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Canine parvovirus type 2 is one of the most common causes of death among puppies. Despite preventive vaccination, the disease continues to be diagnosed. The aim of the study was to provide a molecular characterization of CPV-2 isolates found in southeastern Poland. Genetic CPV-2 material was isolated from the blood (n=10) and feces (n=50) of infected dogs. The presence of CPV-2 was confirmed by amplification of sequences coding both VP1 and VP2 protein. The products of the PCR reaction with primers amplifying VP2 protein were sequenced and used for genotyping. Bioinformatics analysis of the sequenced PCR product was performed to determine the phylogenetic relationships with variants recorded in the public databases. Based on the analysis of polymorphism in the nucleotide sequence 7 nucleotide variants were detected and assigned into four amino acid groups. Representatives of three groups contained asparagine at amino acid position 426 of the VP2 protein, which is characteristic of CPV-2a. The variant from the fourth group belonged to type CPV-2b. CPV-2a is the dominant antigenic type of CPV-2 in Poland. The pathogen’s high degree of polymorphism is manifested not only by the presence of numerous variants within the type, but also by the presence of representatives of type CPV-2b. Further studies of the molecular epidemiology of CPV-2 are necessary to optimize the effectiveness of preventive measures.

Mycoplasma bovis is known to be a cause of chronic pneumonia in cattle. To date, the disease pathomechanism has not been fully elucidated. Leukocytes play a key role in host antimicrobial defense mechanisms. Many in vitro studies of the effect of Mycoplasma bovis (M. bovis) on leukocytes have been performed, but it is difficult to apply these results to in vivo conditions. Additionally, only a few studies on a local immune response in M. bovis pneumonia have been undertaken. In this study, the experimental calf-infection model was used to determine the effect of field M. bovis strains on changes of the peripheral blood leukocyte response, including phagocytic activity and oxygen metabolism by cytometry analyses. An additional aim was to evaluate the lung local immunity of the experimentally infected calves using immunohistochemical staining. The general stimulation of phagocytic and killing activity of peripheral blood leukocytes in response to the M. bovis infection points to upregulation of cellular antimicrobial mechanisms. The local immune response in the infected lungs was characterized by the T- and B-cell stimulation, however, most seen in the increased T lymphocyte response. Post-infection, strong expression of the antigen-presenting cells and phagocytes also confirmed the activation of lung local immunity. In this study&mdash, despite the stimulation&mdash, both the peripheral and local cellular antimicrobial mechanisms seem to appear ineffective in eliminating M. bovis from the host and preventing the specific lung lesions, indicating an ability of the pathogen to avoid the host immune response in the M. bovis pneumonia.

The Aleutian mink disease virus (AMDV) is one of the most serious threats to modern mink breeding. The disease can have various courses, from progressive to subclinical infections. The objective of the study was to provide a comparative molecular characterization of isolates of AMDV from farms with a clinical and subclinical course of the disease. The qPCR analysis showed a difference of two orders of magnitude between the number of copies of the viral DNA on the farm with the clinical course of the disease (105) and the farm with the subclinical course (103). The sequencing results confirm a high level of homogeneity within each farm and variation between them. The phylogenetic analysis indicates that the variants belonging to different farms are closely related and occupy different branches of the same clade. The in silico analysis of the effect of differences in the sequence encoding the VP2 protein between the farms revealed no effect of the polymorphism on its functionality. The close phylogenetic relationship between the isolates from the two farms, the synonymous nature of most of the polymorphisms and the potentially minor effect on the functionality of the protein indicate that the differences in the clinical picture may be due not only to polymorphisms in the nucleotide and amino acid sequences, but also to the stage of infection on the farm and the degree of stabilization of the pathogen-host relationship.

Arabitol is used in the food industry as a low-calorie sweetener. It is produced by yeasts during the biotransformation process of l-arabinose. Genome shuffling was performed in Candida parapsilosis DSM 70125, an efficient producer of arabitol, to obtain fusants with improved arabitol production ability. Four mutants from the parental library were used for the first round of genome shuffling. The best fusants, GSI-1 and GSI-10A, were subjected to a second round of genome shuffling. Finally, two fusants, GSII-3 and GSII-16, produced concentrations of arabitol that were 50% higher than that of the wild-type strain during selection culture. Under the optimal conditions established for C. parapsilosis, the two fusants produced 11.83 and 11.75 g/L of arabitol and were approximately 15–16% more efficient than the wild-type strain. Flow cytometry analysis showed that the ploidy of the new strains did not change.

Introduction Phagocytic activity and oxygen metabolism of peripheral blood granulocytes from rabbits with experimental trichophytosis were assessed by flow cytometry. Material and Methods Virulent species of T. mentagrophytes var. granulosum (Tm-K) isolated from rabbits with natural trichophytosis was used for experimental infection. The phagocytic activity of granulocytes was measured in whole blood by flow cytometry using the commercial Phagotest kit. Oxidative burst was measured in whole blood by flow cytometry using the commercial Bursttest kit. Results It was found that rabbits were susceptible to infection with Trichophyton mentagrophytes under experimental conditions. The analysis of the phagocytic activity indices and oxygen metabolism of granulocytes in peripheral blood of infected rabbits showed that changes of the indices were connected with the progression and regression of the disease. A significant decrease in phagocytic activity and oxygen metabolism was observed during development of fungal lesions and it remained similar throughout the progress of the disease. The highest means of the percentage of activated and ingesting phagocytes and a significant increase in the mean fluorescence intensity (representing the number of ingested bacteria) were observed during spontaneous recovery. Therefore, the decrease or increase in the indices of phagocytic activity and oxygen metabolism of granulocytes from rabbits experimentally infected with T. mentagrophytes is somehow related to the progress of infection and suppressive activity of the fungus, whose elimination during recovery caused significant increases in investigated indices of non-specific cellular immunity. Conclusion The results of the present investigation confirm that the mechanism of oxygen-dependent killing is crucial in infections caused by T. mentagrophytes.

