Polish Journal of Veterinary Sciences

Cytological evaluation of bone marrow smears stained by May-Grünwald Giemsa method was performed. The smears came from 20 fallow deer (Dama dama) 3 days old divided into 2 groups each consisting of 10 animals. The experimental group (E) received intramuscularly selenium and vitamin E at a dose of 3.0 ml (tocopherol acetate – 50 mg, sodium selenite – 0.5 mg, solvent - 1 ml) in the 3rd day of age. The control group (C) did not receive any supplementation or placebo. For hematological analyzes blood was collected three times: on 0, 15th and 25th day of the experiment. Serum concentration of selenium and vitamin E was determined using high perfor- mance liquid chromatography and glutathione peroxidase activity (GSH-Px) by kinetic method. On the 15th day after supplementation, a statistically significant increase in the percentage of erythroblastic cell line was observed in bone marrow smears. At that time, the increase in GSH-Px activity in the E group was also observed, reaching the value of 165.3 U/gHb, which was statisti- cally significant. The percentage of proerythroblasts (8.23% in group E and 5.02% in group C) differed significantly between groups at the 25th day after supplementation. This study revealed that supplementation of selenium and vitamin E resulted in an increase in the number of erythro- cytes to an average of 13.5 (˟ 10¹²/l) in the experimental group on 25th day with a significant increase in hemoglobin to 193 g/l in the experimental group.

There are several infectious agents of domestic cattle that can also be present in free-living ruminant populations. These include bovine herpesvirus 1 (BoHV-1) and bovine viral diarrhea virus (BVDV) which are the causative agents of infectious bovine rhinotracheitis and bovine viral diarrhea, respectively. The study was conducted on serum samples from 59 red deer, 24 roe deer, and 3 fallow deer (86 in total), originating from two geographically separate areas of Poland. The samples were tested with commercially available ELISA tests for BoHV-1 and BVDV. The overall seroprevalence was 5.8% and 3.5%, respectively. All positive samples originated exclusively from red deer. Because of BoHV-1 ELISA cross reactivity with cervid herpesvirus 1 and 2 (CvHV-1 and -2) the nature of alphaherpesviruses infecting the sampled animals could not be assessed.

Perinatal calf mortality in dairy herds has been reported worldwide. The etiology of stillbirth is multifactorial, and can be caused by various species of bacteria and environmental factors. Among them some potential pathogens from the Mollicutes class such as Mycoplasma (M.) spp. and Ureaplasma (U.) diversum can be isolated from the bovine genital tract and other organs of the suspected cattle. The aim of this study was to evaluate if the bacteria belonging to the Molli- cutes class i.e. M. bovis, M. bovigenitalium, M. canadense, M. canis, M. arginini, M. bovirhinis, M. dispar, M. alkalescens and U. diversum could have an impact on perinatal calf mortality in selected Polish dairy farms. The material was: 121 stillborn calves (SB), 21 live born calves (C) and 131 cows (dams) from 30 Polish Holstein-Friesian herds. Samples were examined from all the SB calves’ and six control euthanized calves’ abomasal contents and lung samples collected during necropsy, and from the dams’ serum and placenta. In dams the serological ELISA, and in calves and placenta samples molecular PCR/denaturing gradient gel electrophoresis, methods were used. Screening of dams’ sera for antibodies to M. bovis (ELISA) showed seven dams positive for M. bovis, whereas none of the nine examined Mollicutes microorganisms were detected in the placenta and calves.

The aim of the study was to establish normal ranges for chosen biochemical parameters of haemolymph of snails (Gasropoda: Mollusca), in the light of the use of these animals as experi- mental models in various types of studies. The study was conducted on 100 specimens of Cornu aspersum maxima (CAM) and 100 specimens of Cepaea nemoralis (CN). The haemolymph col- lected from the animals was analysed using colorimetry to assay aspartate transaminase (AST) activity, alanine transaminase (ALT) activity, amylase activity and the concentrations of urea and triglycerides. In the further part of the study, the influence of administering doxycycline with feed on the change of AST and ALT activity in snail haemolymph has been studied. The normal values established for CAM are as follow: AST activity: 26-38 u/l, ALT activity: 0-11 u/l, amylase activity 9-16 u/l, concentration of urea: 3-6 mg/dl, concentration of triglycerides: 16-20 mg/dl. For CN, the following data have been obtained: AST activity: 30-80 u/l, ALT activity: 0-15 u/l, amylase activity 12-15 u/l, concentration of urea: 5-8 mg/dl, concentration of triglycerides: 18-24 mg/dl. It has been shown that doxycycline presents a high workload on the hepatopancreas of snails, which is reflected by a statistically significant (p<0.05) increase of AST and ALT activity in the haemolymph of the specimens which obtained doxycycline in feed, as compared to the groups with antibiotic-free feed. The haemolymph activity of both studied parameters increased together with study time and tetracycline administration time.

