Direct measurement of progesterone in plasma and milk by a simple solid-phase radioimmunoassay

A rapid method for determination of progesterone in bovine, goat and porcine plasma as well as in bovine milk was evaluated. The method employed was a solid-phase (125)I radioimmunoassay equipped with progesterone standards in human serum and called the Coat-A-Count procedure. The dilution curves of bovine plasma samples with high progesterone content were parallel with the standard curve based on human serum. The relation between measurements of progesterone levels in bovine plasma using the reference extraction method and the direct Coat-A-Count procedure was highly significant, resulting in the linear regression equation Y = 1.06x-0.04. In case of goat and porcine plasma, the direct method yielded higher results than the reference extraction method (Y = 1.37x + 1.38 and Y = 1.69x - 6.47, respectively). Progesterone concentration in bovine whole milk was much higher when measured by the Coat-A-Count procedure than by the reference Farmos kit (Y = 1.59x + 1.51). However, when the same samples were assayed by a modified Coat-A- Count procedure, i.e. progesterone standards from Farmos kit, the values were more or less identical (Y = 0.88x - 0.21).     

Clinical signs and hormonal changes in dairy heifers after induction of parturition with prostaglandin F2 alpha

Parturition was induced in 12 dairy heifers with prostaglandin (PG) F2 alpha about 2 weeks before the expected time of calving. Eight animals gave birth after two injections (group 1), three animals needed more than two injections (group 2) and one animal (cow no. 740) required one injection. All animals in groups 1 and 2 had retained foetal membranes and the time needed to induce parturition was 59 +/- 7 and 149 +/- 10 h, respectively. As cow no. 740 did not have retained foetal membranes and calved 24 h after one PGF2 alpha injection, it was excluded from the results. Udder distension and relaxation of the pelvic ligaments could predict the calving to within 12 h. Furthermore, the pre-calving drop of body temperature could predict the time of parturition to within 16 h. The total white blood cells and polymorphonuclear cells were at their highest values on the day preceding parturition whereas mononuclear cells had a tendency to increase 3 days after calving. Increased levels of haemoglobin were found at the time of parturition, whereas, plasma-calcium levels significantly decreased after parturition (P < 0.001). Progesterone levels markedly decreased after the first PGF2 alpha injection and reached 2 nmol/l at the time of parturition. Plasma levels of oestradiol-17 beta reached the peak at the time of parturition, whereas, the highest levels of the PGF2 alpha metabolite and cortisol were recorded 16 h after calving.     

Cardiovascular,endocrine and behavioural responses to suckling and permanent separation in goats

Background

Suckling can be a peaceful or vulnerable event for goats and kids, whereas, separation is suggested as stressful. The aim of this study was to investigate physiology and behaviour in these two different situations in dairy goats.

Methods

Four studies were performed with seven goats kept with their first-born kid in individual boxes. The goats were videotaped and heart rate and arterial blood pressure were recorded every minute by telemetry from parturition until 24 hours after separation. One to two days after parturition, Study 1 was performed with analyses of heart rate and blood pressure around a suckling. In Study 2, performed 3-5 days after parturition, blood sampling was done before, during and after suckling. Study 3 was performed 4-6 days post partum, with blood sampling before and after a permanent goat and kid separation. In addition, vocalisations were recorded after separation. Blood samples were obtained from a jugular vein catheter and analysed for plasma cortisol, β-endorphin, oxytocin, and vasopressin concentrations. Study 4 was performed during the first (N1) and second nights (N2) after parturition and the nights after Study 2 (N3) and 3 (N4). Heart rate, blood pressure and time spent lying down were recorded.

