Effects of oral supplementation with β-carotene on concentrations of β-carotene, vitamin A and α-tocopherol in plasma, colostrum and milk of mares and plasma of their foals and on fertility in mares

In this study, effects of oral β-carotene supplementation to mares (β-carotene group: 1000 mg/day, n = 15; control group: n = 15) from 2 weeks before foaling until 6 weeks thereafter on concentrations of β-carotene, vitamin A and α-tocopherol in plasma, colostrum and milk and plasma of their foals were determined. In addition, effects on fertility were studied. Beta-carotene concentrations increased in plasma and colostrum of β-carotene-supplemented mares compared to control mares (p < 0.05). In mares of both groups, β-carotene concentrations were higher in colostrum than in milk (p < 0.05). In foals, β-carotene concentrations increased with colostrum uptake and were higher in foals born to supplemented mares (p < 0.05; control group: 0.0003 ± 0.0002 μg/ml on day 0, 0.008 ± 0.0023 μg/ml on day 1; β-carotene group: 0.0005 ± 0.0003 μg/ml on day 0, 0.048 ± 0.018 μg/ml on day 1). Concentrations of vitamin A and α-tocopherol were higher in colostrum than in milk (p < 0.05) but did not differ between groups. Concentration of α-tocopherol in plasma of mares decreased over time and in foals, increased markedly within 4 days after birth. All but one mare (control group) showed oestrus within 2 weeks post-partum. Occurrence of oestrus did not differ between groups. More mares of the control group (7/7 vs. 5/12 in the β-carotene group) became pregnant after being bred in first post-partum oestrus (p < 0.05). In conclusion, β-carotene supplementation to mares increased β-carotene concentrations in plasma, colostrum and milk of mares and plasma of their foals but had no positive effects on fertility.     

Evaluation of a fluorescence-polarization assay for the diagnosis of bovine brucellosis in México.

A homogeneous fluorescence-polarization assay (FPA) was used for the serological diagnosis of bovine brucellosis in México. The assay uses O-polysaccharide prepared from Brucella abortus lipoplysaccharide (20-30 kDa) conjugated with fluorescein isothiocyanate as a tracer. To measure the fluorescence polarization, a FPM-1 fluorescence-polarization analyzer was used with the procedure described by Nielsen et al. (1996b). A cut-off value of 90 millipolarization (mP) units was used for testing 560 bovine sera from different areas of México. (305 positive sera and 255 negative sera according to the complement fixation test; CFT.) Some were tested with the Rose Bengal plate (RB) test (n = 490) and some with the rivanol-agglutination (RIV) test (n = 190). Sensitivities were 98.3%, 99.3% and 99.0%, and specificities were 68.8%, 55.4% and 96.9%, respectively, for RB, RIV and FPA. The FPA gave a kappa coefficient of agreement with respect to CFT of 0.96, while RB and RIV (relative to the CFT) gave coefficients of 0.70 and 0.61, respectively. Finally, ROC analysis suggested a cut-off value which agreed with the one recommended in the test procedure. We concluded that FPA is a suitable test to be used instead of the CFT in Mexican conditions.     

Egg‐shell deposition and blood plasma inorganic phosphorus concentration in individual laying hens

1. The relationship between plasma inorganic phosphorus measured at the end of the dark period and the weight of the next egg shell produced was investigated in 30 Hisex Brown hens.

2. Hens received an adequate conventional layers’ diet and the lighting pattern was conventional (17 L:7 D). Observations were made early (31 to 40 weeks) and late (62 to 68 weeks of age) in the laying period.

3. Eggs were classified by position in the clutch sequence and significant negative correlations were found between shell weights of early eggs in the sequence and plasma phosphate at the end of the dark period.

4. No significant trends were found in plasma total calcium.

5. No significant differences were found in bone compositions of birds producing consistently more or less than average shell weight during the laying period.

6. The negative correlations between plasma phosphate and shell weight are consistent with the observations of Sauveur and Mongin (1983) and show that impairment of shell deposition is associated with skeletal mobilisation as indicated by increase in plasma phosphate. This is consistent with the observations on bone composition and indicates that selection for shell quality will tend to select birds which are not dependent on excessive skeletal mobilisation during shell formation.     

Effects of clopidogrel therapy on whole blood platelet aggregation,the Plateletworks® assay and coagulation parameters in cats with asymptomatic hypertrophic cardiomyopathy: a pilot study

Background: Although scientific evidence is limited, clopidogrel is frequently used as prophylaxis for arterial thromboembolism in cats with hypertrophic cardiomyopathy (HCM).

