Feeder cells can promote cell proliferation and help overcome the developmental arrest of early embryos by producing growth factors. The objective of this study was to evaluate the effects of feeder cells on the development of all single porcine parthenogenetic embryos in vitro. Firstly, we showed that the cleavage and blastocyst formation rate of all single procine parthenogenetic embryos co-cultured with feeder cells increased in contrast to those cultured without feeder cells (p<0.05). However, no statistically significant differences were observed between the blastocyst formation rate in the embryos co-cultured with 3 different kinds feeder cells namely oviduct epithelial feeder cells, granulose feeder cells and porcine fetal fibroblast feeder cells (p>0.05). Secondly, highly significant differences were observed between the cleavage and blastocyst formation rate (p<0.05) when the embryos were co-cultured with oviduct epithelial feeder cells in different volume drops ranging from 3 to 20 μL and the cleavage rate were the highest when cultured in 5 μL drops. Thirdly, the tempospacial pattern of the development of single embryos co-cultured with oviduct epithelial feeder cells was consistent with that of traditional multi-embryo culture, indicating that the co-culturing does not affect the developmental competence of the porcine parthenogenetic embryos. Finally, highly significant differences were observed between the cleavage and blastocyst formation rate with and without zona pellucida in vitro (p<0.05). In this study, a new adaption of in vitro co-culture of single porcine parthenogenetic embryos using feeder cells has been successfully established and this will facilitate further investigations to discover the mechanistic mode of developmental arrest of porcine embryos.

This study investigates the effectiveness of intra-mammary ozone administration in the dry period and at the time of delivery for preventing against mastitis in herds with contagious mastitis. The cows were divided into five groups with 10 cows in each. Group 1 was administered an ozone-containing foam preparation via the teat canal into four udder quarters for 5 seconds at the beginning of the dry period; Group 2 was administered ozone at the beginning of the dry period and at the time of delivery; Group 3 was administered ozone at the time of delivery; Group 4 was administered a dry period udder preparation at the beginning of the dry period; and Group 5 was administered only teat seal at the beginning of the dry period. No statistically significant difference was found between the cows with regard to the SCC values at the beginning of the dry period and at the time of delivery (in cows without clinical mastitis, n=25). The SCC values were reported to decrease when the values at the beginning of the dry period and at the time of delivery were compared. All cows except two in Group 1 were detected to have clinical mastitis according to the frequency of microbial isolation in milk at the time of delivery. In conclusion, intra-mammary ozone administration did not prevent mastitis in the dry period or at the time of delivery in herds with contagious mastitis; moreover, it was determined to increase the rate of clinical mastitis in the postpartum period.

Pseudorabies (PR) outbreaks have devastated many swine farms in several parts of China since late 2011. The outbreak-associated pseudorabies virus (PRV) variant strains exhibited some typical amino acid changes in glycoprotein E (gE), a diagnostic antigen used for discriminating between PRV-infected and vaccinated animals (DIVA). To counteract the potential impact of epitope variations on current serological diagnostics of PRV, we produced monoclonal antibodies (mAbs) against gE protein of one representative PRV variant strain and developed a blocking immunoperoxidase monolayer assay (b-IPMA) for DIVA. The b-IPMA was based on the inhibition of binding between PRV-infected cells and mAb by PRV-specific antibodies present in clinical swine sera and was validated by comparison with a commercial PRV gpI Antibody Test Kit (IDEXX Laboratories, USA). The diagnostic sensitivity, diagnostic specificity and agreement were determined to be 99.25%, 98.18% and 99.02% respectively upon testing 509 serum samples. b-IPMA detected only PRV-specific antibodies and showed no cross- -reactivity with antibodies elicited by gE-deleted vaccine or other common swine pathogens. Thus, b-IPMA has the potential to be used for high-throughput screening of PRV-infected animals in veterinary clinics.