In this paper we present the first cases of rabbit haemorrhagic disease virus 2 (RHDV2 – GI.2) in Poland. The virus was detected in liver samples of RHD-suspected rabbits from Lodzkie and west Pomeranian voivodeships. In both cases, the typical clinical symptoms of the disease were observed despite the fact that the rabbits were previously vaccinated against RHD. In order to extend the analysis of the RHDV2 strain infecting the rabbits, the entire VP60 and NSP genes were amplified and sequenced. The results of rRT-PCR assay have shown that tested RHDV samples were positive for the presence of RHDV2. In the phylogenetic analysis of vp60gene the first Polish RHDV isolates (RED 2016 and VMS 2017) clustered together with the reference RHDV2, meaning they represent new evolutionary RHDV linkeages. The first Polish RHDV2 isolates showed about 97% nucleotide sequence identity with the reference RHDV2 strains and approximately 18% difference from classic RHDV and RHDVa variants.

Recent outbreaks of adenovirus (FAdV) infections in poultry flocks have been determined in many countries in Europe, Asia and Australia connected with economic consequences, and loses in poultry production. To better understand the evolution and transmission of FAdV viruses, de- tailed codon usage analysis was performed for 137 recently obtained FAdV strains. A high effec- tive number of codons, and an indication the presence of low codon usage were determined. The presence of mutations, and their influence on codon usage was confirmed by a correlation be- tween nucleotide compositions at the 3rd codon positions, HVRs1-4, and ENCs. This presence indicate some influence of natural selection, and antigenic properties of examined FAdV strains.

Calretinin (CR), a calcium-binding protein from EF-hand family, is localised in non-pyramidal GABA-ergic interneurons of the hippocampus. CR takes part in maintaining calcium binding homeostasis, which suggests its neuroprotective role. Hippocampal neurons contain membrane transient receptor potential vanilloid 1 (TRPV1) which binds to capsaicin (CAP) contained in habanero pepper fruits. Few in vivo studies have revealed the effect of CAP on interneurons containing CR. The aim of the present study was to investigate the CR immunoreac- tivity in interneurons of the hippocampal CA1 field and dentate gyrus (DG) in adult rats after intragastric admin- istration of the habanero pepper fruits. Wistar rats received a peanut oil – control group (C), and oil suspension of habanero pepper fruits at doses of 0.025 g dm/kg b.w. – group I and 0.08 g dm/kg b.w. – group II for 28 days. After euthanasia, the brains were collected and embedded in paraffin blocks using a routine histological tech- nique. Frontal hippocampal sections were immunohistochemically stained for CR by using a peroxidase-antiper- oxidase method. CR immunoreactive (CR-IR) interneurons were morphologically and morphometrically ana- lyzed under a light microscope. The results showed similar shapes and distribution of cells in both areas of the brain in group C and I of animals. However, CR-IR interneurons in the hippocampal CA1 field and in DG were occasionally observed in the group II of rats.

The results of morphometric studies did not reveal statistically significant differences in the surface area and shape index of cells between examined brain regions from groups I and II compared to group C.

Only in group II of rats, an increase in the digital immunostaining intensity of CR-IR interneurons was found in DG. Low number of CR-IR interneurons in the hippocampal CA1 field and in the DG, under the influence of a large dose of habanero pepper fruits containing CAP, may be caused by the activation of TRPV1 receptors and the increase in Ca2+ ions in these cells. This phenomenon may ultimately lead to neuronal death and may disturb neuronal conduction.

