Polish Journal of Veterinary Sciences

The number of human cases of salmonellosis in the EU was 94,625 in 2015. Considering the source of these infections, Salmonella spp. was most frequently detected in broiler chicken meat and Salmonella Enteritidis (SE) was the most commonly reported serovar.

The efficacy of probiotics in limiting Salmonella spp. infection in poultry has been demonstrated in numerous papers. The administration of probiotics at the level of primary production reduces the risk of contamination of poultry food products with Salmonella spp.

A study was carried out in order to determine the potential for reducing the Salmonella spp. population in broiler chickens with the use of the Lavipan (JHJ, Poland) probiotic that comprised selected stains of lactic acid bacteria and Saccharomyces cervisae.

Salmonella spp.-free broiler chickens were divided into two groups and received the same feed with (group L) or without (group C) the probiotic throughout the experiment. All day-old chickens were infected per os with SE. Samples of cecum content were collected 2, 4, and 6 weeks after SE infection and pectoral muscles were collected 6 weeks following SE infection for the evaluation of the SE population number. Serum samples for serological examinations were collected 6 weeks after infection.

Six weeks after infection, the number of SE-positive cecal samples was lower in the L group (12.5% positive) in comparison to the C group (87.5%). Similar results were demonstrated for the muscle samples (25% in contrast to 87.5%). At the same time, in both cases, the SE CFU/g was significantly lower in the L group. The results of our study indicate that Lavipan was capable of reducing the population of SE in the gastrointestinal tract, which eventually improved the hygienic parameters of the pectoral muscles.

Four weeks after infection, SE was not detected in any of the experimental groups. In both groups, no specific anti-SE antibodies were detected.

The aim of this study was to analyse and identify specific buffalo seminal plasma proteins (SPPs) responsible for sperm cryotolerance during low temperature storage. Computer Assisted Sperm Analysis (CASA) of the motility and viability of buffalo spermatozoa was performed before freezing and after thawing. Two sample groups were formed – ejaculates with high cryotol- erance (group A) and low cryotolerance (group B). CASA demonstrated that the initial progres- sive motility after thawing of the spermatozoa in group A is significantly higher than in group B (p<0.001). Group B showed a significant increase in the percentage of static and non-progressive spermatozoa at 240 min, when compared to group A (p<0.05). SPPs, proteins in the cryoprotec- tive medium (PM) and proteins in the mixture of PM and SP were separated by High Perfor- mance Liquid Chromatography (HPLC). Comparative analysis of the chromatographic profiles was performed to identify specific proteins related to sperm cryotolerance. SPPs profiles showed 5 distinct protein peaks in both groups, ranging from 500 kDa to 50 Da. Chromatograms of group A and group B showed quantitative and qualitative differences in protein content. Chromato- grams of proteins in PM showed 11 well-expressed peaks. HPLC analysis of the mixtures of SPPs from the two groups and PM visualized the formation of a new bio-complex structure expressed by a protein peak specific for group A (7.674 min, AU 1.50). This protein peak can be referred as a phenotypic trait for buffalo ejaculates with high sperm cryotolerance.

The study was conducted on 26 male, 30 days-old goats, separated from their mothers, divided into two equal groups: I - control and II - experimental, consisting of 13 animals each. All animals were fed with milk replacer, experimental group received additionally 50 g/kg body weight, additive of HMB, for 60 days. The following features were analyzed: body weight, daily increases of body weight, as well as hematological and biochemical blood features. Differences in body weight were found, between experimental and control group, after 60 days of experiment 0.57 kg (p≤0.01). The daily weight gain of experimental animals was higher in comparison with control group. Significant differences were also noted in results of hematological and biochemical blood parameters. Experimental animals showed a higher level of red blood cells as well as number of lymphocytes in comparison with the control group, (p≤0.01).Significant changes were also observed in the level of triglycerides, inorganic phosphorus and protein between both groups. The acid-base balance parameters and ionogram, showed a higher pH level (p≤0.05) HCO – act., HCO – std., BE, ctCO , O sat, K+, Cl– (p≤0.01), while the anion gap (AG) and Na+ were significantly lower in control group (p≤0.01).

