Altered functional and immunophenotypical properties of neutrophilic granulocytes in postpartum cows associated with fatty liver.

The intention of the study was to analyze the relationship between liver triacyl glycerol content (liver TAG content) and immunophenotypical and functional properties of polymorphonuclear neutrophilic granulocytes (PMN) of dairy cows in the peripartum period. We investigated characteristics of bovine PMN from the blood and uterus of clinically healthy cows in the periparturient period. The numbers of circulating leukocytes and segmented granulocytes continuously increased until parturition and declined afterwards to starting values. This was independent of the liver TAG content and mainly affected neutrophils. The liver TAG content exceeded 40 mg/g liver, the reference value, in 12 of 19 cows in the first two weeks postpartum. Increased liver TAG content, > 40 mg/g, went in parallel with a reduced expression of function-associated surface molecules on blood neutrophils (e.g. CD11b/CD18 = CR3 and CD11c/CD18 = CR4). Moreover, in cows with high liver TAG levels the antibody-independent and -dependent cellular cytotoxicity (AICC, ADCC) of blood PMN was markedly reduced. PMN also were less capable of ROS generation after stimulation with Phorbol Myristate Acetate (PMA). In comparison with contemporarily harvested blood PMN, neutrophils recovered from the uterine lumen showed a decreased expression of 4/6 examined surface structures. Only the expression densities of CR3 molecules and those detected by mAb IL-A110 were enhanced on uterine PMN. The cytotoxic capacity and the ROS generation were significantly lower for uterine PMN than for blood PMN. The results suggest that increased liver TAG content in the first and second week after calving is associated with decreased functional capacities of PMN derived from blood and uterus. This may help to explain why cows who are too fat at calving (who therefore have an increased liver TAG content) have a higher incidence of infectious diseases such as endometritis

Urinary concentration of corticoids in ponies with hyperlipoproteinaemia or hyperadrenocorticism.

The urinary corticoid:creatinine (c:c) ratio was determined in ten pony mares suffering from hyperlipoproteinaemia. The mean (+/- sd) urinary c:c ratio of these ten ponies (47 +/- 31 x 10(-6)) was not significantly different from that of twelve pony mares with a pituitary pars intermedia adenoma (31 +/- 18 x 10(-6). The correlation between the urinary concentration of corticoids and plasma total lipids, and the correlation between the urinary c:c ratio and plasma total lipids in ponies with hyperlipoproteinaemia were not significant (P > 0.05; r = 0.53 and r = 0.008, respectively). Preliminary results favour primary hyperadrenocorticism being associated with hyperlipoproteinaemia. In conclusion, the data presented here suggest that cortisol can contribute to insulin resistance in ponies with hyperlipoproteinaemia.

Insulin resistance: the link between metabolic disorders and cystic ovarian disease in high yielding dairy cows?

A preliminary investigation was performed to examine whether insulin resistance is a factor in the pathogenesis of cystic ovarian disease (COD) in high-yielding dairy cows. In total 30 cows, of which 15 were diagnosed as suffering from COD based on the anamnesis and clinical examination, and the other 15 served as matched controls, were subjected to an intravenous glucose tolerance test (IVGTT). The aim of the study was to investigate whether insulin activity was altered in COD cows. Differences in glucose clearance between the COD cows and their controls were analyzed comparing the fractional turnover rate (k), the glucose half-time (T1/2), and the area under the curve (AUC) 60 and 120 min after infusion. Differences in insulin response were analyzed comparing the insulin increment, the insulin peak concentration, and the AUC 60 and 120 min after glucose infusion. Although insulin resistance, attended by a secondary hyperinsulinemia, is stated to directly contribute to the ovarian abnormalities that characterize the polycystic ovary syndrome (PCOS) in human medicine, this was not observed in COD cows. On the contrary, COD cows appeared to have a low insulin response following an intravenous glucose load as compared with their matched controls. This was illustrated by significantly lower insulin increments (P = 0.04) and lower insulin peak concentrations (P = 0.04). As COD cows had a significantly lower insulin response to a standard glucose load, it was concluded that insulin could be a factor in the pathogenesis of COD in dairy cows.

Reversible translocation of CTP:phosphocholine cytidylyltransferase from cytosol to membranes in the adult bovine liver around parturition.

