Validation of a portable device (iSperm®) for the assessment of stallion sperm motility and concentration

The objective of this study was to evaluate the accuracy of a novel, portable device (iSperm^® Equine for assessing concentration and motility of stallion semen). In the first experiment, semen concentration was determined by the iSperm^® Equine (Aidmics Biotechnology), Androvision^® (Minitube) and NucleoCounter^® SP‐100? (ChemoMetec). The total motility and progressive motility were determined by the iSperm^® Equine and the Androvision^® using the manufacturer's guidelines. Frozen/thawed semen samples (n = 33) at various dilutions were analysed for concentration and motility with the above‐mentioned devices. There was a significant correlation between the concentrations measured with iSperm^® and NucleoCounter^® at all the measured dilutions. Moreover, <10% difference in concentrations was observed between the iSperm^® and NucleoCounter^® using the Bland–Altman test. There was also a significant correlation between iSperm^® and Androvision^® for total and progressive motility. In the second experiment, the parameters used in the Androvision^® were modified to match those of the iSperm^®. Total motility and progressive motility of frozen/thawed semen samples (n = 10) were determined, and the similarity between the Androvision^® and iSperm^® was confirmed by correlation studies and Bland–Altman test. The results of these experiments demonstrate that the iSperm^® offers a reliable and practical alternative for the semi‐automated measurement of concentration and motility of stallion semen in the field. The iSperm^® enables the practitioner to obtain objective and repeatable measurements on a variety of semen types (fresh, cooled and frozen) in the field at the time of insemination and thus acquire more insight into the quantity and quality of the provided insemination doses. This mare‐side diagnostic tool may help practitioners in identifying presumed subfertility problems more rapidly and act accordingly.     

Comparative evaluation of DNA integrity using sperm chromatin structure assay and Sperm‐Ovis‐Halomax during in vitro capacitation of cryopreserved ram spermatozoa

This study aimed to compare the ability of sperm chromatin structure assay (SCSA^®) and Sperm‐Ovis‐Halomax^® to detect DNA fragmentation in frozen‐thawed ram spermatozoa incubated under capacitating conditions in synthetic oviductal fluid (SOF) supplemented with oestrous sheep serum (SOF‐ESS) at multiple time points (0–240 min). Incubation in SOF‐ESS had no significant effects on SCSA^® parameters while the percentage of spermatozoa with fragmented DNA measured by Sperm‐Ovis‐Halomax^® increased after 180 min of incubation. In addition, no correlation or agreement was found between the techniques, suggesting that SCSA^® and Sperm‐Ovis‐Halomax^® may quantify different types of DNA damage in ram spermatozoa under these experimental conditions.     

CONTRAST‐ENHANCED HARMONIC ULTRASONOGRAPHY OF MEDIAL ILIAC LYMPH NODES IN HEALTHY DOGS

Herein, we describe the normal contrast‐enhanced harmonic, color, and power Doppler ultrasonographic characteristics of the medial iliac lymph nodes in healthy dogs. Contrast‐enhanced harmonic ultrasonography of the medial iliac lymph nodes was performed on 14 healthy dogs after intravenous administration of the lipoprotein‐bound inert gas‐filled microbubble contrast media Definity^®. Time–pixel intensity curves were generated for 1‐min postinjection. Quantification of these curves was performed using Philips QLab software. Non‐contrast‐enhanced power and color Doppler examinations were performed in each node to assess vascular patterns subjectively. Normal lymph nodes exhibited a mean contrast wash‐in phase beginning at 6.3 s from the time of injection with mean peak pixel intensity at 12.1 s. Angioarchitecture was best visualized with contrast‐enhanced harmonic ultrasound compared with power and color Doppler. Normal lymph nodes in dogs have a central artery with a centrifugal and uniform branching pattern. Contrast‐enhanced harmonic ultrasonography is a noninvasive examination that demonstrates improved visibility of the intranodal architecture of healthy medial iliac lymph nodes in dogs compared with conventional, non‐contrast‐enhanced Doppler methods that may have future clinical applications.     

Effect of freezing bull semen in two non‐egg yolk extenders on post‐thaw sperm quality