Background The aim of the present study was to assess the effects of subclinical inflammation on specific humoral immunity in dogs vaccinated with Nobivac® DHP based on serum levels of CRP and Hp. Dogs from the group I were administered Nobivac® DHP, the vaccine against distemper, infectious hepatitis and parvovirus whereas group II animals received subcutaneous turpentine oil to induce subclinical inflammation, followed by Nobivac® DHP after 24 h. Animals in group III received only turpentine oil in the way and amount identical to that as in group II. Results Nobivac DHP relatively poorly induced the immune inflammatory response showing good immunogenic properties, which was evidenced by only a double increase in mean CRP and Hp levels associated with antigenic stimulation in group I. In group II, serum neutralization (SN) and haemagglutination inhibition (HI) results were quite closely correlated with serum levels of CPR and Hp. Conclusions Our findings suggest that the efficacy of vaccinations in dogs can be significantly affected by subclinical inflammations, which is indicated by a correlation between serum CRP and Hp levels versus antibody titres for canine distemper and parvovirus in both experimental groups of dogs (group I and II). The correlation of mean CRP and Hp values in dogs with subclinical inflammation and after vaccination with the kinetics of increasing antibody titres against distemper and parvovirus in group II dogs reflects the severity of inflammatory response and the extent of specific humoral immunity. Routine determinations of serum CRP and Hp levels as the indices of inflammation severity can be the essential biochemical markers for assessment of dogs’ health in the period preceding specific immunoprophylaxis and efficacy of the vaccine.

Equine Anoplocephalosis constitute a significant problem in horses worldwide. The aim of this study was to analyse intrinsic (host age and sex) and extrinsic (management type, pasture type and moisture) factors that influence the prevalence and FEC of A. perfoliata infections. Faecal samples were collected from 994 horses managed in studs or individually between 2012 and 2014. The Sedimentation-flotation method was applied for coproscopic analysis, and faecal egg counts were calculated. The overall prevalence was 25.1% (21.4–29.0) with the highest prevalence (36.1% [28.1–44.8]) found in horses 10–20 years old. The individuals kept in studs showed three times higher A. perfoliata prevalence compared to the ones managed individually. The prevalence significantly differed between pasture types, with individuals kept in studs (37.6% [34.3–40.9]) showing four times higher prevalence than horses kept individually (9.2% [4.8–16.5]). More horses kept on watery (42.0% [36.6–47.6]) and semi-watery (35.9% [31.3–40.7]) pastures were infected than those on dry (6.6% [4.6–9.2]) pastures. The overall A. perfoliata FEC in all examined individual was 2.67 and differed within sex, with mares showing 4.3 − times higher FEC of infection than stallions. Horses bred in studs (3.65 ± 0.289) showed higher FEC than these bred individually (1.28 ± 0.198). There was the effect of pasture type on A. perfoliata FEC, with horses kept on joint pastures (4.06 ± 0.29) showing higher FEC than individuals kept individually (0.88 ± 0.23). Pasture moisture significantly affected A. perfoliata FEC with the highest FECs in horses from watery pastures. Horses bred on dry pastures showed 16 times lower FEC than horses bred on watery pastures. Host age also significantly affected A. perfoliata FEC, with the oldest individuals showing the highest mean FEC. The presented analysis of intrinsic and extrinsic factors may help to overcome A. perfoliata infections in horses in different breeding systems. Understanding the role of management and pasture type risk factors that influence this parasitosis may benefit both breeders and veterinary surgeons.

Introduction: The aim of the study was to investigate the mechanical and geometric properties as well as bone tissue and mineral density of long bones in mink dams exposed to deoxynivalenol (DON) since one day after mating, throughout gestation (ca. 46 d) and lactation to pelt harvesting. Material and Methods: Thirty clinically healthy multiparous minks (Neovison vison) of the standard dark brown type were used. After the mating, the minks were randomly assigned into two equal groups: nontreated control group and DON group fed wheat contaminated naturally with DON at a concentration of 1.1 mg·kg-1of feed. Results: The final body weight and weight and length of the femur did not differ between the groups. However, DON contamination decreased mechanical endurance of the femur. Furthermore, DON reduced the mean relative wall thickness and vertical wall thickness of the femur, while vertical cortical index, midshaft volume, and cross-sectional moment of inertia increased. Finally, DON contamination did not alter bone tissue density, bone mineral density, or bone mineral content, but decreased the values of all investigated structural and material properties. Conclusion: DON at applied concentration probably intensified the process of endosteal resorption, which was the main reason for bone wall thinning and the weakening of the whole bone.

Summary The aim of the study was to evaluate the effect of the diet, mother type and sex of the offspring on the mechanical and geometric parameters of long bones as well as bone tissue density in minks. Primiparous and multiparous dams were supplemented with β-hydroxy β-methylbutyrate (a metabolite of leucine, at the daily dosage of 0.02 g/kg of body weight) and/or 2-oxoglutaric acid (a precursor of glutamine, at the daily dosage of 0.4 g/kg of body weight) during gestation. The diet did not influence bone tissue density and the length of the humerus. An increase in the length of the femur was noted in male offspring delivered by multiparous dams. The diet resulted in an increase in the weight of the humerus in males from multiparous dams and a decrease in offspring from primiparous dams. Heavier femora were noted in male offspring delivered by both types of dams. The maximum elastic strength of the humerus was higher in the offspring delivered by multiparous than primiparous dams, irrespective of the offspring sex. The diet resulted in reduction in the ultimate strength of the femur in the male offspring delivered by primiparous dams. Only females born by multiparous dams, irrespective of the diet, showed a significant increase in the cross-sectional area of the humerus, while a significant decline was noted in males delivered by multiparous dams and in all the offspring delivered by primiparous dams. An increase in the cross-sectional area of the femur was noted in the offspring delivered by multiparous dams, while reduction was observed in the offspring delivered by primiparous dams. These results have shown for the first time that the presence of β-hydroxy-β-methylbutyrate or 2-oxoglutaric acid in the diet of pregnant primiparous or multiparous dams unambiguously affects the geometry and mechanical properties of offspring's long bones.

