The relationships between the myocardial bridge and ramus interventricularis paraconalis characteristics in lamb and sheep

The myocardial bridge (MB) is an anomaly that the myocardial fibres cover on a segment of the subepicardial coronary arteries or their branches in domestic animals and humans. The aim of the present study was to determine the relationships between the characteristics of the MB and ramus interventricularis paraconalis at three levels in lambs and adult sheep. Thirty-three hearts (16 lambs and 17 sheep) were used to determine the MB (length, angle and thickness) and vessel (vessel diameter and thicknesses of tunica intima et media of ramus interventricularis paraconalis) characteristics. Independent-samples t test was applied to compare variables between lambs and sheep. Spearman's correlation analysis was conducted to evaluate the relationships between bridge and vessel characteristics at three bridge levels. Length, angle and thickness of myocardial bridges were not significantly different between the lambs and sheep (p > .05). The mean length, angle and thickness were 24.9 ± 16.1 mm, 113.7 ± 11.2° and 1,098 ± 555 µm in 33 hearts, respectively. In lambs, the mean vessel diameters were 1,930 ± 742 µm (1,534–2,325 µm), 1,247 ± 665 µm (893–1,601 µm) and 865 ± 172 µm (774–957 µm) at the pre-bridge, bridge and post-bridge levels, respectively. In sheep, the mean vessel diameters in the same order were 1,861 ± 1,068 µm, 1,337 ± 308 µm and 1,287 ± 549 µm. The bridge prevalence was 100% in the samples examined. In conclusion, coronary arterial diseases related to myocardial bridge should not be expected in sheep for veterinary cardiology practice. It may also be concluded that the cross-breeds of the Awassi and Chios sheep may be useful in experimental studies related to myocardial bridge surgery.     

Anatomical and Histological Organization of the Testes of the Inseminating Catfish Trachelyopterus striatulus (Steindachner, 1877) (Siluriformes: Auchenipteridae)

The testicular morphology, spermatogenesis and occurrence of sperm in the ovarian lumen of Trachelyopterus striatulus were studied using anatomical, histological and biometric techniques. A total of 50 catfish (T. striatulus) were captured, measuring 14.9 ± 2.5 cm of standard length, body weight was 81.2 ± 34.5 g and their testes weighed 16.9 ± 6.1 g. The testes of T. striatulus are paired organs, showing two distinct regions: cranial, which shows a compact medial part and with fringes ventrally, and caudal region, which is formed of the seminal vesicle with fringes laterally and two saculiform expansions. The testes presented a length of 35.2 ± 6.9 mm, and the fringes showed a cranial length of 12.1 ± 3.8 mm and caudal length of 6.4 ± 2.6 mm. Histologically, the cranial fringes are spermatogenic and showed cells with significantly different nuclear diameters, ranging from 8.2 ± 1.5 μm (primary spermatogonia) to 1.88 ± 0.3 μm (spermatid). The seminal vesicles and saculiform expansions showed tubules with a simple prismatic secretory epithelium containing spermatozoa and secretion into the lumen. The caudal fringes are exclusively for secretory flow, consisting of tubules with a simple cuboidal epithelium. The common spermatic duct showed a simple cuboidal epithelium and contained spermatozoa with secretion into the lumen. The secretion of the caudal region is acidophilic, with neutral glycoproteins and sialomucin. T. striatulus ovaries showed free spermatozoa or were organized in spermatozeugmata into the ovarian lumen and between the ovuligers lamellae.     

