Polish Journal of Veterinary Sciences

Different approaches to enhance healing of hard or soft tissues include the use of cytokines and growth factors to modify cellular behaviour. Numerous growth factors are found in autologous blood concentrates – platelet-rich plasma (PRP) and platelet-rich fibrin (PRF). Enamel matrix derivative (EMD) may improve tissue healing via amelogenins. Bilayered collagen matrix (CM) is used for soft tissue augmentation.

The aim of the present study was to assess potential benefits of PRP, PRF and EMD in combination with bilayered collagen matrix or CM alone in treatment of oral mucosal defects in rabbits.

Twenty-seven New Zealand white rabbits were included in this randomized controlled trial. Artificial oral mucosal defects were treated with one of these five approaches: PRP+CM, PRF+CM, EMD+CM, CM alone, or left untreated as a negative control - CO. The animals were euthanized 1 day, 7 days, or 28 days after surgery and necropsies were harvested. Histological and molecular biological analyses were performed.

All defects were healed by day 28. No differences between PRP+CM, PRF+CM, CM alone and CO groups were recorded at any time point. Slower angiogenesis and a higher presence of inflammatory infiltrate were observed in the EMD+CM group 28 days after surgery. Molecular biological analyses did not reveal any statistically significant changes.

In conclusion, no improvement in mucosal healing of wounds covered with a collagen membrane and PRP, PRF, or EMD was observed, compared with CM alone or untreated controls.

Herpesviruses (HV) are pathogens causing infections in humans and animals worldwide. Since it shares many common features with other HV, bovine HV type 1 (BoHV-1) was selected as a model to test the anti-herpesviral activity of medicinal plants.

Fifteen plants were chosen in this study for their medical, antibacterial and antiviral properties. The aim was to investigate ethanolic extracts from the selected medicinal plants for anti-BoHV-1 activity. The virucidal activities were evaluated by comparing the effect of noncytotoxic concentrations of extracts on BoHV-1 strain 1640 replication in Madin-Darby bovine kidney (MDBK) cells. Virucidal activity was determined by means of virus titration after exposure to the extracts. The extract of Desmodium canadense was found to be the most effective virucide – the 50% tissue culture infective dose (TCID50) after exposure was 3.75 log10 and the virus reduction factor was ≥5.0±0.25 log10. The extract of D. canadense was therefore chosen for further studies. Virus yield reduction assays showed that D. canadense extract had time-dependent and dose-dependent effects. It effectively reduced virus titre from 8.33 log10 to 4.67 log10 (p<0.01). The virucidal activity was also confirmed by real-time polymerase chain reaction (real-time PCR), where the number of threshold cycles (Ct) was inversely proportional to the virus titre in TCID50 The virucidal activity was also confirmed by real-time polymerase chain reaction (real-time PCR). This method showed that the number of threshold cycles (Ct) was inversely proportional to the virus titre (direct correlation with exposure time R=0.9321). The extract of D. canadense showed a high virus reduction capacity. In future, such active substances should be identified for the development of effective antivirals.

The aim of the present study was to determine changes in oxidative status in plasma of sows during periparturient period using markers of lipid and protein peroxidation intensity. Plasma from 12 pregnant gilts collected on days 14 and 1 before parturition and on days 1, 7 and 14 postpartum was used in the study. As indicators of alterations in the oxidative status, plasma concentrations of thiobarbituric acid reactive substances (TBARS) and sulphydryl groups (SH groups) were measured spectrophotometrically, as well as plasma concentrations of bityrosine and formylokinurenine were determined by the spectrofluorimetric method. Compared to the 14th day antepartum, the concentration of TBARS increased significantly on day 1 postpartum and then decreased significantly on days 7 and 14 after parturition. The concentration of SH groups increased significantly on the last day before parturition and decreased significantly on day 14 postpartum. The concentration of bityrosine increased significantly on the last day before parturition and decreased significantly on days 7 and 14 postpartum. The concentration of formylokinurenine increased significantly on the last day before parturition, remained at this level on the first day after parturition and then decreased slightly on days 7 and 14 postpartum. These results indicate that during periparturient period in sows, especially around the parturition, the oxidative/antioxidative balance is disturbed, which leads to oxidative stress. Considering that oxidative stress is potentially harmful for mothers and neonates, this may be an important factor in the increased risk of diseases occurring during this period.