Traditionally, extenders for bull semen included egg yolk or milk, but recently there has been a move to avoid material of animal origin. The aim of this study was to evaluate the effects of two commercial extenders (based on soya lecithin and liposomes) on bull sperm quality after cryopreservation. Post‐thaw sperm quality was evaluated by computer‐assisted sperm analysis and flow cytometric assessment of membrane integrity, chromatin integrity, mitochondrial membrane potential, production of reactive oxygen species and tyrosine phosphorylation. Furthermore, an artificial insemination (AI) trial was conducted, and 56‐day non‐return rates were evaluated. Semen frozen in the liposome‐based extender showed similar membrane integrity and higher mitochondrial membrane potential compared to those in the soya lecithin‐based extender. Chromatin integrity and production of live H₂O₂+ reactive oxygen species were similar in both extenders. Less superoxide was produced in the samples extended with liposome‐based extender, with or without menadione stimulation. Chromatin integrity and tyrosine phosphorylation were not affected by either type of extender. No differences in 56‐day non‐return rate between extenders containing soya lecithin and liposomes were observed in the AI trial (66% ± 0.8 and 65% ± 0.8, respectively). In conclusion, the sperm quality of bull semen frozen in the two extenders that do not contain material of animal origin was similar, although the semen frozen in the liposome‐based extender had higher mitochondrial membrane potential. Either extender could be used in situations where extenders containing material of animal origin are to be avoided.     

Thermophysiological,haematological, biochemical and behavioural stress responses of sheep transported on road

The study was conducted to evaluate the thermophysiological, haematological, biochemical and behavioural stress responses of sheep transported on road. A total of 44 Chamarita breed adult ewes were randomly allotted to one of two groups, one control group (untransported) and transported group (journey of 4 h), and blood stress indicators were measured 1 day before transport and at four time points post‐transport (0, 4 and 24 h). Thermophysiological profiles of ewes were measured by temperature buttons (iButton Thermochron^®) and placed in intravaginal sponges. Direct observations, with a combination of scan and behaviour sampling, were carried out to collect information on individual behaviour and the time it took the ewes to drink water, eat and rest after returning to their pen respectively. Transported ewes lost approximately 1 kg live weight compared to controls and had higher body temperatures until 12 h post‐transport. Cortisol, glucose, non‐esterified fatty acid (NEFA) concentrations as well as the neutrophil–lymphocyte ratio (N/L) and other physiological indicators were higher immediately after unloading in transported ewes but mostly returned to normal after 4 h, with complete recovery after 24 h. Behavioural analysis post‐transport demonstrated that transported ewes chose to eat before drinking and spent less time resting than controls in the first 3 h after unloading. The study demonstrates that transportation even under short‐journey conditions induced behavioural, physiological and thermophysiological responses indicative of the induction of significant stress, leading to live weight shrinkage that may jeopardize farmer's incomes. Finally, results of this study validated the use of iButton Thermochron^® data loggers for monitoring the stress response during transport.     

Evaluation of a Computer‐aided Lung Auscultation System for Diagnosis of Bovine Respiratory Disease in Feedlot Cattle

Background

A computer‐aided lung auscultation (CALA) system was recently developed to diagnose bovine respiratory disease (BRD) in feedlot cattle.

Objectives

To determine, in a case–control study, the level of agreement between CALA and veterinary lung auscultation and to evaluate the sensitivity (Se) and specificity (Sp) of CALA to diagnose BRD in feedlot cattle.

Animals

A total of 561 Angus cross‐steers (initial body weight = 246 ± 45 kg) were observed during the first 50 day after entry to a feedlot.

Methods

Case–control study. Steers with visual signs of BRD identified by pen checkers were examined by a veterinarian, including lung auscultation using a conventional stethoscope and CALA that produced a lung score from 1 (normal) to 5 (chronic). For each steer examined for BRD, 1 apparently healthy steer was selected as control and similarly examined. Agreement between CALA and veterinary auscultation was assessed by kappa statistic. CALA''s Se and Sp were estimated using Bayesian latent class analysis.

Results

Of the 561 steers, 35 were identified with visual signs of BRD and 35 were selected as controls. Comparison of veterinary auscultation and CALA (using a CALA score ≥2 as a cut off) revealed a substantial agreement (kappa = 0.77). Using latent class analysis, CALA had a relatively high Se (92.9%; 95% credible interval [CI] = 0.71–0.99) and Sp (89.6%; 95% CI = 0.64–0.99) for diagnosing BRD compared with pen checking.

Conclusions

CALA had good diagnostic accuracy (albeit with a relatively wide CI). Its use in feedlots could increase the proportion of cattle accurately diagnosed with BRD.     

In vitro studies of Norwegian Red bovine semen immobilized and cryopreserved in alginate solid gel network

Development of new semen cryopreservation techniques improving sperm survival and ensuring availability of viable spermatozoa for a prolonged time‐period after AI is promising tools to reduce sensitivity of timing of AI and enhance overall fertility. The SpermVital^® technology utilizes immobilization of bull spermatozoa in a solid network of alginate gel prior to freezing, which will provide a gradual release of spermatozoa after AI. The objective of this study was to compare post‐thaw sperm quality and in vitro sperm survival over time of Norwegian Red bull semen processed by the SpermVital^® (SV) technology, the first commercialized production line of SpermVital^® (C) and by conventional procedure applying Biladyl^® extender (B). Post‐thaw sperm motility was not significantly different between SV, C and B semen (p > .05). However, sperm viability and acrosome intactness were higher for SV than C and B semen (p < .05). Small differences in DNA quality were observed (p < .05). Sperm viability after storage in uterus ex vivo was higher for SV than for C semen (p < .05). Furthermore, sperm survival in vitro over time at physiological temperature was significantly higher for SV semen than C semen as well as B semen during the incubation period of 48 hr (p < .05). In conclusion, the SpermVital^® technology is improved and is more efficient in conserving post‐thaw sperm quality and results in higher sperm viability over time in vitro for SV than for C and B semen.     