Harderian Gland in Canadian Ostrich (Struthio camelus): A Morphological and Histochemical Study

Heads of ten healthy adult ostrich obtained from slaughter house were the constituted materials of the study. The Harderian gland (HG) was dissected out, and all of the gross morphometrical parameters including length, width and thickness as well as weight of left and right glands were recorded. Tissue sections were stained, using haematoxylin and eosin, Masson trichrome, periodic acid‐Schiff and Alcian blue (pH 2.5) techniques. In ostrich, HG was an orbital organ located ventromedially around the posterior part of the eyeball. It was an oval flatted shape, light pink colour with irregular outline and was pointed in the dorsal end. Its mean length was 35.30 ± 2.84 mm and 35.55 ± 3.58 mm in left and right sides, respectively, and mean width 15.30 ± 1.20 mm and 15.65 ± 1.18 mm in left and right sides, respectively. There was no significant difference between length, thickness, weight and width of left and right glands. Histological results showed that the glandular epithelium was multilobular and compound tubuloalveolar. The gland was surrounded by a connective tissue capsule, and the epithelium was lined by simple columnar epithelial cells of varying height. The secretion of HG was mucous and the secretion type was apocrine. Mucosubstance analysis revealed that secretory units contained acidic and neutral glycoproteins. The granules within the epithelial cells lining the intralobular and inter‐lobular excretory ducts of the gland were positive for periodic acid‐Schiff and Alcian blue (pH 2.5).     

Morphological and morphometric characterization of domestic cat epididymal sperm

Sperm morphometry is the tool that confers objectivity to the morphological evaluation by accurately measuring the dimensions of the gamete and its structures. Thus, the aim of the study was to perform a morphometric characterization of the domestic cat sperm. Therefore, sperm samples were collected from twenty pairs of epididymis in a TRIS extender at 37ºC. An aliquot of the sample was used to make a smear with Rose Bengal solution, and afterwards, the morphology and morphometry were analysed. In the morphology, were quantified the percentage of normal sperm cells, morphological changes of head, midpiece and tail. In morphometry, each normal sperm cell was measured for length, width, area and perimeter of head and midpiece, tail length and total length. The parameters ellipticity, elongation, regularity and rugosity were also determined. The percentage of normal sperm was 67.21%. Of the abnormalities, the curled/folded tail, followed by the curved midpiece, abnormal shaped head and detached head were the most quantified. The sperm head presented 5.56 ± 0.01 μm and 3.10 ± 0.01 μm of length and width, respectively. The head area was 16.94 ± 0.05 μm², while the perimeter was 16.16 ± 0.03 μm. In the derived parameters, the values were as follows: ellipticity of 1.81 ± 0.00; elongation of 21.39 ± 0.12; regularity of 0.81 ± 0.00; and rugosity of 0.14 ± 0.00. The midpiece presented length and width of 7.96 ± 0.01 μm and 0.76 ± 0.01 μm, respectively. The mean length of the sperm tail was 45.12 ± 0.06 μm, and the total cell size was 58.67 ± 0.06 μm. Thus, it was concluded that the cat sperm is an elongated cell, with high rugosity and regularity. The spermatic tail represents more than ¾ of the total length of the cell and the midpiece exceeds the length of the head.     

Ultrastructural characteristics of black marsh turtle spermatozoa obtained by electroejaculation

The black marsh turtle (Geoemydidae: Siebenrockiella crassicollis) is a freshwater turtle that occurs in equatorial tropical climates in South East Asia. The semen of S. crassicollis was investigated by electroejaculation. The spermatozoa of S. crassicollis are filiform in shape with curved heads. The entire length, midpiece to tail length, tail width and tail length of the spermatozoa were 71.33 ± 1.55 μm, 49.92 ± 1.13 μm, 0.43 ± 0.02 μm and 48.53 ± 0.25 μm, respectively. The head length, head width across the middle and head width across the base were 14.00 ± 0.38 μm, 0.79 ± 0.03 μm and 0.91 ±0.0.03 μm, respectively. The acrosomal region of the S. crassicollis spermatozoa was narrower than the head, with an acrosomal length and width at the annulus of 2.90 ± 0.13 μm and 0.43 ± 0.01 μm, respectively. The midpiece of the S. crassicollis spermatozoa was narrower than the head and contained 30–40 mitochondrial balls, each with a ball diameter of 0.16 ± 0.002 μm. The midpiece length, midpiece width and tail length were 4.92 ± 0.16, 0.78 ± 0.03 and 48.53 ± 0.25 μm, respectively. This study presents the characteristic appearance of a freshwater turtle spermatozoa in Southeast Asia, as observed under an electron microscope. The spermatozoa of Siebenrockiella crassicollis are morphologically different from those of other freshwater turtles from other regions described in previous studies.     

