Study of Sperm Concentration,Seminal Plasma Composition and their Physiological Correlation in the Persian Sturgeon,Acipenser persicus

The objectives of the present study were to determine ionic and organic composition of seminal plasma, sperm concentration and their relationships in the Persian sturgeon (Acipenser persicus). In this regard, ionic content (Na⁺, K⁺, Cl^?, Ca²⁺ and Mg²⁺) and organic content (total protein, glucose, cholesterol and triglyceride) along with sperm concentration were measured in 17 specimens of the Persian sturgeon. The seminal plasma contained 59.53 ± 2.56 mm /l sodium, 9.1 ± 1.42 mm chloride, 4.72 ± 0.3 mm potassium, 1.45 ± 0.075 mm calcium and 0.7 ± 0.072 mm magnesium. The following organic contents were found: total protein 0.11 ± 0.02 g/dl, glucose 22.18 ± 4.16 mg/dl, cholesterol 6.67 ± 1.04 mg/dl and triglyceride 15.2 ± 0.65 mg/dl. The mean sperm concentration was estimated to be 1.6 ± 0.12 (×10⁹ sperm/ml). A significant relationship was found between sperm concentration and K⁺ of seminal plasma (r = 0.533, p < 0.05). Significant correlations were observed between ionic contents: Na⁺ vs Cl^? (r = ?0.854, p < 0.01) and Mg²⁺ vs K⁺ (?0.583, p < 0.05). Also, level of triglyceride was negatively correlated with Mg²⁺ (r = ?0.503, p < 0.05). Presented data could be considered as a complementary study for developing special extenders and protectant solutions for improving artificial fertilization in this valuable species.     

Influence of Inseminate Components on Porcine Leucocyte Migration In Vitro and In Vivo after Pre- and Post-Ovulatory Insemination

A post‐breeding migration of leucocytes (PMN) into the uterus is considered to be an important reason for sperm losses. Minimizing such effects may be necessary for successful insemination with low sperm numbers, as required with sex‐sorted spermatozoa. We examined the magnitude of PMN influx 3 h after pre‐ or post‐ovulatory insemination with various combinations of seminal plasma (SP), semen extender Androhep? (AH; Minitüb, Tiefenbach, Germany) and sperm preparations (S). Pre‐ovulatory inseminations with preparations containing 98% AH caused a massive influx of PMN, independent of whether spermatozoa were present (628 ± 189 × 10⁶ leucocytes/uterine horn) or not (580 ± 153 × 10⁶). Post‐ovulatory, 98% AH caused a comparable immigration only in the absence of sperm cells (AH: 569 ± 198 × 10⁶, AH+S: 162 ± 102 × 10⁶). The presence of SP significantly dampened the numbers of recruited uterine leucocytes. The reaction to all inseminates containing 98% SP both with and without spermatozoa, used before ovulation (SP: 14 ± 6 × 10⁶, SP+S: 73 ± 27 × 10⁶) and after ovulation (SP: 60 ± 32 × 10⁶, SP+S: 51 ± 33 × 10⁶) did not differ significantly from controls using phosphate buffered saline (PBS) (pre‐ovulatory: 1 ± 1 × 10⁶, post‐ovulatory: 11 ± 9 × 10⁶). Quantitative in vitro transmigration assays with blood‐derived PMN proved that AH‐induced leucocyte migration into the uterus to be not as a result of direct chemotaxis, because, on account of the chelator citrate, AH significantly inhibited the transmigration towards recombinant human Interleukin‐8 (rhCXCL8) (AH: 14 ± 5% migration rate vs controls: 37 ± 6%, p < 0.05). Supernatants of spermatozoa incubated in PBS for 1, 12 or 24 h showed neither chemoattractive nor chemotaxis‐inhibiting properties. SP at ≥0.1% [v/v] significantly inhibited the in vitro transmigration of PMN. With respect to in vivo migration of neutrophils, the striking difference in the results between semen extender and seminal plasma suggests that adaptation of extender composition is needed to reflect more closely the in vivo regulatory potential of natural seminal plasma.     

