Polish Journal of Veterinary Sciences

The main objective of this study was to evaluate the intensity and character of the inflammatory reaction caused by an innovative polyester-polyurethane vascular prosthesis implanted into the abdominal aorta of 9 Beagle dogs aged 1-3 years. At 6 and 12 months post implantation the prostheses were removed and tissues samples were examined using 2 methods: histology and immunohistochemistry (IHC). Histology slides stained with hematoxylin and eosin (H&E) were evaluated for the intensity of inflammation by observing the density of inflammatory cells and graded 1 to 4 (1- light inflammation, 4 – severe inflammation). The pro-inflammatory mediator tumor necrosis factor-alpha (TNFα) and two anti-inflammatory mediators, interleukin 1 receptor antagonist (IL1ra), and interleukin 10 (IL10), were also assessed in the tissue samples by IHC methods. Mean (n=5) inflammation grade in H&E slides at 6 months post-implantation (6Mpost) was 2 and mean (n=4) inflammation grade at 12 months (12Mpost) was almost 3. IHC staining showed that TNFα and IL1ra in tissue samples obtained from 6Mpost dogs were expressed at the same intensity indicating equal pro- and anti-inflammatory cytokine levels. However, in the 12Mpost tissues TNFα was expressed more intensely than IL1ra and IL10. Moreover, in 2 dogs at 12Mpost, there were signs of infection assessed on the basis of neutrophil infiltration in the prostheses. In conclusion, the assessment of pro-inflammatory mediators such as TNFα and anti-inflammatory mediators, such as IL1ra and IL10, can help to interpret the intensity of the inflammatory process directed at synthetic prostheses.

The presence of lipopolysaccharide (LPS) in blood induces an inflammatory response which leads to multiple organ dysfunction and numerous metabolic disorders. Uncontrolled, improper or late intervention may lead to tissue hypoxia, anaerobic glycolysis and a disturbance in the acid -base balance. The effects of LPS-induced toxemia on biological and immunological markers were well studied. However, parameters such as base excess, ions, and acid-base balance were not fully investigated. Therefore, the objective of this study was to examine these blood parameters collectively in LPS-induced inflammatory toxemia in rat’s model. After induction of toxemia by injecting LPS at a rate of 5 mg/kg body weight intravenously, blood was collected from the tail vein of twenty rats and immediately analyzed. After 24 hours, the animals were sacrificed and the blood was collected from the caudal vena cava. The results revealed that the levels of pH, bicarbonate, partial pressure of oxygen, oxygen saturation, Alveolar oxygen, hemoglobin, hematocrit, magnesium (Mg2+), and calcium (Ca2+) were significantly decreased. On the other side, the levels of Base excess blood, Base excess extracellular fluid, partial pressure of carbon dioxide, lactate, Ca2+/Mg2+, potassium, and chloride were significantly increased compared to those found pre toxemia induction. However, sodium level showed no significant change. In conclusion, Acute LPS-toxemia model disturbs acid-base balance, blood gases, and ions. These parameters can be used to monitor human and animal toxemic inflammatory response induced by bacterial LPS conditions to assist in the management of the diagnosed cases.

Myxomatous mitral valve disease (MMVD) is a cardiac condition commonly found in older dogs. The disease process can lead to heart failure (HF). In HF, an increase of reactive oxygen species (ROS) and abnormal mitochondrial activity, as well as apoptosis, have been reported. Humanin (HN) is a polypeptide that has a cardioprotective effect against apoptosis and oxidative stress. The purposes of this study were (1) to investigate the potential role of plasma HN as a cardiac biomarker to predict disease progression of MMVD, and (2) to compare plasma HN concentrations with plasma NT-pro BNP concentrations. Thirty-one dogs were included in the study. The dogs were separated into four groups: Group 1 was healthy dogs (n = 8), Group 2 was MMVD class B (n = 8), Group 3 was MMVD class C (n = 8), and Group 4 was MMVD class D (n = 7). All dogs were given a physical examination, thoracic radiography, echocardiography, and samples of their blood were collected for hematology and blood chemistry analysis. Levels of plasma HN and plasma NT-proBNP were also investigated. The results showed that plasma HN levels were lower in the dogs with MMVD and that lower plasma HN levels were associated with greater severity of MMVD-induced HF. It was possible to observe changes in plasma HN levels at a less severe disease stage than plasma NT-proBNP in dogs with MMVD. These findings sug- gest that a decreased plasma HN level can be used as a biomarker to identify dogs with MMVD -induced HF.