Results

The kids suckled 2 ± 0.2 times per hour and each suckling bout lasted 43 ± 15 s. In Study 1, heart rate and blood pressure did not change significantly during undisturbed suckling. In Study 2, plasma cortisol (P ≤ 0.05 during suckling and P ≤ 0.01 five minutes after suckling) and β-endorphin (P ≤ 0.05) concentrations increased during suckling, but oxytocin and vasopressin concentrations did not change. In Study 3, the goats and kids vocalised intensively during the first 20 minutes after separation, but the physiological variables were not affected. In Study 4, heart rate and arterial blood pressure declined gradually after parturition and were lowest during N4 (P ≤ 0.05) when the goats spent longer time lying down than during earlier nights (P ≤ 0.01 during N1 and N3 and P ≤ 0.05 during N2).

Conclusions

Suckling elevated plasma cortisol and β-endorphin concentrations in the goats. The intensive vocalisation in the goats after separation, earlier suggested to indicate stress, was not accompanied by cardiovascular or endocrine responses.     

Afforestation of a trace-element polluted area in SW Spain: woody plant performance and trace element accumulation

Trace element soil pollution can have ecotoxic effects on plants, which could negatively affect the restoration of a degraded area. In this work, we studied the revegetation success in different sites within a trace element-polluted area (Guadiamar River Valley, SW Spain). We analysed the survival and growth patterns of afforested plants of seven Mediterranean woody species, and their relation to soil pollution, over 3 years. We also analysed the trace element accumulation in the leaves of these species. The area was polluted mainly by As, Cd, Cu, Pb and Zn (soil total concentrations up to 250, 3.6, 236, 385 and 510 mg kg⁻¹, respectively). The woody plant performance was very different between sites and between species; in the riparian sites, plant survival rates were nearly 100%, while in the upland terrace sites species such as Quercus ilex and Ceratonia siliqua showed the lowest survival rates (less than 30%) and also the lowest relative growth rates. There were no significant relationships between plant performance and soil pollution in the riparian sites, while in the upland sites mortality, but not growth, was related to soil pollution, although that could be an indirect effect of different substrate alteration between sites. The accumulation of soil pollutants in the studied plants was low, with the exception of Salicaceae species, which accumulated Cd and Zn in the leaves above 1 and 200 mg kg⁻¹, respectively. We discuss the results with regard to the afforestation of trace-element polluted areas.     

The cannabinoid receptors system in horses: Tissue distribution and cellular identification in skin

BackgroundThe endocannabinoid system (ECS) is composed of cannabinoid receptors type 1 (CBR1) and type 2 (CBR2), cannabinoid‐based ligands (endogenous chemically synthesized phytocannabinoids), and endogenous enzymes controlling their concentrations. Cannabinoid receptors (CBRs) have been identified in invertebrates and in almost all vertebrate species in the central and peripheral nervous system as well as in immune cells, where they control neuroimmune homeostasis. In humans, rodents, dogs, and cats, CBRs expression has been confirmed in the skin, and their expression and tissue distribution become disordered in pathological conditions. Cannabinoid receptors may be a possible therapeutic target in skin diseases.ObjectivesTo characterize the distribution and cellular expression of CBRs in the skin of horses under normal conditions.AnimalsFifteen healthy horses.MethodsUsing full‐thickness skin punch biopsy samples, skin‐derived primary epidermal keratinocytes and dermal‐derived cells, we performed analysis of Cnr1 and Cnr2 genes using real‐time PCR and CBR1 and CBR2 protein expression by confocal microscopy and Western blotting.ResultsNormal equine skin, including equine epidermal keratinocytes and dermal fibroblast‐like cells, all exhibited constant gene and protein expression of CBRs.Conclusions and Clinical ImportanceOur results represent a starting point for developing and translating new veterinary medicine‐based pharmacotherapies using ECS as a possible target.     

Genetic analysis of a possible case of canine X-linked ectodermal dysplasia

In the present report, we describe targeted next-generation sequencing of the EDA gene of a male poodle with a clinical and histopathological diagnosis of X-linked hypohidrotic ectodermal dysplasia. The result was compared with the reference sequence and with the result of the sequencing of a normal dog's EDA gene. No point variant, small deletion or insertion were found in the exons and splice sites, but a transition and a transversion were found in the intron 6 and 3′ UTR, respectively. The cause of the dysplasia of the affected dog in this study is neither a point variant nor a small deletion or insertion in the exons and splice sites of the EDA gene. Therefore, patients with phenotype of XLHED may have other types of variants in the EDA gene or variants in other genes of the EDA signalling pathway.     