Effect of food on the pharmacokinetics of oral cefuroxime axetil in dogs

Cefuroxime axetil pharmacokinetic profile was investigated in 12 Beagle dogs after single intravenous and oral administration of tablets or suspension at a dose of 20 mg/kg, under both fasting and fed conditions. A three-period, three-treatment crossover study (IV, PO under fasting and fed condition) was applied. Blood samples were withdrawn at predetermined times over a 12-hr period. Cefuroxime plasma concentrations were determined by HPLC. Data were analyzed by compartmental analysis. No statistically significant differences were observed between formulations and feeding conditions on PK parameters. Independently of the feeding condition, absorption of cefuroxime axetil after tablet administration was low and erratic. The drug has been quantified in plasma in 3 out of 6 and 5 out of 6 dogs in the fasted and fed groups. For this formulation, the bioavailability (F), peak plasma concentration (C_max), and area under the concentration–time curve (AUC) of cefuroxime axetil were significantly enhanced (p < .05) by the concomitant ingestion of food (32.97 ± 13.47–14.08 ± 7.79%, 6.30 ± 2.62–2.74 ± 0.66 µg/ml, and 15.75 ± 3.98–7.82 ± 2.76 µg.hr/ml for F, C_max, and AUC in fed and fasted dogs, respectively), while for cefuroxime axetil suspension, feeding conditions affected only the rate of absorption, as reflected by the significantly shorter absorption half-life (T_½(a)) and time to peak concentration (T_max) (0.55 ± 0.27–1.15 ± 0.19 hr and 1.21 ± 0.22–1.70 ± 0.30 for T_½(a) and T_max in fed and fasted dogs, respectively). For cefuroxime axetil tablets, T > MIC (≤1 µg/ml) was <2 hr in fasted and ≈4 hr in fed animals, and for cefuroxime axetil suspension, T > MIC (≤1 µg/ml) was ≈5 hr and for T >MIC (≤4 µg/ml) was ≈2.5 hr for fasted and fed dogs, respectively. Cefuroxime axetil as a suspension formulation seems to be a better option than tablets. However, its short permanence in plasma could reduce its clinical usefulness in dogs.     

The intravenous pharmacokinetics of butorphanol and detomidine dosed in combination compared with individual dose administrations to exercised horses

In equine and racing practice, detomidine and butorphanol are commonly used in combination for their sedative properties. The aim of the study was to produce detection times to better inform European veterinary surgeons, so that both drugs can be used appropriately under regulatory rules. Three independent groups of 7, 8 and 6 horses, respectively, were given either a single intravenous administration of butorphanol (100 µg/kg), a single intravenous administration of detomidine (10 µg/kg) or a combination of both at 25 (butorphanol) and 10 (detomidine) µg/kg. Plasma and urine concentrations of butorphanol, detomidine and 3-hydroxydetomidine at predetermined time points were measured by liquid chromatography–tandem mass spectrometry (LC-MS/MS). The intravenous pharmacokinetics of butorphanol dosed individually compared with co-administration with detomidine had approximately a twofold larger clearance (646 ± 137 vs. 380 ± 86 ml hr⁻¹ kg⁻¹) but similar terminal half-life (5.21 ± 1.56 vs. 5.43 ± 0.44 hr). Pseudo-steady-state urine to plasma butorphanol concentration ratios were 730 and 560, respectively. The intravenous pharmacokinetics of detomidine dosed as a single administration compared with co-administration with butorphanol had similar clearance (3,278 ± 1,412 vs. 2,519 ± 630 ml hr⁻¹ kg⁻¹) but a slightly shorter terminal half-life (0.57 ± 0.06 vs. 0.70 ± 0.11 hr). Pseudo-steady-state urine to plasma detomidine concentration ratios are 4 and 8, respectively. The 3-hydroxy metabolite of detomidine was detected for at least 35 hr in urine from both the single and co-administrations. Detection times of 72 and 48 hr are recommended for the control of butorphanol and detomidine, respectively, in horseracing and equestrian competitions.     