Atopic dermatitis (AD) is the most frequent allergic disease in dogs. AD can be treated using allergenspecific immunotherapy as well as symptomatic antipruritic treatment including the use of lokivetmab - caninized anti-interleukine-31 antibody.

The aim of the study was to evaluate the effectiveness of lokivetmab over 12 weeks of treatment. Studies have been carried out in 89 dogs. In all affected animals, the severity of lesions was assessed using the CADESI 04 and the pruritus was assessed using the VAS.

After the first dose of lokivetmab, both CADESI 04 and VAS statistical decreased by 4 weeks from 40.48 to 20.31, and from 7.42 to 2.48, respectively (p = 0.0000001) maintained significantly decresed values during the whole treatment period (CADESI 04 15.64, 15.07 after 8 and 12 weeks, respectively, PVAS 2.03, 1.95 after 8 and 12 weeks, respectively).

Lokivetmab leads to a significant reduction of CADESI 04 and pruritus, within four weeks and maximum effect is achived after the second dose.

In this study, a SYBR Green-based real-time quantitative polymerase chain reaction (qPCR) assay was developed for rapid detection of porcine parvovirus (PPV) 6. Primer pairs targeting the conserved regions of PPV6 Capsid gene were designed. Sensitivity analyses revealed the lowest detection limit of the SYBR Green-based real-time PCR assay to be 47.8 copies/μL, which indicated it was 1000 times higher than that found in the conventional PCR investigations. This assay was specific and showed no cross-species amplification with other six porcine viruses. The assay demonstrated high repeatability and reproducibility; the intra- and inter-assay coefficients of variation were 0.79% and 0.42%, respectively. The positive detection rates of 180 clinical samples with SYBR Green-based real-time PCR and conventional PCR were 12.22% (22/180) and 4.44% (8/180), respectively. Our method is sensitive, specific, and reproducible. The use of SYBR Green-based real-time PCR may be suitable for the clinical detection and epidemiological investigation of PPV6.

The study was carried out in Polish goat population to estimate the prevalence of the nasal cavity infection with various staphylococcal species including methicillin-resistant Staphylococcus aureus (MRSA), investigate the potential permissive role of small ruminant lentivirus (SRLV) infection and determine the level of clonality of S. aureus nasal isolates. Nasal swabs and blood samples were collected from 1300 clinically healthy adult goats from 21 Polish goat herds. Blood samples were serologically screened for SRLV. Staphylococci were isolated from nasal swabs and identified using classical microbiological methods, MALDI-TOF, multiplex-PCR, and their clonality was assessed using PFGE. Antimicrobial resistance was determined on the basis of minimum inhibitory concentration and by demonstration of the presence of the mecA gene encoding the multiplex-PCR PBP2a protein and of the five main types of staphylococcal cassette chromosome mec. The apparent prevalence of staphylococcal and S. aureus infection of the nasal cavity was 29.1% (CI 95%: 26.9%, 31.5%) and 7.3% (CI 95%: 6.1%, 8.8%), respectively. No relationship was found between the SRLV-infection and the presence of any staphylococcal species including S. aureus (p=0.143). Only 9.8% of S. aureus isolates were resistant to amoxicillin/clavulanic acid and 5.9% to chloramphenicol and ciprofloxacin. All tested isolates proved to be phenotypically and genotypically sensitive to methicillin, which yielded the apparent prevalence of MRSA of 0% (CI 95%: 0%, 7.0%). S. aureus isolates show high genetic similarity within goat herds, however vary considerably between herds. Goats do not appear to be an important source of S. aureus for humans in Poland.

The aim of this study was to evaluate the intestinal and cardiac biomarkers in the determination of intestinal and cardiac damage in dogs with parvoviral enteritis. The material of this study consisted of 10 healthy dogs (control group) and 30 dogs with parvoviral enteritis (experimental group) admitted to the Department of Internal Medicine, Faculty of Veterinary Medicine, Selcuk University.

Serum samples were extracted from the collected blood samples taken from vena cephalica venipuncture for analysis of blood gases, haemogram and to measure the levels of intestinal-fatty acid-binding protein (I-FABP), trefoil factor 3 (TFF-3), claudin-3 (CLDN-3), heart-type fatty acid-binding protein (H-FABP), cardiac troponin I (cTnI), and creatine kinase-myocardial band (CK-MB) by enzyme linked immunosorbent assay (ELISA) test kits.