Background: Equine sarcoids are the most common neoplasms in horses. Bovine papilloma- virus type 1 (BPV-1) is the main viral type identified in equine sarcoids in Europe.

Objective: The aim of the present study was to genetically evaluate BPV types based on DNA analyses of the CDS of the L1 gene. The presence of BPV DNA was confirmed by Degenerate Oligonucleotide-Primed Polymerase Chain Reaction (DOP PCR) with FAP59/FAP64 consensus primers.

Results: The DNA was detected in 21/40 (52.5%) of clinically diagnosed sarcoids. More than half of 14 isolates (66.7%) shared 100% homology with BPV-1 Deltapapillomavirus 4 isolate 09 asi UK (Acc. No. MF384289) and 99% nucleotide identity with BPV-1 isolate EqSarc1 (Acc. No. JX678969). A comparison with BPV-1 isolate EqSarc1 revealed one silent mutation in C5827T which did not change the aminoacid codon. The remaining 6 isolates (28.6%) shared 100% nucleotide identity with the BPV-1 (Acc. No. X02346) “wild type” isolate, and 1 isolate (4.8%) demonstrated 99% nucleotide identity with BPV-2 (Acc. No. M20219).

Conclusions: Variants of BPV-1 isolate EqSarc1 (Acc. No. JX678969) constitute the most prevalent type of BPV-1 in Polish horses.

The aim of the study was to compare anaesthesia protocol utilizing combination ketamine/ medetomidine with epidural lidocaine or morphine to orthopaedic surgery in rabbits. All rabbits received intramuscularly medetomidine (0.5 mg/kg) and ketamine (20 mg/kg). In group L (n=13) epidural injection was performed with lidocaine (3 mg/kg) and in group M (n=13) with morphine (0.1 mg/kg) diluted with sterile saline to 0.6 ml. Rabbits underwent bilateral mosaicplasty surgery. Heart rate (HR), systolic (SBP), diastolic (DBP) and mean arterial blood pressure (MAP), arte- rial oxygen saturation (SpO2) and respiratory rate (RR) were measured every 5 minute during surgical procedure. Duration of sensory blockade was measured based on toe pinch reflex and recorded.

Hemodynamic parameters were comparable at baseline in both groups. We did not observe significant differences in HR, RR, SBP, DBP, MAP between group L and M. Intragroup statistical analysis revealed differences at different time points in group M in terms of HR, SBP, DBP, MAP. In rabbits in group M blood pressure dropped while HR increased over time. In both groups a decrease in SpO2 was observed. Mean duration of loss of the toe pinch reflex was similar between group L and M, and amounted to 72.79±34.48 and 82.80±17.85 minutes, respectively.

The present study aimed to evaluate the efficiency of hypertonic saline solution (HSS) as a novel treatment of acute ruminal lactic acidosis (ARLA) in cattle, focusing on urinary excretion of acids. Twelve cannulated steers were submitted to experimentally induced ARLA by adminis- tering sucrose into the rumen. Twenty hours later, the cattle were randomly divided into two equal groups. The first group was treated with 7.5% HSS (5 mL/kg) over 15 min, and isotonic saline solution (ISS; 20 mL/kg) for the subsequent 165 minutes. The control group was administered ISS instead of HSS. Rumen and urine samples were collected at different times during the experiment from the baseline to 64 h post-induction. The induction caused a medium-to-moderate ruminal acidosis, and a moderate degree of systemic acidosis and dehydration. Steers treated with HSS increased by 50% its glomerular filtration rate (1.61 mL/min) compared to ISS group (1.06 mL/ min; p<0.03). The overall volume of urine excreted by HSS group was higher than that in ISS group (1.62 L vs 0.7 L; p<0.02). This increase in total volume of urine provided by HSS favored a greater excretion of H+ ions in urine, which was 3.39-fold higher in HSS group (64.3*10-7 vs 18.9*10-7 Mol) as well as lactate (241.7 vs 181.8 mMol) and P urinary excretion (3.8 vs 1.1 mMol) that reduced the urine pH (5.3 vs 5.7). Only the HSS group decreased significantly blood total lactic acid concentration (20.3 %) throughout the treatment. A positive relationship was found between the excretion of urinary phosphorus and urinary pH (r2=0.562). The results showed that this novel treatment with HSS enhanced renal excretion of acids and may be recommended as an additional treatment for cattle with lactic acidosis.