The aim of this study was to determine whether the serum concentration of the phosphate (Pi) and the Ca x P value correlate with the IRIS stage of chronic kidney disease (CKD) in cats and, thus, whether they can be used as markers of the disease progression. Another aim was to assess whether the concentration of Ca in blood needs to be corrected based on the albumin concentration. The study was performed on 165 cats divided into five groups: the healthy group – C and study groups: I, II, III and IV with cats assigned to the groups based on the IRIS scale. Blood was collected from all the animals. The product of Ca x Pi, Cacorr and the product of Cacorrx Pi were calculated based on the obtained results. Despite no differences between groups I-III, there was a clear upward trend in the Pi concentration, in the Ca x Pi and in the Cacorr x Pi with CKD progression. In group IV, the Pi concentration and the Ca x Pi as well as the Cacorr x Pi value were significantly higher than the other groups. The concentration of Ca and its albumin-corrected serum values did not differ significantly. The serum concentration of Pi and the Ca x P product cannot be used as indicators of CKD progression in cats, but they may be used as additional elements in the diagnosis of stage IV CKD. The results also suggest that the serum calcium concentrations do not need to be albumin-corrected in cats.

Since late 2011, porcine infections with highly virulent and antigenic variant of pseudorabies virus (PRV) cause great economic loss in the swine industry in China, and its emergence leads to variable protection efficacy of the commercially available PRV vaccine.

In the present study, the potential cross-protective efficacy of two live virus vaccines, includ- ing a commercial vaccine, and an attenuated low pathogenic PRV variant (rPRVTJ-delTK/gE/gI) against a PRV variant Tianjing (TJ) was evaluated in piglets. Vaccination of piglets with the live vaccine Bartha-K61 could not reduce the clinical signs, and was partially efficacious in the reduc- tion of viral loads upon PRV variant TJ challenge, indicating that this live vaccine provided limited cross-protection efficacy against the PRV variant infection. Additionally, rPRVTJ-delTK/gE/gI appeared to exert some beneficial efficiency in shortening the period of clinical fever and improv- ing the growth performance of the challenged pigs.

Our findings give a valuable guidance for the choice and use of PRV vaccines to control PRV variant infection in the field.

In this study, the effects of oleic (18:1 cis-9-octadecenoic acid) and linoleic (18:2 (n-6), 9,12-octadecadienoic acid) acids added to the embryo culture media for bovine embryonic development after vitrification were investigated in cattle. Following maturation and fertilization, the oocytes were placed in Charles Rosencrans (CR1aa) culture drops containing 10, 100, 500, and 1000 μM of oleic or linoleic acids. On day 7 or 8 of the culture, the blastocysts and expanded blastocysts were vitrified and warmed to evaluate the viability and development. High doses of oleic acid (1000 μM) in the culture media increased the viability of embryos after vitrification. Similarly, linoleic acid at 1000 μM increased the viability compared to the other linoleic acid doses. It was observed that the addition of essential fatty acids improved the development of embryos. Increasing the concentration of linoleic and oleic acid concentrations in the media proportionally advanced the embryonic development and hatching capability after vitrification/warming. Specifically, the addition of high doses of oleic acid had dramatic effects on the embryonic development after vitrification/warming probably due to the increased lipid storage. In conclusion, the present results suggest that the ratio of unsaturated fatty acids in the culture media affects significantly the embryonic development in vitro.

Asthma is one of the most common non-infectious respiratory diseases in horses. Ultrasound examination is a widely available non-invasive additional diagnostic tool. To date, there are no studies focusing on ultrasonographic findings in horses with asthma. The aim of this study was to analyse the prevalence and severity of ultrasound lesions in lung tissue in horses with asthma. Lung ultrasonography was carried out on six healthy horses (controls) and 12 horses with asthma (six with mild and six with severe asthma). The sonographic changes in three lung sections were assessed using a scoring system. The most common changes present in all the animals were comet- tail artefacts. More advanced lesions were present in horses with severe asthma. Statistically significant differences in the overall average intensity of the ultrasound changes were seen between the controls and the study group and between the horses with mild and severe asthma. The lesions were usually located in the caudal lung regions, but they were also present in other areas as the disease progressed. Ultrasonography is a useful additional diagnostic tool enabling an assessment of the stage of the asthma progression. It is a very sensitive technique that visualizes minor lesions in the lung tissue even in clinically healthy animals. Due to its low specificity, it cannot replace endoscopy and the bronchoalveolar lavage in horses with asthma.

