Postoperative complications and owner assessment of single portal laparoscopic ovariectomy in dogs.

The aim of this retrospective study and owner survey was to record the postoperative complication rate, pain score and owners' satisfaction rates of single portal laparoscopic ovariectomy in dogs using an operating laparoscope and to compare our results with data in the veterinary literature on other laparoscopic methods. Therefore, the medical records of all bitches that were spayed using a laparoscopic single port ovariectomy with an operating laparoscope between November 2007 and November 2014 were reviewed. A minimum follow-up period of 12 months was required for study inclusion. One hundred and thirty two adult, healthy, femal dogs were included in our study. The information collected included breed, bodyweight, age at ovariectomy and postoperative complications detected during follow-up 9-14 days after surgery. Furthermore, an electronic questionnaire was implemented, and the owners of the dogs were asked to report on long-term complications and to retrospectively evaluate postoperative pain and overall satisfaction. Sixty-two different and mixed breed dogs with weights ranging from 1.9 to 50 kg and ages ranging from 0.5 to 10.8 years were included. Short-term complications were detected in 21 out of 132 cases, mostly at the surgical site (15.9 per cent). According to our questionnaire (n=65), 10 dogs developed urinary incontinence categorised as a major complication (15.4 per cent). The owner's evaluated pain score was less than 300 on a 1000-unit visual analogue scale for 56 out of 65 dogs (86.2 per cent) and less than 500 in 63 dogs (96.9 per cent). The dog owners' satisfaction rate was 95.3 per cent (87.5 per cent evaluated the surgery as excellent and 7.8 per cent as good). Altogether, our study suggests that single port access using an operating laparoscope for ovariectomy has minor complication rates comparable to other laparoscopic techniques but has the advantages of a low postoperative pain level as evaluated by the owner and a very high owner's satisfaction rate.

A surgical glove port technique for laparoscopic-assisted ovariohysterectomy for pyometra in the bitch.

The objective of the study was to describe the feasibility of a glove port technique for laparoscopic-assisted surgical treatment of canine pyometra. In this retrospective case series, a total of 10 female dogs (median age 7 years, range 5.5-10.5 years; median weight 37.0 kg, range 12.9-64.0 kg) with pyometra were included. A multiaccess port was created from a surgical glove attached to an Alexis wound retractor and placed in the ventral midline between the middle and caudal third of the distance between umbilicus and pubic rim. A vessel sealing device was used for transection of the ovarian pedicle. The port size was selected on the basis of maximum uterine diameter determined by ultrasound. Median incision length was 5.0 cm (range 3.1-7.7 cm) for a maximum uterine diameter of 4.0 cm (range 2.0-7.0 cm). Median surgical time was 57 minutes (range 48-65 minutes). No case had to be converted to open celiotomy. Complications included one case of minor, self-limiting splenic trauma by the endoscope. In eight dogs, the distended uterine horns endangered safe access to the ovarian pedicle, and the vessel sealing device was inserted through a second cannula placed periumbilically. Extension of the original incision was necessary to exteriorize organs in two dogs. All dogs recovered quickly and were discharged either on the day of surgery or 1 day thereafter. In conclusion, a surgical glove port technique in combination with an Alexis wound retractor is feasible for surgical laparoscopic treatment of canine pyometra up to a diameter of 7 cm.

The PGF2α agonists luprostiol and d-cloprostenol reliably induce luteolysis in luteal phase mares without evoking clinical side effects or a stress response.

