Advanced oxidation protein products are generated by bovine neutrophils and inhibit free radical production in vitro

Despite the recognised importance of oxidative stress in the health and immune function of dairy cows, protein oxidation markers have been poorly studied in this species. The current study aimed to characterise markers of protein oxidation generated by activated bovine neutrophils and investigate the biological effects of advanced oxidation protein products (AOPP) on bovine neutrophils. Markers of protein oxidation (AOPP, dityrosines and carbonyls) were measured in culture medium containing bovine serum albumin (BSA) exposed to neutrophils. The effect of AOPP-BSA on generation of reactive oxygen species (ROS) was assessed by chemiluminescence. Activation of caspases-3, -8 and -9 and the presence of DNA laddering were used as apoptosis markers.Greater amounts of AOPP were generated by phorbol myristate acetate (PMA)-activated than non-activated neutrophils (1.46 ± 0.13 vs. 0.75 ± 0.13 nmol/mg protein, respectively; P < 0.05). Activated neutrophils and hypochlorous acid generated slightly different patterns of oxidized protein markers. Exposure to AOPP-BSA did not stimulate ROS production. Activated neutrophils generated a lesser amount of ROS when incubated with AOPP-BSA (P < 0.001). Activation with PMA induced a loss of viable neutrophils after 3 h, which was greater with AOPP-BSA incubation (P < 0.05). Detectable amounts of active caspases-3, -8 and -9 were found in nearly all samples but differences in caspase activation or DNA laddering were not observed comparing treatment groups. Apoptosis was unlikely to be responsible for the greater loss of PMA-activated neutrophils cultured in AOPP-BSA and it is possible that primary necrosis occurred. The results suggest that accumulation of oxidized proteins at an inflammatory site might result in a progressive reduction of neutrophil viability.     

Plasma Steroid Variations in Bull Calves Repeatedly Treated with Testosterone,Nortestosterone and Oestradiol Administered Alone or in Combination

The aim of this work was to investigate the secretion of dehydroepiandrosterone (DHEA), testosterone (T), dihydrotestosterone (DHT) and oestradiol (E) as biological markers in response to illegal administration of testosterone, 19-nortestosterone (N) and oestradiol, either alone or in combination. Twenty male Friesian calves (age 13-14 months) were allotted to a control group (n = 5), and five experimental groups (n = 3) each. Each experimental animal was repeatedly injected with one of the following hormonal treatments: E, T, N, T+E and N+E. Circulating DHEA, T, DHT and E were determined by radioimmunoassay. The administration of T alone did not induce any variation in plasma DHEA, T, DHT and E, which were similar to those in the control group. In contrast, DHEA, T and DHT were on average significantly lower in the T+E and N-treated groups (p < 0.01), whereas the administration of N+E resulted in the reduction of plasma T and DHT without any modification of plasma DHEA. The administration of E alone or in combination increased circulating levels of E but did not affect androgen plasma profiles. The results indicate that plasma levels of T do not permit detection of illegal treatments because plasma androgens always remained within the physiological range. Illegal E treatment could be detected in blood samples when they were collected at least every 20 days.     

Relationship between oxidative stress and the success of artificial insemination in dairy cows in a pasture-based system

This study was designed to evaluate whether the outcome of artificial insemination (AI) was affected by the metabolic and oxidative status of dairy cows. Seventy-nine inseminations in 40 cows, were classified, on the basis of blood progesterone (P4) and pregnancy-associated glycoprotein (PAG) concentrations and clinical confirmation of pregnancy into, three categories: (1) positive (AI+, resulted in pregnancy, n=26; 33%), (2) negative (AI-, did not result in pregnancy, n=49; 62%), and (3) embryonic mortality (EM, n=4; 5%). Reactive oxygen metabolites, biological antioxidant potential, oxidative stress index, body condition score, glucose, total proteins, albumin, urea, non-esterified fatty acids (NEFAs), cholesterol, triglycerides, haptoglobin and advanced oxidative protein products (AOPPs) were measured on the day of AI (day 0), and 30 and 42days later. Cows with EM had lower BCS scores (2.5) than AI+ (2.8) and AI- (2.9) cows (P<0.05). During the post-partum period, body condition score (BCS) increased and NEFAs decreased (P<0.05) suggesting a recovery from the negative energy balance (NEB). The only significant differences found were that the mean concentration of AOPPs was higher and that of albumin lower in EM cows than in AI+ and AI- (P<0.05) animals. Plasma concentration of reactive oxygen metabolites and biological antioxidant potential were not related to AI outcome. Further studies are required to confirm this finding and to clarify the role of oxidative status on cows' fertility.     