Dendritic cells (DCs) due to their ability to present antigens are essential during the immune response to infections. The aim of the study was to evaluate the influence of bovine leukaemia virus (BLV) infection on DC properties. Cytokine profiles of myeloid, plasmacytoid and mono- cyte derived DCs from BLV infected cattle were analysed. Concentrations of IL-6, IL-10, IL-12, IFN-γ, and TNF-α in DC cultures were measured by flow cytometry. Obtained results indicated activation of pDCs population, where a significant increase in production of the IFN-γ was shown. Meanwhile, a decrease in production of IFN-γ and increase in production of IL-10 were shown in mDCs; the main population responsible for antigens presentation. This may indicate a contribu- tory role of the population during the process of persistent infection. In MoDCs population a significant elevation in secretion of proinflammatory cytokines – IL-6 and TNF-α was noted.

The aim of this study was to measure serum neopterin and C-reactive protein (CRP) concen- trations in female dogs with mammary tumours and evaluate the association between the values of these indicators and some clinical characteristics of the tumour. Fifty three female dogs were used for this study, including 43 dogs with mammary gland tumours (10 benign and 33 malignant) and 10 healthy controls. The concentrations of neopterin and CRP were determined using the ELISA technique and commercial ELISA kits. The mean serum neopterin concentration in fe- male dogs with mammary tumours was lower than in healthy dogs, but significant difference was not found. Similarly, there were no significant differences in neopterin concentrations in female dogs based on tumour size, tumour ulceration and metastasis. The mean CRP concentration was significantly higher (p<0.05) in dogs with malignant tumours compared to dogs with benign tu- mours and control. Furthermore, serum CRP concentration was significantly higher (p<0.05) in dogs with metastatic malignant tumours compared to dogs with non-metastatic mammary tu- mours. The CRP concentration was significantly lower (p<0.05) in dogs with tumours less than 3 cm compared to those with larger tumours, and significantly higher in dogs with ulcerated tu- mours compared to those without ulceration. Our findings suggest that the neoplastic process in the mammary gland does not cause significant changes in serum neopterin concentrations in dogs. Higher concentrations of serum CRP in dogs with advanced stages of malignant tumours may suggest that CRP could be a potential prognostic marker in canine malignant mammary tu- mours, but this hypothesis needs further study.

Mechanical ventilation (MV) is a supportive and life-saving therapy, however, it can cause ventilator-induced lung injury as a common complication. Thus, recruitment manoeuvres (RM) are applied to open the collapsed alveoli to ensure sufficient alveolar surface area for gas exchange. In the light of the fact that positive pressure ventilation is currently the standard treat- ment for improving pulmonary function, extrathoracic negative pressure is considered as an alter- native form of respiratory support. The aim of this study was to estimate the proinflammatory and oxidative response during MV and lung injury as well as the response after RM. All studied parameters were assessed at the following time points: T1-spontaneous breathing, T2- MV, T3- lung injury, T4 –RM. During MV (T2) elastase, MPO, ALP release, nitrite and superoxide generation significantly increased, whereas in later measurements a decrease in these values was noted. The MDA plasma concentration significantly (p<0.05) increased at T2, reaching a level of 13.30±0.87 nmol/ml; at other time points the values obtained were similar to the baseline value of 9.94±0.94 nmol/ml, whereas a gradual decrease in SOD activity at time T2-T4 points in comparison with the baseline value was found. During the study both neutrophil activity and oxi- dative stress indicate exacerbated response after MV and lung injury by bronchoalveolar lavage; however, extrathoracic negative pressure system as the MR ameliorates damaging changes which could further lead to serious lung injury.