Despite the consensus on the role of lung and pleura ultrasound in human medicine, veteri- nary medicine questions credibility of the pulmonary evaluation in ultrasound examination, based on the analysis of artifacts in animals with clinical signs of respiratory failure and possibility of pulmonary edema diagnosis with recognition of the degree of its severity. The study was conduct- ed on 47 animals (29 dogs and 18 cats) of different breeds, age and sex. In all of animals prior to the transthoracic lung and pleura ultrasound examination (TLPUS), all animals were subjected to a clinical examination and hematological blood test as well as chest radiography examination in three projections. Ultrasound imaging of the chest in each animal was performed at designated four defined segments. TLPUS in dogs and cats based on an analysis of artifacts allows recogni- tion of pulmonary edema, to the degree comparable to chest X-ray examination. The number of depicted B-lines artifacts is proportional to the degree of pulmonary edema. These results allow to reduce the number of radiographs and allow the shortening of the diagnostic process for pa- tients in life-threatening condition.

This work reports on cadmium and lead contaminations in the edible snail Helix pomatia harvested in Poland. One hundred and 24 samples of Helix pomatia meat collected from seven provinces (voivodeships) of Poland were analyzed for their trace metal levels by graphite furnace atomic absorption spectrometry (GFAAS). The research was conducted in 2 stages. The 1st stage analyzed snail meat prior to any further technological treatment (raw meat). In the 2nd stage, the trace element levels were measured in meat subjected to technological treatment (processed meat). The trace element contents in raw meat samples ranged from 0.06 mg kg-1 to 0.22 mg kg-1 for Cd and from 0.06 mg kg-1 to 0.18 mg kg-1 for Pb. The analyses revealed an increase in the cad- mium content from 0.12 mg kg-1 to 0.18 mg kg-1 in thermally treated snail meat and no changes in lead concentration during the two-stage heat treatment. Regulation (EC) 1881/2006 does not specify the Cd and Pb residue limits in meat of terrestrial edible snails. The limits are set for in- vertebrate aquatic organisms meat (i.e. shellfish, mollusc, cephalopod) and range from 0.5 mg/kg to 1.5 mg/kg of tissue fresh weight for Pb and from 0.5 mg kg -1 to 1 mg kg-1 for Cd (EU Commis- sion 2006). The results demonstrate that the land snail Helix pomatia has a tendency to bioaccu- mulate trace elements, and the cooking process is likely to affect (increase) the Cd content in the snail meat.

In order to understand infection of avian influenza A virus (AIV) and canine distemper virus (CDV) in the Siberian Tiger in Northeast China, 75 Siberian Tiger serum samples from three cap- tive facilities in northeastern China were collected. AIV and CDV antibody surveillance was test- ed by using hemagglutination inhibition and serum neutralization methods. The results showed that the seroprevalence of H5 AIV, H9 AIV and CDV was respectively 9.33% (7/75), 61.33% (46/75) and 16% (12/75). In the 1<years <2 and > 5 year-old group, the seroprevalence of the H9 AIV was 24% and 80% (P < 0.01), and the CDV seroprevalence was 6% and 36% (P < 0.01), respectively. It was demonstrated that 3 (4%) out of 75 serum samples were AIV+CDV seropos- itive, with 2.67% (2/75) in H9+AIV and 1.33% (1/75) in H5+H9+AIV. To our knowledge, this is the first report of AIV and CDV seroprevalence in Siberian Tigers in China, which will provide base-line data for the control of AIV and CDV infection in Siberian Tigers in China.

Current study was designed to investigate the protective effects of royal jelly on Flunixin me- glumine (FM)-induced spermiotoxicity related to sperm concentration, abnormal spermatozoa count and histopathological changes in mice testis. The subjects were divided into five groups according to FM and/or royal jelly intake: Control group; group 1, FM alone (25 mg/kg, im); group 2, combination of FM (25 mg/kg, im) and royal jelly (200 mg/kg, oral); group 3, FM alone (50 mg/kg, im); and group 4, combination of FM (50 mg/kg, im) and royal jelly (200 mg/kg, oral). The animals were fed once daily for 15 days and they were sacrificed last day. Epididymal sperm concentration and abnormal spermatozoa count were noted. Testicular histological findings were evaluated. On purpose, organization of each animal was graded according to Johnsen’s scoring to assess the spermatogenesis relying on seminiferous tubule cross-section scores. Comparing to controls, FM administration caused a decrease in sperm concentration (p<0.05), an increase in total abnormal spermatozoa rates (p<0.05) and more degenerative changes in testes in mice.