Florfenicol is a broad-spectrum bacteriostatic antibiotic commonly used for the treatment of systemic infections in farm animals. The aim of this study was to determine the effect of florfenicol on the percentage of T lymphocytes (CD3+, CD4+, CD8+, TCRgd+ cells) and B lymphocytes (Bu-1+ cells) and on total serum anti - sheep red blood cell (SRBC) haemagglutinin titer in the peripheral blood of SRBC–immunized broiler chickens. The study included three groups of broiler chickens differentiated by weight (0.5, 1.2, 2.4 kg). Florfenicol was administered orally at a dose of 30 mg/kg. The drug was administered eight times at 24 h intervals. The chickens were immunized with SRBC 24 h after administration of the third dose of florfenicol. Florfenicol increased the percentage of CD3+ blood lymphocytes with a corresponding decrease in the percentage of B lymphocytes in birds weighing 0.5 and 2.4 kg. Florfenicol reduced the production of total anti SRBC-haemagglutinins on day 5 after antigen injection in all three body weight groups of the broiler chickens. In conclusion, florfenicol exerted a modulating effect on the immune response of the birds and this should be taken into consideration when using this antibiotic for certain indications.

The current study is the first phylogenetic and secondary RNA structure analysis of Dactylogyrus species parasitising gill filaments of Iraqi cyprinid fishes. Most previous phylogenetic studies have targeted on primary DNA sequence data. Nevertheless, RNA secondary configuration is principally helpful in systematics since they comprise features that do not appear in the primary sequence and provide morphological information. The primary objective was molecular-based identification of Dactylogyrids species using evolutionary tree and secondary RNA structure prediction. A total of 681 fish were collected from the Lesser Zab River in the northeast of Iraq in the sub-district of Altun-Kopru from August 2016 to September 2017 and brought to the Zoology Research Laboratory, Salahaddin University-Erbil, Iraq. All fish were classified as 18 cyprinid species. The species of Dactylogyrus were identified by the 28S rDNA subunit using PCR and sequencing methods, and the obtained nucleotide sequences were then compared with the available GenBank sequences. Phylogenetic relationships were concluded using Neighbour-Joining (NJ), Maximum Likelihood (ML), and Minimum Evolution (ME) methods. The results justify the validation of 11 Dactylogyrus species (three of them were newly recorded in Iraq). Additionally, out of nine infected fish species, seven of them were regarded as a new host for Dactylogyrus species. Secondary RNA configuration prediction using minimum free energy was considered as a hopeful tool for species identification. This was considered the first comprehensive phylogenetic study in the area. It was concluded that PCR sequencing, phylogenetic and secondary RNA analysis were proper molecular methods for identifying Dactylogyrids species on the gills of fishes.

Progesterone (P4) is responsible for the main reproduction processes. Concentration of P4 varies widely among different determination methods, and interpretation of these values may be difficult. The objective of the current study was to assess the agreement of three different enzyme immunoassays (ELISA) in relation to radioimmunoassay (RIA) of P4 concentration assessment of beef cow serum samples. Samples were collected randomly considering high (pregnant cows) and low (non-pregnant cows) P4 concentrations. Depending on the P4 assessment method, four groups were created as follows: Group 1 – direct samples assessed by ELISA, Group 2 – extracted samples assessed by ELISA, Group 3 – samples assessed by automated ELISA, and Group 4 – samples assessed by RIA.