The use of lactoferrin (LF) and/or lactobacillus sp. (LB) to improve animal health and production has increased recently. However, information regarding the immune-modulatory role of LB supplementations either alone or in combination with LF in sheep remains unclear. Therefore, the present study was designed to evaluate the immune modulating properties and the antioxidant activity of supplementing commercially available LF and/or LB in healthy lambs. For this reason, twenty-four apparently healthy Ossimi lambs were used. After three weeks of acclimatization, the lambs were randomly allocated to four equal-sized groups and assigned to receive one of the following supplements: LB at a dose of ~ 1 g active ingredient/head (group 1), LF at a dose rate of 0.5 gm /head (group 2), a combination of both treatments using the same dosing regimens (group 3), and (group 4) received only 10 mL of isotonic saline and was considered as a control group. All supplements were given orally twice daily for 30 consecutive days. Blood samples were collected from each lamb before starting the experiment (T0) and two weeks (T15), and four weeks (T30) after giving supplements for hematological examinations, serum biochemical analyses, and RT-PCR assays. Our findings demonstrated that lambs receiving LB showed statistically significant (P<0.05) higher values of total leucocytes, lymphocytes and lysozyme activity than those receiving LF. In contrast, lambs that received LF had significantly (P< 0.05) higher values of serum catalase, nitric oxide and GSH with a significantly lower MDA level compared with those supplemented with LB. A combination of LF and LB supplementation elicited maximal up-regulation of Tollip, TLR4, IL-5, and IL-6 gene expression compared with other groups. The results suggest that bovine LF and or LB could be used as useful nutritional supplements to support the immune system in healthy lambs.

Concentrations of four trace elements, copper (Cu), zinc (Zn), manganese (Mn) and seleni- um (Se), have thus far proven to be affected by lentiviral infections in people and rhesus monkeys. As small ruminant lentivirus (SRLV) infection is responsible for one of the most important goat diseases, caprine arthritis-encephalitis (CAE), we evaluated serum and liver concentrations of Cu, Zn, Mn, Se in goats severely affected by symptomatic CAE and compared them with litera- ture reference intervals. Serum and liver samples of dairy goats euthanized due to severe clinical form of CAE were collected and screened for the concentration of Cu, Zn, Mn (54 serum sam- ples, 22 liver samples), and Se (36 serum samples, 22 liver samples) using flame atomic absorption spectrometry for Cu, Zn, Mn and graphite furnace atomic absorption spectroscopy for Se. In both serum and liver samples concentration of Zn was the highest, followed by Cu concentration, and then by Mn and Se. There was no relationship between serum and liver concentrations of trace elements. Liver concentrations of all four trace elements and serum Cu concentration fell within literature reference intervals, although liver Se concentration was mainly in the lower marginal range (between 0.4 and 1.0 mg/L). Serum Zn concentration was elevated (>1.2 mg/L) in all goats, serum Mn concentration was elevated (>0.04 mg/L) in 42 (78%) goats and serum Se concentra- tion was elevated (>1.6 mg/L) in 13 (36%) goats. Concluding, severe symptomatic CAE does not appear to be associated with the level of any of the four trace elements.

Polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its metabolite, dichlorodiphenyldichloroethylene (DDE) can disturb the secretory function of the ovary and both contractions and secretory function of the uterus during the estrus cycle and pregnancy. Additionally, PCBs can pass through the placental barrier into allantoic and amniotic fluid. The presence of PCBs in these fluids is associated with higher frequency of spontaneous abortions and premature births in humans and animals. Therefore, the effect of PCBs, DDT and DDE on the connexins (Cx26, Cx32 and Cx43) and keratin 8 (KRT8) expression in bovine placentomes was investigated. The placentome slices from the second trimester of pregnancy were incubated with PCB153, 126, 77, DDT and DDE (each at doses of 1, 10 or 100 ng/ml) for 48 h. Then, the slices were stained using immunohistochemistry. The density of Cxs staining was measured with Axio- Vision Rel. 4.8 software in fetal-maternal connections and binuclear cells (BNC). None of the tested xenobiotics (XBs) affected the localization of Cxs and KRT8 in the fetal-maternal connection area, but the XBs affected the density of Cxs in fetal-maternal connections and binuclear cells (BNCs). Depend on the doses, in fetal-maternal connections all used PCBs changed the protein expression of different Cxs, while in BNCs, all tested XBs except DDT increased the expression of Cxs. None of investigated XBs affected on KRT8 expression. In summary, used XBs affect the expression of Cxs and change the quantitative relationships between them. Therefore, XBs can unfavorably influence function of the utero-placental barrier in cows.