This study investigated the hypothesis that the antioxidant potential of lamb blood increases with age and depends on lambs being reared and fed by their mothers. The study was performed on lambs (35 ewes and 35 rams) of the synthetic SCP line. From 10 days of age until weaning (at 90 days of life) the lambs received properly-balanced concentrate diets for every fattening stage. All the lambs also received a structural dietary additive of meadow hay during the entire rearing period. The blood samples were collected on 2 h of life and days 14, 56, 84, 112 of life. It was determined several parameters in the blood. Lamb sex did not affect levels of blood antioxidant parameters however rams tested higher in bilirubin content. The lambs younger than 84 days of age displayed higher antioxidant potential and lower oxidative stress compared to older lambs (above 112 days of life) that were weaned from their dams.

The progressive growth and spread of tumour cells in the form of metastases requires an interaction of healthy host cells, such as endothelial cells, fibroblasts, and other cells of mesenchymal origin with immune cells taking part in innate and adaptive responses within the tumour lesion and entire body. The host cells interact with tumour cells to create a dynamic tumour microenvironment, in which healthy cells can both positively and negatively influence the growth and spread of the tumour. The balance of cellular homeostasis and the effect of substances they secrete on the tumour microenvironment determine whether the tumour has a tendency to grow or disappear, and whether the cells remain within the lesion or are capable of metastasis to other regions of the body. Intercellular interactions also determine the tumour’s susceptibility to radiation or other types of cancer treatment. They may also be a rational explanation for differences in treatment outcomes, in which some metastases regress and others progress in response to the same treatment method.

The aim of this study was to determine the serum concentrations of acute phase proteins (APPs), including C-reactive protein (CRP), serum amyloid A (SAA) and haptoglobin (Hp) in dogs with circumanal gland tumours. Investigations were conducted on 39 male dogs of mixed breed. The animals were divided into four groups depending on the degree of tumour malignancy and type of hormones that were dominant in the bloodstream. All measurements of serum APPs were conducted by the use of commercial diagnostic kits. In dogs with benign tumours, the concentrations of each APP decreased during therapy, while in dogs with malignant tumours, despite anti-tumour therapy, concentrations of APPs in serum increased markedly. The results of this study suggest that changes in serum concentrations of CRP, SAA and Hp during anti-hormone therapy may be a reliable marker for differentiating tumour grade and degree of malignancy. Including APPs in routine diagnostics may assist with formulating a prognosis for the outcome of the disease, before implementing anti-tumour therapy. Moreover, monitoring the effectiveness of therapy may be possible based on the determination of serum APP concentrations.

Aleutian mink disease virus is one of the greatest threats to modern mink farming. The disease reduces fecundity and causes high mortality among kits. The aim of this study was to evaluate the effectiveness of methisoprinol in counteracting the effects of Aleutian disease, both by inhibiting replication of the virus and by mitigating the harmful effects of the disease on the fecundity and weight of infected animals. The study included 300 individuals with confirmed infection, divided according to antibody titres into three experimental groups, which received a 20% methisoprinol solution, and three control groups, which did not receive the immunostimulant. In the mink from the experimental groups, the number of copies of the genetic material of the virus in the spleens and lymph nodes was one order of magnitude lower than in the case of the control groups. Mink receiving the supplement also showed higher fecundity (on average 5.83 in the experimental groups and 4.83 in the control groups), and the weight of their offspring before slaughter was over 200 g higher. Given the lack of effective methods for immunoprophylaxis and treatment, methisoprinol supplementation can be an effective means of counteracting the effects of AMDV on persistently infected farms. Electronic supplementary material The online version of this article (10.1007/s00705-019-04375-x) contains supplementary material, which is available to authorized users.

The Aleutian mink disease (AD) is one of the most serious threats to modern mink breeding. In farms where AD is endemic, a significant effect on breeding and reproductive effects is caused by permanent AMDV infection. The aim of this study was investigating the influence of subclinical infection of AMDV on the functional immune status and on the state of postvaccinal immunity against mink distemper. The analyses involved 30 brown mink females aged 12 weeks and allocated into an experimental group (group I) and control group (group II), with 15 animals in each. The minks from group I originated from the farm with an endemic form of Aleutian disease. The control minks from group II originated from the AMDV-uninfected farm. The dynamics of the growth of specific antibodies was examined after vaccination of the minks from both groups against distemper. Serum levels were determined in both groups with a seroneutralisation assay (SN). Obtaining positive values in the SN test in both groups of animals confirmed the induction of a specific immune response. There were significant differences in the magnitude of the positive SN assay values between the minks from group I and group II at all days of determination. The significantly lower SN values noted in the group minks with subclinical course of AMDV reveal that Aleutian disease can cause a suppressive effect on humoral response after vaccination.

The aim of the study was to determine the mechanical and geometric properties as well as bone tissue density of long bones in primiparous and multiparous dams of minks supplemented with β-hydroxy β-methylbutyrate (HMB) and/or 2-oxoketoglutarate (2-Ox) during gestation. Powdered 2-Ox was given at the daily dosage of 0.4 g/kg b.w. separately or simultaneously with HMB, which was administered at the daily dosage of 0.02 g/kg b.w. The study demonstrates for the first time that administration of 2-Ox and/or HMB to dams markedly influences bone tissue density and the mechanical and geometrical properties of mother`s bones in minks. Moreover, it was demonstrated that the supplementation was more effective in the thoracic limb, which was comprehensively used in contrast to the pelvic limb. The mechanical parameters and bone tissue density significantly increased in the humerus in multiparous minks. Only such diet may provide satisfactory production results in the animals. Nutritional deficiencies occurring during pregnancies may trigger body`s own reserves to cover the bone mass increase in developing foetuses and support milk production. This can prevent regeneration of dams’ organisms, which negatively affects their reproductive performance. 2-Ox or HMB may be regarded as a protective metabolite when administered orally to minks, counteracting the negative influences of pregnancy and lactation periods on bones condition. Both simultaneous treatment with 2-Ox and HMB and their separate administration were equally effective.

