Tomasz Janowski’s research while affiliated with University of Warmia and Mazury in Olsztyn and other places

The aims of our study were to evaluate (1) the prevalence of subclinical endometritis (SE) in clinically cured cows after treatment of clinical endometritis (CE) with uterine cephapirin infusion or with systemic application of PGF2α and after self-healing, (2) the prevalence of SE in clinically cured cows in relation to the type of vaginal discharge. The study was conducted on 222 Polish Holstein Friesian cows with CE diagnosed by vaginoscopy and ultrasound. The animals were randomly allocated to one of three groups: Group 1 (n = 72): intrauterine cephapirin infusion; Group 2 (n = 73): single intramuscular injection of dinoprost; and Group 3 (n = 77): left untreated as a control. All cows were reexamined two weeks later (Exam 1). If there was still CE at this examination, these cows were treated using the same method as previously and were examined 2 weeks later (Exam 2). Cows still having CE at the second examination were examined two weeks later (Exam 3). Cows without signs of CE at Exams 1, 2 and 3 were considered cured, and endometrial samples from the uteri were collected by cytobrush to diagnose SE using cytological evaluation of PMN percentage. The threshold for SE was set at ≥ 5% PMNs. Two cows each from Groups 1 and 3 and three cows from Group 2 were not clinically recovered at Exam 3 and were excluded from the analysis. In total, SE was diagnosed in an average of 40.9% of clinically cured cows. The prevalence of SE in Group 1, 2 and 3 was 35.0%, 47.1% and 40.0%, respectively (p > 0.05). There were significantly higher (p < 0.05) SE cases after treatment of CE associated with purulent vaginal discharge than with mucopurulent discharge. In relation to the method of treatment, the prevalence of SE was significantly higher (p < 0.05) in cows treated with cephapirin with purulent vaginal discharge than with mucopurulent vaginal discharge. In conclusion, the study showed a high prevalence of SE in cows clinically cured after treatment of clinical endometritis with cephapirin or PGF2α and after self-healing. The prevalence of SE was significantly higher in cows with purulent vaginal discharge than with mucopurulent vaginal discharge. Therefore, in cows clinically recovered from clinical endometritis, the prevalence of SE should always be considered a negative consequence, and the resulting management of this uterine disease should be implemented at the herd level.

Simple Summary The role of preovulatory progesterone for LH release and ovulation in the bitch is not clear. The aim of the study was to evaluate the influence of administration of aglepristone in mid-proestrus on progesterone concentration, LH release, and occurrence of ovulation in the bitch. Experimental bitches (n = 7) were treated on days 4 and 5 of proestrus with aglepristone (Alizin®, Virbac) at the dose of 10 mg/kg body weight s.c. (i.e., the two treatments were 24 h apart). The progesterone concentration showed a similar pattern in both groups. The LH peak value and area under the curve for LH in bitches treated with aglepristone were significantly lower than those in control bitches. The ovulation occurred in all animals in both groups. The presented study showed that withdrawal of progesterone by administration of aglepristone in the mid-proestrus significantly reduced the preovulatory LH surge, but it had no effect on periovulatory progesterone concentration or the occurrence of ovulation. Abstract The aim of the study was to evaluate the influence of administration of aglepristone in mid-proestrus on progesterone concentration, LH release, and occurrence of ovulation in the bitch. Experimental bitches (n = 7) were treated on days 4 and 5 of proestrus with aglepristone at the dose of 10 mg/kg body weight s.c. (i.e., the two treatments were 24 h apart). Control animals (n = 7) received s.c. injections of saline. For progesterone determination, blood was collected daily until the first day of cytological diestrus. For LH determination, blood was collected daily and in the periovulatory phase every 8 h. The progesterone concentration showed a similar pattern in both groups. The LH peak value in bitches treated with aglepristone was significantly lower (p < 0.05) than that in control bitches (4.83 ± 1.20 vs. 13.66 ± 1.21 ng/mL). The area under the curve (AUC) for LH was significantly (p < 0.05) lower in treated than in control animals (6.85 ± 1.21 ng/mL/d vs. 12.25 ± 1.35 ng/mL/d). The ovulation occurred in all animals in both groups. The study showed that administration of aglepristone in the mid-proestrus significantly reduced the preovulatory LH surge, but it had no effect on progesterone concentration and the occurrence of ovulation.