The mean progesterone concentration was 4.50 ng/mL, 1.24 ng/mL, 4.07 ng/mL and 4.39 ng/mL from Group 1 to Group 4, respectively. The mean difference (MD) between Group 1, Group 2 and Group 3 individually compared with Group 4 was −0.10 ± 1.24 ng/mL, 3.15 ± 3.58 ng/mL and 0.33 ± 1.42 ng/mL, and the 95% confidence interval (CI) for the differences (s) was from −0.99 to 0.78 ng/mL, from 0.59 to 5.71 ng/mL, and from −0.69 to 1.34 ng/mL, respectively. The confidence interval for the lower and upper limit of the agreement ranged from −4.12 to −1.05 ng/mL and from 0.84 to 3.91 ng/mL between Group 1 and Group 4, from −8.45 to 0.42 ng/ mL and from 5.88 to 14.75 ng/mL between Group 2 and Group 4, from −4.29 to −0.76 ng/mL, and from 1.41 to 4.94 ng/mL between Group 3 and Group 4.

Our findings show that the best agreement with RIA was observed for Group 1 and Group 3, while the agreement in the extraction method was least accurate.

The objective of this study was to determine the association between subclinical acidosis (SARA) and subclinical ketosis (SCK) with biomarkers from an automatic milking system (AMS) measuring in relation to rumination time (RT), milk yield (MY), bodyweight (BW), milk temperature, the milk fat-to-protein ratio, and the electrical conductivity of milk at the udder quarters-level which can be read in fresh dairy cows. During the course of the study, all of the fresh dairy cows (n=711) were examined according to a general clinical investigation plan. The cows were selected for 1-30 days of milk (DIM) and were milked using Lely Astronaut® A3 milking robots with free traffic. Rumination time shows a statistically significant positive correlation with milk yield (milk temperature) and is negatively correlated with the fat and protein ratio. Healthy cows demonstrated the highest level of rumination time and the lowest milk temperature. The average BW for these cows was 1.64% lower than for the SARA group and the BW kg was 2.10% higher than SCK cows. MY was 14.01% lower in comparison with SARA and 6.42% higher in comparison with SCK. According to these results, some biomarkers from the AMS have an association with SARA and SCK. However, further research with a higher number of cows is needed to confirm this conclusion.

Insulin receptor substrate 2 (IRS-2) modulates the phosphatidylinositol 3-kinase (PI3K)/Akt pathway, which controls the suppression of gluconeogenic genes; IRS-2 is also a critical node of insulin signaling. Because of the high homology between pig and human IRS-2, we investigated the expression pattern and function of porcine IRS-2. QPCR and immunoblotting were used to detect the IRS-2 expression level in different tissues. There were high IRS-2 levels in the cerebral cortex, hypothalamus, and cerebellum in the central nervous system. In peripheral tissues, IRS-2 was expressed at relatively high levels in the liver. Immunohistochemistry analysis revealed that IRS-2 was mainly distributed in the hypothalamus and cerebral cortex. Furthermore, IRS-2 knockdown porcine hepatocytes and porcine aortic endothelial cells (PAECs) were generated. The IRS-2 knockdown induced abnormal expression of genes involved in glycolipid metabolism in hepatocytes and reduced the antiatherosclerosis ability in PAECs. In addition, we disrupted IRS-2 in porcine embryonic fibroblasts (PEFs) using the CRISPR/Cas9 genome editing system, before finally generating IRS-2 knockout embryos by somatic cell nuclear transfer (SCNT). Taken together, our results indicate that IRS-2 might be a valuable target to establish diabetes and vascular disease models in the pig.