The aim of the study was to determine the effect of the perinatal period on redox status indicators in the blood of ewes before and after lambing and during lactation. The study was performed on 12 ewes of the synthetic SCP line. Blood for testing of redox parameters was collected seven times: before pregnancy, 1.5 months and 24 h before lambing, 2 and 24 h after lambing, and in the fourth and eighth weeks of lactation. The following blood indices were determined by spectrophotometry: lipid peroxides, malondialdehyde, superoxide dismutase, catalase, plasma total antioxidant capacity, uric acid, urea, bilirubin, and creatinine. The tests showed that during the perinatal period reactions are generated which lead to oxidative stress. Oxidative stress in pregnant ewes was found to increase during the period before lambing and may persist even up to weeks 4-8 of lactation.

Vector-borne infection constitutes a significant health issue in dogs worldwide. Recent reports point to an increasing number of canine vector-borne disease cases in European countries, including Poland. Canine babesiosis caused by various Babesia species is a protozoal tick-borne disease with worldwide distribution and significant veterinary importance. The development and application of molecular methods have increased our knowledge about canine babesiosis, its prevalence, and clinical and pathological aspects of the infection. Parasitologists and veterinary surgeons need an accurate description of the species responsible for canine babesiosis to improve diagnostic and therapeutic methods, as well as predictions for the course of the disease. Therefore, we decided to summarise recent knowledge concerning Babesia species and B. canis.

The purpose of this study was to determine concentrations of IL-2, IL-10, TGF-β1 in serum and T regulatory cell (Treg) percentage in peripheral blood of dogs with perianal tumours. Investigations were conducted on 32 male dogs of mixed breed. The animals were divided into 4 experimental groups and control group. The groups were established depending on the tumour malignancy degree and the type of dominant hormones. All measurements of serum cytokine concentrations were conducted by the use of commercial diagnostic ELISA kits. Treg lymphocyte percentage was measured by flow cytometry. In both groups with benign tumours cytokine levels decreased during therapy, whilst in groups with malignant tumors, in spite of applying anti-tumour therapy, concentrations of cytokines in serum markedly increased. The mean percentage of Treg lymphocytes in dogs with benign tumours (group I and II) was significantly lower than the mean percentage of these cells in control group at all time points, but after applying of anti-hormonal therapy, the significant increase of Treg percentage was observed compared to baseline values. By contrast, in both groups with malignant tumours (group III and IV), the mean percentage of Treg lymphocytes was significantly higher at the beginning of the experiment comparing with the control group as well as both groups with benign tumours and this percentage increased during anti-tumour therapy. The results of this study suggest that monitoring changes in cytokine serum concentrations and Treg percentage in the bloodstream during anti-hormonal therapy may constitute a subsidiary marker in the monitoring of therapy effectiveness, in prognosis the outcome of a disease or in differentiating tumour degree of malignancy.

The aim of the study was to determine total antioxidative capacity (TAC) and zinc concentration in serum of dogs suffering from perianal tumours just before the start of the antihormonal treatment (AHT) and one and six months later. The study was performed on 45 dogs divided into two groups: control group suffering from non-malignant tumours (N = 24) and a group with malignant neoplastic changes (N = 21). Serum TAC and zinc concentrations were measured using photometric and atomic absorption spectrophotometric methods. Six months after the start of the AHT, TAC was significantly lower by 10.6% in dogs with malignant tumours when compared to controls (P = 0.03). In the non-malignant group, serum zinc concentration was higher before the treatment than in the malignant group, while the opposite results were observed six months later (P < 0.001). In the non-malignant group, gradually decreasing values of serum zinc concentration at each stage of the investigation were observed, while the opposite results were obtained in the malignant group (P < 0.05). The obtained results indicate that malignant neoplastic process is associated with significantly reduced TAC. Determination of serum zinc concentration in dogs with non-malignant and malignant perianal tumours may have practical diagnostic and prognostic values and may serve towards increasing the effectiveness of AHT monitoring.

The aim of the study was the evaluation of changes in the percentage profile of CD4+, CD8+, and CD25+ T lymphocytes, and their predictive value with respect to the course of experimental skin burns and early necrectomy in pigs. Thirty Large White Landrace pigs of both genders, weighing 50 kg (±2 kg), were used. Burns to their skin were performed with the use of a computer-controlled heating plate, applied to the animal’s body and heated to 2000°C, using 2.5 kg pressure for 10 s. It produced a burn of 30% (±2%) of body surface with a range of damage between II b° and III°. In animals of each experimental group fascial necrectomy was performed, according to the testing module. Blood from experimental and non-treated control animals was collected from the external jugular vein before the beginning of the experiment (hour 0) and at 12, 24, 36, 48, 60, 72, 84, 96, 108, 120, 132, 144, 156, 168, and 180 h of the experiment. An immune response profile was evaluated using flow cytometry analysis of the level and expression dynamics of CD4+, CD8+, and CD25+ particles on the surface of T lymphocytes. The study demonstrated that experimentally-induced burns in pigs caused cell-mediated immune response reflected in the changes in the percentage of CD4+, CD8+, and CD25+ T lymphocytes, and that early necrectomy in burnt pigs acted in a protective manner for the organism, based on the immunological index values. The study also proved that the dynamics of cell-mediated immunological response intensification determined on the basis of the percentage of CD4+, CD8+, and CD25+ T lymphocytes is conditioned by the size of the burnt surface and the time of necrectomy procedure.