Statistically significant decreases in the blood gas hydrogen ion concentration (pH), partial pressure of oxygen (pO2), sodium (Na), bicarbonate (HCO3), and oxygen saturation (SatO2) levels and significant increase in the levels of I-FABP, TFF-3, CK-MB, cTnI and also in the haemogram, a decrease in leukocyte (WBC) level and an increase in platelet (THR) level were detected in parvoviral dogs compared to the control group (p<0.05). Also ROC analysis revealed on 0th hour for the utility of I-FABP and on 48th hour for TFF-3 in differentiating in the experimental group between the survivor and non-survivor dogs. Other intestinal-related biomarker (CLDN-3) and none of the cardiac-related biomarkers (H-FABP, CK-MB and cTnI) are not high enough for prediction of mortality.

In conclusion, it was determined that I-FABP and TFF-3 for the intestinal injury and mortality prediction, and CK-MB and cTnI for the cardiac injury were useful and reliable biomarkers to determine the damage caused by parvovirus in dogs.

The aim of this study was to determine if reticulorumen ph, temperature and cow activity registered before calving can serve as indicators of diseases after calving.

The cows were selected according to those fitting the profile of having had two or more lactations (on average 2.9±0.13 lactations), from 60 to 0 days before and the first 30 days after calving, and being clinically healthy. The clinical examination (identification of diseases after calving) was performed from 60 days before calving to 60 days after calving. Diseases after calving were diagnosed based on clinical symptoms specific to these diseases. The pH and temperature of the contents of the cow reticulorumens and cow activity were measured using specific smaX-tec boluses manufactured for animal care.

We found that the highest pH and temperature before calving can serve as biomarkers of healthy cows after calving. The lowest reticulum temperature before calving can serve as an indicator of MF after calving. A positive correlation of reticulum pH and temperature before calving can serve as biomarkers of PR. Decreasing cow activity before calving can serve as an indicator of diseases after calving. For calving prognosis, temperature of the reticulorumen can be used; it decreased 6–7 days before calving.

A disease survey was conducted between 2014 and 2018 in the Polish goat population to determine the seroprevalence of pestiviral infection. Blood samples from 910 goats (782 females and 128 males) were collected in 62 goat herds and tested for bovine viral diarrhea virus (BVDV) infection with a competitive ELISA in a serial fashion. Between 10 and 13 adult female goats were randomly selected from each herd, assuming individual-level seroprevalence of pestiviral infection in a herd of at least 30% and a level of confidence of 95%. In each herd, all males were tested. At least one seropositive goat was found in each of the 4 herds. However, in one herd, the only positive goat tested negative in serial retesting. Finally, 3 herds were considered as seropositive which yielded apparent herd-level seroprevalence of 4.8% (CI 95%: 1.7%, 13.3%). After adjusting for the ELISA herd-level sensitivity and specificity, the true herd-level prevalence was 3.9% (CI 95%: 1.2%, 12.0%). No males tested positive. In 2 out of 3 seropositive herds, goats regularly shared pastures with cattle.

Emerging researches in humans, pigs and mice, highlighted that estrogen plays a pivotal role in self-renewal and differentiation of bone marrow mesenchymal stem cells (BMSCs). The present study aimed at evaluating effects of 17 beta-estradiol (E2) on proliferation and apoptosis of canine-derived bone marrow mesenchymal stem cells (cBMSCs) in vitro. The results showed that E2 supplementation at the concentration of 10-11 M promoted the proliferation of cBMSCs by CCK-8 assay and RT-qPCR analysis for the proliferation-related genes, with proliferating cell nuclear antigen (PCNA), cyclin-D1 (CCND1) being up-regulated and cyclin-dependent kinase inhibitor 1B (CDKN1B) being down-regulated. Contrarily, analysis of fluorescence-activated cell sorting (FACS) and RT-qPCR demonstrated that E2 supplementation above 10-11 M had inhibitory effects on the proliferation of cBMSCs and induced apoptosis. Intriguingly, cBMSCs still possessed the capability to differentiate into osteoblasts and adipocytes with 10-11 M E2 addition. Taken together, this study determined the optimal culture condition of cBMSCs in vitro, and has important implications for further understanding the regulatory effect of E2 on the self-renewal of cBMSCs, which are helpful for the clinical application of BMSCs.