Zusammenfassung Ein Rüde mit benigner Prostatahyperplasie und mehreren kleineren intraprostatischen Zysten wurde mit einem GnRH-Agonist-Implantat mit 4,7 mg Deslorelin (Suprelorin®) behandelt. Innerhalb von 2 Wochen nach der Implantation kam es zur Verstärkung der zuvor aufgetretenen Blutung aus der Harnröhre. Sonografisch ließ sich eine große intraprostatische Zyste darstellen. Der Rüde wurde mit Osateronacetat (0,4 mg/kg p. o. 1-mal täglich über 7 Tage) und Enrofloxacin (5 mg/kg p. o. 1-mal täglich über 21 Tage) behandelt. Im Verlauf eines Monats bildete sich die Zyste vollständig zurück. Die Mechanismen der Zystenvergrößerung werden diskutiert.

The failure of the maternal immune system to recognize fetal antigens and vice versa due to MHC similarity between the foal and its dam might result in the lack of placental separation during parturition in mares. The aim of the study was to investigate the influence of MHC similarity between a mare and a foal on the incidence of retained fetal membranes (RFM) in post-partum mares. DNA was sampled from 43 draft mares and their foals. Mares which failed to expel fetal membranes within three hours after foal expulsion were considered the RFM group (n = 14) and mares that expelled fetal membranes during the above period were the control group (n = 29). Nine MHC microsatellites of MHC I and MHC II were amplified for all mares and foals. MHC compatibility and MHC genetic similarity between mares and their foals was determined based on MHC microsatellites. The inbreeding coefficient was also calculated for all horses. The incidence of RFM in the studied population was 33%. Compatibility in MHC I and MHC II did not increase the risk of RFM in the studied population of draft mares (P>0.05). Differences in MHC similarity at the genetic level were not observed between mare-foal pairs in RFM and control group (P>0.05). We suspect that RFM in draft mares may not be associated with MHC similarity between a foal and its dam. Despite the above, draft horses could be genetically predisposed to the disease.

Bacteriophages and the associated endolysins have been proposed as an alternative to antibiotic treatment of mastitis and metritis in cows. Many bacteriophages have been isolated and characterized with a large amount of lytic potential against the bacteria causing mastitis and metritis in cows. Several endolysins with marked lytic activity against mastitis pathogens in vitro were also produced from staphylococcal and streptococcal bacteriophages. In the few clinical studies, however, there has been marginal efficacy of bacteriophages in the therapy of mastitis caused by Staphylococcus aureus. Similarly, lytic bacteriophages have marked antimicrobial activity in vitro against E. coli strains from the uteri of postpartum dairy cows. In clinical studies, however, neither administration of bacteriophages early postpartum nor prepartum was effective in the prevention of metritis in cows. More clinical studies on the effectiveness of bacteriophages and the associated endolysins in the prevention and therapy of mastitis and metritis in cows, therefore, are needed.

Despite their importance in mammalian reproduction, substances in the oxytocin-prostaglandins pathways have not been investigated in the horse placenta during most of pregnancy and parturition. Therefore, we quantified placental content of oxytocin (OXT), oxytocin receptor (OXTR), and prostaglandin E2 and F2 alpha during days 90–240 of pregnancy (PREG), physiological parturition (PHYS), and parturition with fetal membrane retention (FMR) in heavy draft horses (PREG = 13, PHYS = 11, FMR = 10). We also quantified OXTR and prostaglandin endoperoxide synthase-2 (PTGS2) mRNA expression and determined the immunolocalization of OXT, OXTR, and PTGS2. For relative quantification of OXT and OXTR, we used western blotting with densitometry. To quantify the prostaglandins, we used enzyme immunoassays. For relative quantification of OXTR and PTGS2, we used RT-qPCR. For immunolocalization of OXT, OXTR, and PTGS2, we used immunohistochemistry. We found that OXT was present in cells of the allantochorion and endometrium in all groups. PTGS2 expression in the allantochorion was 14.7-fold lower in FMR than in PHYS (p = 0.007). These results suggest that OXT is synthesized in the horse placenta. As PTGS2 synthesis is induced by inflammation, they also suggest that FMR in heavy draft horses may be associated with dysregulation of inflammatory processes.

Major histocompatibility complex (MHC) is a suggested criterion of reproductive mate choice in many species. MHC-dissimilarity was suggested to play role in male selection by female horses (mares) in experimental settings. In our project, we asked whether under natural conditions mares choose breeding stallions guided by a certain MHC-type, either MHC-dissimilar or MHC-diverse. We observed the reproductive behavior of 26 mares and five stallions of semi-feral Konik Polski horses for eight years from which only 4 stallions and 16 mares could be used in analysis. Between-group (harems) migrations of mares and stallions and the composition of harems were noted. The choice of a reproductive partner was confirmed by observation (a mare present in the harem of a chosen stallion) and by the actual reproductive outcome (parentage of foals). The MHC of mares and stallions was tested using 11 MHC microsatellites. Genetic similarity in MHC was measured by pairwise relatedness and allele sharing between mares and stallions. MHC heterozygosity of every stallion was calculated. For all mares the choice of reproductive partner was the harem stallion. No extra-harem matings were observed. Allele sharing did not differ between mating pairs and between all of the possible combinations of mares and stallions (0.36 vs. 0.33; P = 0.66). Genetic similarity in MHC between mares and stallions did not influence the choice of reproductive partner by mares (P > 0.05), but mares tended (P = 0.08) to stay in harems of MHC-diverse stallion. The present findings suggest that mares in semi-feral conditions do not actively search for MHC dissimilar stallions, however they might favour MHC diverse ones. However, because the choice of reproductive partner was the current harem stallion, in the case of every mare we speculate that in feral conditions selection of reproductive stallion by mares might be either restricted or influenced by variety of factors other than MHC of a stallion.