The aim of the study was to evaluate the concentrations of proinflammatory cytokines: tumor necrosis factor (TNF-α) and interleukin-6 (IL-6), anti-inflammatory cytokine interleukin-10 (IL-10), and acute phase proteins (APPs)—haptoglobin (Hp) and serum amyloid A (SAA) in serum and uterine washings of cows with subclinical endometritis, and compare them to healthy animals. The study was performed on 24 cows on day 60 after delivery. The cows were divided into two groups based on the results of cytological tests: 12 cows with subclinical endometritis and 12 healthy cows. Experimental material consisted of blood serum and uterine washings. The levels of the following cytokines in the study material were determined with ELISA: TNF-α, IL-6, IL-10 and APPs - Hp and SAA. The results show that the levels of TNF-α (p

Pyometra is one of the most common diseases of the reproductive system in bitches. The inflammatory processes occurring in the uterus result in changes in concentrations of numerous serum biomarkers, which are used for monitoring the postoperative period. The aim of the present report was to study the evolution of serum levels of insulin-like growth factor-1 (IGF-1) and C-reactive protein (CRP) in bitches suffered from pyometra and after ovariohysterectomy in comparison with the control group of healthy dogs submitted to the surgery. Our results indicate that the serum level of IGF-1 is decreased (101.6 ng/mL [73.8-177.8 ng/mL]), whereas CRP is increased (114.6 μg/mL [95.3-131.6 μg/mL]) during severe inflammation. When inflammation caused by pyometra and surgery is resolved, the level of IGF-1 is increased (186.0 ng/mL [94.6-344.3 ng/mL], P 0.05) and CRP decreased (23.3 μg/mL [9.9-77.0 μg/mL], P 0.01), suggesting that these markers could be useful for assessment of the postoperative period in bitches.

The aim of this study was to assess the impact of asymptomatic infections of mink with Aleutian mink disease virus (AMD on reproductive performance and the morphology of the internal organs. The study drew its subjects from two farms where the disease was endemic and included 450 clinically healthy female mink divided into three equal groups: two AMD positive groups and one AMD negative control group. The mink were subjected to clinical observations throughout the period of reproduction, and then for the first seven weeks after birth. Data collected during this period were evaluated using selected breeding and reproductive indices. All reproductive indices including conception rate, litter size, and litter weight at birth as well as neonatal mortality were substantially poorer. Histopathological examination revealed changes typical for AMD, particularly in the spleen, kidneys, liver, and lymph nodes. The results highlighted and proved the frequently underestimated extent of economic losses on farms where AMD is endemic yet the mink stock is apparently healthy.

Pyometra is a serious problem in dairy cow herds, causing large economic losses due to infertility. The development of pyometra depends mainly on the immunological status of the cow. The aim of the study was a comparative evaluation of selected indicators involving non-specific and specific immunity in cows with pyometra and in cows without inflammation of the uterus. The study was performed in 20 cows, which were divided into two groups: pyometra group and healthy group, each comprising 10 cows, based on the results of cytological and ultrasonographic tests. A flow cytometric analysis was performed for the surface molecules CD4, CD8, CD14, CD21, CD25 and CD4(+) CD25(+) on leucocytes, and the phagocytic activity was determined from granulocytes and monocytes/macrophages in the peripheral blood and uterine washings, respectively. It was demonstrated that the percentage of phagocytic granulocytes and monocytes/macrophages in both the peripheral blood and uterine washings was significantly lower in cows with pyometra compared with the healthy group (p

Endometritis in dairy cow herds is a serious economic problem all over the world due to the large economic losses. The aim of the study was a comparative evaluation of selected indicators of non-specific and specific immunity in cows with subclinical endometritis and in cows without inflammation of the uterus. The study was performed on 40 cows on day 65 after delivery. Based on the results of cytological tests, the cows were divided into two groups: experimental (subclinical endometritis) and control (20 cows in each group). A flow cytometric analysis was performed for the leukocyte surface molecules CD4, CD8, CD14, CD21, CD25. Moreover the phagocytic activity of granulocytes and monocytes/macrophages in peripheral blood and uterine washings was determined. It has been demonstrated that the percentage of phagocytic granulocytes and monocytes/macrophages in both the peripheral blood and uterine washings was significantly lower for cows with subclinical endometritis when compared to cows undergoing a normal puerperal period (p

This study was a comparative evaluation of selected immunological parameters in peripheral blood and uterine wash samples from cows with a normal postpartum period compared with cows with endometritis. We aimed to determine the usefulness of these parameters in monitoring the puerperium. In total, 40 cows were included in the study: 20 had endometritis (experimental group), and 20 did not have uterine inflammation (control group). Animals were chosen on the basis of cytological and bacteriological test results. The tests were conducted 5, 22, and 40 days postpartum. In both groups, flow cytometric analysis of the surface molecules CD4, CD8, CD21, CD25, and CD14 in the peripheral blood and uterine washings was performed. Granulocyte and monocyte phagocytic activity was determined using a commercial Phagotest kit that was adapted for flow cytometry. The percentage of phagocytic granulocytes and monocytes in both the peripheral blood and the uterine washings was significantly lower for cows in the experimental group compared with the control group (P < 0.01). A significant decrease (P < 0.01) in the percentage of CD4+, CD25+, CD14+, and CD4 + CD25(high) leukocyte subpopulations was also observed in the peripheral blood of cows with endometritis. A significant decrease (P < 0.01) in CD21+ lymphocytes and an increase in CD8+ lymphocytes was detected in uterine washings. The results of this work indicate that cell immunity dysfunction may be the main factor causing advanced inflammation of the uterus in endometritis. Knowledge of the immunological mechanisms observed in cows with endometritis might aid in choosing the correct immunomodulating agent-based adjuvant therapy.

There were two aims of this study. One was to evaluate the postpartum state of the reproductive system in cows based on ultrasonography, bacterial culture, and cytological examination of the uterus. The other was to determine whether postpartum endometritis affects the subsequent state of the endometrium, and, in consequence, selected reproductive parameters in cows. The study was conducted on 60 cows: the experimental group of 30 cows with endometritis, and 30 cows free of uterine inflammation (control). The percentage of leukocytes in both groups was similar only on day 5 of postpartum. In all subsequent tests (26, 40, 61 d postpartum), the percentage of leukocytes in the experimental group was statistically significantly higher than in the control (P < 0.001), both in samples collected with a brush and in lavage samples. Involution of the uterus in the experimental group was also slower (P < 0.001). The analysed reproductive parameters were markedly less favourable in the experimental group than in the control. The study showed that postpartum inflammation of the uterus can persist for a long time in the form of endometritis, causing substantial deterioration of reproductive parameters in cows. The authors suggest that cytological evaluation of the uterus, preferably using a brush, should be performed as soon as possible after parturition, even before day 21, up to which time puerperal metritis may still persist. Evaluation of the inflammatory process based on the number of leukocytes and the quality of endometrial cells is important.