Pharmacokinetics of sodium baicalin following intravenous and intramuscular administration to piglets

The purpose of this study was to determine the pharmacokinetics of baicalin after intravenous and intramuscular administration of sodium baicalin at 50 mg/kg to piglets. Plasma baicalin levels were determined by high‐performance liquid chromatography. The plasma concentration–time data of baicalin for both administration routes were best described by two‐compartmental open model. The area under the plasma concentration–time curve and the elimination half‐lives were 77.47 ± 6.14 µg/ml × h and 1.73 ± 0.16 hr for intravenous and 64.85 ± 5.67 µg/ml × h and 2.42 ± 0.15 hr for intramuscular administration, respectively. The apparent volume of distribution and body clearance were 1.63 ± 0.23 L/kg and 2.74 ± 0.30 L h^?1 kg^?1 for intravenous and 0.51 ± 0.10 L/kg and 0.78 ± 0.08 L h^?1 kg^?1 for intramuscular routes, respectively. An intramuscular injection of sodium baicalin in piglets resulted in rapid and complete absorption, with a mean maximal plasma concentration of 77.28 ± 7.40 µg/ml at 0.17 hr and a high absolute bioavailability of 83.73 ± 5.53%.     

Prostasomes of Canine Seminal Plasma – Zinc‐Binding Ability and Effects on Motility Characteristics and Plasma Membrane Integrity of Spermatozoa

Prostasomes are small lipid membrane‐confined vesicles that are involved in various fertilization‐related processes. The aim of this study was to demonstrate canine seminal plasma prostasomes' ability to bind zinc ions, as well as examining their effects on sperm motility characteristics and plasma membrane integrity during cold storage. Ejaculates, collected from five cross‐bred dogs (n = 50), were subjected to ultracentrifugation followed by gel filtration (GF) on a Superose 6 column. Prostasomes appeared as a single fraction in the elution profile. Transmission electron microscopy (TEM) analysis of canine prostasomes revealed the presence of membrane vesicles with diameters ranging from 20.3 to 301 nm. The zinc‐affinity chromatography on a Chelating Sepharose Fast Flow – Zn^{2 +} showed that from 93 to 100% of the prostasome proteins bind zinc ions (P⁺Zn). SDS‐PAGE revealed that canine P⁺Zn comprised four protein bands, with low molecular weights (10.2–12 kDa). We have also shown a positive effect of prostasomes (p < 0.05), especially variant B (2% of total seminal plasma protein) on canine sperm motility parameters after 2 h storage at 5°C (TMOT%, 44.75 ± 5.18) and PMOT%, 12.42 ± 1.59) and VAP, VSL, VCL, when compared with Control (TMOT%, 7.30 ± 1.41 and PMOT%, 1.70 ± 0.42). Higher percentage of spermatozoa with intact plasma membrane (SYBR/PI dual staining) and intact acrosome (Giemsa stained), after 2 h storage at 5°C, was showed, in variant A (1.5% of total seminal plasma protein) and B, when compared with Control and variant C (2.5% of total seminal plasma protein). The prostasomes' effect on motility and plasma membrane integrity of canine cold‐stored spermatozoa may be related to their ability to bind zinc ions and regulate their availability to the sperm.     

Does Coenzyme Q10 Exert Antioxidant Effect on Frozen Equine Sperm?

During semen cryopreservation, the sensitivity of equine sperm to oxidative stress is increased by the eliminated seminal plasma. Thus, antioxidant addition to the semen extender can be helpful to the sperm survival after freezing and thawing. This work aimed to test whether coenzyme Q10 (CoQ10) added in different concentrations to the INRA 82 freezing extender has antioxidant function on equine sperm to improve its fertilizing ability. Semen samples from five stallions were frozen with the extenders: (T1) INRA 82, control, (T2) T1+ 5 μM CoQ10, (T3) T1+ 25 μM CoQ10, and (T4) T1+ 50 μM CoQ10. After sample thawing, sperm motility and kinetics characteristics were evaluated using a computer-assisted sperm analysis and sperm membrane functionality and integrity were evaluated with a hypo-osmotic swelling test and an epifluorescence microscopy, respectively. The nitrite (NO₂^-) and hydrogen peroxide (H₂O₂) concentrations of the semen samples were measured with spectrophotometry. There was no difference on the sperm characteristics among all treatments (P > .05). However, the 25 μM CoQ10 (T3) decreased NO₂⁻ concentration (6.7 ± 2.2 μM/μg protein) compared with the treatments T1, T2, and T4 (64.3 ± 3.7, 59.4 ± 5.3, 45.1 ± 8.6 μM/μg protein), respectively, as well H₂O₂ concentration (1.8 ± 0.3 μM/μg protein) compared with the control (4.6 ± 0.4 μM/μg protein) and 5 μM CoQ10 treatments (4.8 ± 0.2 μM/μg protein, P < .05). In conclusion, 25 μM CoQ10 plays a significant role as antioxidant to the frozen equine sperm, decreasing NO₂⁻ and H₂O₂ concentrations. Thus, its addition to the INRA 82 freezing extender may be beneficial to the fertilizing ability of equine semen.     