The aim of this study was to investigate the effect of the addition of fungal solid-state fermented product (FP) enriched with gamma-linolenic acid (GLA) and β-carotene to feed on the haematological and immunological parameters of broiler chickens. Eighty 1-day-old COBB 500 broiler chickens were divided into two groups. The control group was fed with basic diets and chickens of the experimental group received 10% addition of FP, while the amount of basic diet was reduced. FP was produced during a solid-state fermentation (SSF) process using Umbellopsis isabellina CCF2412 as a producer of GLA and β-carotene. After 38 days of feeding, blood samples were collected and analyzed. Lower total and LDL-cholesterol values were measured in blood samples of the experimental animals (p<0.05). However, the triacylglycerol content was higher in the experimental group (p<0.05). Significantly higher levels of hematocrit and hemoglobin, and lower eosinophil and basophil content in the experimental group were recorded (p<0.05). The experimental group showed higher numbers of B lymphocytes and greater phagocytic capacity (p<0.05). The results indicate that a fermented product produced by SSF, using the fungal strain Umbellopsis isabellina, is a good source of GLA and β-carotene, which can influence the biochemical, hematological and immunological parameters of broiler chickens.

This study investigated the utility of patent ductus arteriosus (PDA) closure with hemostatic clip by comparing with traditional PDA closure. Medical records of 51 dogs with surgical closure of PDA were reviewed and retrospective study was conducted. 29 dogs were treated by procedure with hemostatic clip (Group HC), and 22 dogs were treated by surgical ligation (Group SL). Data pertaining to breed, sex, age and body weight at the time of surgery, echocardiographic minimal ductal diameter, duration of surgery, hemostatic clip size, echocardiographic findings, hemorrhage, residual ductal flow and recanalization were collected from records. The results showed that procedure with hemostatic clip had been selected in lighter dogs than traditional PDA closure. Duration of surgery performed only hemostatic clip technique was significantly shorter than that in group SL. Preoperative LVIDd, E-wave and FS were significantly lower than postoperative ones. As regard all parameters, the differences between pre- and postoperative periods were not significantly different between group HC and group SL. Hemorrhage, residual ductal flow, and recanalization were not significantly different in both groups. The present study showed that procedure with hemostatic clip is beneficial in that it is available in smaller dogs and can make shorter operation duration than traditional PDA closure. Moreover, the procedure is effective for the resolution of volume overload of the left atrium and ventricle in short-term outcome. Complications including hemorrhage, residual ductal flow and recanalization were not significantly different with both techniques.

Lumbosacral transitional vertebrae (LTV) are abnormally formed vertebrae of congenital origin. Dogs with LTV are predisposed to premature degeneration of the lumbosacral spine and hip dysplasia due to a weakened sacroiliac attachment. Moreover, LTV has been discussed as a cause of cauda equina syndrome. To date, LTV remain poorly understood and a diagnostic standard is yet to be established. This study examines prevalence, types and breed predispositions for LTV in the canine population in Berlin. The diagnostic value of laterolateral radiographs of the lumbosacral region, in addition to ventrodorsal radiographs, was also evaluated. The prevalence of LTV was assessed by reviewing ventrodorsal pelvic radiographs of 1030 dogs. LTV were detected in 95 (9.2%) dogs. The prevalence was higher in Pugs (63.6%) and Jack Russel Terriers (27.6%) than in the other breeds. The most common type of LTV was type II (37.9%), showing separation of the first sacral segment from the sacrum, the presence of a rudimentary intervertebral space between the first sacral segment and the rest of the sacrum, and symmetrically formed transverse processes. Laterolateral radiographs were available for 66 of 95 dogs with LTV and provided evidence of a rudimentary intervertebral disc space between the first and second sacral vertebrae in all cases of LTV type II and III (100%). The results of this study contribute to a better understanding of the condition. Furthermore, they demonstrate that laterolateral radiographs are a valuable addition to standard ventrodorsal radiographs and should be included in routine LTV screening protocols to provide a complete evaluation

The research was conducted on 40 young alpine goats (kids) divided into two groups. First group consisted of 20 kids demonstrating clinical signs of muscular dystrophy. Second group was a control and consisted of 20 animals that received intramuscular injection (2ml per animal) of vitamin E and selenium preparation containing in 1ml 50 mg of tocopherol acetate, 0.5mg of sodium selenite and solvent on 2nd day of life. The kids were clinically examined and blood for laboratory analyses was sampled three times from day 5 of their life in 10 day intervals. In addition, six 24 days old kids demonstrating clinical signs of muscular dystrophy and six control kids were subjected to biceps femoris biopsy.