The key regulatory enzyme of phosphatidylcholine (PC) synthesis, CTP:phosphocholine cytidylyltransferase (CT), is known to be activated in vitro by translocation from soluble to particulate fractions of the cell. In the present study the periparturient cow was chosen as a model to investigate whether translocation of CT can contribute to the regulation of PC synthesis in vivo. Between parturition and 1.5 weeks post-partum, the cytosolic CT activity in the liver of the adult animal decreased 1.9-fold, and this correlated with a 1.8-fold increase in microsomal CT activity. At that time, microsomal CT activity started to decline again whereas the cytosolic activity rose concomitantly until both activities reached their pre-partum values at 8 weeks post-partum. The activities of soluble and membrane-bound CTP:phosphoethanolamine cytidylyltransferase (ET), the analogous enzyme in the CDP-ethanolamine pathway, did not change significantly throughout this period. Whereas hepatic PC concentrations declined until about 2 weeks post-partum and thereafter gradually returned to pre-partum levels, the PC levels in very-low-density-lipoproteins, started to rise 2 weeks after the partus reaching a maximum of 219% of the original value at 8 weeks post-partum. These results strongly suggest that there is a reversible redistribution of CT between cytosol and membranes in a physiologically relevant animal model, supporting the concept that translocation of CT is occurring in vivo.

Effects of pentoxifylline and polymyxin B on the acute-phase-response to Escherichia coli endotoxin in dwarf goats.

The purpose of this study was to assess whether polymyxin B together with pentoxifylline, had beneficial effects on the acute-phase-response to E. coli endotoxin in the dwarf goat (n = 6). Polymyxin B partly neutralizes E. coli endotoxin by forming inactive polymyxin B-lipopolysaccharide (LPS) complexes; pentoxifylline has been reported to suppress the LPS-induced production of tumour necrosis factor (TNF-alpha). E. coli LPS (0.0067 microgram/kg/min over 30 min) induced fever, tachycardia, inhibition of rumen motility, a decline in WBC, lymphopenia, and decreases in plasma zinc and iron concentrations. Most of the haematological, blood biochemical and clinical effects of E. coli LPS were significantly reduced by polymyxin B pretreatment (0.1 mg/kg/min over 30 min, i.v.). Pentoxifylline (0.3 mg/kg/min over 30 min, i.v.) did not reduce the clinical and blood biochemical effects of E. coli LPS, however, it modulated the number of circulating neutrophils. No synergistic effects were observed after i.v. infusion of polymyxin B with pentoxifylline. The lack of synergy may be due to the production and release of pro-inflammatory cytokines other than TNF-alpha.

Lipid metabolism in horses with hyperadrenocorticism.

Lipid metabolism was studied in 21 horses with hyperadrenocorticism. To be included in the study, horses had to have histologic evidence of a pars intermedia adenoma found at necropsy (n = 9), a baseline ACTH concentration greater than 400 pg/ml (n = 6), or a plasma cortisol concentration 2 hours after i.v. administration of 25 IU of ACTH greater than 413 nmol/L (n = 16). Mean +/- SD baseline plasma cortisol concentration was 338 +/- 261 nmol/L (n = 20), mean +/- SD plasma insulin concentration was 97 +/- 54 microU/ml (n = 15), mean +/- SD plasma beta-hydroxybutyrate concentration was 1.8 +/- 1.2 mg/dl (n = 21), and mean +/- SD plasma nonesterified fatty acids concentration was 6.2 +/- 6.4 mg/dl (n = 21). None of the horses had hyperlipemia. Compared with clinically normal horses, horses with hyperadrenocorticism had increased lipolysis and increased ketogenesis. It was concluded that cortisol cannot be the sole factor contributing to insulin resistance in horses with hyperadrenocorticism.

Laboratory diagnosis of equine pituitary pars intermedia adenoma.

The objective of the study was to determine the sensitivity with which an adenoma of the pars intermedia of the pituitary gland can be predicted in horses by measuring the basal concentrations of glucose, cortisol, adrenocorticotropin (ACTH), and insulin, the urinary concentration of corticoids, the urinary corticoid:creatinine ratio, and the plasma cortisol concentration after the administration of 25 IU of ACTH intravenously. The records of 24 cases of histologically confirmed equine pituitary pars intermedia adenomas were used. An adenoma of the pars intermedia of the pituitary gland in horses was predicted with a sensitivity of 100, 100, and 92% for basal plasma ACTH (n = 16), the urinary corticoid:creatinine ratio (n = 7), and basal plasma insulin (n = 12), respectively. The sensitivity of the ACTH-stimulation test was 79% (n = 14).

Hyperlipoproteinaemia associated with atherosclerosis and cutaneous xanthomatosis in a cat.