Postoperative adhesion (POA) is a common and well-known complication with an estimated risk of 50-100%. The antioxidant effect of n-acetyl-cysteine (NAC) can increase intracellular glutathione levels, thereby reducing adhesion. This study was conducted to compare the outcomes of NAC nanoparticles (Nano-NAC) on intra-abdominal adhesion (IAA) after laparotomy in rat. A total of 25 male Wistar rats were randomized into five groups: 50 mg/kg Nano-NAC, 75 mg/kg Nano-NAC, 150 mg/kg Nano-NAC, NAC and control. During the surgical procedure, some sections (2×2cm) were collected through abdominal midline incision to ensure the infliction of peritoneal damage by a standard adhesion. Macroscopic evaluation was performed on the 14th and 28th day and blood samples were collected to evaluate the inflammatory factor (C-reactive protein) on days 0, 14 and 28. According to the serologic results (CRP test), C-reactive protein was at highest level in 150 mg/kg Nano-NAC and control groups and at lowest level in 50 mg/kg Nano-NAC and 75 mg/kg Nano-NAC groups (p<0.001). The macroscopic evaluation results showed that frequency of adhesion bands was significantly lower in 50 mg/kg Nano-NAC group than the control at the intervals. Results showed that the intraperitoneal administration of lower Nano-NAC dosages (50 and 75 mg/kg) had a major role in the management of postoperative inflammation. Nano-NAC administration was proved feasible, safe and effective in reduction of the C-reactive protein level.

Early lactation period in dairy cows could be harmful to their health since it is challenging and demanding. Proinflammatory cytokine concentrations are increased in the early phase of the inflammatory response and during the early lactation period in cows. The aim of this study was to determine if ketoprofen treatment in the first days following parturition would decrease proinflammatory cytokine concentration and their correlation between lipid mobilization, ketogenesis and metabolic parameters in cows. The study was conducted on 30 cows divided into two groups of 15 cows each. The experimental group was treated with 3 mg × kg.bw.-1 ketoprofen for three consecutive days after parturition. The blood samples were collected on the first day of treatment and in the first and second week postpartum and they were analyzed for biochemical parameters such as non-esterified fatty acids (NEFA), beta-hydroxybutyrate (BHBA), glucose, cholesterol and total bilirubine and inflammatory parameters such as tumour necrosis factor-α (TNF-α), interleukin-1α (IL-1α) and interferon-γ (IFN-γ). The results suggested that ketoprofen- treated cows had a significantly lower concentration of TNF-α, IL-1α, IFN-γ, NEFA and BHBA in the first and second postpartum week compared to the control group. Ketoprofen administration increased glucose levels (the first week, p<0.05), increased cholesterol levels (the second week, p<0.01) and decreased serum total bilirubin levels (second week, p<0.01) compared to the control group of cows. A positive correlation was found between TNF-α and NEFA and total bilirubin, significantly more expressed in the control than in experimental group of cows (p<0.01) and it was also found between IL-1α and NEFA (p<0.01). A negative correlation was found between TNF-α and glucose and cholesterol, significantly more expressed in the control than in experimental group of cows (p<0.01). A positive correlation was also found between IL-1α and glucose (p<0.01). Ketoprofen given parenterally to Holstein cows immediately after calving could reduce inflammation and decrease the relation between inflammatory response and lipogenesis and ketogenesis in postpartum cows.

Reference intervals (RIs) are one of the essential elements in the procedure of disease diagnosis. This is especially true for feline species in which RI is less available than in canine species. RIs are affected by biological, geographical and instrumental factors, yet published RIs with incomplete background are popularly used. Inappropriate interpretations of RIs may affect classification of disease and subsequent treatment. In this study, we demonstrated the step-by-step establishment of feline RIs following the American Society for Veterinary Clinical Pathology (ASVCP) reference interval guideline. A total of 51 parameters were examined, including 20 hematology and 31 biochemistry parameters, and the results were compared to one local RI and two foreign RIs. Overall, about 29% (10/35) of tested parameters were different form local RIs and 60% (30/50) were different from the two foreign RIs, highlighting geographical variations. A higher upper reference limit (URL) in red blood cell count (RBC), hematocrit (Hct), Hemoglobin (Hgb), albumin, creatinine and lower URL in potassium and white blood cell count (WBC) were identified, which may impact the interpretation. In addition, statistical analysis of age and gender were factored separately and indicated that 10 parameters were significantly higher in the adult group. For the impact of gender, percentage of basophil and total iron-binding capacity (TIBC) were lower in female and male cats, respectively. In conclusion, we have demonstrated that it is desirable to establish in-house RIs or RIs of local sources. An age specific RI for the geriatric feline population is advisable for better diagnosis and monitoring the disease.