Morphometry,Morphology and Ultrastructure of Ring‐tailed Coati Sperm (Nasua nasua Linnaeus, 1766)

The ring‐tailed coati (Nasua nasua) is a procyonid whose population is in sharp decline. Therefore, studies are needed to better understand the reproduction of this animal. For this reason, this study aimed to evaluate the morphology, morphometry and sperm ultrastructure of ring‐tailed coati sperm. Four captive adult males were used for this study. Slides stained with Bengal Rose were used for the morphometric and morphologic analyses. The length and width of the head were measured, as well as the length of the midpiece and tail and the total length of the sperm. Scanning electron microscopy and transmission electron microscopy were used for the ultrastructural analyses. The most obvious morphological abnormalities observed were coiled tails (6.1 ± 8.7%) and the lack of acrosomes (5.4 ± 4.4%). Regarding the morphometry, the measurements of the head (length × width), midpiece (length) and tail (length) were (mean ± SD) 6.2 ± 0.4 × 8.1 ± 0.6 μm, 14.1 ± 0.5 and 63.9 ± 4.1 μm, respectively, and the total length of the sperm was 86.1 ± 4.3 μm. Through electron microscopy, the presence of electron‐lucent points in the nucleus and the presence of approximately 55 mitochondrial spirals in the midpiece were identified. The data obtained in this study provide detailed information on the sperm characteristics of coatis and may inform future research on germplasm conservation, both for this species and other threatened procyonids.     

The comparison of two different extenders for the improvement of large‐scale sperm cryopreservation in common carp (Cyprinus carpio)

In our study, a traditionally used (Grayling, already used in cyprinid species) and a newly tested (Pike) extender was tested to avoid sperm agglutination phenomenon following thawing during carp sperm cryopreservation. A large‐scale (elevated volume of sperm) freezing method in a controlled‐rate freezer using 5 ml straw and 10 ml cryotube was also systematically established. In all experiments, the sperm cryopreserved in using Grayling extender (except only one sample) showed an agglutination phenomenon (damaged and intact cells adhered to each other) after thawing where Pike extender resulted the regular cell suspension. No significant difference was observed between the two cryopreserved groups (Pike and Grayling extender) in all motility parameters using the 0.5 ml straw and the polystyrene box. Similarly, motility parameters did not show a significant difference in the two frozen groups with the 5 ml straw, also in the polystyrene box. A significantly higher progressive motility (pMOT, Grayling: 54% ± 8%, Pike: 37% ± 5%), straight line velocity (VSL, Grayling: 50 ± 5 µm/s, Pike: 39 ± 4 µm/s) and beat cross frequency (BCF, Grayling: 20 ± 1 Hz, Pike: 17 ± 1 Hz) was observed in the case of the grayling extender by the 5 ml straw cryopreserved in a controlled‐rate freezer (CRF) compare to the pike extender. A significantly higher VSL (Grayling: 45 ± 3 µm/s, Pike: 38 ± 4 µm/s) was observed by the grayling extender using the 10 ml cryotube than with the pike extender. Despite the randomly occurring differences in a few parameters, our new controlled freezing method using the newly tested Pike extender, the 5 ml straw or the 10 ml cryotube can be a good solution for the preservation of elevated volume of carp sperm.     