Royal jelly supplementation ameliorated both sperm concentration and abnormal spermato- zoa (p<0.05) comparing to the control group. In conclusion, we suggested that royal jelly might have protective effects in the FM-induced reductions in epididymal sperm concentration and in- crease in abnormal spermatozoa rate.

In humans, iron deficiency represents a relevant occurrence in heart failure (HF), with or without anaemia, and is associated with the worst outcome. Moreover, chronic kidney disease (CKD) is a well-known comorbidity of HF and is strongly associated with the risk of developing anaemia. The most common cause of HF in dogs is myxomatous mitral valve disease (MMVD). To the best of our knowledge, no studies have examined the iron status in dogs with HF, with and without CKD. The aim of this retrospective study was to evaluate the iron status in dogs affected by MMVD and how strong is the relation with HF.

The retrospective study included 54 dogs with complete case records, echocardiography and laboratory analyses. Iron status was evaluated by measuring serum iron concentration (SIC), un- saturated iron binding capacity (UIBC), total iron binding capacity (TIBC), and percentage of saturation (%SAT).

The prevalence of dogs showing low serum iron concentration (SIC) was 18% in the whole population, 33% in symptomatic patients, 100% in dogs with acute decompensated HF. No signif- icant differences in SIC, UIBC, TIBC and %SAT median values were found among dogs classi- fied in different ACVIM (American College of Veterinary Internal Medicine) classes, between symptomatic and non-symptomatic patients, and among IRIS (International Renal Interest Soci- ety) classes. Azotemic and non-azotemic patients presented a significant difference in SIC mean values (p=0.02). Generalised linear model (GLM) revealed that dogs with low SIC were at high- er risk of being included in a higher ACVIM class (OR=6.383, p-value=0.014).

Log-rank analysis showed shorter survival in dogs with low SIC (p=0.020), multivariate Cox analysis revealed that only HF symptoms can affect survival.

The present study was aimed to investigate oxidative stress, DNA damage, and histopatholog- ical alterations in hepatic tissues of splenectomized Wistar rats experimentally infected with Ba- besia bigemina. Rats were challenged with 5x106 infected erythrocytes. Babesia infection was con- firmed both with Giemsa’s staining blood smears and nested-PCR amplified region of apical membrane antigen-1 (AMA-1) gene. Parasitemia reached approximately 10 % at day 5 post-in- fection. Livers of infected rats were enlarged and darker in color, became extremely brittle with marked congestion. Microscopic evaluation showed cytoplasmic clearing of hepatocytes and se- vere hydropic changes with significantly dilated sinusoids containing macrophages and also intra- sinosoidal parasitized erythrocytes. Severe infiltration of lymphoplasma cells was also present throughout the liver parenchyma. Furthermore, Kupffer cells were enlarged and, occasionally, containing Babesia-parasitized erythrocytes. The activity of Glutathione (GSH) and catalase (CAT), and total antioxidant capacity (TAC) were also significantly decreased (p < 0.05) after infection of rats with B. bigemina. B. bigemina infection also induced a significant increase (p < 0.05) in hepatic malondialdehyde (MDA) and nitric oxide-derived products (NOx) concentra- tions as well as amount of endogenous hepatocytes DNA damage. Hepatic damage was also re- flected through the measurement of lactic acid dehydrogenase (LDH) and protein carbonyl con- tent (PCO) in liver cells. These two indices of liver injury were also significantly elevated (p < 0.5) during B. bigemina infection. Evaluation of correlation between assayed variables in infected rats revealed that MDA levels were positively correlated with PCO, NOx, LDH and DNA damage in the infected group and negatively correlated with GSH, CAT and TAC. There was also an inverse relationship between the antioxidant enzymes activities of GSH, CAT and TAC with PCO, NOx and DNA damage in infected rats. However, NOx showed positive correlation with PCO and DNA damage in infected rats. On the basis of the above results it can be concluded that the Ba- besia infection increases oxidative stress markers, protein carbonyl content and DNA damage and decreases antioxidant enzymes activities in the liver. These results suggest that B. bigemina infec- tion could alter the liver histopathology and causes DNA damage following oxidative stress in hepatic tissue. Further studies are needed to precisely define how hepatic tissue damage takes place in B. bigemina infection.