Cryopreservation of Nili‐Ravi buffalo (Bubalus bubalis) semen in AndroMed® extender; in vitro and in vivo evaluation

The study was designed to evaluate AndroMed^® for the freezability and fertility of Nili‐Ravi buffalo semen. Semen was collected from four adult Nili‐Ravi buffalo (Bubalus bubalis) bulls for 3 weeks (replicate). Semen ejaculates from each buffalo bull were divided into three aliquots. One aliquot was used for evaluation of motility, plasma membrane integrity, livability, viability, DNA integrity and normal apical ridge. Remaining two aliquots were diluted (37°C; 50 × 10⁶ spermatozoa/ml) in tris‐citric egg yolk or AndroMed^® extender and cryopreserved in 0.5 ml French straws. After thawing, per cent post‐thaw motility (47.9 ± 0.8, 49.2 ± 1.7), plasma membrane integrity (44.4 ± 1.2, 46.8 ± 1.8) and normal apical ridge (81.4 ± 0.3, 83.2 ± 0.3) were recorded similar (p > .05) in tris‐citric egg yolk and AndroMed^® extender. Higher (p < .05) percentage of sperm livability (70.5 ± 1.4 and 64.4 ± 1.0), viability (67.5 ± 1.5 and 61.5 ± 0.6) and DNA integrity (97.0 ± 0.3 and 93.4 ± 0.21) were recorded in AndroMed^® compared to tris‐citric egg yolk post‐thaw. Values for all the aforementioned spermatozoal quality parameters were observed lower (p < .05) in frozen‐thawed compared to fresh semen irrespective of the experimental extenders. Fertility rates of buffalo semen did not differ (p > .05) either cryopreserved in tris‐citric egg yolk or AndroMed^® extender (45.5% vs. 49%). It is concluded that AndroMed^® is capable in protecting the buffalo bull sperm during freeze‐thawing process and can be adopted safely for routine use replacing the tris‐citric egg yolk extender in artificial insemination programme.     

INTRAOPERATIVE CONTRAST‐ENHANCED ULTRASONOGRAPHY OF NORMAL CANINE JEJUNUM

Nine normal juvenile dogs were evaluated with direct jejunal contrast‐enhanced ultrasonography via midline celiotomy. Three different doses of ultrasound contrast medium (Definity^®) were injected through a peripheral venous catheter. Time‐intensity curves were used to calculate baseline, time to initial rise, inflow slope, time‐to‐peak, peak intensity (PI), and outflow slope for each administered dose. PI was directly proportional to dose. Outflow slope was similar for all patients, independent of dose. The most favorable images were acquired with a dose of 0.030 ml/kg given as a rapid intravenous manual bolus. The technique and normal jejunal perfusion pattern described herein may provide useful data for evaluation of intestinal vascular, inflammatory, and neoplastic disease in the dog.     

Ileal digestibility of amino acids,phosphorus, phytate and energy in pigs fed sorghum‐based diets supplemented with phytase and Pancreatin®

The effects of phytase supplementation on the apparent ileal digestibility (AID) of amino acids (AA) have been inconsistent. Two experiments evaluated the effect of providing a mixture of pancreatic enzymes (Pancreatin^®) to growing pigs fed sorghum–soybean meal diets supplemented with phytase on the AID of AA, energy, and phosphorus (P), as well as the ileal digestibility (ID) of phytate; there were four periods per experiment. In Experiment 1, eight pigs (BW 22.1 ± 1.3 kg) were fitted with a T‐cannula at the distal ileum. Each period consisted of 9 days; 7 days for diet adaptation, and 2 days for digesta collection. Treatments (T) were: (i) basal sorghum–soybean meal diet, (ii) basal diet plus Pancreatin^®, (iii) basal diet plus phytase and (iv) basal diet plus phytase and Pancreatin^®. Phytase increased the digestibilities of phytate and P (p < 0.001), but did not affect the AID of AA and energy (p > 0.10). Except for methionine (p = 0.07), Pancreatin^® did not affect the AID of AA. Phytase and Pancreatin^® did not interact (p > 0.10). Experiment 2 was similar to Experiment 1, but Pancreatin^® was infused into duodenum. Pancreatin^® infusion did not affect the AID of AA (p > 0.10); and tended to reduce (p = 0.09) the AID of lysine. Phytase × Pancreatin^® interactions were not observed (p > 0.10). In conclusion, phytase and Pancreatin^® did not improve the AID of AA in growing pigs fed sorghum–soybean meal diets indicating that phytates did not affect AA digestibility.     