The aim of this study was to characterise the genetic variability of the Aleutian mink disease virus (AMDV) circulating among mink farmed in Poland and to compare Polish isolates with AMDV variants available in the GenBank database. For this purpose PCR amplification and analysis of the 429 bp DNA fragment of the AMDV NS1 gene from 13 randomly selected AMDV infected mink was performed. A comparison showed that all tested amplicons were closely related to the sequence of the NS1 gene of AMDV and showed high (94%-97%) homology to virus variants from American mink (Neovison vison) isolated in Canada in 2007-2008. Eleven samples showing a high percentage (95%-97%) of sequence similarity together with three similar isolates originating from Canada formed one clade (monophyletic group). Two variants showing a lower percentage (about 94%- 95%) of sequence similarity to isolates from Canada formed a separate clade. Polish viruses can be subdivided into two main groups with a putative ancestor common to both Polish and three Canadian isolates. This result confirms the literature data indicating the occurrence of American mink in Eastern Europe (including Poland) from the 1950s when the animals were imported for breeding purposes. In conclusion, we provide for the first time a report on the genetic characteristics of the AMDV variants circulating in the Polish population of farmed mink and their relationship with previously known AMDV variants isolated and described abroad.

The experiment was conducted on 30 Holstein-Friesian cows: 10 cows in the follicular phase of the cycle and in the luteal phase 10 d later, 10 cows with follicular cysts, and 10 with luteal cysts. The presence of the ovarian structures was confirmed by ultrasonography. Serum levels of progesterone and 17β-oestradiol were tested with ELISA. Samples for cytological examination were collected from the uterus of all cows using a cytological brush. Following staining, the smears were evaluated in terms of quality and percentages of endometrial cells. In the follicular phase of the oestrous cycle, cells of type A - superficial cells (64.6 ± 4.48) were proportionally the largest group of cells. Cells of type C - basal cells (19.8 ± 2.75) were also present. In the luteal phase, the highest percentage of cells was of type B - intermediate cells (76.9 ± 4.26). When follicular cysts were present on the ovaries, the cytology resembled the follicular phase of the cycle, but with many younger type C cells (33.1 ± 4.11). In the case of luteal cysts on the ovaries, the cytology was similar to that of the luteal phase of the cycle, however with a lower percentage of type B cells (58.1 ± 5.71), and a slightly higher percentage of the other types. The differences in the cytological image of the uterus when different ovarian structures are present, depend on the hormonal activity of those structures. Due to the lack of literature data, the results of the study are important as a model, and may substantially facilitate identification of phases of the oestrus cycle, or the pathologies described, as well as indicate the current status of the endometrium

Expression of CD4, CD8, and CD25 surface markers on T lymphocytes and levels of IFNγ, IL-10, and TNF-α in colostrum and milk were determined in sows vaccinated against Trueperella pyogenes in the final stage of pregnancy. The autovaccine, prepared from Trueperella pyogenes, administered twice to pregnant sows six and three weeks before the anticipated delivery significantly increased the percentages of TCD4+, TCD8+, and TCD25+ as well as levels of IFNγ, TNF-α, and IL-10 in colostrum and milk. The enhanced immune potential of colostrum effectively protected the piglets against T. pyogenes infections during weaning and thus reduced the economic losses on the particular farm concerned, where T. pyogenes infections occur endemically. Knowledge of the profile of cellular and humoral immune response in colostrum and milk of vaccinated sows will enable the design of a T. pyogenes infection prophylactic programme for suckling pigs and weaners, which are most susceptible to infections.

The objective of the study was to evaluate cytometrically the percentage of apoptotic and necrotic spermatozoa in fresh semen of silver foxes in the breeding season. In males F3 and F4 with high percentages of early apoptotic (A+Pi−), late apoptotic (A+Pi+) and necrotic (A−Pi+) spermatozoa as well as 56–65% of living spermatozoa (A−Pi−) with progressive motility, the semen was characterised by reduced fertility. In males F1 and F2 with spermatozoa showing the motility and viability of 89–90% and high percentages of living cells that do not bind Annexin V and propidium iodide, the semen was assessed as valuable and useful for artificial insemination. Amongst 16 females of group I and II inseminated with semen from F1 and F2 males, 15 (93.75%) had multi-cub litters – on average 6.1 and 4.8, respectively. In contrast, amongst 16 females of group III and IV inseminated with semen from F3 and F4 males, only 10 (62.5%) had litters with few cubs (on average 2.6 in group III and 2.1 in group IV). Our findings explicitly indicate that semen of farm male foxes should be evaluated before the breeding season, as one of the causes of reproduction failures is likely to be a high percentage of apoptotic and necrotic spermatozoa. Thanks to flow cytometry, fresh ejaculates can be speedily evaluated and their usefulness for artificial insemination determined.

Parvoviruses exhibit extreme genetic plasticity. The VP2 protein, containing a hypervariable region, is of particular importance. A single nucleotide change in this part of the genome and its effect on the amino acid sequence may significantly affect the range of infected hosts, tropism for specific tissues, or virulence. The high polymorphism in the hypervariable region can be exploited for phylogenetic analysis. The aim of this study was to analyze the polymorphism of the VP2 hypervariable region in isolates of the Aleutian mink disease virus (AMDV) infecting Polish mink farms and to determine the phylogenetic relationships between the Polish isolates and genetic variants of the pathogen occurring in other countries. The study compares farms from two regions of Poland. The isolates contained five changes in the amino acid sequence, which had not previously been recorded in the NCBI database. There were 21 changes noted between the genotypes obtained and the sequence of the reference strain [GenBank NC_001662.1], of which 8 were in the hypervariable region. The isolates identified in our study exhibit a high degree of similarity within the farms, but between farms there is considerable variation in the amino acid sequence of the VP2 protein fragment. Because variants characteristic for farms were obtained, it will be possible to trace the movement of the virus between farms, and in the longer term to use the characteristic sequences as a marker of the origin of infected animals.

