Michael Hässig

Seasonal changes in semen quality and freezability in the Warmblood stallion

The objective of this study was to investigate seasonal changes in stallion semen quality and to determine the best time for semen cryopreservation. Experiments were performed using 10 Warmblood stallions from the National Stud Farm in Avenches (Switzerland). Ejaculates were collected and frozen every other week during 1 year from January to December 1999. Volume, concentration, and motility, and the number of morphologically normal sperm and sperm with major defects (abnormal heads, acrosome defects, nuclear vacuoles, proximal droplets, abnormal midpieces) were evaluated. For all frozen-thawed semen samples motility as well as viability (SYBR-14/PI) was tested, and the hypoosmotic swelling test (HOS) was performed. To analyze seasonal differences 4 periods of 3 months each were defined: autumn (September, October, November), winter (December, January, February), spring (March, April, May) and summer (June, July, August). During the 1 year experiment all semen quality parameters showed a clear seasonal pattern. The volume, total sperm count and motility in fresh semen were significantly higher (P<0.05) in summer than in winter, while sperm concentration was significantly lower in summer compared to the other seasons. Regarding morphology, normal sperm was significantly lower (P<0.05) in summer than at any other time of the year and higher values (P<0.05) were found for major defects in summer than in spring and autumn. In frozen-thawed semen motility was significantly (P<0.05) improved in autumn when compared to spring and summer. Viability was lowest in summer and differed significantly (P<0.05) from other seasons. The HOS test revealed significantly more (P<0.05) membrane damaged spermatozoa in winter than in spring, summer and autumn. Our results demonstrate that in our climatic conditions clear seasonal differences occur in semen quality of fresh and frozen-thawed semen and that cryopreservation of stallion semen should preferably be performed in autumn.

Evaluation of a rebound tonometer (Tonovet) in clinically normal cat eyes.

OBJECTIVE To determine the accuracy of and to establish reference values for a rebound tonometer (Tonovet) in normal feline eyes, to compare it with an applanation tonometer (Tonopen Vet) and to evaluate the effect of topical anesthesia on rebound tonometry. PROCEDURES Six enucleated eyes were used to compare both tonometers with direct manometry. Intraocular pressure (IOP) was measured in 100 cats to establish reference values for rebound tonometry. Of these, 22 cats were used to compare rebound tonometry with and without topical anesthesia and 33 cats to compare the rebound and applanation tonometers. All evaluated eyes were free of ocular disease. RESULTS Both tonometers correlated well with direct manometry. The best agreement with the rebound tonometer was achieved between 25-50 mmHg. The applanation tonometer was accurate at pressures between 0 and 30 mmHg. The mean IOP in clinically normal cats was 20.74 mmHg with the rebound tonometer and 18.4 mmHg with the applanation tonometer. Topical anesthesia did not significantly affect rebound tonometry. CONCLUSIONS As the rebound tonometer correlated well with direct manometry in the clinically important pressure range and was well tolerated by cats, it appears suitable for glaucoma diagnosis. The mean IOP obtained with the rebound tonometer was 2-3 mmHg higher than that measured with the applanation tonometer. This difference is within clinically acceptable limits, but indicates that the same type of tonometer should be used in follow-up examinations in a given cat.

Seasonal changes of semen quality and freezability in Franches–Montagnes stallions