The canine luteal phase exhibits several peculiarities compared with other species. In early diestrus, the corpus luteum (CL) is, at least in part, independent of gonadotropins, and prostaglandins (PGs) appear to be among its main regulators. This was also observed with the inhibition in vivo of COX2, when also transcriptional capacity, vascularization and immune-related factors were affected. Here, we aimed to further investigate the potential effects of PGs withdrawal on the CL transcriptome by performing deep RNA sequencing (RNA-Seq). Samples from a previous in vivo study were used; bitches were treated for 5, 10, 20, or 30 days after ovulation with firocoxib (Previcox®), a PTGS2/COX2 inhibitor, or a placebo. Analysis of results was performed with SUSHI (framework from FGCZ) and with pathways and functional networks analyzers. Time-dependent effects were also investigated and used for quality control. More highly represented differentially expressed genes (DEGs, P < 0.01, FDR < 0.1) in the early CL (days 5 and 10) referred to proliferation and immune system, while in the mature CL (days 20 and 30) they were related with steroidogenesis. The absence of genes concomitantly affected by the treatment at all time-points suggested stage-dependency in the observed effects. Little effect was observed on days 5 and 10. Day 20 had the highest number of DEGs (n = 1,741), related with increased immune response. On day 30, DEGs found (n = 552) referred to decreased steroidogenesis and vascularization. Our results suggest the presence of strong compensatory effects in the early CL and multidirectional effects toward gonadotropin-dependency of the CL after COX2 inhibition.

Background With the increasing popularity of planned caesarean section, the need for knowledge regarding this surgery has become increasingly important. The reported death and survival rates for caesarean sections vary widely. Another important aspect is the fertility rate in subsequent oestrous after caesarean section. The aim of this study was to investigate the mortality and survival rate of bitches during caesarean section. Additionally, the fertility of bitches after caesarean sections was determined. Methods Caesarean sections which were performed in the years 1997–2009 at two university clinics were evaluated retrospectively. A distinction was made between bitches in which a conservative caesarean section was performed and bitches with a caesarean section followed by an ovariohysterectomy. Results A total of 482 caesarean sections were included in the study. The overall mortality rate was 3.11 per cent, with 2.59 per cent during or after a conservative caesarean section and 4.19 per cent during or after caesarean section with ovariohysterectomy. The reason for ovariohysterectomy was the owner’s preference in 63 bitches (47.01 per cent); in 71 (52.98 per cent) bitches, ovariohysterectomy was performed due to a medical indication. The fertility rate after caesarean section was 100 per cent. Conclusion The results show a high mortality rate during and after caesarean section. On the other hand, caesarean section does not seem to have a big impact on further fertility. Further studies are needed to investigate possible reduction of litter sizes and the suitability of caesarean section in subsequent pregnancies.

Approaches to downregulate ovarian function in the sexually mature bitch by applying slow release GnRH agonist implants are hampered by the initial stimulation of folliculogenesis (flare up) and the resulting side effects. The present pilot study was designed to test to what extent these effects can be suppressed by simultaneous treatment with the 3ß-hydroxysteroid-dehydrogenase (HSD3B) blocker trilostane (T). Treatment with T in 6-h intervals completely blocked adrenal cortisol production. However, in parallel and concomitant with the increase of LH, progesterone and estradiol levels increased, ending up in pro-estric steroid levels in two of the three dogs. Hormonal changes were reflected in the respective clinical symptoms. During the whole observation period the course of LH concentrations did not indicate downregulation of pituitary function as a result of treatment with the GnRH-agonist Suprelorin®, 4.7 mg. The incomplete inhibitory effect of T on the follicular production of sex steroids could be explained by an insufficient transfer of T into the follicular compartment or the existence of a HSD3B isoform in the dog ovary different from the adrenal enzyme. Concerning the lack of downregulation and when accounting for published data different pharmacodynamics/pharmacokinetic activities of GnRH-agonists should be taken into account.