Bilateral uveitis and multiple xanthomas (fat deposits in the skin) are described in a 3.5-year-old ovariohysterectomized female Persian cat. The cat had been treated for 2 years with corticosteroids. Examinations included a routine blood chemistry profile, radiographic examination of the thorax and abdomen, histopathological examination of multiple skin punch biopsies, and analysis of blood lipid components by cellulose-acetate electrophoresis and by preparative ultracentrifugation studies. Total lipid values were 23 g/l. Ultracentrifugation studies indicated strongly elevated VLDL and LDL fractions and a decreased concentration of the HDL fraction. Because of sudden blindness the cat was euthanized at the request of the owner. Autopsy revealed massive atherosclerotic changes in the large abdominal vessels, the wall of the aorta, and the coronary vessels. Although the exact pathogenesis remains uncertain, these unusual findings might be explained by a primary hyperlipoproteinaemia, complicated by long-term use of corticosteroids.

Undernutrition in relation to changed tissue composition in wild boar (Sus scrofa).

The effect of long term dietary restriction on the composition of liver, kidney, and several muscles was studied in wild boar (Sus scrofa) in order to evaluate the tissue protein content as an indicator for the nutritional status. Extreme undernutrition was associated with a relatively large weight reduction in liver and kidney. In liver and kidney, total protein content and the protein/DNA ratio were reduced through a reduction in both cell number and cell size. The lower protein/DNA ratios and the increased DNA concentration in the muscle of food restricted animals suggested only a reduction of cell size. The response to undernutrition varied considerably between two muscles. It was concluded that the DNA concentrations and the protein/DNA ratio in several tissues provides an additional tool for assessing nutritional status in living or shot wild boar.

The effect of undernutrition on haematological and serum biochemical variables in wild boar (Sus scrofa).

The effect of induced long term undernutrition on the blood composition of wild boar was studied. During the first weeks, the activity of plasma alkaline phosphatase decreased, while creatinine concentrations increased, and the ratio urea/creatinine decreased, suggesting a renal mechanism as well as recycling of nitrogenous waste products to conserve nitrogen. During the last weeks, haemoglobin and packed cell volume decreased, suggesting protein or mineral deficiencies, and the ratio urea/creatinine increased, indicating substantial endogenous protein catabolism.

Sedation of wild boar (Sus scrofa) and red deer (Cervus elaphus) with medetomidine and the influence on some haematological and serum biochemical variables.

Medetomidine, an alpha-2 agonist, was tested for the sedation of wild boar (Sus scrofa) (n = 6) and red deer (Cervus elaphus) (n = 7). The effectivity of this drug and the dose required for handling, blood sampling, and minor surgery were established for both species. In addition the effect of medetomidine sedation on haematological and serum biochemical variables was studied. All animals used were clinically normal females aged approximately 1.5 years. The minimal dose required to sedate a wild boar was 80 micrograms/kg (i.m.). The induction time was 35-50 min, and the sedation time was between 40 and 55 min; however, the sedation was insufficient. The minimal dose required for red deer was 80 micrograms/kg (i.m.). Blood sampling and minor surgery could be carried out without any problem. The induction time was 21-29 min and the sedation time was between 120 and 210 min. The alpha-2 antagonist atipamezole (300 micrograms/kg) was effective in reversing sedation, with recovery occurring between 3 and 9 min. Medetomidine sedation influenced the blood composition of wild boar. Haemoglobin and packed cell volume decreased by about 20% (P < 0.002), total serum protein content decreased slightly (P = 0.011), and the glucose concentration nearly doubled (P = 0.002).

Equine pituitary neoplasia: a clinical report of 21 cases (1990-1992).

The records of 21 horses with an adenoma of the pars intermedia of the pituitary gland were reviewed. The animals comprised nine Dutch warmblood horses and 12 ponies. They ranged in age from 12 to 30 years (mean +/- sem 21 +/- 1.3 years) for 20 of the horses, and comprised 13 geldings and eight mares. All the animals showed hirsutism. In 1991, nine horses were diagnosed as having the adenoma, equivalent to 0.5 per cent of the horses examined in 1991. The mean +/- sem survival time of four of the horses (from discharge to the development of complications which required euthanasia) was 192 +/- 59 days (range 120 to 368 days). Routine biochemical measurements were within their normal ranges except for a high alkaline phosphatase activity (768 +/- 487 iu/litre) and a high plasma glucose concentration (7.8 +/- 0.9 mmol/litre) in 16 of the horses; only six had a glucose concentration above 9.0 mmol/litre. The basal plasma concentration of adrenocorticotrophic hormone (ACTH) in the diseased horses was at least 191 pg/ml. Eight of the horses had mean plasma cortisol concentrations before and after the administration of 25 iu of synthetic ACTH1-24 of 168 +/- 21 (range 74 to 248) and 478 +/- 58 nmol/litre (range 248 to 771), respectively. Analysis of the urine of five of the horses revealed a mean specific gravity of 1.034 +/- 0.0034 (range 1.025 to 1.045).(ABSTRACT TRUNCATED AT 250 WORDS)

Mucosal uptake, mucosal transfer and retention of iron in veal calves.