The effect of astaxanthin supplementation on the post-thaw quality of dog semen

Astaxanthin is a member of the carotenoid family well known for its anti-cancer, anti-diabetic, anti-inflammatory and antioxidant nature. This study was designed to investigate the effects of astaxanthin supplementation of the extender (buffer 2) on post-thaw dog semen quality. Semen from four healthy dogs was collected by digital manipulation twice a week. The ejaculates were pooled, washed, divided into four equal aliquots, diluted with the extender supplemented with different concentrations of astaxanthin (0, 0.5, 1 and 2 µM) and cryopreserved. The results showed that 1 µM astaxanthin was the optimum concentration that led to significantly higher (p < .05) post-thaw motility, kinematic parameters and viability than the other groups. In comparison with the control group, sperm samples supplemented with 1 µM astaxanthin showed significantly higher (p < .05) sperm counts with intact membranes (55.7 ± 0.6% vs. 51.3 ± 0.9%), intact acrosome (58.4 ± 0.7% vs. 53.5 ± 0.6%), active mitochondria (54.9 ± 0.5% vs. 42.6 ± 0.6%) and normal chromatin (67.6 ± 0.9% vs. 61.7 ± 0.6%). Furthermore, astaxanthin-supplemented samples showed significantly lower expression levels (p < .05) of pro-apoptotic (BAX), oxidative induced DNA damage repair (OGG1), oxidative stress-related (ROMO1) genes and higher expression levels of anti-apoptotic (BCL2), and sperm acrosome-associated (SPACA3) genes compared to the control. Thus, supplementation of 1 µM astaxanthin in semen extender results in improved freeze-thaw sperm quality of the dog.     

Pharmacokinetic–pharmacodynamic relationship of marbofloxacin for Escherichia coli and Pasturella multocida following repeated intramuscular administration in goats

The pharmacokinetics (PK) and pharmacodynamics (PD) of marbofloxacin (MBF) were determined in six healthy female goats of age 1.00–1.25 years after repeated administration of MBF. The MBF was administered intramuscularly (IM) at 2 mg kg^?1 day^?1 for 5 days. Plasma concentrations of MBF were determined by high‐performance liquid chromatography, and PK parameters were obtained using noncompartmental analysis. The MBF concentrations peaked at 1 hr, and peak concentration (C_max) was 1.760 µg/ml on day 1 and 1.817 µg/ml on day 5. Repeated dosing of MBF caused no significant change in PK parameters except area under curve (AUC) between day 1 (AUC_0–∞D1 = 7.67 ± 0.719 µg × hr/ml) and day 5 (AUC_0‐∞D5 = 8.70 ± 0.857 µg × hr/ml). A slight difference in mean residence time between 1st and 5th day of administration and accumulation index (AI = 1.13 ± 0.017) suggested lack of drug accumulation following repeated IM administration up to 5 days. Minimum inhibitory concentration (MIC) demonstrated that Escherichia coli (MIC = 0.04 µg/ml) and Pasturella multocida (MIC = 0.05 µg/ml) were highly sensitive to MBF. Time‐kill kinetics demonstrated rapid and concentration‐dependent activity of MBF against these pathogens. PK/PD integration of data for E. coli and P. multocida, using efficacy indices: C_max/MIC and AUC_0–24hr/MIC, suggested that IM administration of MBF at a dose of 2 mg kg^?1 day^?1 is appropriate to treat infections caused by E. coli. However, a dose of 5 mg kg^?1 day^?1 is recommended to treat pneumonia caused by P. multocida in goats. The study indicated that MBF can be used repeatedly at dosage of 2 mg/kg in goats without risk of drug accumulation up to 5 days.     