The aim of this study was to obtain reference values for diastolic cardiac function parameters in healthy dogs and to ascertain if significant differences exist between dogs of various age, weight and sex. The study was performed on 82 healthy dogs of different age and breed. Eleven param- eters were analyzed: peak velocity during early diastolic filling, acceleration time of early diastol- ic filling, deceleration time of early diastolic filling, total time of early diastolic filling, peak veloc- ity during late diastolic filling, acceleration time of late diastolic filling, deceleration time of late diastolic filling, total time of late diastolic filling, total time of early and late diastolic filling, ratio of peak velocities during early and late diastolic filling, isovolumetric relaxation time. The Dop- pler measurements used for general assessment of diastolic function in healthy dogs were signifi- cantly influenced by body weight, heart rate and age. No significant differences were found be- tween males and females. This study described the value of non-invasive echocardiographic assessment of diastolic function in healthy dogs.

In individual dogs, despite good quality of raw sperm, some parameters are significantly changed after thawing, which cannot be predicted. We therefore investigated whether motility parameters objectively obtained by CASA, membrane integrity (MI), cell morphology or a combination are suitable to improve the prediction of bad post-thaw quality. For this purpose 250 sperm analysis protocols from 141 healthy stud dogs, all patients introduced for sperm cryopreservation, were evaluated and a Classification and Regression Tree (CART) -analysis performed. The sperm was routinely collected, analysed, and frozen by using a modified Uppsala system. After thawing, data were routinely examined by using CASA, fluorescent microscopy for membrane integrity (MI) and Hancock’s fixation for evaluation of cell morphology. Samples were sorted by post-thaw progressive motility (P) in good (P > / = 50%, n=135) and bad freezers (P<50%, n=115). Among bad freezers, 73.9% showed in addition post-thaw total morphological abberations of >40% and/or MI <50%.

Bad freezers were significantly older than good freezers (p<0.05). Progressive motility (P), velocity curvilinear (VCL), mean coefficient (STR), and linear coefficient (LIN) were potential predictors for post-thaw sperm quality since specifity was best (85.8%) and sensitivity (75.4 %) and accuracy (80.4 %) good. For these objectively measured raw sperm parameters, cut-off values were calculated allowing prediction of bad post-thaw results with high accuracy: P = 83.1 % VCL = 161.3 µm/sec, STR = 0.83 %, and LIN = 0.48 %. Raw sperm samples with values below these cut off values will have below average post-thaw quality with a probability of 85.8%. We conclude that VCL, P, STR and LIN are potential predictors of the outcome of sperm cryopreservation, when combined.

Bacteria from the Simkaniaceae family are intracellular parasites belonging to the Chlamydiales order, detected in surface waters, drinking water, chlorine water, and in wastewater. Its main representative, Simkania negevensis, is pathogenic to humans and animals, especially fishes, as it principally causes respiratory tract diseases. Bacteria from this family are also capable of surviving and existing in free-living amoebas, omnipresent in the natural environment, which makes them an additional risk for human and animal health. The aim of the present study was to search for representatives of this family in freshwaters from the Odra River and two municipal lakes (Rusałka and Goplana). Out of 100 water samples analysed, the sequence of bacteria of Simkaniaceae family was found just in 1 percent, because phylogenetic analysis revealed that the obtained OdraWCh30 sequence shows 93% similarity to Simkania negevensis strain Z as well as 87% similarity to Candidatus Syngnamydia salmonis isolate Ho-2008 and Candidatus Syngnamydia salmonis isolate VS10102006 and 84-85% similarity to endosymbiont of Xenoturbella westbladi, Simkaniaceae bacterium clone SM081012-5s and Candidatus Syngnamydia venezia strain Pi3-2. This is the first case of detecting sequence of bacteria of Simkaniaceae family in the aquatic environment in Poland.