Effect of caspase inhibitor Z-VAD-FMK on bovine sperm cryotolerance

The aim of this study was to evaluate the treatment of bovine semen with the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-FMK), before or after freezing on semen quality. After the initial assessment, sperm from 4 bulls were pooled (Experiment 1) and cryopreserved in BioXcell containing 0, 20 and 100 μM Z-VAD-FMK. After thawing semen viability, motility, membrane integrity, as well as DNA fragmentation and ΔΨm were evaluated. In Experiment 2, bovine frozen/thawed sperm were incubated for 1 hr with 0, 20 and 100 µM Z-VAD-FMK before assessing the semen quality. The treatment with Z -VAD-FMK before cryopreservation improved post-thawing sperm motility compared to the control group (p < .05), while no differences were recorded in sperm viability and membrane integrity among groups (on average 86.8 ± 1.5 and 69.1 ± 1.4, respectively). Interestingly, at the highest concentration, DNA fragmentation decreased (p < .05), while the percentage of spermatozoa with high ΔΨm increased (p < .05). The results of Experiment 2 showed that 1-hr treatment with Z-VAD-FMK did not affect sperm motility and viability (on average 63.4 ± 5.8 and 83.7.1 ± 1.2, respectively). However, Z-VAD-FMK improved sperm membrane integrity (p < .05) and at the highest concentration tested decreased the proportion of sperm showing DNA fragmentation (p < .05). No differences were recorded in the percentage of spermatozoa with high ΔΨm (on average 57.0 ± 11.4). In conclusion, the treatment with 100 µM of the caspase inhibitor Z-VAD-FMK before freezing increased bovine sperm mass motility and ΔΨm, while decreasing sperm DNA fragmentation. Treatment of semen after thawing with 100 µM Z-VAD-FMK improved sperm membrane integrity and reduced DNA fragmentation.     

Correlation of testicular artery Doppler velocimetry with kinetics and morphologic characteristics of epididymal sperm in dogs

Sperm quality can be affected by a reduction in testicular blood flow, which can be measured by Doppler ultrasonography. The aim of this study was to correlate the Doppler velocimetry of the testicular artery with kinetics of the epididymal spermatozoa in dogs. Twenty-two dogs (44 testicles) were evaluated by Doppler ultrasonography in five regions of the testicular artery before orchiectomy. Spermatozoa were recovered by the epididymal tail compression technique and analysed for kinetics on a computer-assisted semen analysis (CASA system). Morphology (modified Karras) and sperm membrane integrity were analysed by eosin–nigrosine staining. Data were analysed by Pearson's correlation test (p < .01). The mean total motility was 69.0% ± 17.7, progressive motility was 43.7% ± 14.7, average path velocity (VAP) was 127.0 µm/s ± 20.7, curvilinear velocity (VCL) was 221.0 µm/s ± 31.1, and sperm velocity index (SVI) was 389.9 ± 56.1. There were positive correlations between the peak systolic velocity (PSV) in the proximal supratesticular region with the SVI (r = .529), VCL (r = .555) and VAP (r = .473), and a negative correlation with the percentage of slow spermatozoa (r = −.463). The results suggest that the testicular artery blood flow velocity can positively affect the speed of spermatozoa movement. For the first time, we have correlated sperm kinetics with the Doppler evaluation of the testicular artery in dogs.     

Correlation of testicular ultrasonography,testicular biometry,serum testosterone levels and seminal attributes in pre and post-pubertal age for breeding soundness evaluation in Osmanabadi bucks