In the present study we have evaluated a possible stress reaction in response to two different PGF2α analogs-luprostiol and D-cloprostenol--and their effects on estrous cycle characteristics. In a cross-over-design eight mares received in alternating order either luprostiol (Treatment LUP; 3.75 mg im), D-cloprostenol (Treatment CLO; 22.5µg im) or saline (Treatment CON; NaCl 0.9% 0.5ml im) on day 8 after ovulation. Injection of either LUP or CLO, but not of CON resulted in a significant decline of progesterone concentration in plasma to baseline concentrations within two days (time: p<0.001, treatment: p<0.01, time × treatment: p<0.05). The treatment to ovulation interval was significantly shorter in LUP and CLO than in CON cycles (LUP: 9.4 ± 0.4 d; CLO: 9.4 ± 1.3 d; CON: 16.1 ± 0.8 d; p<0.001). Injection of either LUP or CLO, but not of CON significantly increased salivary cortisol concentration (immediately before injection: CON 1.3 ± 0.2, LUP 1.4 ± 0.3, CLO 1.4 ± 0.3 ng/ml; 60 min after injection: CON 1.0 ± 0.3, LUP 8.0 ± 1.4, CLO 4.2 ± 0.7 ng/ml; time: p<0.01, treatment: p<0.001, time × treatment: p<0.001). Heart rate decreased over time (p<0.05) independent of treatment and no changes in heart rate variability were detected. Injection of the PGF2α analogs CLO and LUP reliably induced luteolysis and apart from a transient increase in salivary cortisol concentration no signs of a physiological stress response or apparent side effects occurred.

Development of the genital microflora in stallions used for artificial insemination throughout the breeding season.

An important factor influencing stallion fertility is the microbial contamination of semen. Aims of this study were to investigate changes in the microbiological population of the genital mucosa and semen in artificial insemination stallions (n=16) from before to after one breeding season (February-August). MALDI-TOF-MS (matrix assisted laser desorption/ionization time-of-flight mass spectrometry) was used for identification of microbial agents. For bacteriology, swabs from the urethral opening, urethral fossa and penile sheath as well as semen were collected at 4-week-intervals. For semen motility and percentage of morphologically normal spermatozoa, changes over time (P<0.001) occurred. In 14.3% of genital swabs and 25.0% of ejaculates no microbial growth was found. Intensity of total microbial growth increased throughout the breeding season (P<0.001). From the penile sheath, between 1.4±0.1 microbial species in February and 3.3±0.4 in August were identified. From semen, 1.1±0.3 microbial species in February and 2.9±0.6 in August were obtained. The number of microbial species isolated from the sheath of the penis (2.0±0.1) and urethral fossa (2.0±0.1) was greater (P<0.01) compared with the urethral opening (1.6±0.1) and semen (1.5±0.1). The microbial flora consisted of aerobic and anaerobic bacteria, dominated by coagulase-negative staphylococci, alpha-haemolytic streptococci and coryneforms. Only occasionally potentially pathogenic agents (E. coli, Ps. aeruginosa) were found. The microbial flora was not related to seminal characteristics.

Changes in faecal bacteria and metabolic parameters in foals during the first six weeks of life.

Many foals develop diarrhoea within the first two weeks of life which has been suggested to coincide with postpartum oestrus in their dams. To analyse the pathogenesis of this diarrhoea we have determined faecal bacteria in foals and their dams (n=30 each), and serum IGF-1 and γ-globulins for 6 weeks after birth. In addition, effects of ß-carotene supplementation to mares (group 1: 1000 mg/day, n=15, group 2: control, n=15) on diarrhoea in foals were studied. Diarrhoea occurred in 92 and 79% of foals in groups 1 and 2, respectively, but was not correlated with oestrus in mares. Beta-carotene supplementation was without effect on foal diarrhoea. In mares, bacterial flora remained stable. The percentage of foals with cultures positive for E. coli was low at birth but increased within one day, the percentage positive for Enterococcus sp. was low for 10 days and for Streptococcus sp. and Staphylococcus sp. was low for 2-4 weeks. By 4 weeks of age, bacterial flora in foals resembled an adult pattern. Concentration of serum IGF-1 was low at birth (group 1: 149 ± 11, group 2: 166 ± 17ng/ml), increased after day 1 (day 7 group 1: 384 ± 30, group 2: 372 ± 36) but at no time differed between groups. Serum γ-globulin concentration in foals was low before colostrum intake and highest on day 1 (p<0.001 over time). In conclusion, neonatal diarrhoea in foals does not coincide with postpartum oestrus in their dams but with changes in intestinal bacteria and is not influenced by ß-carotene supplementation given to mares.

