Sławomir Zduńczyk

The diagnosis and prevalence of subclinical endometritis in cows evaluated by different cytologic thresholds

The aims of our study were to determine (1) how the prevalence of cytologically determined subclinical endometritis varies when using three different cytological threshold ratios to categorize cows as either with or without endometritis, (2) how the number of animals categorized as having endometritis changes from the fourth to the sixth wk postpartum when using each threshold, (3) how subclinical endometritis influences the number of days open, and (4) how the results of cytological and bacterial examinations correlate. To answer these questions, 222 clinically healthy cows in two herds were examined in the fourth (Exam 1) and the sixth wk (Exam 2) postpartum, when endometrial surface scrapings for bacteriologic and cytologic examination were collected by cytobrush from their uterine horns. After each examination, all cows were categorized using three different thresholds: (1) > 18% polymorphonuclear leucocytes in Exam 1 and > 10% in Exam 2, (2) > 8% in both exams, and (3) > 5% in both exams. It was found that: (1) The number of cows categorized as having endometritis increased as the threshold was lowered, and ranged from 18.9% to 75.4% according to herd, time of examination, and the threshold used; (2) with all three thresholds and in both herds, the number of cows categorized as having endometritis in Exam 1 was approximately double that in Exam 2; whereas depending on the herd and the threshold used, 6.1% to 17.0% of the cows that were negative in the first exam were positive in the second, and 7.4% to 33.3% were positive in both exams; (3) cows were open for a significantly greater number of days if categorized as having endometritis with the first threshold in Exam 1 (mean ± SEM 151.5 ± 9.5 vs. 115.9 ± 7.8; P < 0.01), or with either the first or the second threshold in Exam 2 (mean ± SEM 155.0 ± 15.0 vs. 125.1 ± 6.6; P < 0.05); and (4) the most common bacteria were Streptococcus acidominimus and Escherichia coli, and the correlation between cytologic and bacteriologic findings was low (Φ = 0.08 to 0.17 for different tested thresholds). Subclinical endometritis seems to be associated more with the postpartum recovery of the endometrium than with bacterial infection.

Mammary secretion of oestrogens in the cow

Two experiments in vivo and one experiment in vitro were conduced to examine the mechanisms involved, which lead to mammary secretion of oestrogens and its importance for milk production and udder health in cows. In experiment 1 in six cows of the White-Black breed on day 268 of pregnancy catheters were inserted into uterine vein of pregnant horn, the abdominal aorta and the caudal superficial epigastric (milk) vein. Blood samples for estimation of oestrone, oestrone sulphate, oestradiol-17alpha and -17beta by RIA were obtained daily from day 7 pre-partum until day 1 post-partum. Only the concentration of oestradiol-17beta was statistically higher (P< or =0.01) in mammary venous plasma than in aortal and uterine plasma. In experiment 2 forty late-pregnant cows were divided into two groups according to their milk production in the previous lactation: group 1 (n=20) high-yielding cows (>6500kg milk per lactation), and group 2 (n=20) low-yielding cows (<3700kg milk per lactation). Blood samples for measurement of oestradiol-17beta by RIA were collected from milk and tail veins every fourth day during a period from day 20 prior to parturition to day 4 post-partum. The concentration of oestradiol-17beta was significantly higher (P< or =0.01) in the milk vein than in the peripheral plasma from day 12 pre-partum to parturition. In high-yielding cows the level of oestradiol-17beta in mammary venous blood was significantly higher (P< or =0.01) than in low-yielding cows. In six cows with pathological udder oedema ante-partum the concentration of oestradiol-17beta in milk vein was significantly higher (P< or =0.05) than in control cows. There were no statistically significant differences in the levels of oestradiol-17beta in cows with clinical mastitis (n=10) during 2 weeks after parturition and without it (P> or =0.05). In an in vitro experiment, homogenates of mammary tissue collected on day 7 pre-partum from two cows were incubated with 3H-androstendione. After incubation the samples were extracted and 3H-oestradiol-17beta was separated by HPLC. 3H-oestradiol-17beta was formed in a total yield of 37%. These results indicate that oestrone, oestrone sulphate and oestradiol-17alpha are not secreted by bovine mammary gland. Furthermore, the secretion of oestradiol-17beta starts about day 12 pre-partum and is associated with milk yield and udder oedema. Preliminary in vitro study suggests the synthesis of oestradiol-17beta by mammary tissue.

In vivo investigations on luteotropic activity of prostaglandins during early diestrus in nonpregnant bitches.