The aim of this study was to determine the effects of factors such as sex, eye-side and diurnal variation on intraocular pressure (IOP) values measured by Tono-Pen Vet® in healthy calves. Twenty four (12 males, 12 females) calves were used in this study. IOP measurements were per- formed in the morning (6:00 a.m.) and at night (8:00 p.m). Average IOP values in all calves were measured as 17.67±2.64 mmHg in the morning and 15.52±2.05 at night and the difference between these two time points was found to be statistically significant (p<0.0001). Average IOP values were measured as 16.04±2.82 mmHg in males and 17.15±2.23 mmHg in females with a statistically significant (p<0.05=0.023) difference between them. Average IOP values were 16.81±2.85 mmHg in right eyes and 16.37±2.23 mmHg in left eyes and the difference between these values were statistically insignificant (p>0.05). At the end of study, overall average IOP in all calves was measured as 16.59±2.59 mmHg. The present data showed a significant difference in terms of sex and diurnal variations and a non-significant difference in eye-side. In addition, this study is the first research article in which the intraocular pressure in calves was measured by Tono-Pen Vet®.

In the current study, twenty lambs, aged 4 months, half male and half female, were classified into four groups, with five in each group. The experimental three groups of lambs were given intravenous (IV), intramuscular (IM) and subcutaneous (SC) administrations of recombinant ovine interferon-τ (roIFN-τ). The fourth group (normal control) of lambs was given normal saline injections in the same way. After administrations, blood samples were collected from the tested animals at different time points post injection, and the serum titers of roIFN-τ were measured using cytopathic effect (CPE) inhibition bioassay. The results of calculating pharmacokinetic (PK) parameters using DAS software showed that the PK characteristics of roIFN-τ through IV injection conformed to the two-compartment open model, whose half-life of distribution phases (T1/2α) was 0.33±0.034 h and the elimination half-life(T1/2β) was 5.01±0.24 h. However, the PK features of IM injection and SC injection of roIFN-τ conformed to the one compartment open model, whose Tmax were 3.11±0.26 h and 4.83±0.43 h, respectively, together with an elimination half life(T1/2β) of 9.11±0.76 h and 7. 43±0.58 h, and an absorption half-life (T1/2k(a)) of 1.13±0.31 h and 1.85±0.40 h, respectively. The bioavailability of roIFN-τ after IM administration reaches 73.57%, which is greater than that of SC administration (53.43%). These results indicate that the drug administration effect can be preferably obtained following a single dose IM administration of the roIFN-τ aqueous preparation. This study will facilitate the clinical application of roIFN-τ as a potential antiviral agent in future work.

The purpose of the study was to study the activity of the phytoestrogen genistein (GEN) act- ing on FSHR and LHR in rat ovaries with polycystic ovary syndrome (PCOS). Sixty rats were di- vided into six groups. Rats in the dose group received genistein at a concentration of either 5 (low genistein dose group, L-gen), 10 (middle genistein dose group, M-Gen) or 20 (high genistein dose group, H-Gen) mg per kg of body weight per day. Estrogen group (EG, received 0.5 mg/kg Dieth- ylstilbestrol). Concentration of sex hormones in serum was quantified by enzyme-linked immuno- sorbent assay (ELISA). Expressions of follicle-stimulating hormone receptor (FSHR) and lutein- izing hormone receptor (LHR) protein were determined by immunohistochemistry. Treatment with genistein resulted in a strong stimulation of the concentration of sex hormone in serum. The concentration of progesterone and FSH was significantly higher in H-Gen when compared to the PCOS model control group (MG) (P < 0.01). In contrast, the concentration of testosterone, LH and the ratio of LH/FSH decreased in GEN treatment groups compared to MG, the effect was statistically significant, tested by the ANOVA test (p<0.01). For hormone receptor activity, treat- ment with genistein resulted in an improvement of ovarian function with LHR protein expression being enhanced and FSHR protein expression being suppressed. Our results demonstrate that Genistein played a significant role in regulating FSH and LH receptor expression to improve perimenopausal ovarian and uterine function.