Serum total protein, glucose, triglycerides, cholesterol as well as AST, CK and LDH were determined in all the animals. In addition, the activity of glutathione peroxidase (GSH-Px) was determined in whole blood and serum concentrations of selenium and vitamin E were determined in 6 kids from each group. Total lactate dehydrogenase activity and its separation into isoenzymatic fractions were determined in the collected biopsy material. The muscle samples collected were additionally subjected to histopathological examination consisting of HE staining and HBFP staining to detect necrotic muscle fibers.

Symptoms of muscular dystrophy began to appear in the first group between 17 and 23 days of age and included tremors of the limbs, poor posture, stilt gait and increased time of laying. The control animals did not show any symptoms of the disease during the experiment. Hypoproteinemia, hypoglycemia, cholesterol reduction and elevated triglycerides level associated with lipolysis of adipose tissue have been found in the sick kids. A significant decrease in selenium, vitamin E and activity of glutathione peroxidase levels was observed in the kids with symptoms of muscular dystrophy. The activity of AST, CK and LDH was significantly higher in the animals with symptoms of the disease as well. Five isoenzymes were obtained in the electrophoretic separation of lactate dehydrogenase into isoenzymatic fractions in the muscle tissue. LDH4 and LDH5 isoenzymes were dominating, and a significant increase in LDH5 fraction of the sick animals was also observed. Histopathological examination of muscle samples from sick animals revealed changes characteristic for the presence of Zenker necrosis.

The aim of the study was to compare the effects of corneal healing in case of application of stem cells in various forms, in relation to the antibiotic-assisted procedures. Rabbits were divided into 4 groups in the first stage of the experiment. Group 0 (negative control group) was not subjected to any actions, which would cause damage to the cornea. The remaining three groups had their cornea damaged. Group 1 (positive control group) – no drugs were administered during the experiment. Rabbits in group 2 were administered with ointment containing stem cells to the lesion, while group 3 – with ofloxacinum. The stem cells were administered during the first five days, twice a day, onto the corneal surface. The further course of the experiment consisted of observing the rate of healing of the injured cornea and assessment of its transparency, size of lesion, hyperaemia, eyelid spasm and outflow from the conjunctival sac after 5, 10 and 20 days.

In the second stage the animals were euthanised after clinical examination on the twentieth day of the experiment, in order to analyse the corneal reparative processes on the same day. The studies revealed that the application of antlerogenic stem cells had a positive effect on the healing process of corneal defects. The application thereof not only shortened the healing time, but also weakened or arrested the development of side effects. The results have demonstrated that the epithelial proliferation in each group was different. The longest was maintained in the group with stem cells, the shortest – in the group with chemotherapeutics. The use of antlerogenic stem cells had a positive effect on the healing process of corneal lesions. The use of stem cells helped to maintain high transparency of the cornea.

Conventional methods for determining the reproductive performance of sheep bred either after estrus synchronization during the breeding season or after induction of estrus/ovulation during the non-breeding season take a long time and may give misleading results due to the effect of environmental factors. Laparoscopic observations allow real-time monitoring of ovarian activity around estrus or ovulation. This study was aimed at assessing the superovulatory effects of follicle-stimulating hormone (FSH) and equine chorionic gonadotropin (eCG) treatments by laparoscopy during breeding (September-November, n=12) and non-breeding (April-June, n=12) seasons in Akkaraman sheep. In both seasons, after CIDR withdrawal, the ewes were injected either with 600 IU eCG or 300 μl (20 mg/ml) FSH twice at 12 hour intervals. Plasma P4, E2 and LH concentrations were determined at the time of intra-vaginal CIDR insertion (day 0) and then at its withdrawal (day 12), followed by 3 and 6 days of eCG or FSH injections. After 3 (first observation) and 6 (second observation) days of hormone injections, laparoscopy was performed to record ovarian activity in both seasons. The eCG increased (p<0.05) the numbers of large follicles (first observation) and CL (first and second observations) in the breeding season compared to FSH treatment. CL, small-moderate and large follicle numbers of eCG treated ewes were higher (p<0.05) than those of FSH at both observations in the non-breeding season. In the breeding season, eCG treated ewes had higher (p<0.05) plasma P4 (3 and 6 days after hormones injections) and E2 (3 days after hormones injections) concentrations than those of FSH. In conclusion, the results of the present study indicate that treatment with eCG during the non-breeding season can support ovarian activity, and thus increase ovulation rate and plasma hormone concentrations around induced estrus/ovulation in Akkaraman ewes.