The normotensive (Wistar) and spontaneously hypertensive (SHR) rats were examined to assess the response of the organism to selenium (Se) overdose. Moreover, the effect of zinc (Zn) and vitamin E, i.e. dietary components interacting in many biochemical processes with Se, on the Se uptake was evaluated. The control group was fed an untreated diet, and the diets of two other groups were overdosed with Se in the form of sodium selenite (9 mg/kg) and supplemented with Zn (13 mg/kg). Two experimental groups were fed a diet supplemented with Zn (13 mg/kg) and Se at an adequate level (0.009 mg/kg); a half of the animals was supplemented with vitamin

E. The results showed significant differences in the Se contents between the rat strains in case of Se-overdosed groups, where in the liver and kidney tissue Se contents of SHR rats exceeded 3- and 7-fold the normotensive ones. The Se uptake was altered by the vitamin E; no effect of Zn was observed. Activities of antioxidant enzymes were determined in the animal tissues indicating different patterns according to rat strain, tissue analysed, and administered Se dose. Thus, Se overdose, for instance, via an incorrectly prepared dietary supplement, can result in serious imbalances of the biochemical status of the animals.

Early recognition of altered lactate levels is considered a useful prognostic indicator in dis- ease detection for both human beings and animals. It is reasonable therefore to hypothesize that a portable, point of care (POC) spectrophotometric device for analysis of lactate levels, may have an application for field veterinarians across a range of conditions and diagnostic procedures. In this study, a total of 72 cattle in the transition period underwent POC spectrophotometric lactate measurement with a portable device (The Vet Photometer) in the field, with a small portion of blood used for comparative ELISA evaluation. Lactate measurements were compared using a of Passing-Bablok regression analysis and Bland-Altman plots. The Vet Photometer lactate mea- surement results were in agreement with those generated by the ELISA method. Values for the agreement were derived, in a 95% CI between -1.3 and 0.99, and a positive correlation (r=0.71) between the two measurements. The equation y= 0.68x + 0.60 was achieved using a Pass- ing-Bablok regression analysis. There were no statistical differences in mean values between the measurement methods. In conclusion, a novel veterinary POC spectrophotometric device “Vet Photometer” is an accurate device for evaluation of lactate levels in healthy transition cows.

Enterococcus hirae belongs in the Enterococcus faecium group within the genus Enterococcus. This species occurs naturally in the environment, commensally in the alimentary tracts of animals, and pathologically for example in humans with urinary infections. Some strains of E. hirae possess virulence factors, including biofilm formation. Biofilm growth protects bacteria against host de- fences; biofilm can be a source of persistent infection. Testing bacterial strains for their ability to form biofilm might therefore facilitate their treatment or prevention. This study focuses on bio- film formation by E. hirae strains derived from various animals. This kind of testing has never been done before. A total of 64 identified E. hirae from laying hens, ducks, pheasants, ostriches, rabbits, horses and a goat were tested by means of three methods; using Congo red agar, the tube method and microtiter plate agar. The majority of strains were found to form biofilm. 62.5% of strains were biofilm-forming, four categorized as highly positive (OD570 ≥1); most strains were low-grade biofilm positive (0.1 ≤ OD 570 < 1). Related to poultry, 55 E. hirae strains were tested nd found to produce biofilm; 24 strains did not form biofilm, 31 strains were biofilm-forming; 27 strains showed low-grade biofilm formation, and four strains were highly biofilm-forming. Four strains from hens and ostriches reached the highest OD570 values, more than 0.500. Rabbit-derived E. hirae strains as well as strains isolated from horses and the goat were low-grade bio- film-forming. Microtiter plate assay proved to be the best tool for testing the in vitro biofilm for- mation capacity of E. hirae strains from different species of animals.