Animals as a source of organs and tissues for xenotransplantation could become a backup solution for the growing shortage of human donors. The presence of human xenoreactive anti- bodies directed against Galα1,3Gal antigens on the cell surface of a pig donor triggers the activa- tion of the complement leading to a hyperacute reaction. The development of genetic engineer- ing techniques has enabled the modification of genomes by knocking in and/or knocking out genes. In this paper, we report the generation of modified pigs with ZFN mediated disruption of the GGTA1 gene encoding the enzyme responsible for synthesis of Galα1,3Gal antigens. ZFN plasmids designed to target the exon 9 region of the pig GGTA1 gene encoding the catalytic domain were injected into the pronuclei of fertilized egg cells. Among 107 piglets of the F0 gene- ration analyzed, one female with 9-nt deletion in exon 9 of the GGTA1 gene was found. 13 of 33 piglets of the F1 generation represented the +/- GGTA1 genotype and 2 of 13 F2 piglets repre- sented the -/- GGTA1 genotype. No changes in the animals’ behavior, phenotype or karyotype were observed. Analysis confirmed heredity of the trait in all animals. A complex functional analysis of the modified animals, including flow cytometry, human serum cytotoxicity test and immunohistochemical detection, was performed to estimate the phenotype effect of genetic modification and this indicated an efficient GGTA1 knock-out in modified pigs.

European beaver (Castor fiber), the largest rodent species inhabiting a wide area of Eurasia, feeds mainly on dry parts of plants, bark or wood. Such kind of nourishment needs to be properly digested in each part of the gastrointestinal tract. The time of stomach digestion, which directly influences all the following steps of the digestion process, is precisely controlled by the pylorus and its innervation. However, virtually no data is available on the organization of the enteric nervous system in most of the wild animal species, including beavers. On the other hand, a pecu- liar diet consumed by beavers, suggests that the arrangement of their stomach intramural nerve elements can be atypical. Therefore, the present study investigated the distribution and chemical coding of neurons and nerve fibers in the pylorus of the European beaver.

The experiment was performed on stomachs obtained from a group of 6 beavers caught in Northeastern region of Poland (due to beaver overpopulation). Pyloric wall tissue cryosections were double immunostained with a mixture of antibodies against pan-neuronal marker PGP 9.5 (to visualize enteric neurons) and ChAT (cholinergic marker), nNOS (nitrergic marker), SP, CGRP, Gal (peptidergic markers).

Confocal microscopy analysis revealed that the majority of enteric nerve cells were clustered forming submucosal and myenteric ganglia and all the studied substances were expressed (in various amounts) in these neurons.

We conclude, that the anatomical arrangement and chemical coding of intramural nerve elements in the beaver pylorus resemble those found in other mammalian species.

Introduction: Platelet-rich plasma (PRP) preparations can be used in bone tissue healing but there are numerous doubts among clinical orthopedists about effectiveness of this method.

Materials and methods: The studies were carried out in 12 rabbits of white termond breed. In operating room we operationally generated cylindrical, unicortical defects of the diameter of 4 mm in the middle of the shafts of both femurs. The defects in the left bones were left without filling and served as controls, and 0.7 ml of the ready-to-use PRP was administered to the defects in the right bones (experimental group). We evaluated the usefulness of the diagnostic methods applied: biomechanical tests, micro-CT tests, densitometry, typical radiology, macroscopic measurements, histopathological examinations.

Results: The macroscopic measurements showed a statistically significant increase in the dimension in the area of the right defect filled with PRP in relation to the control group. In experimented group, the assessment of the X-ray images showed the formation of a callus cuff around the defects. Densitometric examinations showed no statistically significant differences between defects in the experimental and control group. The analysis of the micro-CT examina- tions showed an increase in the total volume of the tissue examined (Vb) and the low density tissue fraction (Vb2) in the experimental group. The biomechanical examinations revealed signi- ficant decrease in the maximum breaking force (F max) necessary to break the bone in the experi- mental group in relation to the control group.

Conclusions: Platelet-rich plasma (PRP) stimulates bone formation in the area of bone defects and may accelerate bone regeneration.

The application of immune serum is one of the most efficient method used formerly in the protection of raised piglets’/weaners’ health . The objective of the study was to determine specific antibody response during hyperimmunization of fatteners with a self-prepared subunit vaccine, and to propose production method of immune serum against Gram-negative bacteria antigens. The vaccine was administered every two weeks, 4 times. Individual and pooled serum samples were assayed for IgM, IgG and IgA antibodies against Histophilus somni recombinant Hsp60, H.somni rOMP40 and Pasteurella multocida LPS. Additionally total serum IgG and haptoglobin concentrations were measured.

