities of sorbitol dehydrogenase (SD), glutamate dehydrogenase (GD) and aspartate aminotransferase (AST) in plasma during the course of the disease. Liver specimens from naturally infected sheep were obtained from the slaughter house. The activities of aminopyrine N-demethylase and aniline 4-hydroxylase were significantly decreased in sheep either naturally infected or during the acute stage of experimental fascioliasis (killed 5 weeks post-infection). The activity of UDP-glucuronyltransferase was decreased in naturally infected sheep and those killed 9 or 13 weeks post-experimental infection.     

Methimazole-mediated modulation of netobimin biotransformation in sheep: a pharmacokinetic assessment

The effects of modulation of liver microsomal sulphoxidation on the disposition kinetics of netobimin (NTB) metabolites were investigated in sheep. A zwitterion suspension of NTB was given orally at 7.5 mg/kg to sheep either alone (control treatment) or co-administered with methimazole (MTZ) orally (NTB + MTZ oral treatment) or intra-muscularly (NTB + MTZ i.m.) at 3 mg/kg. Blood samples were taken serially over a 72 h period and plasma was analysed by HPLC for NTB and its major metabolites, i.e. albendazole (ABZ), albendazole sulphoxide (ABZSO) and albendazole sulphone (ABZSO2). Only trace amounts of NTB parent drug and ABZ were detected in the earliest samples after either treatment. There were significant modifications to the disposition kinetics of ABZSO in the presence of MTZ. ABZSO elimination half-life increased from 7.27 h (control treatment) to 14.57 h (NTB + MTZ oral) and to 11.39 h (NTB + MTZ i.m.). ABZSO AUCs were significantly higher (P less than 0.05) for the NTB + MTZ oral treatment (+55%) and for the NTB + MTZ i.m. treatment (+61%), compared with the NTB alone treatment. The mean residence times for ABZSO were 12.66 +/- 0.68 h (control treatment), 18.85 +/- 2.35 h (NTB + MTZ oral) and 17.02 +/- 0.90 h (NTB + MTZ i.m.). There were no major changes in the overall pharmacokinetics of ABZSO2 for the concomitant MTZ treatments. However, delayed appearance of this metabolite in the plasma resulted in longer ABZSO2 lag times and a delayed Tmax for treatments with MTZ.(ABSTRACT TRUNCATED AT 250 WORDS)     

Maturational development of drug-metabolizing enzymes in sheep

A qualitative and quantitative assessment was made of the development of hepatic drug-metabolizing enzymes (DME) in sheep as part of a study of the ability of the food-producing species to metabolize drugs. The following DME and components were measured in this study: cytochromes P-450 and b5 and NADPH and NADPH-dependent reductases associated with each of these cytochromes; cytochrome P-450-mediated reactions, including aniline and coumarin hydroxylases, aminopyrine N-demethylase, and 7-ethoxycoumarin 0-deethylase; a uridine diphosphoglucuronic acid glucuronyl transferase with 4-methylumbelliferone as substrate; and glutathione-S-transferase with dinitrochlorobenzene and dichloronitrobenzene as substrates. Amounts or activities of most of these components and enzymes increased up to and beyond the time of weaning. Amount of cytochrome b5 and uridine diphosphoglucuronic acid transferase activity were not affected by age, whereas NADPH cytochrome c (P-450) reductase activity actually decreased after weaning. In some instances (eg, coumarin hydroxylase, cytochrome P-450, and dinitrochlorobenzene-glutathione-S-transferase), differences from preweaning DME values were observed only after sheep were greater than or equal to 6 months old. All other DME activities were definitely increased, compared with the values in lambs before weaning (0 to 12 weeks old). Approximately a third of the sheep studied had some type of clinical disease that might have affected the DME activities. Diseases were classified as sore mouth, pneumonia, foot rot, parasitism, and systemic bacterial infections. Except in a few instances, these diseases had minimal effect on DME activities measured in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)