The present research work entitled “Correlation of testicular ultrasonography, testicular biometry, serum testosterone levels and seminal attributes in pre- and post-pubertal age for breeding soundness evaluation in Osmanabadi bucks” was undertaken in 18 healthy Osmanabadi bucks from the Instructional Livestock Farm Complex, Bombay Veterinary College, Mumbai, Maharashtra. The body weight (kg), scrotal circumference (cm) and testicular biometry (cm) of post-weaning 18 Osmanabadi male kids was recorded every 15 days from weaning, i.e., 120?±?10 days along with serum testosterone (ng/ml) by radioimmunoassay method at monthly intervals for the next 6 months. Semen was collected six times on the seventh month onward during post-pubertal age at 15-day interval from 18 bucks. The semen was evaluated for macroscopic and microscopic tests. The body weight increased from 14.45?±?0.67 to 19.57?±?0.70 kg from four to nine and a half months of age. The average daily body weight gain was 31.27 g. Maximum body weight gain was 01.19?±?0.16 kg from 5 to 6 followed by 01.15?±?0.16 kg from 4 to 5 months of age. The scrotal circumference increased from 17.22?±?0.56 to 19.03?±?0.55 cm from four to nine and a half months of age with maximum increased between 4 and 5 followed by 6 and 7 months of age. The testicular length, width and thickness of right and left testicles were recorded by ultrasonography method. There was increase in mean right and left testicular length, width and thickness from 5.25?±?0.19 to 5.84?±?0.18 and 5.49?±?0.21 to 6.16?±?0.20; 2.99?±?0.12 to 3.32?±?0.12 and 3.10?±?0.13 to 3.44?±?0.12 and 2.97?±?0.12 to 3.16?±?0.12 and 3.06?±?0.12 to 3.31?±?0.11 cm, respectively by ultrasonography, between four to nine and a half months of age. Testicular length, width and thickness gain was at maximum in 5 to 6 months of age. Left testicular length was more than the right testis. Before puberty, there was sudden gain in body weight, testicular length and width. However, scrotal circumference showed significant increase after puberty. Body weight had highest correlation with ultrasonographic left testicular thickness (r?=?1) followed by scrotal circumference, ultrasonographic right and left testicular width, left testicular length, right testicular length and thickness and least by right testicular thickness (r?=?0.95). The semen was thin to thick in consistency and average semen density was 3.10?±?0.05. Average semen volume was 0.81?±?0.02 ml, mass activity, initial motility, live and dead sperm count, abnormal sperm count and sperm concentration were 3.45?±?0.13, 76.16?±?1.16 and 75.16?±?1.28% and 24.84?±?1.28, 12.30?±?0.50% and 2631.04?±?45.74 million/ml, respectively in 18 bucks in six collection at 15 days. There was significant rise in semen volume, mass activity, initial motility and concentration at 8.5 months and live count, density at 9 months of age which indicates the age of sexual maturity is 8.5 to 9 months in Osmanabadi bucks. The body weight had highest positive correlation with mass activity (r?=?98) followed by initial motility, live sperm count and total sperm concentration, semen volume (r?=?76). The scrotal circumference had highest positive correlation with initial motility (r?=?98) followed by live sperm count, total sperm count, mass activity, semen volume (r?=?86). On the other hand, body weight and scrotal circumference were negatively correlated with abnormal and dead sperm count. The mean testosterone concentration increased from 0.02?±?0.004 to 5.75?±?0.80 ng/ml between four and half to nine and half months of age, respectively. There was significant rise (p?<?0.01) up to 1.38?±?0.28 ng/ ml at 6.5 months, i.e., age of puberty and up to 5.75?±?0.80 ng/ml at 9.5 months, i.e., age of sexual maturity. Testosterone had highest positive correlation with testicular length followed by testicular width, length, body weight and scrotal circumference, mass activity, live sperm count, initial motility, while it had highest negative correlation with dead and abnormal sperm count. From the present research work, it was concluded that the scrotal circumference, testicular length, width and thickness increased with increasing body weight. Before puberty, there was sudden gain in body weight, testicular length and width. However, scrotal circumference increased significantly at post-pubertal age. So testicular length, body weight, testicular width in pre pubertal age and scrotal circumference post-pubertal age can be used as indicator for selection of Osmanabadi bucks for breeding purpose. On the other hand, the semen parameters should consider only after 8.5 to 9 months of age for selection of Osmanabadi bucks for breeding.