Sperm-mediated gene transfer (SMGT) is based on the ability of spermatozoa to bind exoge- nous DNA and transfer it into oocytes by fertilization. However, SMGT is still undergoing opti- mization to improve its efficiency to produce transgenic animals. The acrosome reaction is neces- sary for spermatozoa to carry the exogenous DNA into oocytes. In this study, the effect of the acrosome reaction on the efficiency of spermatozoa carrying exogenous DNA was evalua- ted. The results showed that the efficiency of the acrosome reaction was significantly higher (p<0.05) after incubation with 50 μmol/L progesterone compared to incubation without proges- terone. It was significantly higher (p<0.05) in the 20, 40, and 60 min of progesterone treatment groups than in the 0 min treatment group. The spermatozoa were further incubated with cyanine dye Cy5 labeled DNA (Cy5-DNA) for 30 min at 37°C, and positive fluorescence signals were detected after the acrosome reaction was induced by progesterone at concentrations of 0 and 50 μmol/L for 40 min. The percentage of positive Cy5-DNA signals in spermatozoa was 96.61±2.06% and 97.51±2.03% following exposure to 0 and 50 μmol/L progesterone, respective- ly. The percentage of partial spermatozoa heads observed following combination with Cy5-DNA was 39.73±3.03% and 56.88±3.12% following exposure to 0 and 50 μmol/L progesterone, respec- tively. The ratio of positively stained spermatozoa combined with exogenous DNA showed no reduction after the acrosome reaction. These results suggest that the acrosome reaction might not be the key factor affecting the efficiency of SMGT.

This study investigated changes in the real-time measured levels of milk β-hydroxybutyrate according milk yield, lactation number and status of reproduction in dairy cows. A total of 378 cows were selected. According to their reproductive status the cows were classified as belonging to the following groups: Fresh (1 – 44 days after calving. n=43). Open (45 – 65 days after calving. n=78), Inseminated (1 – 35 days after insemination. n=133). Pregnant (35 – 60 days after insemination and pregnant (relatively pregnant) (n=124). The cows were milked with DeLaval milking robot (DeLaval Inc., Tumba, Sweden) in combination with a Herd Navigator (Lattec I/S. Hillerød. Denmark) analyser. We observed that milk β-hydroxybutyrate (BHB) had a tendency of increasing with an increase of lactation number. The average BHB in multiparous cows was 11.111% higher in comparison with primiparous cows (p<0.001). We found higher BHB concentration in the multiparous cows in all reproduction status groups (p<0.001). A strong positive statistically significant (p<0.001) relationship has been found between BHB and the average milk yield within all groups of primiparous cows although we found a statistically unreliable coefficient of correlation (from -0.202 to 0.057) between highest milk yield and BHB in primiparous and multiparous cows.

Disseminated intravascular coagulation (DIC) is a complex, dynamic and hemostatic disorder which develops secondarily to a disease characterized with an imbalance in the pro-coagulant and anti-coagulant components of hemostasis. The aim of the study is to evaluate hemostatic dysfunc- tion and the DIC syndrome in cattle with displaced abomasum (DA), with using the hematologic analyses and an extensive coagulation profile in the 96 hour-period including before and after surgery. The animal material of the study consisted of 12 dairy cows diagnosed with displaced abomasum (9 LDA and 3 RDA without volvulus) in the 2-4 week period after parturation and with no other post-partum disease. In dairy cows diagnosed with DA, hematological, coagulomet- ric (PT, APTT, Fibrinogen) and coagulation factor analyses [D-Dimer, TAT (thrombin-anti- thrombin complex), ATIII (antithrombin III), PAI-1 (plazminogen activator inhibitor-1] were performed in blood samples obtained before the operation as well as 30 minutes, 60 minutes and 2, 5, 10, 24, 48, 72 and 96 hours after the operation. In the DA cases, abnormalities were found in 6 of the 8 coagulation parameters. In the LDA and RDA groups, prolonged PT (sec), PT (INR) and APTT, hypofibrinogenemia, an increase in serum D-Dimer concentration at 72 and 96 hours after the operation and an increase in serum ATIII concentrations before and 30, 60 minutes and 2, 5, 72 and 96 hours after the operation was found (p<0.05). Hemostatic dysfunction and the risk of DIC developing in DA cases and continuing in the post-operative period was determined.

The aim of this study was to establish reference values for 2D and M-mode measurements in Dachshunds. Basic echocardiographic data, including M-mode, 2D and spectral Doppler measurements, was collected, analyzed and compared between 41 healthy Dachshunds and 50 other healthy dogs of similar weight. Echocardiographic reference intervals were prepared for Dachshunds. Dachshunds had a smaller left ventricular diameter in diastole and systole and a thicker septum than other dog breeds. Male Dachshunds had larger diastolic and systolic left ventricular diameter than females. Reference intervals for 2D and M-mode measurements in healthy Dachshunds differ from other dogs of similar weight and should be used for this breed to assess chamber enlargement.