The aim was to explore the feasibility of using bamboo vinegar powder as an antibiotics substitute in weaning piglets. Forty-five healthy Duroc × Landrance × Yorshire piglets (weight 6.74 ± 0.17 kg; age 31 days) were randomly divided into the control group (basic diet), ANT group (basic diet + 0.12% compound antibiotics), BV1 group (basic diet + 0.1% bamboo vinegar powder), BV5 group (basic diet + 0.5% bamboo vinegar powder) and BV10 group (basic diet + 1% bamboo vinegar powder). MyD88 and CD14 expression in immune tissues was examined using real-time PCR. MyD88 expression in the control group were significantly lower than that in other groups in all tissues (p<0.05), while CD14 expression showed the opposite trend. MyD88 expression was significantly higher in the BV10 group than in other groups in lung tissue (P<0.05), significantly higher in the ANT group than in the BV1 group in the kidneys (P<0.05), significantly higher in the BV10 group than in the BV1 group in the thymus (P<0.05), and signifi- cantly higher in the BV1 group than in the BV10 group in the lymphatic tissue (P<0.05). These differences between experimental groups were not observed for the CD14 gene (P>0.05). Thus, adding bamboo vinegar powder to the basic diet of weaning piglets had immune effects similar to antibiotics and the effect was dose-dependent. Moreover, the MyD88 and CD14 genes appear to play a role in these immune effects

MDAP-2 is a new antibacterial peptide with a unique structure that was isolated from house- flies. However, its biological characteristics and antibacterial mechanisms against bacteria are still poorly understood. To study the biological characteristics, antibacterial activity, hemolytic activi- ty, cytotoxicity to mammalian cells, and the secondary structure of MDAP-2 were detected; the results showed that MDAP-2 displayed high antibacterial activity against all of the tested Gram-negative bacteria. MDAP-2 had lower hemolytic activity to rabbit red blood cells; only 3.4% hemolytic activity was observed at a concentration of 800μg/ml. MDAP-2 also had lower cytotoxicity to mammalian cells; IC50 values for HEK-293 cells, VERO cells, and IPEC-J2 cells were greater than 1000 μg/ml. The circular dichroism (CD) spectra showed that the peptide most- ly has α-helical properties and some β-fold structure in water and in membrane-like conditions. MDAP-2 is therefore a promising antibacterial agent against Gram-negative bacteria. To deter- mine the antibacterial mechanism(s) of action, fluorescent probes, flow cytometry, and transmis- sion electron microscopy (TEM) were used to study the effects of MDAP-2 on membrane perme- ability, polarization ability, and integrity of Gram-negative bacteria. The results indicated that the peptide caused membrane depolarization, increased membrane permeability, and destroyed membrane integrity. In conclusion, MDAP-2 is a broad-spectrum, lower hemolytic activity, and lower cytotoxicity antibacterial peptide, which is mainly effective on Gram-negative bacteria. It exerts its antimicrobial effects by causing bacterial cytoplasm membrane depolarization, increas- ing cell membrane permeability and disturbing the membrane integrity of Gram-negative bacte- ria. MDAP-2 may offer a new strategy to for defense against Gram-negative bacteria.

Wild ruminants are an interesting topic for research because only limited information exists regarding their microbiota. They could also be an environmental reservoir of undesirable bacteria for other animals or humans. In this study faeces of the 21 free-living animals was sampled (9 Cervus elaphus-red deer, adult females, 12 Capreolus capreolus-roe deer, young females). They were culled by selective-reductive shooting during the winter season of 2014/2015 in the Strzałowo Forest District-Piska Primeval Forest (53° 36 min 43.56 sec N, 21° 30 min 58.68 sec E) in Poland. Buttiauxella sp. is a psychrotolerant, facultatively anaerobic, Gram-negative rod anaerobic bacte- rial species belonging to the Phylum Proteobacteria, Class Gammaproteobacteria, Order Entero- bacteriales, Family Enterobacteriacae and to Genus Buttiauxella. Buttiauxella sp. has never previ- ously been reported in wild ruminants. In this study, identification, antimicrobial profile and sensitivity to enterocins of Buttiauxella strains were studied as a contribution to the microbiota of wild animals, but also to extend knowledge regarding the antimicrobial spectrum of enterocins. Five strains were identified using the MALDI-TOF identification system (evaluation score value was up to 2.224) and allotted to the genus Buttiauxella including the species Buttiauxella gaviniae,

B. ferragutiae, B. agrestis. Strains were DNase negative, and they hydrolysed esculin; fermentation of L-arabinose, D-mannitol and D-mannose was positive. Dulcitol, inositol reaction, urea and indol were negative. Buttiauxella strains did not form biofilm. They were resistant to at least one of the 13 antibiotics tested. B. agrestis 2/109/1 was resistant to amdinocillin, clindamycin and pen- icillin. However, Buttiauxella strains were sensitive to the enterocins used (inhibition activity ranged from 100 to 25 600 AU/ml).