Inotropic and lusitropic effects of incremental doses of dobutamine in dogs with right ventricular apical pacing‐induced cardiac dysfunction

This study aimed to assess the effects of incremental doses of dobutamine on diastolic function in healthy and rapid ventricular apical pacing (RVAP)‐induced cardiac dysfunction anesthetized dogs. Inotropic and lusitropic effects of dobutamine (2, 4, 8, and 12 μg kg^?1 min^?1) were assessed through left ventricle (LV) pressure–volume relation and Doppler echocardiography in six female dogs before and after 8 weeks of RVAP. Peak rate of LV pressure fall (?dP/dt_min) improved with doses >4 μg kg^?1 min^?1 in healthy (4,490 ± 970 vs. 3,265 ± 471 mmHg/s, p < 0.05) and >8 μg kg^?1 min^?1 in RVAP dogs (3,385 ± 1,122 vs. 1,864 ± 849 mmHg/s, p < 0.05) while the time constant of relaxation (tau) reduced with doses >4 μg kg^?1 min^?1 in both groups (healthy: 24.0 ± 3.7 vs. 28.2 ± 4.9 ms; RVAP: 32.6 ± 8.5 vs. 37.5 ± 11.4 ms, p < 0.05) comparing with baseline. Indices of relaxation (?dP/dt_min and tau) suggested preserved lusitropic response in contrast with markedly reduced indices of contractility in the RVAP group compared with healthy group at same infusion rates. Doppler echocardiography showed significant reduction of elastic recoil in failing hearts. The results of this study demonstrated maximal positive lusitropic effects of dobutamine at a dose of 8 μg kg^?1 min^?1 in ventricular pacing‐induced cardiac dysfunction without further impairment of ventricular filling.     

Coenzyme Q10 and α‐Tocopherol Prevent the Lipid Peroxidation of Cooled Equine Semen

Biotechnology applied for equine semen increases the levels of reactive oxygen species and reduces the natural antioxidant defence, by both dilution and removal of seminal plasma. Therefore, the aims of this study were to evaluate the effect of adding coenzyme Q10 (CoQ10) and α‐tocopherol (α‐TOH) to the cooling extender, singly or in combination, on sperm parameters, and their effectiveness in preventing lipid peroxidation (LPO) of equine semen during cooling at 5°C for 72 h. Ten adult stallions of proven fertility were used, using two ejaculates each, subjecting them to the treatments with the following concentrations: α‐TOH: 2 mm ; CoQ10: 40 μg/ml; and CoQ10 + α‐TOH: 40 μg/ml + 2 mm for control (C) without the addition of antioxidants and for vehicle control (EtOH) with 100 μl ethanol. The CoQ10 group had a higher percentage of total motility (69.1 ± 16.2%) compared to control (62.1 ± 16.2%) and EtOH (58.1 ± 18.6%). CoQ10 + α‐TOH and α‐TOH groups were most effective in preventing LPO compared to controls (1765.9 ± 695.9, 1890.8 ± 749.5, 2506.2 ± 769.4 ng malondialdehyde/10⁸ sptz, respectively). In conclusion, CoQ10 and α‐TOH were effective during the cooling process of equine semen at 5°C for 72 h, providing increased levels of total motility, as well as lower LPO.     

RELATIONSHIP BETWEEN AGE,PLASMA RENIN ACTIVITY,AND RENAL RESISTIVE INDEX IN DOGS

The renal resistive index (RI) value of 0.73 has been proposed as the upper limit in normal adult dogs. In humans, changes in RI with age are associated with plasma renin activity. There are relatively few equivalent reference data for dogs. We obtained reference RI data from 22 clinically healthy dogs <4 months of age and 33 healthy dogs between 4 months and 7 years of age. An association between the RI and plasma renin activity was investigated. The mean RI in the older dogs was 0.65 ± 0.05 vs. 0.75 ± 0.05 in dogs <4 months of age. The mean plasma renin activity in the older dogs was 1.18 ± 1.03 vs. 4.23 ± 3.09 ng/ml/h in dogs <4 months of age. There was a weak linear relationship between the RI and plasma renin activity (r²=0.280, P<0.01) in dogs <4 months of age. Also in these younger dogs, RI was negatively correlated with age (r²=0.682, P<0.01). The RI was higher in dogs <4 months of age than in older dogs. Therefore, the mean renal RI is slightly higher in young dogs than reported for an older population and interpretation of the RI must include an assessment of patient age.     