The objective of this study was to investigate seasonal changes of semen quality parameters in Franches-Montagnes stallions and to compare the freezability of ejaculates collected in autumn and winter. Experiments were performed using 15 stallions from the National Stud Farm in Avenches (Switzerland). Ejaculates were collected and evaluated every month during 1 year as well as cryopreserved in autumn and winter (September to February). In fresh semen the gel-free volume, concentration, motility and morphology (normal sperm, major defects, vacuoles and acrosome defects) were evaluated and in frozen-thawed semen the motility as well as the viability (SYBR-14/PI) were performed. To analyse seasonal differences four periods of 3 months each were defined as autumn (September, October, November), winter (December, January, February), spring (March, April, May) and summer (June, July, August). During the 1-year experiment all fresh semen quality parameters demonstrated a clear seasonal and individual pattern. The gel-free volume was significantly (P<0.05) higher in spring and summer compared to autumn and winter while sperm concentration was significantly (P<0.05) lower in spring than at any other time of the year. Total sperm number was significantly (P<0.05) higher and sperm motility significantly (P<0.05) lower in summer than in other seasons. Regarding sperm morphology, normal sperm was significantly (P<0.05) higher in autumn than in winter and summer and major defects were lowest (P<0.05) in autumn. In frozen-thawed semen motility was significantly (P<0.05) improved in the ejaculates collected in autumn compared to winter, while viability showed no obvious differences. Our results clearly demonstrate that individual and seasonal differences occurred in semen quality of Franches-Montagnes stallions. Ejaculates collected in autumn (September, October, November) demonstrated good quality, especially regarding sperm morphology, and were more suitable for cryopreservation because of better motility in frozen-thawed semen collected during autumn than in winter.

Waddlia, Parachlamydia and Chlamydiaceae in bovine abortion.

The etiology remains unknown in many cases of bovine abortion in Switzerland. Bacteria of the Chlamydiales order are known abortive agents, therefore cases of bovine abortion from three representative regions of Switzerland were investigated in this study. Particularly Chlamydiaceae as well as the Chlamydia-like organisms Waddlia and Parachlamydia were of interest, especially because of their possible zoonotic potential. Placenta samples (n=343) were tested for these bacteria by different PCR-methods, immunohistochemistry and serology for Chlamydia abortus. Additionally an attempt for the isolation of Waddlia and Parachlamydia was made by co-cultivation in amoebae. In 67.3% of the 343 cases a necrotizing and/or purulent placentitis was found histologically. By real-time PCR, 0.9% (3/343) of the cases were positive for Waddlia, 13.4% (46/343) positive for Parachlamydia and 14.6% (50/343) positive or questionable positive for Chlamydiaceae. Of these samples, confirmation by immunohistochemistry was possible in 2/3 cases for Waddlia, 25/46 for Parachlamydia and 4/50 for Chlamydiaceae. Of the 50 cases positive or questionable positive for Chlamydiaceae, species-identification by ArrayTube Microarray or 16S rRNA PCR resulted in 41 cases positive for C. abortus whereas the presence of Chlamydia suis was confirmed in four and Chlamydia pecorum in one case. This study brought evidence for the importance of different members of Chlamydiales in different regions of Switzerland although Waddlia is not occurring in a high prevalence. On the other hand mixed infections with different Chlamydiales as well as with other abortigenic agents could be found.

Effect of tibial tuberosity advancement on femorotibial shear in cranial cruciate-deficient stifles An in vitro study

OBJECTIVES This in vitro study compares the femorotibial shear in canine stifles with intact and transected cranial cruciate ligaments (CrCL), before and after tibial tuberosity advancement (TTA) by measuring the distance between origin and insertion of the cruciate ligaments. METHODS Radiodense markers were inserted into bones at the attachment sites of the cruciate ligaments in sixteen cadaveric stifles of adult dogs. Each stifle was then mounted in a testing apparatus. The distances between the markers were measured on mediolateral radiographs, performed on each stifle under three different situations: intact, after CrCL transection, and after performing a TTA. Stifles were loaded to create a constant tibiofemoral reaction force by maintaining the load parallel to the patellar ligament. Radiographs were taken in a preloaded and loaded state in the intact stifle and only in a loaded state after CrCL transection, and after performing a TTA. RESULTS Loading the stifle joints after transection of the CrCL resulted in a mean lengthening of the CrCL marker distance of 22.4%. Loading the transected CrCL stifles after performing a TTA resulted in a mean shortening of the CrCL marker distance by 3.0% compared to the loaded intact condition. CLINICAL SIGNIFICANCE This study demonstrates that, in loaded stifles with transected CrCLs, TTA causes a caudal shift in the cranial shear force, counteracting cranial subluxation of the tibia.