The aim of this study was to test for the postulated luteotropic effect of prostaglandin E2 during early diestrus in the dog in an in vivo study. This study was performed on 30 bitches which were randomly assigned to a treatment group (TG) and a control group. Starting on the day of ovulation (Day 0), dogs of the TG were treated for 5, 10, 20, or 30 days with 10 mg firocoxib/kg body weight per day (Previcox, a selective PTGS2 inhibitor) and ovariohysterectomized for collection of corpora lutea on the last day of treatment. Similarly, dogs of the control group were ovariohysterectomized on Days 0, 5, 10, 20, and 30. Blood samples for progesterone measurement were collected every second day; additionally, the area of luteal cell nuclei and the expression of 3β-hydroxysteroid-dehydrogenase at the mRNA and the protein levels were assessed. Mean P4 concentrations were lower in TGs; however, a significant difference was only observed on Day 10. This observation is in line with the finding that treatment with firocoxib reduced expression of 3β-hydroxysteroid-dehydrogenase mRNA and protein (P < 0.05) and the area of luteal cell nuclei (P < 0.05). The results of this study further point to the postulated luteotropic function of prostaglandin E2.

Effect of Selenium and Vitamin E Supplementation on Semen Quality in Dogs with Lowered Fertility

Abstract Thirty clinically healthy dogs with poor semen quality were used in the study. Fifteen dogs were supplemented daily with selenium (0.6 mg/kg organic selenium from yeast) and vitamin E (5 mg/kg) per os for 60 d. The control group (15 dogs) was not supplemented. Semen was collected from all dogs by manual manipulation on days 0, 30, 60, and 90. The sperm concentration and motility parameters were evaluated with a Hamilton Thorne sperm analyser, version IVOS 12.3. For the assessment of sperm morphology, Diff-Quik stain was used. The percentage of live and dead spermatozoa was estimated on dried smears stained with eosin-nigrosin. The concentration of spermatozoa, most motility parameters determined (PMOT, VSL, VCL, ALH, BCF, RAPID, MEDIUM, SLOW, and STATIC), and the percentage of spermatozoa morphologically normal and live increased significantly (P < 0.05) after 60 d of supplementation. In the control group, there were no changes in motility parameters while the concentration and total sperm count decreased over the duration of the study. In conclusion, supplementation with selenium and vitamin E for 60 d can improve the quality of semen in dogs with lowered fertility.

Assessment of semen quality in infertile dogs using computer-assisted sperm analysis by the Hamilton-Thorne Semen Analyser

Abstract Semen quality parameters of infertile and fertile dogs were compared. Sperm concentration and semen motility parameters were measured by the Hamilton-Thorne Semen Analyser IVOS 12.3. The spermatozoal morphology and the percentage of live spermatozoa were examined microscopically. Forty-six dogs of various breeds were examined. Twenty dogs had a conception failure within last year. These dogs had a history of prior normal fertility. Twenty six fertile dogs served as control. All animals underwent a clinical examination as well as ultrasonography. Sperm concentration was significantly lower in infertile dogs than in fertile dogs. For most determined motility parameters (MOT, PMOT, VAP, VSL, VCL, BCF, RAPID, STATIC) significant differences between infertile and fertile dogs were found. The percentage of spermatozoa with normal morphology also significantly differed between both groups. Ultrasonography of the prostate gland and testes revealed no pathological conditions. The testicular degeneration was assumed to be a possible cause of infertility in these dogs. The present study showed that the most sperm CASA motility parameters were significantly lower in infertile dogs in comparison to the fertile ones, and confirmed the usefulness of the Hamilton-Thorne Semen Analyser for a quick and objective analysis of sperm concentration and motility in dogs.

Stenotrophomonas maltophilia isolated from prostatic fluid as an infertility factor in a male dog

The aim of this case was to describe very rare infection of canine prostate gland with Stenotrophomonas maltophilia which had influence on male fertility. The bacterium was cultured from third fraction of the ejaculate collected by manual manipulation. The sperm concentration and motility parameters were evaluated by Hamilton‐Thorne Sperm Analyser, version IVOS 12.3, sperm morphology by Diff‐Quick staining and live/dead spermatozoa by eosin/nigrosin staining. After 3 weeks of treatment with targeted antibiotic, ciprofloxacin, there was no bacterial growth in prostate fluid. Semen parameters were improved after 60 days from the end of treatment, and females were successfully mated.