Objectives: Evaluating effects of clopidogrel therapy in asymptomatic cats with HCM on (1) conventional whole blood aggregation (WBA), (2) alternative platelet aggregation assessed with tubes of the Plateletworks® assay and (3) standard coagulation parameters.

Animals and methods: Prospective, randomized, double-blind, placebo-controlled pilot study. Fourteen asymptomatic HCM cats were randomly allocated to receive placebo (n = 5) or clopidogrel (18.75 mg/cat q24h, n = 9) as part of a larger study. Aggregation responses (to 20 µM adenosine diphosphate (ADP) and 10 µg/ml collagen) in WBA and the Plateletworks® assay and standard coagulation parameters were evaluated at baseline and after seven days of therapy.

Results: Clopidogrel therapy significantly reduced aggregation responses to ADP and collagen in the Plateletworks® agonists tubes (ADP and collagen: P < 0.001), but did not significantly reduce aggregation responses to ADP and collagen in the WBA technique (ADP: P = 0.07, collagen: P = 0.30). Clopidogrel therapy did not show a significant effect on prothrombin time, activated partial thromboplastin time, antithrombin, D-dimers and fibrinogen concentrations.

Conclusion and clinical importance: Clopidogrel therapy at a dose of 18.75 mg/cat q24h for seven days causes a significant decrease in in vitro platelet aggregation evaluated with the Plateletworks® assay, without affecting standard coagulation parameters in cats with asymptomatic HCM.     

Determination of decisional cut-off values for the optimal diagnosis of bovine tuberculosis with a modified IFNγ assay (Bovigam®) in a low prevalence area in France

The Bovigam(?) gamma interferon (IFNγ) assay was used to complement official skin-test screening in a low bovine tuberculosis (bTB) prevalence region in France. The aim of our work was to determine decisional cut-off values for protein purified derivatives (PPD) and ESAT6-CFP10 antigens (R) in order to optimize the efficacy of the modified Bovigam(?) test, in this low-prevalence area, for optimal classification of infected or non-infected herds following positive skin tests. The sensitivity of the IFNγ assay relative to post-mortem bTB-positive animals (Se(r)) was studied in 60 cattle from 20 bTB-infected herds. Its absolute specificity (Sp) was studied in 492 cattle from 25 bTB-free herds from a bTB-free zone. Its operational specificity (relative to the positive skin test) (Sp(r)) was also studied in 547 skin-test positive cattle from 172 bTB-free herds from an infected zone. Using normalized interpretations for individual (PPD or R) results, the cut-off values at 0.02 for PPD and 0.01 for R were obtained with a view to employ them in low prevalence areas with no previously observed non-specific reactions to SITT. Concerning its use after positive skin tests, cut-off values were set at 0.05 for PPD and at 0.03 for R. The choice of an interpretation method considering positive results with PPD and/or R (PPDUR), justified in a high risk context, provided a test Se(r) of 93% [84-98] and Sp(r) of 71.8% [67.9-75.6]. Analysis of positive results with PPD and R (PPDUR), ideal for low-risk contexts, provided a test Sp(r) of 94.3% [92.0-96.1] and Se(r) of 77% [64-87]. Thus, adapting the criteria to the region's infection status and to the conditions for its application is essential for the appropriate use of the IFNγ assay.     

Relationship between embryo collection results after superovulation treatment of Japanese Black cows and their plasma β-carotene and vitamin concentrations

The objective of this study was to investigate the relationship between the plasma concentrations of vitamin A (VA), vitamin E (VE) and β-carotene (BC) during embryo collection in Japanese Black cows that had undergone superovulation treatment and the embryo collection results. Following superovulation treatment in 116 Japanese Black cows, we collected 1317 embryos by nonsurgical means seven days after artificial insemination. The collected embryos were classified into transferable embryos, unfertilized oocytes and degenerated embryos. After embryo collection, we collected blood samples from the cows and measured the plasma concentrations of VA, VE and BC. The cows were then divided into 2 groups depending on the plasma concentration of VA (L and H: < 80 IU/dl and ≥ 80 IU/dl), VE (L and H: < 150 μg/dl and ≥ 150 μg/dl) and BC (L and H: < 150 μg/dl and ≥ 150 μg/dl). As a result, the number of collected embryos in the H group of VE was significantly (P<0.01) higher than that in L groups. Furthermore, the number of transferable embryos was higher (P<0.05) in all VA, VE and BC H groups than in the L groups. The H group for BC showed a high ratio of transferable embryos compared with the L group (P<0.05). Consequently, the present study suggests that the plasma VE and BC concentrations are positively correlated with the embryo collection results.     