Isoflurane and sevoflurane provide equally effective anaesthesia in laboratory mice

Isoflurane is currently the most common volatile anaesthetic used in laboratory mice, whereas in human medicine the more modern sevoflurane is often used for inhalation anaesthesia. This study aimed to characterize and compare the clinical properties of both anaesthetics for inhalation anaesthesia in mice. In an approach mirroring routine laboratory conditions (spontaneous breathing, gas supply via nose mask, preventing hypothermia by a warming mat) a 50 min anaesthesia was performed. Anaesthetics were administered in oxygen as carrier gas at standardized dosages of 1.5 minimum alveolar concentrations, which was 2.8% for isoflurane and 4.9% for sevoflurane. Both induction and recovery from anaesthesia proceeded quickly, within 1–2 min. During anaesthesia, all reflex testing was negative and no serious impairment of vital functions was found; all animals survived. The most prominent side-effect during anaesthesia was respiratory depression with hypercapnia, acidosis and a marked decrease in respiration rate. Under anaesthesia, heart rate and core body temperature remained within the normal range, but were significantly increased for 12 h after anaesthesia. Locomotor activity, daily food and water consumption and body weight progression showed no abnormalities after anaesthesia. No significant difference was found between the two anaesthetics. In conclusion, isoflurane and sevoflurane provided an equally reliable anaesthesia in laboratory mice.

Effect of dog breed and body conformation on vertical ground reaction forces, impulses, and stance times.

OBJECTIVES To assess whether fully normalised vertical ground reaction forces and stance times obtained at a trot depend on dog breed or body conformations. METHODS Peak vertical forces (PVF), vertical impulses (VI), stance times (ST), and ratio of forelimb impulse to total impulse (RVI) of 54 dogs of seven different breeds were normalised to body weight and body size according to the theory of dynamic similarity, and were tested for differences between breeds. Breeds were Borzoi, Bernese Mountain dog, Great Dane, Labrador Retriever, Landseer, Rhodesian Ridgeback, and Rottweiler. Body length ratio (BLR) and body mass index (BMI) were also compared between breeds. RESULTS Significant differences between breeds were found for the normalised forelimb PVF, VI and ST, and hindlimb PVF. Looking at individual breeds, it was most evident that Borzois had a lower forelimb VI, and a higher hindlimb PVF than the other breeds. This resulted in Borzois having a lower RVI compared to other dogs, indicating a more caudally located centre of gravity. Only a few differences in gait parameters were found between other dog breeds. The BMI was significantly lower in Borzois than in other breeds, but was otherwise not associated with gait parameters. CLINICAL SIGNIFICANCE Force plate data of dogs of different breeds are not necessarily comparable, even after full normalisation to body weight and body size. Group comparisons should only be made when the groups consist of breeds with similar body conformations.

Histomorphological and immunohistochemical findings in testes, bulbourethral glands and brain of immunologically castrated male piglets.

The aim of this study was the histological and immunohistochemical evaluation and comparison of testicular, bulbourethral and brain tissue in immunized and intact control boars. Fourteen male piglets, aged between 10 and 16 weeks, were vaccinated twice subcutaneously 4 to 5 weeks apart with Improvac, an anti-GnRH vaccine. The pigs were sacrificed 1 to 16 weeks following the second injection. Testicular weight was recorded and various tissue samples were collected and fixed in formalin and Bouins fixative for histological examination. In addition, 2 boars were immunized five times and slaughtered 60 weeks after the last injection. Histological and immunohistological studies performed on testes and epididymes showed clear signs of atrophy in the immunized animals and a significant reduction in paired testes weight was seen in treated boars. Microscopically, the mean diameter of the seminiferous tubules was markedly reduced. Spermatogonia as well as few spermatocytes were visible between the Sertoli cells and Leydig cells were atrophic. None or only few spermatozoa were detected in the epididymis. The bulbourethral glands of immunocastrated pigs were smaller than in control pigs and showed histological evidence of atrophy. Immunohistological detection of LH and FSH in the pituitary gland of treated and control boars showed no quantifiable difference in the amount of these two gonadotropins and no lesions were visible in the hypothalamus and the pituitary gland. From our findings it can be concluded that the anti-GnRH vaccine Improvac induces severe atrophy of testes and bulbourethral glands in immunized pigs. This effect appears to be reversible, depending on the immune response of each animal and the time elapsed after the last booster injection.