The Forkhead genes (transcription complex Fox) play many important roles in the maintenance and determination of biological processes underlying carbon-based life. The expression of Fox genes occurring as a result of reciprocal interactions at the transcriptional level influences the correct function of the immune system as regards the context of general activity of immunological parameters, as well as exposure to aetiological agents. In the case of model organisms and species, in which the knowledge about genomic sequences is incomplete, understanding the above-mentioned transcriptional complex is still insufficient, despite the existence of numerous scientific publications. It is worth noting that Fox genes exist in amphioxus Branchiostoma lanceolatum and in the majority of species they were not characterised. Among many coding sequences included in the transcriptional complex Forkhead, Foxp3, Foxn1, and Foxj1 genes are of a great importance from immunological point of view, being partially jointly responsible for determining features related to productivity. The paper provides general characterisation of selected Fox genes and is an introduction to the presentation of new data, which can be applied at the level of biological differentiation of the populations of domestic and wild animals.

Pyometra is a life-threatening reproductive disorder that affects the uterus of female dogs. This study was designed to identify the possible indicators of uterine inflammation by comparing C-reactive protein (CRP), serum amyloid A (SAA), and haptoglobin (Hp) concentrations in uterine arterial and peripheral venous blood in bitches with open- and closed-cervix pyometra. CRP, SAA, and Hp concentrations were higher in bitches with closed-cervix pyometra irrespective of the site of blood collection. Higher acute-phase protein concentrations were observed in peripheral compared with uterine arterial blood in bitches with closed-cervix pyometra, whereas the levels were comparable in dogs with open-cervix pyometra. Our results indicate that mean acute-phase protein concentrations differ according to pyometra type/severity and blood source and suggest the possible use of peripheral blood levels of CRP, SAA, and Hp to monitor inflammation during the course of pyometra.

Deep crusty dermatophytosis was demonstrated in 10-week-old rabbits naturally and experimentally infected with T. mentagrophytes. Moreover, specific cell-mediated immune response in T. mentagrophytes-infected rabbits occurrd in two phases. During the presence of clinical symptoms of trichophytosis, slightly positive results of leukocyte migration inhibition factor (LMIF) assay were noted, whereas the highest values were observed during spontaneous regression of fungal lesions and complete recovery. The slightly positive LMIF values in rabbits with fungal lesions reflect the mechanism of cell-mediated immune response during T. mentagrophytes infection. On the other hand, the highest positive LMIF values during spontaneous recovery indicate that regression of fungal lesions is attributed to acquired specific cellular immunity. The findings also confirm that the used vaccine against trichophytosis quite strongly stimulates the cell-mediated immune mechanisms, which is confirmed by positive LMIF values in immunised rabbits.

Radiographic examination of the distal radius and ulna of dogs showed changes in metaphyseal areas of both bones characteristic of hypertrophic osteodystrophy (HOD). The serum of the patients showed increased activity of alkaline phosphatase (ALKP) and elevated concentrations of Ca and P. Oral application of vitamin C and injection of sodium salt of hyaluronic acid in the bolus caused a reduction in swelling, disappearance of pain symptoms around the joints, and increased appetite. Radiographic examinations performed on the 60th d of treatment revealed the withdrawal of the changes characteristic for HOD. The chosen biochemical parameters tested during the treatment showed a decrease in comparison to the values before treatment, namely: ALKP by 27.7%, Ca by 22.2%, P by 12.3%, and Mg by 2.4%. Clinical condition of the animals was defined as good or very good. The use of hyaluronic acid and vitamin C in the treatment of HOD in the first stage of the disease proved to be highly successful, resulting in the inhibition of the disease course.

Contents The objective of the study was to assess apoptosis and DNA defragmentation in equine semen diluted and chilled to +4°C. Semen was collected from nine fertile stallions, including four Arabian thoroughbreds and five coldbloods. Examinations were carried out immediately after semen collection (0) and at five storage times (24, 48, 72, 96 and 120 h). The basic semen evaluation was performed in terms of volume, sperm concentration, viable sperm percentage, progressive motility and morphology. Using flow cytometry, DNA defragmentation and cell membrane integrity of spermatozoa were determined. The results of basic tests did not demonstrate significant differences amongst stallions, except for progressive sperm motility, which was significantly higher (p

The paper assesses the treatment of cutaneous fungal infection in young beef cattle caused by Trichophyton verrucosum using orally administered zinc sulphate for 28 d at 2 g per animal per day and subcutaneous application of 2.5 mg/kg levamisole administered three times every 7 d. The study was conducted on animals at the age of 110 to 122 d, which were divided into two experimental groups: group I (animals suffering from ringworm with low levels of zinc in serum) and group II (animals suffering from ringworm with normal levels of zinc in serum). The remaining animals (control group) were divided into three subgroups: K (clinically healthy animals, which had normal serum levels of zinc), KI (animals infected with ringworm and showing a reduced level of zinc), and KII (animals suffering from ringworm with normal serum zinc levels). During the study the following parameters were investigated: clinical signs of fungal infection, zinc content in serum, percentage of phagocytic cells, and phagocytic index. The symptoms of the disease were observed only in group II on day 56. In group I, an increase in the percentage of phagocytic cells was observed, as compared to the original values, on days 7, 21, and 56 and a fall in this parameter was noted on days 14 and 28. In group II, there was a decrease on day 14 and an increase in comparison to the original values, followed by a gradual fall till day 56 of the experiment. In group I, phagocytic index remained on a stable level till day 56 and in group II until day 28, and subsequently began to increase gradually. It was found that oral supplementation of zinc sulphate combined with nonspecific immunostimulation may be applied in fighting skin fungal infection in cattle, and that zinc and levamisole may exhibit synergism affecting the evolution of nonspecific immunity.