     

Variability in the eruption of the permanent incisor teeth in sheep

The incisor teeth of 176 sheep of six breeds were inspected every two to three months for a year to record the shedding of the deciduous teeth and the eruption of the permanent teeth. In all the breeds the permanent central incisors erupted at between 12 and 18 months of age. In 96 per cent of the sheep the permanent middle incisors erupted at between 18 and 26 months; and in 92 per cent the permanent lateral incisors erupted at between 24 and 36 months of age. The permanent corner teeth erupted at between 32 and 44 months in 96 per cent of the sheep. The gingival redness and swelling accompanying the eruption of a permanent tooth disappeared within two months. In 14 cases two pairs of incisors erupted during the year, in 18 cases the incisors erupted asymmetrically, and in 22 cases no incisors erupted. Rotation of one incisor was observed in five sheep and was combined with dental deviation in one.     

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.     

Combining ultrasound and lactobacilli treatment for high‐fat‐diet‐induced obesity in mice

Chronic systemic lipopolysaccharide‐induced inflammation can cause obesity. In animal experiments, lactobacilli have been shown to inhibit obesity by modifying the gut microbiota, controlling inflammation and influencing the associated gene expression. A previous study found that high‐fat‐diet‐induced (HFD) obesity was suppressed by lactobacilli ingestion in rats via the inhibition of parasympathetic nerve activity. This study explored the combined use of lactobacilli ingestion and ultrasound (US) to control body weight and body fat deposition in HFD mice over an 8‐week experimental period. Male C57BL/6J mice received an HFD during treatment and were randomly divided into four groups: (i) control group (H), (ii) lactobacilli alone (HB), (iii) US alone (HU) and (iv) lactobacilli combined with US (HUB). The US was targeted at the inguinal portion of the epididymal fat pad on the right side. At the 8th week, body weight had decreased significantly in the HUB group (15.56 ± 1.18%, mean ± SD) group compared with the HU (26.63 ± 0.96%) and H (32.62 ± 5.03%) groups (p < 0.05). High‐resolution microcomputed tomography (micro‐CT) scans revealed that the reduction in total body fat volume was significantly greater in the HUB group (69%) than in the other two experimental groups (HB, 52%; HU, 37%; p < 0.05). The reductions in the thickness of the subcutaneous epididymal fat pads were significantly greater in the HUB group (final thickness: 340 ± 7 μm) than in the H (final thickness: 1150 ± 21 μm), HB (final thickness: 1060 ± 18 μm) and HU (final thickness: 370 ± 5 μm) groups (all p < 0.05). Combination therapy with lactobacilli and US appears to enhance the reduction in body weight, total and local body fat deposition, adipocyte size and plasma lipid levels over an 8‐week period over that achieved with lactobacilli or US alone in HFD mice. These results indicate that US treatment alone can reduce hyperlipidemia in HFD mice.     

Morphometry of cervical spinal cord in cat using design-based stereology

The spinal cord harbours nerve fibres that facilitate reflex actions and that transmit impulses to and from the brain. The cervical spinal cord is an area of particular interest in medicine and veterinary due to frequent pathologic alterations in this region. This study describes the morphometric features of the cervical spinal cord in cat using design-unbiased stereological methods. The cervical spinal cords of four male cats were dissected and samples were taken according to systematic uniform random sampling. Each sample was embedded in agar and cut into 60-µm thick sections and stained with cresyl violet 0.1% for stereological estimations. The total cervical spinal cord volume obtained by the Cavalieri estimator was 2,321.21 ± 285.5 mm³. The relative volume of grey matter and white matter was 23.8 ± 1.3% and 76.1 ± 1.3%. The dorsal horn and ventral horn volume were 12.3 ± 1.2% and 11.4 ± 0.7% of the whole cervical spinal cord. The volume of central canal was estimated to 3.8 ± 1 mm³. The total number of neurons was accounted 3,405,366.2 ± 267,469.4 using the optical disector/fractionator method. The number of motoneurons and interneurons was estimated to be 1,120,433.2 ± 174,796.7 and 2,284,932.9 ± 127,261.5, respectively. The average volume of the motoneurons and interneurons was estimated to 1980 µm³ and 680 µm³, respectively, using the spatial rotator method. This knowledge of cat spinal cord findings may serve as a foundation as a translational model in spinal cord experimental research and provide basic findings for diagnosis and treatment of spinal cord disorders.     