A method for studying iron absorption in humans was adapted to veal calves. Three 10-week-old calves with moderate (calves 1 and 2) or severe (calf 3) iron deficiency were given an abomasal injection of 59Fe and 51Cr and all their faeces were collected over 15 days in order to measure mucosal uptake, mucosal transfer and retention of iron. The mucosal uptake was 62.2, 53.4 and 71.8% in calves 1, 2 and 3, respectively. The iron retention measured 14 days after administration of the test dose was 57.4, 52.3 and 56.4% in calves 1, 2 and 3, respectively. Maximal plasma activity was found in all three calves between 1 1/2 and 2 h after injection of the test dose. The plasma activity decreased rapidly, with a slight increase between the 5th and the 10th hour. After 21 h, less than 0.25% of the injected dose was still present in 1 litre of plasma. Not all the 51Cr was recovered in the faeces. No 59Fe was found in the urine but some 51Cr could be detected. The results of this study show that the method described is useful for measuring the different steps of iron absorption in iron-deficient veal calves.

Fever and acute phase response induced in dwarf goats by endotoxin and bovine and human recombinant tumour necrosis factor alpha.

Tumour necrosis factor (TNF), a polypeptide produced by mononuclear phagocytes, has been implicated as an important mediator of inflammatory processes and of clinical manifestations in acute infectious diseases. To study further the potential role of TNF in infectious diseases, recombinant Escherichia coli (E. coli) derived human (r.HuTNF-alpha) and bovine TNF (r.BoTNF-alpha) were intravenously (i.v.) administered in dwarf goats. Rectal temperature, heart rate, rumen motility, plasma zinc and iron concentrations, and certain other blood biochemical and haematological values were studied and compared with the changes seen after E. coli endotoxin (LPS) was administered (dose: 0.1 microgram/kg i.v.). Following a single injection of 4 micrograms/kg of r.BoTNF-alpha, shivering and biphasic febrile response were observed, accompanied by tachycardia, inhibition of rumen contractions, drop in plasma zinc and iron concentrations, lymphopenia, and neutropenia followed by neutrophilia. The i.v. administration of a single injection of 4 micrograms/kg r.HuTNF-alpha induced shivering and biphasic febrile responses, accompanied by anorexia and a similar drop in plasma trace metal concentrations when compared with r.BoTNF-alpha-treated goats. The TNF-alpha-induced symptoms were essentially the same as those that occurred after LPS administration. However, the time of onset of these changes after the injection of TNF-alpha was significantly shorter than after LPS. Moreover, the r.BoTNF-alpha induced a longer lasting neutrophilic leucopenia, less neutrophilia, and a more persistent lymphopenia than after LPS injection. Neither r.BoTNF-alpha nor LPS caused severe haemo-concentration. Furthermore, no cross-tolerance between r.BoTNF-alpha and LPS could be demonstrated. We conclude that both r.BoTNF-alpha and r.HuTNF-alpha induce many of the physiologic, haematologic and metabolic changes that characterize the acute phase response to LPS. The overlapping biological activities of r.BoTNF-alpha, r.HuTNF-alpha and LPS in dwarf goats may indicate that both recombinant tumour necrosis factors have some homology with caprine TNF-alpha.

[Toxicity of cadmium in cattle]. / Toxiciteit van cadmium voor rundvee.

After having investigated the metabolism of cadmium in plants and animals, the Animal Health Service of North Brabant in 1982 instigated research into the level of cadmium in the livers and kidneys of beef cattle kept in the vicinity of a zinc processing factory. Cadmium levels in these cattle were more than twice as high as those of cattle kept in control areas. Clinical abnormalities as a result of the increased cadmium levels were not observed, although haemoglobin and iron levels, iron-binding capacity and iron saturation were lower in these animals than in control animals. Cadmium therefore appears to have an adverse effect on iron absorption. Administration of high doses of cadmium for a limited period to 14 heifers did not cause discernable clinical abnormalities. These animals retained cadmium to a much smaller extent than anticipated on the basis of data for monogastric (mainly rats) animals.