Concentrations of platelet α2-adrenoceptors,lymphocyte muscarinic receptors,and blood monoamines in dogs (Canis familiaris) affected by canine cognitive dysfunction syndrome

Canine cognitive dysfunction syndrome (CDS) is a neurodegenerative disorder of aged dogs characterized by a progressive decline in cognitive function. In humans and laboratory animals, a variety of neurotransmitter abnormalities have been described in patients affected by age-related dementia. Specifically, the regulatory role of the catecholaminergic, serotonergic, and cholinergic systems has been outlined. The aim of the present study was to measure blood monoamine levels, platelet α₂-adrenergic receptors, and lymphocyte muscarinic receptors in healthy adult and aged dogs and in dogs affected by canine cognitive dysfunction. Based on clinical and behavioral examination, 40 dogs were divided into 3 groups: healthy adults (n = 14), aged dogs (n = 17), and aged dogs affected by canine cognitive dysfunction (n = 9). A significant reduction in plasma levels of norepinephrine and dopamine was observed both in aged dogs (0.16 ± 0.02 ng/mL, P < 0.01; 0.11 ± 02 ng/mL, P < 0.01, respectively) and in CDS dogs (0.14 ± 0.03 ng/mL, P < 0.05; 0.10 ± 00.005 ng/mL, P < 0.01, respectively) compared with adults (0.29 ± 0.04 ng/mL and 0.15 ± 0.02 ng/mL, respectively). No significant differences were observed among groups for α₂-adrenergic receptor concentrations. Canine lymphocytes express 2 distinct classes of muscarinic receptors, characterized by high (HA) and low affinity (LA) for [³H]-N-methyl-scopolamine. A significant age-dependent decrease in HA muscarinic receptors was observed. However, no differences were found between aged dogs (87.65 ± 11.08 sites/cell × 10²) and in CDS dogs (90.17 ± 6.75 sites/cell × 10² ) for HA muscarinic receptor concentrations. As far as LA muscarinic receptors are concerned, CDS dogs showed a significant increase (393.48 ± 63 sites/cell × 10²; P < 0.05) with respect to healthy adult dogs (188.84 ± 16.50 sites/cell × 10²). Our results suggest that the reduction in HA muscarinic receptor-binding sites could be representative of the physiological aging process, whereas the increase in lymphocyte LA muscarinic receptor levels could be related to the cognitive decline.     

Effect of FMD vaccination on semen quality parameters in Karan Fries and Murrah buffalo bulls

Effect of Foot and Mouth disease (FMD) vaccination was studied on semen quality parameters of 19 Karan Fries (KF) and eight Murrah (MU) breeding bulls during the period 2002 to 2004 at Artificial Breeding Complex, NDRI, Karnal. A total of non-vaccinated 155 KF and 72 MU bulls' ejaculates were taken as control, while 169 KF and 51 MU bulls' ejaculates, collected after vaccination, were used to study the effect of vaccination stress. The results showed that FMD vaccination had no significant (P > 0.05) effect on ejaculate volume and total volume per day of semen in both KF and MU bulls. Volume of semen increased slightly during post-vaccination period in both the breeds. After FMD vaccination, there was significant (P < 0.01) decrease in mass activity (2.27 ± 0.06 vs. 1.67 ± 0.07 and 2.49 ± 0.09. vs. 1.75 ± 0.10, for KF and MU, respectively), initial motility (56.89 ± 0.03% vs. 44.62 ± 0.02% and 62.26 ± 0.04% vs. 47.08 ± 0.05%, for KF and MU, respectively), sperm concentration (754.19 ± 23.96 vs. 554.14 ± 22.95 × 10⁶/ml and 848.61 ± 33.65 vs. 571.57 ± 39.99 × 10⁶/ml, for KF and MU, respectively), and total sperm output per ejaculate (3,685.94 ± 158.40 vs. 2,781.54 ± 151.70 × 10⁶ and 2,218.75 ± 133.14 vs. 1,582.84 ± 158.20 × 10⁶, for KF and MU, respectively). Application of FMD vaccine had significantly (P < 0.05) adverse effect on most of the seminal attributes during post-vaccination in KF and MU buffalo bulls. So, the spermiograms affected following vaccination suggest that in bovines, the semen collection and preservation should be suspended till normal fertility of sperm is restored to avoid the failure of conception from artificial insemination using such semen.     