Comparison of an extender containing defined milk protein fractions with a skim milk-based extender for storage of equine semen at 5 degrees C.

A problem of semen extenders based on milk or egg yolk is the fact that these biological products consist of a variety of substances. Extenders containing only components with clearly protective effects on spermatozoa would thus be an advantage. In this study, we have compared the effects of an extender containing defined caseinates and whey proteins only (EquiPro, defined milk protein extender) with skim milk extender on equine spermatozoa during cooled storage. The defined milk protein extender was used with and without the antioxidant N-acetyl cysteine (NAC). In a second experiment, semen was diluted with PBS or defined milk protein extender and was either stored directly or 90% of seminal plasma was removed by centrifugation and replaced by defined milk protein extender before storage. In both experiments, eight stallions were available for semen collections. Motility, velocity and membrane integrity of spermatozoa were determined by CASA immediately after semen processing and after 24, 48 and 72 h of storage at 5 degrees C. Total motility after 24 h of storage was lowest in semen diluted with PBS (p<0.05 versus all extenders). At 48 and 72 h, motility of spermatozoa in defined milk protein extender was significantly (p<0.05) higher than in PBS or skim milk extender. Velocity of spermatozoa after storage was highest in defined milk protein extender. Membrane integrity after storage was significantly (p<0.05) lower in semen diluted with PBS than in semen diluted with both extenders. Addition of NAC was without effect on the examined parameters. Centrifugation further increased the percentage of motile and membrane-intact spermatozoa in the defined milk protein extender (p<0.05). Velocity of spermatozoa in this extender was not negatively affected by centrifugation.

Immunoglobulins in nasal secretions of dog puppies from birth to six weeks of age.

In order to investigate local immune defence mechanisms in the dog, the concentration of immunoglobulins (Ig) G, A and M in nasal secretions (NS) and serum of 42 healthy, neonatal Rottweiler puppies was determined. Ig were measured with a commercially available, dog-specific ELISA during the first six weeks of life. On average, IgG was the predominant Ig isotype during the first three days of life. The IgA:IgG ratio changed between weeks 1 and 3 due to markedly decreasing IgG concentrations. Between the fourth and sixth week, IgG predominated again. During the first week, only 21-39% of puppies had measurable amounts of IgM in NS, in week 2, this percentage increased to 69%. Marked differences between litters and between individual puppies within litters were found. No puppy diseased during the observation period and all developed normally.

Steroid receptors in canine endometrial cells can be regulated by estrogen and progesterone under in vitro conditions.

The expression of estrogen (ER) and progesterone receptors (PR) in the endometrium is regulated by steroid hormones. An increase in plasma estrogen leads to upregulation of the number of both steroid receptors, whereas a decrease in both receptors population is due to high concentration of plasma progesterone. To study the exact effect of different concentrations of beta-estradiol and progesterone on canine epithelial and stromal endometrial cells an in vitro model from dog uterus was developed and kept for 20 days. Material was obtained from healthy dogs, undergoing ovariohysterectomy. Endometrial epithelial and stromal cells were gained after collagenase treatment, followed by filtration steps. Electron microscopy and immunolabeling were used to study cell morphology and differentiation. Immunocytochemistry was used to determine proliferation rate (Ki-67), ER and PR status on Days 3, 8, 10, 13, and 20. Mitotic activity of both cells was stimulated with different concentrations of steroids and revealed high values until cells reached confluency. ER and PR expression in confluent layer from epithelial and stromal cells was upregulated with beta-estradiol. In addition progesterone significant downregulated both receptors population in stromal cells, whereas the reduction was less pronounced in epithelial cells. Results showed that our in vitro system is a useful tool to study the influence of beta-estradiol and progesterone on cell proliferation rate, ER and PR expression. The primary cell culture model helps to avoid experiments on living animals.

Semen parameters and level of microsatellite heterozygosity in Noriker draught horse stallions.