In our recent study we demonstrated that the holding of fresh semen in fractionated seminal plasma (SP1, >40 kDa; SP2, <40 kDa), obtained by gel filtration chromatography, significantly improved the sperm quality characteristics following cryopreservation (Wasilewska-Sakowska et al. 2019). In this study we investigated the effect of post-thaw (PT) supplementation of fractionated SP (SP1 and SP2) on the survival of spermatozoa from boars with good and poor semen freezability, GSF and PSF, respectively. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) analysis showed distinct differences in the protein profiles of SP1 and SP2 from boars with GSF or PSF regarding the number of protein spots. Sperm motility characteristics and the motion patterns, assessed using the computer-assisted sperm analysis (CASA) system, were markedly higher in PT semen supplemented with SP1 and SP2 from boars with GSF. Post-thaw supplementation of either SP1 or SP2 from boars with GSF significantly improved mitochondrial function, plasma membrane and acrosome integrity, and viability during storage. The findings of this study have confirmed that the presence of protective protein components in varying abundance in either fractionated SP from boars with good freezability ejaculates significantly improved the sperm survival following PT storage.

The study was aimed to develop an indirect enzyme-linked immunosorbent assay (ELISA), which can detect specifically Feline herpesvirus type 1 (FHV-1). The primers were designed based on the conserved sequence of FHV-1 glycoprotein B gene. The recombinant protein with reactogenicity was purified as coating antigen of the assay. The indirect ELISA, characterized by high sensitivity showed no cross-reaction with two types of feline virus, had detection limit at 1:2000 dilution. The positive rate of the assay, according to the determined cutoff value (0.25), was basically consistent with Feline Herpes Virus Antibody ELISA kit. In conclusion, the indirect ELISA with high repeatability and reproducibility can be used for detecting FHV-1, and can provide necessary support to related research.

The aim of the study was to find out whether carriers of new lethal mutation in SDE2 gene occur in the population of Polish Holstein-Friesian bulls. Eighty seven bulls were included in the analysis. Bulls were selected as having in the pedigree known carrier of SDE2 mutation (bull Mountain USAM000002070579). All bulls were diagnosed by PCR amplification of 524 bp fragment of SDE2 gene followed by digestion of Bcc I restriction enzyme. Heterozygotes (carriers) were confirmed by sequencing. Each new carrier was used to trace another potential carriers among its offspring available in Polish Holstein Bull Repository Database. Among 87 bulls, 50 new SDE2 carriers were found. The study has shown that mutation in SDE2 gene causing early embryo mortality is already transmitted to Polish Holstein-Friesian cattle. The results are sufficient to initiate the screening program to reveal new carriers and to avoid further spreading of SDE2 lethal mutation.

Therefore, the aim of the present study was to evaluate the possible effect of bilberry fruit (Vaccinium myrtillus L.) supplement in a daily diet on the cognitive behaviour of the rats and the expression of paravalbumin (PV) in populations of hippocampal neurons. It has been postulated that the antioxidants present in bilberry fruit may act as neuroprotective factors playing also a significant role as memory enhancements. Forty Wistar rats with a similar average body weight (460 ± 0.4 g) were divided into four groups (n=10 per group). The control group received standard feed (210 g/week), whereas animals of experimental groups received standard feed supplemented with bilberry (per os) at consumed doses of 2 g (group I), 5 g (group II), and 10 g/kg b.w./ /day (group III). After three months of feeding with bilberry, the modified elevated plus-maze test (mEPM) was performed. After 32 weeks of feeding, brains were collected and PV-immunoreactive (ir) neurons were immunohistochemically visualized. In the modified elevated plus-maze test, transfer latency examined 2 h and 24 h after the acquisition session was significantly shorter (p<0.05) in the group II in comparison with the control group. In CA1 and CA2/CA3 hippocampal fields as well as dentate gyrus of all experimental groups, a significant (p<0.05) decrease in number of PV-ir neurons were found. In relation to the control group, the mean subpopulation of PV-ir neurons found in groups II and III were significantly reduced. The subpopulations of PV-ir neurons found in DG of all experimental groups were significantly reduced in comparison to the control. In conclusion the in the present paper we demonstrated a relationship between the diet rich in a bilberry fruit and process of memory as well as numbers of calcium- binding protein-expressing hippocampal neurons. Our results may be source of basic knowledge for further research aiming at neuroprotective role of the bilberry fruit.