Two weeks after the first vaccination IgM antibody raised significantly against H.s. rOMP40 and LPS, whereas after 4 weeks it increased against rHsp60 antigens. Anti-LPS IgM antibody raised up stepwise till the end of the observation, but IgM antibody against H.s. rHsp60 and H.s. rOMP40 decreased in further samplings. A significant raise in IgG class H.s. rHsp60-

-antibody was found 4 weeks after the first immunization and a similar raise against two remain- ing antigens after 6 weeks. The intensity of the reaction increased till the end of the experiment. The raise in IgA antibody level was observed only for H.s. rHsp60 antigen. Clinically observed, proper animal health and welfare were confirmed by haptoglobin concentration, which remained in physiological range. At least 4 booster doses were necessary to obtain hyperimmune serum containing a high level of antibodies against examined antigens. The number of immunizations influenced response profiles for specific IgM, IgG, IgA antibodies.

Early castration of male small ruminants is regarded as a risk factor for urolithiasis, although the underlying correlations are still unclear. One possible reason is a deferred development of the penis and the urethra after castration. Therefore, we examined the penis and urethra of castrated and intact lambs by ultrasonography to determine the correlation between urethral area and pe- nile cross-sectional area. Ultrasonography was performed in 6-month-old Lacaune crossbred lambs (early castrated, late castrated, and intact; each group, n = 11). Sectional images at 5 loca- tions (glans penis, penile urethra, distal and proximal sigmoid flexure, and ischial arch) were ob- tained to determine the urethral and penile diameters. Urethral and penile cross-sectional areas were calculated. Grey-scale analysis of ultrasound images was performed to evaluate possible differences in the penile texture between the groups. Correlation analyses between both cross-sectional areas showed a significant general correlation for location 2 in all lambs (R = 0.52; P = 0.003), for location 3 in late-castrated lambs, and for location 5 in early-castrated lambs. Statistically significant correlations between the penile and the urethral area of castrated and intact lambs were not evident. Therefore, measurement of the penile cross-sectional area alone does not allow for accurate estimation of urethral size. Statistically significant differences con- cerning the grey-scale analysis between the groups were also not detectable. Thus, simplification of the formerly presented ultrasonographic examination of the urethra is not recommended. In animals at a risk of obstructive urolithiasis, complete urethral examina- tion is essential.

Characterisation of copy number variation (CNV) and loss of heterozygosity (LOH) has pro- vided evidence for the relationship of this type of genetic variation with the occurrence of a broad spectrum of diseases, including cancer lesions. The role of CNVs and germinal or somatic LOHs in canine mammary tumours is still unknown. Therefore, the aim of this study was to identify CNVs and LOHs in canine mammary tumours. Forty-eight samples obtained from normal (n=24) and tumour (n=24) tissues of dogs were analysed. In the study, we used CanineHD BeadChip assay (Illumina) and OncoSNP software to identify copy number alternations in genomes of dif- ferent dog breeds and in different mammary cancer types occurring in this species. The analyses revealed that, in the case of CNV, the amplification-type variants were longer and more frequent than deletions. Based on the analysis of the frequency of different types of aberrations in the in- dividual parts of the genome, regions that are particularly susceptible to structural aberrations were indicated. The fraction of genes identified within these regions was associated with major processes of neoplastic transformation. Association analysis of such traits as tumour grading as well as the size and age of dogs demonstrated that structural aberrations were more frequent in dogs diagnosed with tumour malignancy grade II and III, in dogs with a larger body size, and in large dogs aged 7-8. The promising results of these pioneering investigations prompt continuation thereof to analyse other types of cancer.