Cell culture transplantation is very promising in the treatment of various diseases. Cells obtained from a number of sources have been analysed to provide a basis for further studies in the area of regenerative medicine. The objective of the study was to compare morphological and phenotypic changes in cat adipose tissue and bone marrow cell cultures from the first to fifth passages. Adipose tissue and bone marrow were used to obtain cell cultures (coming from 3 cats) using standard methods with own modification. Phenotype changes were monitored by CD-marker identification and CD pan-keratin. The cytogenetic analysis was performed on 50 metaphase plates of cell cultures from the first to fifth passage. Cytogenetic assays showed that the adipose tissue cell culture (ATCC) at all passages was more stable than the bone marrow cell culture (BMCC).

The aim of this study was to assess the biological effect of the product Jodis Concentrate (JC) on the rabbit ovaries by evaluating the folliculogenesis and expression of oocyte-specific growth differentiation factor 9 (GDF9).

The experiment was conducted with 30 female two month old New Zealand rabbits that were the F1 offspring born to mothers differently treated with Jodis concentrate. The control group (n=10), consisted of F1 offspring born to mothers without iodine treatment, and was not supple- mented with JC. The first experimental group (n=10), consisted of F1 offspring born to mothers treated with JC during pregnancy and the suckling period, and was supplemented with JC daily at a dose of 2 ml/L drinking. The second experimental group (n=10), consisted of F1 offspring born to mothers without iodine treatment, and was also supplemented daily with the same dose of JC - 2 ml/L drinking. All groups were fed with total mixed ration for growing rabbits. The trial lasted 48 days. The ovaries were weighed and prepared for histological examination. The GDF9 protein expression in the ovary was determined by immunohistochemical analysis. The addition of JC to the drinking water of female rabbits led to more active development of the ovarian follicles from primordial to tertiary stage in both experimental groups. More intensive GDF9 protein expression in the oocytes and cumulus cells of rabbits, supplemented with JC was observed.

The aim of this research was to determine rumination time (RT) and the subsequent milk yield, along with trait changes during lactation dependent on the reproductive status of dairy cows. 728 cows were selected for evaluation in regards to 1–150 days of milk production (DIM). According to their period of lactation and reproductive status, the cows were selected for the following groups: Inseminated (1–35 days after insemination, n=182), Open (45–90 days after calving, n=126), Fresh (1–44 days after calving, n=45); Not-pregnant (>35–60 days after inse- mination and not-pregnant, n=55); Pregnant (35–60 days after insemination and pregnant (n=320). The animals were milked with Lely Astronaut® A3 milking robots. The daily milk yield, rumination time, bodyweight, milk composition (fat, protein, lactose, somatic cell count and gynecological status date) were collected from the Lely T4C management program for analysis. We estimated the lowest productivity in the pregnant cows, where the average milk yield was 28.72 kg and the highest productivity in the fresh cow (p<0.001) (Table 1). The longest rumina- tion time was determined for the inseminated cows, statistically significantly higher at 9.92% (p<0.001) than in the non-pregnant cows, whose rumination time was the shortest. The statisti- cally reliably RT positively correlated with productivity (r=0.384, p<0.001) of the cows (from r=0.302 in the second lactation and r=0.471 in the first lactation to r=0.561 in multiparous cows; p<0.001). Rumination time, according to groups of cows by milk yield, had a tendency to increase (2.14 times) from 202.0± 87.38 (in cows with a productivity of less than 10 kg milk) to 431.6±33.91 (in cows with a milk yield higher than 50 kg) by the linear regression equation: y = 38.02x + 232, R² = 0.721 (p<0.001). The relation between the gynecological status and milk fat-protein ratio of the cows was statistically significant (χ2=2.974, df= 8, p <0.0001). The longest rumination time was determined for the inseminated cows (1 – 35 days after insemination), and the shortest for the not-pregnant cows (>35 – 60 days after insemination and not-pregnant). We can conclude that rumination time, subsequent yield, and milk trait change depends on the period of lactation and reproductive status of a dairy cow.