Effects of dietary fish oil and alpha-tocopherol supplementation on selected blood parameters and fatty acid profiles in mares and their foals

The effects of fish oil (40 ml/day) supplementation, with or without synthetic all-rac-alpha-tocopherol-acetate (2,500 IU/day), during the last 65 days before expected parturition were investigated in 15 adult mares (553 ± 24 kg BW) and their foals. Mares were assigned to one of three diets: control (n = 5), control plus fish oil and alpha-tocopherol (n = 4; FO + AT) or control with just fish oil (n = 6; FO). Blood samples were obtained from the mares before a 15-day dietary adaptation period (T1) and from mares and foals the first (T2) and fifth (T3) days post-partum. Colostrum was collected at T2 and milk at T3. Routine haematological, biochemical and alpha-tocopherol analyses were undertaken on all blood samples. Fatty acid concentrations were determined in the foal serum and alpha-tocopherol concentrations measured in the milk and colostrum. Diet had no effect on haematology or biochemistry in the mares. Alpha-tocopherol concentrations were significantly higher at T2 & T3 in the FO + AT mares. Foal WBCs were higher in FO (11.33 ± 2.59 × 10⁹/l), comparing to FO + AT and control groups (9.18 ± 1.24 × 10⁹/l and 7.26 ± 1.03 × 10⁹/l, respectively), at T3 (p < .05). There was no significant effect of the fish oil supplementation on the foal's serum fatty acid profile. In the FO + AT group, both colostrum and milk alpha-tocopherol concentrations (2.56 ± 0.36 and 1.36 ± 0.22 µg/ml, respectively) were higher compared than those of the FO group (1.33 ± 0.39 and 0.72 ± 0.31 µg/ml, respectively; p < .05). Additional 2,500 IU/day of synthetic alpha-tocopherol in the last 65 days of pregnancy increased alpha-tocopherol concentrations in colostrum and milk and the foal's serum. 40 ml/day fish oil, however, did not significantly increase serum eicosapentaenoic acid and docosahexaenoic acid concentrations in the foals.     

Plasma and serum concentrations of cytarabine administered via continuous intravenous infusion to dogs with meningoencephalomyelitis of unknown etiology

The objective of this study was to evaluate the plasma and serum concentrations of cytarabine (CA) administered via constant rate infusion (CRI) in dogs with meningoencephalomyelitis of unknown etiology (MUE). Nineteen client‐owned dogs received a CRI of CA at a dose of 25 mg/m²/h for 8 h as treatment for MUE. Dogs were divided into four groups, those receiving CA alone and those receiving CA in conjunction with other drugs. Blood samples were collected at 0, 1, 8, and 12 h after initiating the CRI. Plasma (n = 13) and serum (n = 11) cytarabine concentrations were measured by high‐pressure liquid chromatography. The mean peak concentration (C_MAX) and area under the curve (AUC) after CRI administration were 1.70 ± 0.66 μg/mL and 11.39 ± 3.37 h·μg/mL, respectively, for dogs receiving cytarabine alone, 2.36 ± 0.35 μg/mL and 16.91 + 3.60 h·μg/mL for dogs administered cytarabine and concurrently on other drugs. Mean concentrations for all dogs were above 1.0 μg/mL at both the 1‐ and 8‐h time points. The steady‐state achieved with cytarabine CRI produces a consistent and prolonged exposure in plasma and serum, which is likely to produce equilibrium between blood and the central nervous system in dogs with a clinical diagnosis of MUE. Other medications commonly used to treat MUE do not appear to alter CA concentrations in serum and plasma.     