Several human studies have reported that capsaicin has anti-pruritic effects. Moreover, sever- al concentrations of topical capsaicin have been used to alleviate itch. The aim of this study was to investigate the anti-pruritic effect of capsaicin against histamine-induced pruritus compared with that of topical steroid or vehicle in 15 healthy beagles. Fifteen dogs were divided into three groups (n = 5 each), and treated topically with one of the following on the left side of the neck: capsaicin, positive control (steroid), or negative control (vehicle). Each treatment was performed twice daily for 8 days. All dogs were injected with histamine intradermally before treatment and on the 2nd, 4th, 6th, and 8th days of the treatment to evoke itch. Pruritus, wheal, and erythema intensity were assessed at each evaluation; cutaneous temperature was also recorded. On the final day, skin biopsy was conducted for histopathological evaluation for all dogs. The severity of pruritus was lesser in the capsaicin-treated group compared with the negative control group on day 8 (p<0.05). In the capsaicin and steroid groups, wheal size, erythema index, and cutaneous temperature also decreased compared with pretreatment. Histopathological evaluation showed that the capsaicin-treated group had a higher number of inflammatory cells in the dermis com- pared to the vehicle control group; however, the steroid-treated group showed less severe inflam- matory reactions than the vehicle control group. These results suggest that capsaicin cannot reduce inflammation but may play a helpful role in reducing pruritus in dogs.

A clinical trial was performed to evaluate the therapeutic efficacy of osaterone acetate (OSA) in the treatment of benign prostatic hyperplasia (BPH) in dogs. Osaterone acetate (Ypozane, Virbac) was administered orally at a dose of 0.25 mg/kg body weight once a day for seven days to 23 dogs with BPH. During the 28-day trial, the dogs were monitored five times for their clinical signs and prostate volume. The OSA treatment promoted rapid reduction of clinical scores to 73.2% on day 7 and to 5.9% on day 28 (p<0.05). Osaterone acetate induced the complete clinical remission in approximately 83.0% of the dogs on day 28. The prostate volume regressed to 64.3% of the pretreatment volume after two weeks of the treatment (p<0.05) and to 54.7% at the end of the trial (p<0.05). In conclusion, OSA quickly reduced clinical signs and volume of the prostate glands in dogs with BPH.

The aim of the study was to develop new laparoscopic technique for repeated recovery of sheep oocytes. Oocytes were aspirated with specifically designed catheter. It allowed to recover oocytes without ovary damage and to preserve very good quality of recovered oocytes. Fifteen ewes were oocytes donors. Oocytes were collected: one time (group I, n=15), two times (group II, n=15), three times (group III, n=10), four times (group IV, n=5). The endoscope was inserted into the abdominal cavity. Two trockars for putting the manipulators were inserted 15 cm cranial from the udder. Oocytes were collected by aspiration of the follicular fluid from the ovarian follicles. The observed clinical complications were: ovary bleeding and cicatrix at place of needle insertion, the fragmentary adhesion of infundibulum and ovary, adhesions of omentum and peri- toneum near the place where the grasping forceps were inserted and adhesion of ovary and uterus. Ovarian follicles (n=204) were aspirated, 130 (63.8%) oocytes were obtained. Out of 130 obtained oocytes, 112 were qualified for in vitro maturation. The remaining 18 oocytes (13.8%) were rejected due to cytoplasmic changes. The proposed technique allows for the collecting oocytes of good quality that can be used for IMV/IVF techniques and cloning.

The aim of our study was to determine the influence of L-carnitine (L-CAR) on the cellular parameters of hen erythrocytes during a 48 hour exposure to L-CAR at concentrations of 25, 50 and 100 mg/mL in nutrient-deficient medium. Cell morphology, haemolysis, caspase 3/7 activity and glucose uptake (GU) were determined. The results showed a lower percentage of apoptotic cells and decreased haemolysis of erythrocytes treated for 48 hours at all the concentrations of L-CAR. The amino acid at 50 mg/mL inhibited the activity of proapoptotic caspase 3/7; however, it increased GU. In contrast, caspase 3/7 level was increased but GU was decreased in erythrocytes treated with 100 mg/mL of L-CAR when compared to the control. It may be hypothesized that reduction of apoptotic changes in hen erythrocytes may result from increased GU.