The lumbosacral region of the spine is the most susceptible to pathology in large breed dogs. The most common pathologies of this segment include intervertebral disc disease, distortion of vertebrae, narrowing of the lumbosacral canal and congenital defects of the spine. The aim of this study is to develop a mathematical model describing the height of each lumbosacral vertebra in the dog in relation to the position of the vertebra. For the mathematical analysis we used the results of two measurements for each lumbar vertebra. The first measurement was made from the top of the spinous process to the center of the spinal cord. The second measurement was made from the center of the body of one vertebra to the center of the body of the next one. It is possible to determine an approximate mathematical model that would be uniform for the entire species and would connect the height of the lumbar vertebrae with their location for every breed of the domestic dog. Despite the considerable differences in the constitutional type (small, medium and large breeds), the morphology of the lumbosacral region of the spine exhibits similar proportions. Therefore, it is possible to assess an anomaly of this spinal region objectively. These findings suggest that it is possible to determine an approximate mathematical model that would be uniform for the entire species. The present study was carried out as part of a larger project. This particular work is a pilot study.

In the present study, the influence of the infection with equine herpesvirus type 1 (non-neuropathogenic and neuropathogenic strains of EHV-1) on the morphology and distribution of mitochondrial network in equine dermal cell line was investigated. Our results indicate that EHV-1-infection caused changes in the mitochondrial morphology manifested mostly by fission and reactive oxygen species generation.

In our study, we estimated the frequency of haplotype for cholesterol deficiency (HCD) carriers in the Russian Holstein cattle population. We studied 1817 random samples of cows born in 2010-2017 from ten herds and 331 cows and heifers from the other three herds born in 2016-2019, fathers or fathers of mothers of which were HCD carriers. The method of AS-PCR was used for animals genotyping. In the first group of animals, the incidence of HCD carriers was 8.09%, and in the second one - 23.26%. Our results demonstrated the necessity to test cows for the carriage of the HCD genetic defect in the Russian population of Holstein cattle.

A recent study found that an agarose gel electrophoresis (AGE) method yielded two distinct major bands corresponding to the hepatic and bone ALP isoenzymes (ALP2 and ALP3, respectively) in bovine serum treated with protease and neuraminidase (PN-treatment), although there were concerns that the intestinal ALP isoenzyme (ALP5) often overlapped with ALP3 in human serum treated with neuraminidase. Because ALP5 was separated from ALP3 in bovine serum treated with protease alone (P-treatment), we used a modified method employing both P- and PN-treated bovine sera to measure the activities of the three ALP isoenzymes in 53 lactating Holstein cows: 24 primiparous and 29 multiparous. Upon electrophoresis, 51 of 53 samples (96.2%) subjected to P-treatment yielded a distinct fraction corresponding to ALP5, as did the control serum. All PN-treated sera yielded a definite ALP2 fraction. The ALP3 fraction was calculated as the remainder after excluding ALP2 and ALP5. The activities of total ALP (t-ALP) and ALP3 in primiparous cows were higher than those in multiparous cows (p < 0.001) at early-to-peak [10–110 days in milk (DIM)] and mid (111–220 DIM) lactation. In the multiparous cows, the ALP3 activity at late lactation (221−477 DIM) was significantly higher than that at early-to-peak lactation. Thus, the modified AGE method described here is able to discriminate three fractions of ALP isoenzymes in the sera of lactating cows. The AGE pattern of circulating ALP isoenzymes will contribute to the understanding of the physiological bone metabolism status in lactating cows.