Pharmacokinetics of tolfenamic acid in green sea turtles (Chelonia mydas) after intravenous and intramuscular administration

The present study aimed to evaluate the pharmacokinetic features of tolfenamic acid (TA) in green sea turtles, Chelonia mydas. Green sea turtles were administered single either intravenous (i.v.) or intramuscular (i.m.) injection of TA, at a dose of 4 mg/kg body weight (b.w.). Blood samples were collected at preassigned times up to 168 hr. The plasma concentrations of TA were measured using a validated liquid chromatography tandem mass spectrometry method. Tolfenamic acid plasma concentrations were quantifiable for up to 168 hr after i.v. and i.m. administration. The concentration of TA in the experimental green sea turtles with respect to time was pharmacokinetically analyzed using a noncompartment model. The C_max values of TA were 55.01 ± 8.34 µg/ml following i.m. administration. The elimination half-life values were 32.76 ± 4.68 hr and 53.69 ± 3.38 hr after i.v. and i.m. administration, respectively. The absolute i.m. bioavailability was 72.02 ± 10.23%, and the average binding percentage of TA to plasma protein was 19.43 ± 6.75%. Based on the pharmacokinetic data, the i.m. administration of TA at a dosage of 4 mg/kg b.w. might be sufficient to produce a long-lasting anti-inflammatory effect (7 days) for green sea turtles. However, further studies are needed to determine the clinical efficacy of TA for treatment of inflammatory disease after single and multiple dosages.     

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.     

Pathological studies of cheek teeth apical infections in the horse: 2. Quantitative measurements in normal equine dentine

Measurements of primary, regular and irregular secondary dentine and pulp dimensions were made on transverse, sub-occlusal and mid-tooth sections, of 40 maxillary and 42 mandibular control equine cheek teeth (CT) of different ages. Maxillary and mandibular CT primary dentine in different age groups had a mean thickness of 922–1065 μm and 1099–1179 μm, respectively, on the lateral aspects, and 1574–2035 μm and 1155–1330 μm, respectively, on the medial aspects of pulp horns. Surprisingly, some increase in thickness was found in some mandibular CT primary enamel in the first few years following eruption.Regular secondary dentine thickness increased with age, for example at mid-tooth level in mandibular CT from 124 μm at 3 years dental age to 290 μm at >7 years dental age on the lateral aspect of pulp horns, and from 166 μm to 509 μm on the medial aspects of pulp horns, indicating a deposition rate of 0.5–10 μm/day. This type of dentine was thicker sub-occlusally than in the mid-tooth region. Maxillary dentinal dimensions showed a similar age-related increase in thickness. Maxillary CT dentine was significantly thicker (72% in primary, 43% in regular secondary dentine) on the medial compared to the lateral aspects of pulp horns, but mandibular CT dentine was just 15% and 14% thicker in primary and regular secondary dentine thickness, respectively, on the their medial as compared to their lateral aspects. Dentinal and pulp dimensions varied between individual pulp horns, Triadan tooth position, and dental age, with complex interactions between these variables for some parameters.     