Effects of ascorbic acid 2‐glucoside and alpha‐tocopherol on the characteristics of equine spermatozoa stored at 5°C

The aim of this experiment was to evaluate the effects of adding ascorbic acid 2‐glucoside (AA2G), a water‐soluble antioxidant and stable derivative of ascorbate, to the semen extender and compare it to the addition of vitamin C (Vit. C) and the fat‐soluble antioxidant α‐tocopherol (α‐Toh), both individually and in combination, on the seminal variables of equine sperm submitted to cooling for 72 h. We used two ejaculates from 10 stallions and evaluated them for motility, membrane integrity, chromatin fragmentation, mitochondrial activity and lipid peroxidation. In the analysis of lipid peroxidation, the control group showed 2506.2 ± 796.4 ng malondialdehyde/10⁸ sperm, which was higher (P < 0.05) than the groups treated with antioxidants. The average value of motility in the AA2G group was 68.4 ± 18.1%, which was higher (P < 0.05) than that observed in the control group (62.1 ± 16.2%). The variables membrane integrity, chromatin fragmentation and mitochondrial activity did not show significant difference (P > 0.05) between treatments. It was concluded that the antioxidants protected sperm cells from lipid peroxidation and that AA2G was effective during the cooling process of equine semen at 5°C for72 h, providing increased levels of total motility.     

Retracted: Study of enzyme activities and protein content of beluga (Huso huso) semen before and after cryopreservation

Knowledge gained regarding the biochemical processes that occur during sperm collection, processing and freezing‐thawing might improve current sperm cryopreservation techniques. In our present study, we determined the effect of cryopreservation on the total protein concentration (TP) and the activities of certain enzymes in semen samples from the beluga (Huso huso). The TP content of the seminal plasma of fresh semen was 0.47 ± 0.026 g/l, and the TP after cryopreservation was 1.86 ± 0.6 g/l. The activities of acid phosphatase (0.82 ± 0.042 U/l), lactate dehydrogenase (234.4 ± 19.4 U/l), arylsulfatase (143.1 ± 32.5 U/l) and β‐N‐acetylglucosaminidase (58.39 ± 4.14 U/l) in the seminal plasma of fresh semen were significantly lower than those in the supernatant of frozen‐thawed semen samples (7.43 ± 0.64, 3224.6 ± 167.2, 422.6 ± 21.3 and 90.2 ± 5.37 U/l respectively). These parameters may be useful as biomarkers for estimating damage to the cell membrane of spermatozoa caused by freezing‐thawing.     

Feed and water deprivation has a negative but transient effect on the rumen kinetics of Bos indicus steers

The effects on rumen kinetics after feed and water had been deprived for 72 hr were studied using four fistulated Bos indicus steers. The animals were assigned in a 2 × 4 crossover design with two treatments: feed and water ad libitum (control) and no feed and water for 72 hr (deprived) with four steers per treatment over two time periods. Feed and water deprivation caused decreases in the numbers of cellulolytic bacteria (1.4 vs. 0.4 cfu × 10⁶/ml; p = .001), live (23.7 vs. 0.8 × 10⁹/ml; p = .001), dead (12.7 vs. 0.5 × 10⁹/ml; p = .001) and total bacterial counts (36.4 vs. 1.4 × 10⁹/ml; p = .001) at day 0, compared with the control treatment. However, the deprived group had greater numbers of cellulolytic bacteria (2.7 vs. 50.1 cfu × 10⁶/ml; p = .001), live (18.3 vs. 42.2 × 10⁹/ml; p = .001), dead (6. 5 vs. 19.1 × 10⁹/ml; p = .001) and total bacterial counts (24.8 vs. 61.3 × 10⁹/ml; p = .001) from rumen fluid on day 4, compared with the control treatment. The numbers of protozoa in rumen fluid from the deprived group were less than (551.2 vs. 2.4 × 10³/ml; p = .001) the control group on day 0. However, the deprived treatment had fewer protozoa in rumen fluid than the control treatment on day 4 (p = .001) and day 9 (p = .001). Volatile fatty acids and in vitro gas production as functional measurements of rumen fluid followed the same trend as the bacterial and protozoa populations. These results indicate that feed and water deprivation would have a negative but transient effect on the rumen kinetics of Bos indicus steers.