The aim of this research is to investigate amphotericin B (AmB)–Cu2+ complexes in aqueous solution at different pH values. Electronic absorption, circular dichroism (CD), Raman and FTIR spectroscopies were used in this study. We found that different concentrations of AmB and Cu2+ ions in solution leads to formation of complexes with stoichiometry of 2:1 and 1:1. The formation of AmB–Cu2+ complexes at physiological pH values is accompanied by changes of the molecular organization of AmB especially disaggregation. These observed effects might be significant from a medical point of view.

Although a large diversity of mastitis-causing bacteria (150 species, subspecies or biovars) have been documented, an adequate and simple mycological screening for yeast-related mastitis has not been developed. Since yeast incubation normally lasts longer than the majority of bacteriological ones, usually a treatment is administered before the results of microbial cultures are obtained. Therefore, a simple test has been needed for detection of yeast-related mastitis, the results of which can be read in parallel to bacteriological screening. Application of Microstix®-Candida assay was first checked for its specificity in standard Candida albicans (Oxford strain) cultures (phase I) and then in the other 36 strains of yeasts isolated from clinical mastitis cases during field investigations (second phase). Next, a test was implemented for identification of pathogens in 9 cohorts (1,200 cows) from red-white PolishFriesian breed in which the dynamics of mastitis were traced by bacteriological and mycological assays along year-long observations. The data obtained showed that apart from frequent infections (~50-70%) proved by using pure microbial cultures, also mixed (bacterial-fungal) infections (0-25%) appeared as well as negative sieves (in the range of 0-45%). Mycological assays revealed a significant fraction of mixed infections during both lactation and the dry period. Strepto-staphylococcal infection was thus identified, however, its proportion strongly decreased after the introduction of antibiotics (after sensitivity to microbial agents was determined). Microbial monitoring was improved in these herds by parallel application of bacteriological and mycological (particularly Microstix®Candida) tests. Microstix®-assay appears to be useful for mycological diagnosis in field conditions of infected (mastitis) herds. Its advantages include easy performance and short incubation time that is only 24 h at 37 °C (72-96 h at room temperature). This allowed demonstrating the cow’s udder yeast infection simultaneously with the first reading of bacterial cultures. Its specificity was confirmed against standard Candida Oxford strain, and sensitivity (100%) was confirmed in mycological cultures (36 strains) and biochemical assays. Bovine mastitis, mycotic mastitis, bacteriological mastitis, cow mastitis diagnosis, Candida assay The presence of bovine mastitis in herds is a natural consequence of interaction among many constitutional and environmental factors taking place within the cow’s mammary gland. Mastitis control and prevention requires an effort in improving the milking system, cattle feeding and hygiene, housing conditions and breeding policy. Currently in Poland, there is no public or centrally managed mastitis control scheme but the economic advantage of improving cow’s udder health is emphasized by dairy farmers, which often motivates them to take their own uncoordinated measures. An epidemiological study by Malinowski (2001) indicated that nearly 50% of milking cows in Poland might have been affected by a subclinical (SCM) or clinical (CM) mastitis, sometimes causing extreme economic losses in cattle farming. Our current knowledge on the aetiology and pathway of this disease marks contagious cases and constitutional non-infectious factors. Primarily, only bacterial infections were considered, and very rarely viral or mycotic ones were recognized.

Contents The development of pyometra in cows depends largely on the state of local immunity of the uterus. The objective of the study was to evaluate the concentration of the following proinflammatory cytokines: tumour necrosis factor (TNF-α) and interleukin-6 (IL-6); anti-inflammatory cytokine: interleukin-10 (IL-10); and acute-phase proteins (APPs): haptoglobin (Hp) and serum amyloid A (SAA), in serum and uterine washings in cows with pyometra and healthy animals. The study was performed on 20 cows divided into two groups based on the results of cytological and ultrasonographic tests: a pyometra and a healthy group (10 cows per group). Experimental material consisted of blood serum and uterine washings. The levels of the following cytokines, TNF-α, IL-6, IL-10 and APPs – Hp and SAA, in the study material were determined by ELISA. The results showed that the values of TNF-α, IL-6, IL-10 as well as SAA and Hp were significantly higher in serum of cows with pyometra compared to controls (p

The aim of the study was to evaluate the level of proinflammatory cytokines (tumor necrosis factor-α [TNF-α], interleukin-6 [IL-6]), anti-inflammatory cytokine (interleukin-10 [IL-10]), and acute-phase proteins (haptoglobin [Hp] and serum amyloid A [SAA]) in serum and uterine washings in cows that developed endometritis during the early postpartum period. The study was carried out on 40 cows. The experimental group consisted of 20 cows with subclinical endometritis and the control group of 20 cows without endometritis. Analyses in both groups of cows were carried out at 5, 22, and 40 days postpartum (DPP). Experimental material consisted of the blood serum and uterine washings. The levels of the following cytokines: TNF-α, IL-6, IL-10 and acute-phase proteins: Hp and SAA were determined using ELISA. Our study reported that the levels of TNF-α, IL-6, IL-10, Hp, and SAA at 22 DPP were higher in cows with subclinical endometritis (P < 0.001). The levels of TNF-α (P = 0.01), IL-6 and IL-10 (P = 0.001), and Hp (P < 0.001) at 40 DPP were higher in cows with subclinical endometritis compared to healthy cows. The level of IL-10 in uterine washings at 5 DPP was higher (P = 0.001), whereas of SAA was lower (P = 0.01) in cows with subclinical endometritis. At 22 DPP, the levels of IL-6, IL-10, and Hp were higher (P < 0.001) in cows with endometritis. At 40 DPP, the level of TNF-α was lower, whereas these of IL-10 and Hp were elevated (P < 0.001) in cows with endometritis compared to healthy cows. The results indicate that the evaluation of the levels of cytokines and Hp in serum, but primarily in uterine washings, can be an important diagnostic indicator in cows that developed subclinical endometritis. High levels of IL-10 in cows with subclinical endometritis may contribute to the weakening of local resistance mechanisms of the uterus and lead to the persistence of the inflammation in the postpartum period. The present study also shows that the simultaneous examination of selected parameters of antagonistic interactions allows for better assessment of the current state of local immunity in the uterus.