Morphometric Changes in the Dog Trochlear Nerve with Growth

The objective of this work was to analyse changes in morphometric characteristics related to growth in the trochlear nerve in dogs. Twenty beagles, split into four dog age groups (A, 7 days; B, 21 days; C, 35 days; D, 49 days and E, 4 years), were used. The right intracranial portion of the nerve was analysed by light and electron microscopy. The nerve cross‐sectional area was calculated. Number, diameter and cross‐sectional area of unmyelinated and myelinated fibres were also calculated. In myelinated fibres, the corresponding axon area and diameter and myelin sheath thickness were also calculated. The number of myelinated and unmyelinated fibres was 1070.25 ± 112.07 and 592.25 ± 467.53 in group A, 1367 ± 57.98 and 143.67 ± 54.37 in group B, 1574.20 ± 299.50 and 151.67 ± 51.73 in group C, 1340.33 ± 151 and 127 ± 48.75 in group D and 1476 ± 260.71 and 284 ± 101.82 in group E. The mean diameter for myelinated and unmyelinated fibres was 4.37 ± 0.17 μm and 0.41 ± 0.08 μm for group A; 6.21 ± 0.12 μm and 0.30 ± 0.03 μm for B; 6.90 ± 0.91 μm and 0.32 ± 0.03 μm for C; 7.86 ± 1.19 μm and 0.32 ± 0.02 μm for D; 10.63 ± 0.50 μm and 0.30 ± 0.01 μm for E, respectively. This nerve possesses similar structural and ultrastructural features to the same nerve in other species and modifies its morphometry with growth. Results could enhance the understanding of pathological disorders.     

Morphometrical study of the cat cerebellum using unbiased design-based stereology

The objective of the present study was to investigate the morphometrical features of the cat cerebellum using design-based stereology. Cerebellar hemispheres from four male cats were examined. Isotropic, uniform random sections were obtained and processed for light microscopy. Cerebellar total volume (V), white (WM) and grey matter (GM) volume fractions, and the volumes of the molecular and granular layers were measured using the Cavalieri's estimator and the point counting system. Cerebellar surface area was estimated using test lines, and Purkinje cellular and nuclear volumes were analysed using the nucleator technique. The volume of the cat cerebellar hemispheres was 2.06 ± 0.29 cm³. The relative volume fractions of the GM and WM were 70.6 ± 2.6% and 29.3 ± 2.6%, respectively. The surface area of the cerebellar hemisphere was 68.2 ± 17.8 cm². The volumes of the molecular and granular layers were estimated at 0.89 ± 0.16 cm³ and 0.56 ± 0.1 cm³, respectively. The Purkinje cell volumes were found to be ranging from 1,717 to 28,489 µm³, of which cells with a perikaryon volume of 6,994 µm³ had a higher incidence. The Purkinje nuclear volume was estimated at 440–3,561 µm³, and nuclei with a volume of 1,252 µm³ were the most frequently occurring ones. Our data might contribute to the veterinary comparative neuroanatomy knowledge, help develop experimental studies in this field, and possibly lead to advancement in the diagnosis and treatment of nervous diseases in the cat.     

Chilled and post‐thaw storage of sperm in different goldfish types

The effective storage time of sperm after stripping (for 48 hr in 6‐hr intervals) and after thawing (for 6 hr in 2‐hr intervals) in Black moor, Oranda and Calico goldfish types was investigated. Variations in sperm density were also measured in all lines. The efficiency of a sperm cryopreservation method formerly developed for common carp was recorded in all three goldfish lines. Motility parameters ((pMOT, %), curvilinear velocity (VCL, μm/s) and straightness (STR, %)) of Black moor sperm did not decrease significantly during 48 hr of storage. A significant reduction in the Oranda type compared to the fresh control was observed in pMOT after 42 (23 ± 2%) and VCL after 36 (94 ± 12 μm/s) hours (pMOT 84 ± 5%, VCL 150 ± 11 μm/s). In the Calico type, pMOT decreased significantly already after 18 (42 ± 26%) and VCL after 6 (105 ± 8 μm/s) hours (fresh: pMOT 92 ± 5%, VCL 151 ± 6 μm/s). A high pMOT immediately following thawing was measured in Oranda (46 ± 12%) and Calico (55 ± 15%) types, whereas a reduced pMOT was recorded in Black moor (24 ± 19%). In Calico, pMOT showed a significant reduction after 6 hr (19 ± 11%) in comparison with the initial value, with no changes observed in VCL and STR. None of the parameters changed in the Black moor and Oranda types. Evidence was found that different goldfish lines have different sperm quality and characteristics. Further studies can investigate the possible effects of chilled and post‐thaw storage on the fertilizing capacity of sperm in the Black moor, Oranda and Calico goldfish types.