Evaluating the use of plasma hematocrit samples to detect ketones utilizing urine dipstick colorimetric methodology in diabetic dogs and cats

Objective: To determine whether plasma from a heparinized hematocrit tube placed on a urine dipstick would accurately reflect (positive or negative) urine ketone results in diabetic dogs and cats. Design: Prospective study, 37 dogs and 43 cats, with a known history of diabetes or hyperglycemia, glucosuria, and symptoms of undiagnosed diabetes mellitus were tested. Setting: Veterinary Referral Hospital. Animals: Client owned dogs and cats. Interventions: None. Measurement and main results: Heparinized plasma and urine ketone results were recorded using urine reagent strips. Plasma dipstick results were compared to urine dipstick results as the standard. Results were recorded based on the color chart provided by the manufacturer. Two individuals were responsible for verifying the results of the colorimetric test. Test efficiency was 97% (sensitivity = 96%, specificity = 100%) for the canine population, 93% (sensitivity = 100%, specificity = 83%) for the feline population, and 95% (sensitivity = 98%, specificity = 91%) for the total population. Four of 80 animals were found to have discordant results (1 dog and 3 cats). Conclusion: Plasma from heparinized hematocrit tubes is clinically useful for detecting the presence or absence of ketonuria, and therefore ketosis, in diabetic dogs and cats using urine dipstick colorimetric methodology.     

EFFECTS OF GADOXETATE DISODIUM (EOVIST®) CONTRAST ON MAGNETIC RESONANCE IMAGING CHARACTERISTICS OF THE LIVER IN CLINICALLY HEALTHY DOGS

Gadoxetate disodium (Gd‐EOB‐DTPA; gadolinium‐ethoxybenzyl‐diethylene triamine penta‐acetic acid) is a newly developed paramagnetic contrast agent reported to have a high specificity for the hepatobiliary system in humans. The purpose of this prospective study was to describe effects of Gd‐EOB‐DTPA contrast administration on MRI characteristics of the liver in eight clinically healthy dogs. Precontrast dorsal and transverse T1‐weighted spin echo, T2‐weighted fast spin echo, and transverse T1‐weighted 3D gradient echo (VIBE; volume‐interpolated body examination) pulse sequences were acquired for each dog. Dogs were assigned to four groups based on contrast dose administered (0.0125 mmol/kg or 0.025 mmol/kg), and pulse sequences acquired after contrast administration (T1‐weighted spin echo and T1‐weighted 3D gradient echo). Liver signal intensity ratios were calculated and compared between the two contrast dose groups and two postcontrast pulse sequence groups using ANOVA. No adverse effects of contrast administration were observed. All dogs exhibited homogeneous contrast enhancement of the liver with no statistical difference in enhancement between the two different contrast doses. Contrast enhancement in all dogs peaked between 1 and 10 min after intravenous injection. There was a significant difference in mean signal intensity ratios between sequences (P = 0.035) but not between doses (P = 0.421). Postcontrast signal intensities of the liver parenchyma were significantly higher for the T1‐weighted 3D gradient echo images when compared to the T1‐weighted spin echo sequences. Findings indicated that Gd‐EOB‐DTPA contrast administration is safe in healthy dogs and causes homogeneous enhancement of the liver that is more pronounced in T1‐weighted 3D gradient echo MRI pulse sequences.     

Ahmed glaucoma valve implantation with Ologen® Collagen Matrix for the surgical treatment of feline glaucoma

An Ahmed valve implantation with an Ologen^® Collagen Matrix (Ologen^® CM, Aeon Astron, Leiden, the Netherlands) was performed for the treatment of uncontrolled glaucoma in a cat. This cat was a 5‐year‐old castrated Russian Blue male with a 12‐week history of conjunctival hyperemia and mydriasis of the left eye. During the ophthalmic examination, the intraocular pressure (IOP) oculus sinister (OS) was 52 mmHg, and a narrow iridocorneal angle (ICA) was detected by gonioscopy. Medical treatment with Cosopt^® (2% dorzolamide and 0.5% timolol) failed to decrease the IOP. The left eye still had vision, and an Ahmed valve implantation was performed. During the gonioimplantation, Ologen^® CM was used to inhibit scar formation around the valve. Following the operation, the IOP was stable at an approximate average of 15 mmHg during the 7‐month follow‐up period, and vision in the left eye was retained without medication. An adequate subconjunctival filtering bleb was formed after 140 days. This is the first case report in which an Ahmed valve gonioimplant with an Ologen^® CM has been used for the surgical treatment of glaucoma in a cat.