Endometrial mRNA expression of prostaglandin synthase enzymes PTGS 2, PTGFS and mPTGES 1 in repeat-breeding cows with cytologically determined endometritis

Little is known about the inflammatory response of the endometrium in repeat-breeding cows with subclinical endometritis (SE). The objective of this study was to evaluate the mRNA expression of prostaglandin-endoperoxide synthase 2 (PTGS 2), prostaglandin F2α synthase (PTGFS) and prostaglandin E2 microsomal synthase 1 (mPTGES 1) in the endometrium of repeat-breeding cows with and without SE. SE was diagnosed cytologically using the cytobrush method, with the threshold being set at 5% polymorphonuclear neutrophils. Biopsy samples were obtained from the endometrium of repeat-breeding cows with SE (n = 10) and without SE (n = 10). The mRNA expression of the synthases was evaluated using qRT-PCR. Significantly higher (P < 0.05) expression of the PTGS 2 gene was detected in the repeat breeders with SE, whereas there was no significant difference in the expression of PTGFS and mPTGES 1 mRNAs between repeatbreeding cows with SE and those without it (P > 0.05). Our study confirms that increased endometrial expression of the PTGS 2 gene is involved in the inflammatory response in repeat breeders.

Selenium and vitamin E supplementation enhances the antioxidant status of spermatozoa and improves semen quality in male dogs with lowered fertility

Studies showed a beneficial effect of supplementation with selenium (Se) and vitamin E on semen quality. The aim of the study was to investigate the effect of Se and vitamin E supplementation on the antioxidant status of spermatozoa and semen quality in dogs with lowered fertility. Ten dogs were supplemented daily with Se (6 μg/kg organic Se yeast) and vitamin E (5 mg/kg) per os for 60 days. Control group consisted of 10 males without the supplementation. Semen was collected on day 0, 30, 60 and 90. Sperm quality parameters were evaluated using CASA and a microscope. Concentrations of Se and vitamin E in blood as well as glutathione peroxidase (GSH‐Px) activity and total antioxidant capacity (TAC) in the spermatozoa were determined. After 60 days of supplementation the concentration of spermatozoa, the majority of motility indicators and the percentage of normal morphology and live spermatozoa increased significantly (p < .05). An increase (p < .05) in concentration of Se and vitamin E in blood and GSH‐Px‐activity and TAC in the spermatozoa was detected. The study results indicate that Se and vitamin E supplementation for 60 days enhances the antioxidant status of spermatozoa and improves the quality of the semen in dogs with lowered fertility.

Elastography as a diagnostic tool in the prostate tumour detection in Labrador retriever

Prostate tumours are hard to detect in dogs. Early diagnosis is a key to maintain fertility and to protect against metastasis and death. Elastography is a diagnostic tool to obtain very detailed results of changes in tissue stiffness of this organ. There are no studies about the application of elastography to diagnose the prostate tumours in dogs. In our case, in male Labrador retriever there was state a prostate lesion by both scanning—grey ultrasonogram and elastography. Fine needle aspiration biopsy (FNA) confirmed the adenocarcinoma only in elastography‐changed area. Thus, this method is helpful to diagnose prostate tumours and should be the first additional survey in routine examination of the prostate in the dog.

Treatment of Ovarian Cysts in Dairy Cows with Simultaneous Administration of GnRH and PGF2α has no Clear Advantage Over the Use of GnRH Alone

Abstract The aim of the study was to determine the therapeutic efficacy o f simultaneous administration of GnRH and PGF2α in dairy cows with ovarian cysts. Ovarian cyst-affected dairy cows were divided into two experimental groups: 54 cows treated with GnRH and PGF2α, and 42 cows treated with GnRH alone, whereas 22 untreated cows served as the control group. Clinical response and reproductive performance were evaluated. The cumulative disappearance was better in treated cows than in the control group; however, there were no differences between the treatment groups (92.6; 95.2% vs. 72.3%). The mean interval from calving to conception was not significantly shorter (being so by 29 d) in the GnRH/PGF2α group than in the cows treated with GnRH alone (P > 0.05). The intervals from treatment to conception were also similar in these groups. The pregnancy rate in both treated groups was similar (62%) and higher than in the control cows (53%). In the cows with luteal cysts, the total pregnancy rate was higher in all experimental groups; however, only in GnRH-treated cows was this difference statistically significant (77.8% vs. 50.0%, P < 0.05). With time after parturition, the pregnancy rate decreased in all groups. In general, the cows treated with GnRH and PGF2α simultaneously displayed a good clinical response and slight improvement in reproductive performance compared to the single-therapy GnRH group; however, this was not fully convincing.