Repair of long-bone fractures in cats and small dogs with the Unilock mandible locking plate system

OBJECTIVES To retrospectively evaluate stabilisation of long-bone fractures in cats and small dogs using the Unilock system. METHODS Medical histories and radiographs of consecutive patients with long-bone fractures stabilised with the Unilock system were reviewed. Cases with follow-up radiographs taken at least four weeks postoperatively were included. Signalment of the patient, fracture localisation and type, primary fracture repair or revision surgery, single or double plating, and complications for each patient were noted. Additionally, implant size, number of screws, number of cortices engaged with screws, and number of empty holes across the fracture were evaluated in fractures where a single plate had been applied. RESULTS Eighteen humeral, 18 radial, 20 femoral, and 10 tibial fractures were treated. The Unilock system was used for primary repair in 44 fractures and for revision surgery in 22 fractures. Two plates were applied in 17 fractures, and a single plate was applied in 49 fractures. Follow-up radiographs were taken four to 109 weeks postoperatively. Complications were seen in 12 animals and 13 fractures (19.7%). Fixation failure occurred in seven fractures (10.6%). Cases with a single plate that suffered fixation failure had thinner screws in relation to bone diameter than cases with double plates, and more screws in a main fragment than those without fixation failure. CLINICAL SIGNIFICANCE The Unilock system is a suitable implant for fracture fixation of long bones in cats and small dogs.

A study of cardiovascular function under controlled and spontaneous ventilation in isoflurane-medetomidine anaesthetized horses

OBJECTIVE To determine, in mildly hypercapnic horses under isoflurane-medetomidine balanced anaesthesia, whether there is a difference in cardiovascular function between spontaneous ventilation (SV) and intermittent positive pressure ventilation (IPPV). STUDY DESIGN Prospective randomized clinical study. ANIMALS Sixty horses, undergoing elective surgical procedures under general anaesthesia: ASA classification I or II. METHODS Horses were sedated with medetomidine and anaesthesia was induced with ketamine and diazepam. Anaesthesia was maintained with isoflurane and a constant rate infusion of medetomidine. Horses were assigned to either SV or IPPV for the duration of anaesthesia. Horses in group IPPV were maintained mildly hypercapnic (arterial partial pressure of carbon dioxide (PaCO(2)) 50-60 mmHg, 6.7-8 kPa). Mean arterial blood pressure (MAP) was maintained above 70 mmHg by an infusion of dobutamine administered to effect. Heart rate (HR), respiratory rate (f(R)), arterial blood pressure and inspiratory and expiratory gases were monitored continuously. A bolus of ketamine was administered when horses showed nystagmus. Cardiac output was measured using lithium dilution. Arterial blood-gas analysis was performed regularly. Recovery time was noted and recovery quality scored. RESULTS There were no differences between groups concerning age, weight, body position during anaesthesia and anaesthetic duration. Respiratory rate was significantly higher in group IPPV. Significantly more horses in group IPPV received supplemental ketamine. There were no other significant differences between groups. All horses recovered from anaesthesia without complications. CONCLUSIONS There was no difference in cardiovascular function in horses undergoing elective surgery during isoflurane-medetomidine anaesthesia with SV in comparison with IPPV, provided the horses are maintained slightly hypercapnic. CLINICAL RELEVANCE In horses with health status ASA I and II, cardiovascular function under general anaesthesia is equal with or without IPPV if the PaCO(2) is maintained at 50-60 mmHg.