Blastocystis is a common enteric protozoan of humans and various species of animals. Culture and microscopic examination of fecal samples is the conventional method for identifying four major forms of Blastocystis (vacuolar, granular, non-vacuolar or cystic). In this article, we compared eight liquid media for cultivation of Blastocystis spp. Study material included fecal samples from clinically healthy pigs. Significant differences in the growth of Blastocystis on individual media were observed.

Due to the unrecognized effect of tigecycline (TIG) on CD4+ and CD8+ T cells, the present study has been undertaken in order to determine whether the drug can affect these cells in respect of their counts, and the production of IFN-γ, IL-17 (pro-inflammatory and immune-protective cytokines), IL-4 (anti-inflammatory and immune-protective cytokine), IL-10 and TGF-β (anti-inflammatory and immune-suppressive cytokines). Murine lymphocytes were treated with TIG for 48 and 96 h at concentrations reflecting its plasma levels obtained in vivo at therapeutic doses, and at 10-fold lower concentrations. It was found that TIG neither affected substantially the percentage and absolute counts of entire CD4+ and CD8+ T cell populations nor influenced the Foxp3+CD25+CD4+ regulatory/suppressive T cell subset. Furthermore, the percentages of IL-4-, IL-10-, IL-17- and TGF-β-producing CD4+ T cells were not altered following the exposure to TIG. Similarly, TIG did not influence IFN-γ production by CD8+ T cells. Thus, with respect to the parameters evaluated, TIG does not seem to exert immune-suppressive and anti-inflammatory effects.

This study analysed the influence of montelukast (MON; 10-8 - 10-4 M), a cysteinyl leukotriene receptor 1 (CysLTR1) antagonist, on the contractility of the porcine uterine smooth muscle in the luteal phase of the oesterous cycle (n=8) and in early pregnancy (n=8). Stimulation of uterine strips in the luteal phase with MON has been shown to significantly reduce the amplitude of con- tractions, but not to affect the tension or frequency of contractions. A statistically significant tension increase and decrease in the frequency and amplitude of contractions was observed in pigs in early pregnancy. This suggests that MON has a different effect on the parameters under study in cyclic and pregnant pigs.

The reliable and rapid diagnosis of infectious animal diseases presents an exceptionally im- portant aspect when considering their control and prevention. The paper describes the compara- tive evaluation of two rapid isothermal amplification methods for diagnosis of African swine fever (ASF). The robustness of loop-mediated isothermal amplification (LAMP) and the cross-priming amplification (CPA) were compared using samples obtained from ASF confirmed animals. Both assays were evaluated in order to define their diagnostic capabilities in terms of ASF diagnosis and reproducibility of the results. Investigations showed no cross-reactivity for other pig patho- gens and no significant differences in the specificity of both assays. The sensitivity of LAMP reached 90%, while that of CPA was 70%. In conclusion, both methods are suitable for imple- mentation in preliminary ASF diagnosis but further improvements are required to enhance their diagnostic sensitivity.

In this study, we propose a possible way of obtaining reproductive and respiratory syndrome virus (PRRSV) free offspring from genetically valuable lines of Přeštice black-pied boars com- ming from PRRSV-positive pig breeding herds with the use of artificial insemination (AI). The ejaculates were collected from 4 different lines of boars. Samples of fresh semen were not detected with the virus and 12 sows were inseminated. Blood samples of sows and their offspring were repeatedly tested for the virus but the results were negative. We managed in this way to maintain the endangered population of this breed and obtain PRRSV-free offspring.

Four and a half years of African Swine Fever (ASF) in population of free-ranging wild boars and domestic pigs revealed a number of novel insights into the disease epidemiology. Until November 20th, 2018, in total 3048 cases in wild boars and 213 outbreaks in domestic pigs have been confirmed. In spite of low contagiosity as well as low rate of ASF spread in wild boars the disease has an enormous socio-economical impact on the production of pigs in Poland. One of the most important aspects which directly influences the dynamics of ASF spread is the unpredictable hu- man activity. Another important factor responsible for continuous ASF spread is fast recovery of wild boar population in spite of efforts taken by hunters. Assuming our scientific opinion ASF seems to be present in wildlife for the incoming few or several years. Therefore, extraordinary measures should be prepared and undertaken to limit the risk of the occurrence of future out- breaks in domestic pigs. One of the most crucial issues is implementation of strict biosecurity measures in all domestic pigs holdings.