It was the aim of the present study to determine physiological values for different semen parameters in an endangered draught horse breed, the Austrian Noriker. Because small population size is often believed to cause a decrease in fertility and/or semen quality through inbreeding and a reduction in genetic variation, the general genomic heterogeneity of the breed was estimated on the basis of microsatellite variation and correlated to semen parameters. Semen could be collected from 104 of 139 stallions with semen collection being more often successful in younger stallions. Mean volume of ejaculates was 90.8+/-55.1 ml, density 243+/-114 x 10(6)ml(-1), total sperm count 21.0+/-23.7 x 10(9), percentage of morphologically normal spermatozoa 38+/-18% and total motility 50+/-23%. Total sperm count and semen motility were significantly affected by age. Blood samples of 134 stallions were analysed for 12 microsatellite DNA markers. Genotypes of 110 stallions with at least 11 successfully typed markers were used for calculation of heterozygosity. A total of 82 alleles was identified with a mean of 6.8 alleles per marker. Heterozygosity varied between 35 and 76% for the different markers, mean heterozygosity was calculated to 63%. No correlation between heterozygosity and semen parameters was found.

Somatostatin treatment affects testicular function in stallions.

This study investigated the regulation of growth hormone (GH) release in stallions and tested the hypothesis that the somatotrophic axis influences testicular function. Basal plasma GH concentrations, effects of an experimental decrease of GH release on testicular function and an opioidergic regulation of GH release were investigated in Shetland stallions (n=6). No seasonal variations in plasma GH concentrations were found over a 12-month period. Treatment with the somatostatin analogue octreotid (100mg twice daily over 10 days) caused a decrease in semen motility from 38.7+/-8.4% progressively motile spermatozoa before treatment to 18.3+/-5.4% on day 3 after end of treatment (P<0.05). Values returned to 35.0+/-8.5% on day 5 after treatment. On the last day of octreotid treatment, a hCG stimulation test was performed (3000IU hCG i.v.). The hCG-induced testosterone release was significantly higher in saline treated than in octreotid pretreated animals (P<0.05). Neither plasma GH concentrations nor volume and density of ejaculates, total sperm count, or semen morphology were different between saline and octreotid treatments. Injection of the opioid antagonist naloxone (0.5mg/kg) significantly increased GH release in June (from 1.1+/-0.3ng/ml before to 3.7+/-2.2), while a minor and not significant increase occurred in January. In conclusion, our results indicate a non-seasonal basal GH release with a fine-modulation by season-dependent opioidergic mechanisms in the male horse. A transient decrease in semen motility and hCG-induced testosterone release following ocreotid treatment indicate a role of GH in the regulation of testicular function in stallions.

High prevalence of mycoplasmas in the genital tract of asymptomatic stallions in Austria.

Mycoplasma equigenitalium and M. subdolum have been implicated in genital disorders and infertility of horses. The reported cytopathic effects of M. equigenitalium observed in vitro underscore its potential pathogenic role in reproductive dysfunction in mares. This study was initiated to determine the prevalence of mycoplasmas in the genital tract of stallions in relationship to age, clinical signs, geographic location and semen quality. For this purpose the mycoplasma flora of the genital tract of 116 stallions of the Noric breed was determined by isolation and colony immunoblotting and by polymerase chain reaction (PCR) assays. Of 438 swabs from the genital tract, pre-ejaculatory fluid and semen samples, 352 (80%) samples were positive by PCR and 125 (29%) were positive by culture. Mycoplasmas were isolated predominantly from the fossa glandis and urethra and less frequently from the penis shaft and from semen. M. equigenitalium (89 isolates) and M. subdolum (70 isolates) were the predominant species identified. M. equirhinis and M. felis were detected in 27 and 8 samples, respectively. Comparison of these isolations with clinical signs, semen quality, and age of the stallions revealed no significant correlation. However, geographical location of the stallion significantly correlated with mycoplasma detection. These results suggest that mycoplasmas are present as commensals in the genital tract of stallions. Thus, clinically healthy stallions may present a permanent reservoir for infection of mares via venereal transmission.