Osteocalcin is a major non-collagenous component of the bone extracellular matrix and is considered to be an indicative factor of osteoblast differentiation. In the present study, we detected osteocalcin expression in different antler areas and growth phases by immunohisto- chemistry. Osteocalcin was highly expressed in all areas during the mineralization period and in mesenchymal cell and chondrocyte areas during the rapid growth period. The nucleotide sequence of the osteocalcin gene in sika deer antler was determined. The open reading frame was 303 bp encoding a protein of 100 amino acids. The estimated molecular mass of osteocalcin was 10.38 kDa and the theoretical isoelectric point was 5.37. The osteocalcin gene with a 6× His-tag at the C-terminus was cloned into the pGEX-4T1 vector and expressed in Escherichia coli under optimal conditions. The recombinant soluble protein fused with GST was purified with Ni-NTA resin. The purified osteocalcin protein exhibited a significant increase in HA adhesion and promoted antler chondrocyte proliferation. Osteocalcin is an important factor in regulating the rapid growth and differentiation of deer antlers.

The present study investigated the expression of androgen receptor (AR) in neurons of the anterior pelvic ganglion (APG) and celiac-superior mesenteric ganglion (CSMG; ganglion not involved in the innervation of reproductive organs) in the male pig with quantitative real-time PCR (qPCR) and immunohistochemistry. qPCR investigations revealed that the level of AR gene expression in the APG tissue was approximately 2.5 times higher in the adult (180-day-old) than in the juvenile (7-day-old) boars. Furthermore, in both the adult and juvenile animals it was sig- nificantly higher in the APG than in CSMG tissue (42 and 85 times higher, respectively). Immu- nofluorescence results fully confirmed those obtained with qPCR. In the adult boars, nearly all adrenergic (DβH-positive) and the majority of non-adrenergic neurons in APG stained for AR. In the juvenile animals, about half of the adrenergic and non-adrenergic neurons were AR-posi- tive. In both the adult and juvenile animals, only solitary CSMG neurons stained for AR. The present results suggest that in the male pig, pelvic neurons should be considered as an element of highly testosterone-dependent autonomic circuits involved in the regulation of urogenital func- tion, and that their sensitization to androgens is a dynamic process, increasing during the prepu- bertal period.

The objective of this study was to evaluate the effect of a second prostaglandin F2α (PGF2α) treatment during Ovsynch on luteal regression and fertility in dairy cows, compared with standard Ovsynch. The study was conducted on 111 Holstein Friesian multiparous cows on commercial dairy farm. The cows in the experimental group (n=48) received two treatments of PGF2α 24 hours apart during Ovsynch. The cows in the control group (n=63) were synchronized with standard Ovsynch. To assess the progesterone (P4) concentration blood samples were collected at the day of PGF2α treatment and at the 2nd GnRH treatment. Pregnancy was evaluated by ultrasound examination 37-40 days after timed artificial insemination (TAI) by ultrasound. Cows diagnosed pregnant were re-examined between days 70-80 after TAI. The percentage of cows with complete corpus luteum (CL) regression (P4<0.5 ng/ml at the time of the 2nd GnRH treatment) was 89.6 % after two PGF2α treatments and 88.9 % after one PGF2α treatment. There were no statistically significant differences (p>0.05) in the pregnancies per artificial insemination (P/AI) between the experimental and control group (P/AI). However, the pregnancy loss rate was lower in cows receiving two PGF2α treatments than in the control animals (0.0 % vs. 6.4 %; p<0.05). In conclusion, the second PGF2α treatment during Ovsynch protocol had no significant effect on CL regression and P/AI in dairy cows. The pregnancy losses until days 75-80 after TAI were significantly lower after two PGF2α treatments than after one PGF2α treatment.

Duck viral hepatitis (DVH) is an acute and fatal disease of young ducklings characterized by rapid transmission and damages. The most important agent of DVH is duck hepatitis virus 1 (DHV-1). The effective control of DVH was achieved by active immunization of 1-day-old duck- lings with an attenuated DHV-1 virus vaccine. However, the attenuated virus might reverse to virulence. In this study, a DHV-1 strain, Du/CH/LBJ/090809, was identified and its genomic se- quences were determined. The genome of Du/CH/LBJ/090809 is composed of 7,692 nt excluding poly A and the virus was clustered into genotype A by comparing with other referenced DHV-1 strains. Du/CH/LBJ/090809 could lead to 30% mortality of 10-day-old specific pathogen free (SPF) ducklings. The virus was passaged serially in SPF chicken embryonated eggs and three vi- ruses, passage 16 (P16), P29 and P40, were selected for genomic analysis. P29 and P40 were used to evaluate the attenuation in duckling by inoculating the virus to 10-day-old SPF ducklings. Re- sults of vaccination-challenge assay showed that the inactivated virus P40 could evoke protection against the pathogenic parent virus. Nucleotide and amino acid sequences of the genomes of Du/ CH/LBJ/090809, P16, P29 and P40 were compared. Changes both in nucleotides and amino acids, which might be contributed to the decreasing in virulence by chicken embryo-passaging of DHV- 1, were observed. We speculated that these changes might be important in the adaption and at- tenuation of the virulent virus. Additionally, strains obtained in this study will provide potential candidate in the development of vaccines against DHV-1.