Relationship between blood hemoglobin, plasma and tissue iron, muscle heme pigment, and carcass color of veal.

In 41 veal calves divided into three groups and fed different levels of dietary iron, blood hemoglobin, plasma iron, liver, spleen, and muscle iron, muscle heme pigment, and carcass muscle color at slaughter were studied. At 45 min postmortem, total carcass color was visually evaluated in the 41 carcasses. In different muscles of the carcasses the color was measured instrumentally using an invasive color measurement method at 45 min postmortem (MCDI score) and a surface color measurement method at 20 h postmortem (Minolta L*, a*, b*, and Chroma scores). Among the three groups, differences (P less than .05) in muscle iron concentrations, muscle heme pigment concentrations, and Minolta a*, b*, and Chroma scores were found. Most striking were the differences in mean iron concentrations in the longissimus thoracis muscles between Groups A (29 micrograms/g DM) and B (44 micrograms/g DM) and in the semimembranosus muscles between Groups A (31 micrograms/g DM) and C (45 micrograms/g DM). The correlations found between Minolta L*, a*, or Chroma score and the iron and heme pigment concentrations in the semimembranosus muscles were high in comparison with those found in the longissimus thoracis and rectus abdominis muscles. Compared with the plasma iron concentration, the blood hemoglobin concentration showed higher correlations with muscle iron and muscle heme pigment concentrations. It can be concluded that different iron concentrations in the milk replacer during the first 7 wk of fattening influence, to some extent, muscle iron and muscle heme pigment at slaughter. However, these differences were not measurable in the overall visual color evaluation of the carcass surface muscles.

Effect of ileo-rectal anastomosis and post-valve T-caecum cannulation on growing pigs. 2. Blood variables and mineral balances.

In a long-term study nine ileo-rectally anastomosed (IRA) and seven post-valve T-caecum (PVTC)-cannulated pigs were compared with six intact pigs with regard to different blood variables, sodium and potassium retention and weights of selected organs. After surgery, apart from urea and K measured 13 weeks post-surgery, there were no differences in the blood variables between the PVTC-pigs and intact pigs. In IRA-pigs concentrations of creatinine (P < 0.01), Na (P < 0.001), base excess (P < 0.001), pH (P < 0.01) and bicarbonate (P < 0.001) in blood were lower than those in intact pigs. At 13 weeks after surgery the blood K concentration in IRA-pigs was higher (P < 0.001) than that in PVTC-pigs or intact pigs. At 6 weeks after surgery the blood urea concentration in IRA-pigs was higher (P < 0.001) than that in intact and PVTC-pigs. At 13 weeks after surgery the urea concentration in PVTC-pigs was higher (P < 0.001) than those in IRA-pigs or intact pigs. The Na (P < 0.01 11 weeks after surgery) and (P < 0.05 and P < 0.01 5 and 11 weeks after surgery respectively) balances in IRA-pigs were lower than those in intact animals. Na retention was negative for IRA-animals 11 weeks after surgery. Na and K retentions were similar in PVTC-pigs and in intact pigs. The urinary: faecal excretion of Na differed slightly between PVTC-animals and intact animals. At 13 weeks after surgery there were no differences in organ weights between the PVTC-pigs and intact animals. In the IRA-pigs the weights of the liver (P > 0.05), the kidneys (P > 0.05) and the adrenal glands were higher (P < 0.001) than those in the intact animals.

Intestinal permeability in pigs during rotavirus infection.

Macromolecular permeability of the small intestine was tested in four 3-week-old gnotobiotic pigs inoculated with porcine rotavirus strain RV277 (group A). Pigs were administered 125I-labeled polyvinylpyrrolidone (molecular weight [mol wt], 40,000) orally 1 day before and 2 and 24 hours after virus inoculation, and blood samples were obtained every 6 hours. Eight hours after rotavirus inoculation, pigs had watery diarrhea. Increased permeation of 125I-labeled polyvinylpyrrolidone was not observed after clinical signs of infection had developed. Serum total protein and urea nitrogen concentrations increased slightly at the end of the study, probably as a consequence of dehydration. Differences in blood glucose concentration were not seen. At 48 hours after viral inoculation, macromolecular permeability was tested morphologically by injecting horseradish peroxidase (mol wt, 40,000) into the jejunal lumen just distally to the ligamentum colicoduodenale. After an incubation period of 20 minutes, small segments of jejunum were obtained for stereomicroscopic, histologic, and ultrastructural investigations. Moderate hyperregenerative villus atrophy was found. Ultrastructural changes of the villus epithelium were minor, and increased macromolecular permeation was not observed.