The effect of tannic acid and polyethylene glycol on the absorption capacity of chicken intestine for d-xylose and β-carotene

In an experiment, the possible influence of tannic acid (TA) and polyethylene glycol (PEG) on the absorption capacity of intestine for d-xylose and β-carotene in broiler chicken was investigated. Four groups of nine 28-day-old broiler cockerels received d-xylose (500 mg) and β-carotene (52 μg) solutions (Group 1 to 4) with TA (1 g, Group 2 to 4) and PEG (500 mg Group 3 and 1 g Group 4), orally. One blood sample prior to, and four others after the administration of test materials, were collected from wing vein on 40 min basis, for 160 min and the concentration of plasma d-xylose was determined. The concentration of β-carotene was also measured in plasma of blood samples taken prior to and 160 min post-administration of the test materials. Plasma d-xylose concentration of all groups showed quadratic correlations with time (p < 0.001, r(2) = 0.84, 0.60, 0.70 and 0.74 for Group 1 to 4, respectively). Administration of TA reduced the plasma concentration of d-xylose in Group 2. However, feeding PEG after TA raised the concentration of d-xylose in Group 4 to the level that there was no difference in that variable between this group and Group 1. Although the plasma concentration of β-carotene was increased in 160 min post-ingestion of the test material, no difference was found in that variable among the experimental groups. In conclusion, TA and its interaction with PEG have impacts on the absorption capacity of intestine for d-xylose and highly likely other simple sugars, but TA or PEG have no influence on the absorption of β-carotene and most probably other fat soluble vitamins.     

Evaluation of Precision Xceed® meter for on-site monitoring of blood β-hydroxybutyric acid and glucose concentrations in dairy sheep

The accuracy of the Precision Xceed® hand-held meter as an on-site method for measuring blood β-hydroxybutyric acid (BHBA) and glucose concentrations, for the diagnosis of pregnancy toxemia and ketosis in dry and lactating dairy sheep, was assessed. Five to eight hours after the start of the morning feed, blood was collected once from 193 clinically healthy sheep (143 dry and 50 lactating). BHBA and glucose analyses were performed with serum in the laboratory, and with whole blood with the Precision Xceed®. Overall, BHBA and glucose determinations by the two methods were not statistically different (P > 0.05). Strongly significant positive correlations were found for glucose and BHBA concentrations between the Precision Xceed® and laboratory results (r = 0.76, n = 150, P < 0.01 and r = 0.99, n = 193, P < 0.01, respectively). The Precision Xceed® was highly sensitive (98.6%) and specific (98.2%), and had excellent test agreement for the detection of pregnancy toxemia and ketosis.     

Development and application of a real-time TaqMan(®) qPCR assay for detection and quantification of 'Candidatus Mycoplasma haemolamae' in South American camelids

Two alpacas from a herd in southwest Switzerland died for unknown reasons. Necropsy revealed chronic weight loss and pale mucous membranes. Infection with hemotropic mycoplasmas was suspected and subsequently confirmed by molecular methods. In order to investigate the epidemiological situation in this herd, a real-time TaqMan(?) qPCR assay for the specific detection and quantification of hemoplasma infection in South American camelids was developed. This assay was based on the 16S rRNA gene and amplified 'Candidatus Mycoplasma haemolamae' DNA, but not DNA from other hemoplasmas or non-hemotropic mycoplasma species. The lower detection limit was one copy/PCR, and the amplification efficiency was 97.4%. In 11 out of 24 clinically healthy herd mates of the two infected alpacas, 'Candidatus M. haemolamae' infection was confirmed. No correlation was found between bacterial load and clinical signs or anemia. The assay described herein enables to detect and quantify 'Candidatus M. haemolamae' and may be used in future studies to investigate the prevalence, pathogenesis and treatment follow-up of hemoplasma infections in South American camelids.     

Relationship between contractions of the uterus and concentration of PGF2α in uterine venous blood after luteolysis in gilts

The origin and physiological significance of high pulses of prostaglandin F2α (PGF2α) in uterine venous blood that occur 2-3 days after luteolysis are not well understood. We studied the relationship between contractions of the uterus evoked by exogenous oxytocin (OT) and PGF2α concentration in uterine venous blood on day 17 of the porcine oestrous cycle. The infusion of OT into the uterine artery produced an immediate increase in the uterine intraluminal pressure (UIP) (p < 0.001) and a simultaneous elevation in PGF2α concentration in uterine venous blood (p < 0.0001). The infusion of indomethacin (IND) into the uterine artery slightly decreased PGF2α concentration in uterine venous blood, but it did not suppress uterine contraction or the rapid increase in PGF2α concentration in uterine venous blood just after OT infusion (p < 0.0001), which was lower that in gilts not treated with IND. We conclude that the spikes of PGF2α concentration in uterine venous blood occurring after OT infusion on day 17 of the porcine oestrous cycle are mainly caused by the excretion with venous blood from the remodelled uterus and that PGF2α synthesis may contribute to this. These results suggest that the high spikes in PGF2α concentration that occur 2-3 days after luteolysis in pigs, sheep, cows and mares all have a similar origin.     