The aim of this study was to investigate interactions between conventional antifungal drug and essential oils against isolates of Malassezia pachydermatis. Antifungal activity of Cinnamomum cassia, Melaleuca alternifolia, Mentha piperita, Origanum vulgare and Syzygium aromaticum essential oils were tested against 19 strains of M. pachydermatis isolated from healthy dogs and reference strain M. pachydermatis CBS 1879. The checkerboard assay was used to search for in- teractions. Synergism was observed for the combination of clotrimazole with Melaleuca alternifolia essential oil, Mentha piperita and Origanum vulgare. The combinations of Cinnamomum cassia and Syzygium aromaticum essential oils with clotrimazole showed indifferent effect. Additive antimicrobial activity was observed for the combination of clotrimazole with Syzygium aromaticum and Melaleuca alternifolia essential oils against reference strain. The obtained results showed synergistic interactions between essential oils and clotrimazole which could improve effectiveness of this antifungal drug.

The aim of the study was to investigate the effect of soybean lecithin as a substitute for egg yolk in milk and tris based extenders in ram semen cryopreservation. Twenty ejaculates were col- lected from four healthy, mature Wrzosówka rams (2-3 years of age). Each ejaculate was divided into four equal aliquots and diluted with four different extenders: 1) milk extender containing 5% egg yolk, 2) milk extender containing 1.5% soybean lecithin, 3) tris extender containing 20% egg yolk, 4) tris extender containing 1.5% soybean lecithin. Extended semen was loaded into 0.25 ml French straws, cooled and frozen in liquid nitrogen vapor. Total motility, curvilinear velocity, plasma membrane integrity and fertilizing ability of sperm were assessed after thawing. Total mo- tility was lower (p<0.05) in tris-soybean lecithin extender when compared to other extenders. Curvilinear velocity was higher (p<0.05) for spermatozoa cryopreserved in milk-soybean lecithin extender compared to other extenders tested. For the percentage of live sperm no significant difference was observed between extenders. The lambing rate were higher (not statistically signifi- cant) in ewes inseminated with semen doses frozen in milk-soybean lecithin extender (42.9%) than in the tris-egg yolk extender (16.7%). In conclusion, replacing the egg yolk with soybean lecithin was effective in milk but not in tris extender.

In this study the quality of total RNA, isolated from fresh spermatozoa, was compared between boars with good and poor semen freezability (GSF and PSF, respectively). Semen from 3 boars with GSF exhibited significantly higher total motility, mitochondrial function, plasma membrane integrity and reduced lipid peroxidation compared with 3 boars with PSF after cryo- preservation. There were variations in the quality of RNA isolated from spermatozoa of boars with GSF and PSF. Boars with GSF exhibited mainly full-length, intact RNA, whereas substantial amounts of degraded RNA were detected in spermatozoa from boars with PSF. Further under- standing of the biological relevance of RNAs in sperm function is critical to improve the freezabil- ity of boar semen.

Senecavirus A (SVA) the only member of the Senecavirus genus within the Picornaviridae family, is an emerging pathogen causing swine idiopathic vesicular disease and epidemic transient neonatal losses. Here, SVA strain (CH-HNKZ-2017) was isolated from a swine farm exhibiting vesicular disease in Henan Province of Central China. A phylogenetic analysis based on complete genome sequence indicated that CH-HNKZ-2017 was closely related to US-15-40381IA, indica- ting that a new SVA isolate had emerged in China.