αB-crystallin is a member of a small family of thermal shock proteins that protects cells from stress. Because of lack of its expression in peripheral blood leukocytes, it was proposed as a molecular marker of circulating tumor cells in canine mammary gland tumors. The aim of the present study was to determine if αB-crystallin shows stability of expression, what is the requirement for this type of marker. It was also assessed whether there is co-expression of αB-crystallin with the basal marker, cytokeratin 17. For this purpose, samples of various types of canine mammary gland tumors of epithelial origin, were selected. Using RT-qPCR, we have found αB-crystallin and cytokeratin 17 co-expression in benign and malignant canine mammary gland tumors. It has been demonstrated that the expression of αB-crystallin in tested neoplastic samples is not stable in comparison to the control group. Furthermore αB-crystallin overor down- expression was associated witch the same cytokeratin 17 pattern. αB-crystallin can be a marker of circulating tumor cells in the bloodstream, but for cancers in which basal marker expression occurs and thus not universal for all cancers originating from the mammary gland tissue.

The aim of this study was to evaluate the influence of the newly isolated Lactobacillus plantarum LUHS135 and Lactobacillus paracasei LUHS244 strains grown in potato juice (with a cell count of 8.0-9.0 log10 CFU/ml) on the blood and faeces parameters of exercising horses. The horses were classified into four different groups: a control group (which received no probiotics); the first group (which received 200 ml of L. plantarum culture in potato juice); the second group (which received 200 ml of L. paracasei culture in potato juice); and the third group (which received an L. plantarum and L. paracasei mix (with the mix consisting of 100 ml of each). Indices for the blood and faeces microflora were obtained before and after treatment of horses (on days zero and thirty). It was observed that the count for lactic acid bacteria (LAB) in the faeces was significantly higher on day thirty, whereas it was lower when it came to the total enterobacteria count (TCE). Despite the ambiguous influence of any treatment on blood parameters, the L. plantarum × L. paracasei mixture increased the concentration of HGB and O2 saturation in blood samples which were taken from the horses. L. paracasei significantly decreased the lactate concentration levels in horse blood samples. As a result of the present study, it can clearly be seen that the strains being used revealed their potential application as probiotics; however, further studies are required to prove the survival and action mechanisms of the newly isolated strains.

The Intrauterine fetal development process is complicated and affected by many regulating factors such as maternal nutritional status, transcription factors and adipokines. Adipokines are kinds of active substances secreted by adipose tissue, including more than 50 kinds of molecules. To explore the correlation between calf birth weights and adipokines including adiponectin, leptin, visfatin, and IGF-1 in cows venous and venous cord blood. Fifty-four healthy multiparous Chinese Holstein cows were used; in which, cows with a calf weight less than 40 kg were included in group A (n=9); those with a calf weight between 40 kg~45 kg were included in group B (n=25) and ≥45 kg were included in group C (n=20), venous blood and cord venous blood was collected. An ELISA kit was used to evaluate the concentration of adiponectin, leptin, visfatin, and IGF-1, correlations between index-index and index-calf birth weight were analysed. In both cows venous and cord venous blood, adiponectin, leptin, visfatin, and IGF-1 levels were significantly correlated with each other (p<0.01), and levels of these adipokines in venous blood were significantly higher than cord venous blood (p<0.01). Adiponectin, leptin, visfatin, and IGF-1 in venous cord blood were positively correlated with calf birth weights, and significantly correlated with calf birth weights respectively (p<0.01). Our study showed that adiponectin, leptin, and IGF-1 were found in venous blood and cord venous blood, and adiponectin, leptin, and IGF-1 in venous and cord venous blood potentially inter-regulated each other; adiponectin, leptin, and IGF-1 in venous blood were not significantly correlated with calf birth weights, while adiponectin, leptin, visfatin, and IGF-1 in venous cord blood were significantly correlated with calf birth weights, respectively.