Accuracy of capillary blood 3-β-hydroxybutyrate determination for the detection and treatment of canine diabetic ketoacidosis

In human medicine, diagnosis of diabetic ketoacidosis (DKA) is usually based on measurement of capillary 3-β-hydroxybutyrate (3-HB) with a hand held ketone sensor. This study was conducted to determine if measurement of capillary 3-HB could be useful for the diagnosis and monitoring of canine DKA. Fifteen dogs with diabetic ketosis and 10 with DKA were evaluated. Paired measurements of 3-HB of capillary and venous blood samples were analysed by the electrochemical sensor and reference method. Use of capillary 3-HB measurement during DKA management was then evaluated through simultaneous measurements of capillary 3-HB, urinary AcAc and venous blood gas analysis. Good agreement between capillary and venous 3-HB measurement was detected by the electrochemical sensor and reference method. Monitoring treatment of DKA revealed a significant correlation between capillary 3-HB and acidosis markers, while no significant correlation was observed between AcAc and acidosis markers. A cut-off value of capillary blood 3-HB >3.8 mmol/L for diagnosis of DKA resulted in 70% and 92% sensitivity and specificity. The electrochemical sensor accurately measures 3-HB concentration in both capillary and venous blood samples, is accurate in diagnosing canine DKA, and appears to reflect the patient''s metabolic status during DKA treatment.     

Protein heterogeneity of bovine β-casein in Danish dairy breeds and association of rare β-casein F with milk coagulation properties

A proteomic approach was applied to characterize genetic variation and relative protein content of β-casein (β-CN) in milk from two minor native dairy breeds in Denmark; Red Danish 1970 and Jutland cows in comparison to the major dairy breeds; Danish Holstein and Danish Jersey. Genetic β-CN variants were related to the relative concentration of β-CN. Furthermore, a very high frequency of variant F in the native breeds was found. This was remarkable as this variant is only found in a very low frequency in the major Danish breeds. Novel associations were found between β-CN variant F and milk coagulation properties in the native breeds, which is an important finding for understanding the genetic background for non- and poor coagulating milk and potential exploitation of β-CN variants in selective breeding programs. Furthermore, in Red Danish 1970 a very high prevalence of non-coagulating milk was found (18%).     

Evaluation of two cocktails containing ESAT-6, CFP-10 and Rv-3615c in the intradermal test and the interferon-γ assay for diagnosis of bovine tuberculosis

The intradermal tuberculin tests and the interferon-gamma (IFN-γ) assay are the principal tests used worldwide for the ante-mortem diagnosis of bovine tuberculosis. The conventional reagent currently in use in these tests is purified protein derivative (PPD) tuberculin obtained from Mycobacterium bovis culture. The components of PPD are poorly characterized and difficult to standardize. To overcome this issue, antigens specific to the Mycobacterium tuberculosis complex are being studied. Here we have assessed the biological potency of ESAT-6, CFP-10 and Rv-3615c presented as peptide or recombinant protein cocktails in comparison with the standard bovine PPD used routinely in Spanish eradication campaigns. The study was performed in cattle (n=23) from a herd with natural M. bovis infection. Animals were simultaneously injected with PPD and the peptide and protein cocktails. The percentages of cattle reacting positively to single intradermal test were 60.9% (bovine PPD), 47.8% (peptide cocktail) and 60.9% (protein cocktail), with no significant difference between the actual skin fold thickness increases (p>0.05). The IFN-γ assay detected 60.9% of animals when stimulation was performed with bovine PPD, but decreased to 52.2% when stimulation was performed with the peptide cocktail and to 47.8% when stimulation was performed with the protein cocktail. However, no significant differences were found between IFN-γ responder frequencies (p>0.05). These results show a potential use of these defined reagents for in vivo tuberculosis diagnosis.     

Validation of noninvasive blood pressure equipment: which peripheral artery is best for comparison studies in dogs?

Objectives

1) To determine which peripheral artery commonly used for invasive arterial blood pressure (IBP) monitoring yields the least bias when compared with noninvasive blood pressure (NIBP) values obtained at the antebrachium of the dog, and 2) to identify and describe differences in systolic (SAP), mean (MAP) and diastolic arterial pressures (DAP) among different anatomical locations.