The pharmacokinetics and antiparasitic activity of ivermectin in Hutsul and Toric horses

The aim of this study was to compare the pharmacokinetics of ivermectin and its antiparasitic activity in two horse breeds. Eight Hutsul and 14 Toric horses were administered ivermectin orally at a dose of 0.2 mg/kg body weight. Blood samples were collected for 96 hr, and faecal samples were collected one day before and on days 14 and 21 after drug administration. Ivermectin concentrations in plasma samples were determined by high‐performance liquid chromatography. Ivermectin concentration was significantly higher in Toric than in Hutsul horses 90 min after ivermectin administration and was maintained at higher level for up to 96 hr. The area under the concentration versus the time curve from 0 to the last sampling point (AUC_0→t) and the maximum plasma concentration (C_max) were significantly higher in Toric than in Hutsul horses (1792.09 ± 246.22 μg × hr/L vs. 716.99 ± 255.81 μg × hr/L and 62.72 ± 17.97 ng/ml vs. 35.34 ± 13.61 ng/ml, respectively). No parasitic eggs were found in the faecal samples collected from both groups of horses on days 14 and 21 after drug administration. The obtained results indicate that although the pharmacokinetics of ivermectin may differ significantly between horse breeds, these differences do not affect the effectiveness of therapy.     

Cocomposting of Beet Vinasse and Grape Marc In Windrows and Static Pile Systems

Two composts were obtained by cocomposting a concentrated depotassified beet vinasse and grape marc using an aerated static pile and a windrow system. The composting mixtures comprised grape marc (83%) and vinasse (17%) for the aerated static pile system and grape marc (77%), vinasse (20%) and phosphate rock (3%) for the windrow. Changes in temperature followed a similar path for both mixtures, however the thermophilic phase was longer in the aerated static pile (25 days) than in the windrow (10 days). This fact caused differences in both organic matter degradation, weight losses (21% for static pile and 10% for windrow) and gas losses during the process. Nevertheless, the composts obtained by the two systems had a high fertilizer nutrient value (18.2 g kg^?1 N; 3.1 g kg^?1 P; 13.6 g kg^?1 K, C/N 16.1 for compost obtained in static pile and 20.6 g kg^?1 N; 13.7 g kg^?1 P; 13.1 g kg^?1 K; C/N 18 for compost obtained in windrow). A high degree of stability was reached in both composting systems (124 cmol_c kg^?1 CEC for static pile and 153 cmolc kg^?1 CEC for windrow at 80 days of composting). The chemical and physical properties of both vinasse composts suggest their possible use as soil conditioner.     

Using a Second-Order Polynomial Model to Determine The Optimum Vinasse/Grape Marc Ratio For In-Vessel Composting

This paper evaluates the influence of operating conditions (pH, Kjeldahl-N losses, NO₃^?-N, NO₄⁺-N,C/N ratio and biodegradation) used in the vinasse/grape marc incubation on the properties of products obtained to determine the best incubation conditions (time and amount of vinasse proportion) to aid in the design of an in-vessel composting system. A second-order polynomial model consisting of two independent process variables was found to accurately describe, with <10% differences between experimental values and model prediction, the vinasse-grape marc incubation. The dependent variables measured were pH, Kjeldahl-N losses, NO₃-^?N, NO₄⁺-N, C/N ratio and biodegradation and the independent process variables were operation time and amount of vinasse. Results determined products with acceptable chemical properties, high biodegradation and germination index and minimum Kjeldahl-N losses entails operating at medium-to-short operation time (20-35 days) and medium-to-low vinasse percentages (0-20%).