Newcastle disease (ND) is a highly contagious and economically important disease in the poultry industry caused by avian avulavirus-1, historically known as Newcastle disease virus (NDV). Control of ND primarily relies on prophylactic vaccination of flocks, and many vaccines are available on the market, both conventional and more recently introduced new generation recombinant types. To assess the protection level achieved by vaccination ELISA tests are typically used, they also are to track an infection with field strains in non-vaccinated flocks. Special modifications of ELISA can be used as a screening tool to detect infection in flocks vaccinated with new generation vaccines. In this study, we have developed an ELISA test for the detection of antibodies against the nucleoprotein (NP) of NDV and for differentiation of chickens vaccinated with commercial and prototype in-house recombinant vector vaccines from those infected with field NDV strains. The NP gene of LaSota NDV strain expressed in a baculovirus vector was used as a coating antigen in the ELISA. The developed test was optimized, validated and compared to other serological tests. The sensitivity, specificity and accuracy of recombinant NP protein-based ELISA were respectively 96.1%, 96.3%, and 96.2%. Inter-rater (kappa) agreement between the NP-ELISA and the gold standard HI test was calculated to be 0.995. In our comparisons, commercially available ELISA tests revealed different specificities ranging from 95.5–100% and sensitivities at variance, ranging from 90.1 to 99.0%. A high level of maternally derived antibodies was measured in the serum of 1-day-old broilers in the NP-ELISA assay. These antibodies had disappeared and were undetected at 3, 5 and 6 weeks post-vaccination but birds became positive again at 2 weeks after control infection with a velogenic NDV strain. In SPF chickens, antibodies against NP protein were detected only after a challenge. The recombinant NP protein-based ELISA test is sensitive, specific and accurate when compared to the gold standard HI test and commercially available kits. Moreover, the method could be also used for the differentiation between vaccinated and infected birds.

A negative energy balance is a common condition in high yielding dairy cows causing the production of ketone bodies (KB), including beta-hydroxybutyrate (BHB), defined as subclinical ketosis (SCK) if clinical signs are missing. The aim of the present study was to evaluate a handheld electronic device for the detection of SCK (BHB-concentration > 1.2 mmol/l), in capillary blood and venous whole blood in cows (WellionVet BELUA, MED TRUST Handels GmbH, Marz, Austria) as well as the feasibility of the puncture of the external vulva with a single use lancet. For this purpose, the blood BHB-concentration was tested in 250 venous and capillary blood samples and compared to the results of a certified laboratory. The majority (76.3%) of the animals displayed no signs of discomfort related to the puncture and in 74.2% the procedure was successful on the first attempt. The BHB-concentrations detected in capillary blood showed good agreement with the reference method, both in capillary (correlation coefficient 0.94 (p<0.001), Kappa-value 0.89) and venous whole blood (correlation coefficient of 0.95 (p<0.001), Kappa-value 0.89). Altogether, 98% of all the samples were correctly classified as SCK or non-SCK by the handheld device in capillary blood (sensitivity 0.96, specificity 0.98) and 97.4% in venous whole blood (sensitivity 0.889, specificity 0.991), respectively. An increase in the correlation by the adaptation of the cut off level could not be achieved for both sampling sites.