Evaluation of Cabergoline and Buserelin Efficacy for Oestrous Induction in the Bitch

The purpose of this work was to compare two different protocols of oestrous induction, using either a dopamine agonist (cabergoline) or a GnRH agonist (buserelin) in anoestrus bitches. The clinical trial involved 22 Beagle bitches, randomly allotted to two treatment groups: group A (n = 12) was orally administered cabergoline (Galastop^®; Centralvet‐Vetem, Milan, Italy; 5 μg/kg SID), until the onset of cytological oestrus or for a maximum of 30 days and group B (n = 10) was treated with buserelin acetate, (Suprefact^®; Aventis Pharma, Milan, Italy), administered subcutaneously t.i.d., at 1.5 μg/kg for 11 days and 0.75 μg/kg for the following 3 days. Blood samples were collected twice a week to measure progesterone and prolactin concentration. Both cabergoline and buserelin produced a significant early decline in prolactin concentration (p < 0.01), but the effect of cabergoline lasted longer. Progesterone concentration was significantly affected by buserelin administration, showing a significant increase (p < 0.01) from day 3 to day 6 of treatment. Cabergoline confirmed its effectiveness in inducing oestrus as 10 of 12 bitches responded to the treatment, were mated and whelped. On the contrary, oestrus was observed in only three of 10 buserelin‐treated bitches and in two of them 7 and 13 days after the end of treatment. These same two bitches accepted mating and conceived. The results suggest that in a clinical setting, dopaminergic treatment is the treatment of choice as it yields more consistent results and involves a much easier administration protocol.     

Glucose or essential amino acid infusions in late pregnant and early lactating Simmenthal cows failed to induce a leptin response

To assess the leptin response to metabolic challenges, three Italian Simmental cows were infused for 6 h: with (a) saline (control); (b) glucose; and (c) amino acid solutions according to a 3 x 3 Latin square experiment. The infusions were carried out at the 36th week of pregnancy, and the second and 12th week of the following lactation. At each of the three infusion periods, blood samples were collected from the jugular vein before and 15, 30, 120, 180, 240, 300 and 360 min after the beginning of each infusion. All samples were analysed for leptin, insulin, glucagon, growth hormone (GH), glucose, non-esterified fatty acids (NEFA) and urea. The physiological phase of the cows significantly affected the basal concentrations of insulin, glucagon, urea and NEFA. The infusion of both glucose and the amino acid solutions did not affect leptin concentrations. Insulin response was significantly increased when animals were infused with the glucose solution and, within treatment, the greatest response was observed at the 12th week of lactation. The greatest glucagon response was observed when infusing the amino acid solution. Urea response to all treatments increased from the dry period to the 12th week of lactation. The GH and NEFA responses were not affected by treatments. The Multi Species radio-immunoassay used in this study showed a lower sensitivity for ruminant leptin which may partially explain the lack of significant leptin variations. However, it can be hypothesized that leptin variations around parturition can be affected by the negative energy balance, and leptin release is not acutely affected by glucose and amino acid availability. In addition, no short-term relationship were found between insulin, glucagon and GH and leptin release in Italian Simmental cows during the dry period and early lactation.     

Effect of age and sex on plasma cortisol and dehydroepiandrosterone concentrations in the dog (Canis familiaris)

Limited data exist on age-related physiological variations in plasma concentrations of cortisol and dehydroepiandrosterone (DHEA) in dogs, despite their potential role in the pathophysiology of ageing. This study examined plasma cortisol and DHEA concentrations and cortisol/DHEA ratio variations, according to age and sex in 311 dogs, aged from two months to 16 years. Before adulthood, DHEA concentrations were higher in peri-pubertal males. During adulthood, cortisol and DHEA were higher in males than females. Among females, DHEA was lower in older dogs, but the decrease was observed at an older age in intact than ovariectomised females. Variations in the cortisol/DHEA ratio inversely reflected those of DHEA. Results indicate that testicles are an important source of DHEA in males, and that DHEA is mainly secreted by the adrenal glands in females. The ovaries’ contribution to circulating DHEA appears to be limited, although it may partially compensate an age-related decrease in adrenal secretion.     

Effect of Aging and Reproductive Condition on Dehydroepiandrosterone Plasma Levels in the Bitch

Veterinary Research Communications - Marinelli, L., Gabai, G., Simontacchi, C. and Bono, G., 2007. Effect of aging and reproductive condition on dehydroepiandrosterone plasma levels in the bitch....     

Apoptosis and Expression of Related Proteins in Mammary Gland of Heifers during Early Lactation

Veterinary Research Communications -     

Relationship between plasma progesterone and pregnancy-associated glycoprotein concentrations during early pregnancy in dairy cows

The relationship between the concentration of plasma progesterone (P4) during embryo attachment or at recognition of pregnancy, and that of pregnancy-associated glycoprotein (PAG) was assessed in dairy cows. The outcome of artificial insemination (AI) was classified as positive (AI+), negative (AI?), or late embryonic mortality (EM) by measuring circulating PAG concentrations and by ultrasonography. Based on P4 concentrations at either day 21 or day 15, AI+ and EM cows were classified into ‘low’ (P4 concentrations < mean) and ‘high’ (P4 concentrations > mean) P4 groups. In both experiments, the threshold of P4 concentration between the ‘low’ and ‘high’ groups was approximately 6 ng/mL. PAG concentrations were lower in the ‘low’ group only when P4 concentrations were below the threshold. The study findings suggest that a possible P4 threshold exists below which PAG secretion may be impaired.