Sapelovirus A (SV-A) is a positive-sense single-stranded RNA virus which is associated with acute diarrhea, pneumonia and reproductive disorders. The virus capsid is composed of four proteins, and the functions of the structural proteins are unclear. In this study, we expressed SV-A structural protein VP1 and studied its antigenicity and immunogenicity. SDS-PAGE analysis revealed that the target gene was expressed at high levels at 0.6 mM concentration of IPTG for 24 h. The mouse polyclonal antibody against SV-A VP1 protein was produced and reached a high antiserum titer (1: 2,048,000). Immunized mice sera with the recombinant SV-A VP1 protein showed specific recognition of purified VP1 protein by western blot assay and could recognize native SV-A VP1 protein in PK-15 cells infected with SV-A by indirect immunofluorescence assay. The successfully purified recombinant protein was able to preserve its antigenic determinants and the generated mouse anti-SV-A VP1 antibodies could recognize native SV-A, which may have the potential to be used to detect SV-A infection in pigs.

The present study was conducted to characterize the infectious bursal disease virus (IBDV) circulating in clinically diseased broiler chicken flocks with previous vaccination history during 2015-2016 in Egypt. IBDVs were isolated from 48 out of 63 of the investigated bursae from 10 flocks onto embryonated chicken eggs (ECEs) and verified by reverse transcriptase-poly- merase chain reaction (RT-PCR). Histopathologically, bursae lesions revealed some lymphocytes depletion as well as the presence of vesicles in the lining epithelium. The hyper variable region (HVR) of VP2 and VP1 genes of the 10 isolates (1 isolate/flock) were partially sequenced and subjected to comparative alignment and phyologenetic analysis. Phylogenetically, IBDV isolates were clustered into two distinct genetic lineages: variants of classical virulent (cv) and very viru- lent (vv) IBDV strains based on VP1 and VP2 amino acid (aa) sequences. Alignment analysis of HVR-VP2 aa sequences has demonstrated that the vvIBDV isolates have the conserved residues of the vvIBDV pathotype (A222, I242, and I256), while, the cvIBDV isolates have the same aa sequences of the classical attenuated vaccine strain (D78). Expected single point mutation occurred at position 253 (H253N). All previously characterized isolates were re-subjected to molecular analysis with VP1 protein due to its correlation with virulence and pathogenicity of IBDVs. vvIBDV isolates have the conserved tripeptide (TDN), while, the cvIBDV isolates have aa substitutions at conserved tripeptide including NEG at 145-147 amino acid. The present study has demonstrated that variants of classical virulent and very virulent IBDV circulated among vaccinated flocks in Egypt during 2015-2016.

Objective: This study aimed to investigate developmental changes of the thymus and intra- thymic IL-1β, IL-6 and TNF-α expression in weaned Sprague-Dawley rats induced by lipopolysac- charide.

Methods: Forty healthy weaned rats aged 26 days and weighing 83±4 g were randomly and equally divided into two groups. The lipopolysaccharide group was treated daily with a single injection of lipopolysaccharide for 10 consecutive days, and the saline group was treated with an equal volume of sterilized saline. On the 1st, 4th, 7th and 10th day, histological changes and distribu- tion of IL-1β-, IL-6- and TNF-α-positive cells were detected in the thymus by hematoxylin-eosin and immunohistochemistry staining, respectively. Subsequently, the expression levels of IL-1β, IL-6 and TNF-α were evaluated in the thymus by the ELISA method.

Results: Thymus weight and index were significantly smaller in lipopolysaccharide-treated rats than in saline-treated rats (p<0.05), but no substantial changes were found in the thymus microstructure after lipopolysaccharide induction. Moreover, a large number of IL-1β-, IL-6- and TNF-α-positive cells were observed with brownish-yellow color and mainly distributed in the thy- mus parenchyma, both integrated optical density and average optical density increased signifi- cantly in lipopolysaccharide-treated rats than those in saline-treated rats. Compared with the saline group, most of the thymic homogenates had higher levels of IL-1β, IL-6 and TNF-α in the lipopolysaccharide group on different days.

Conclusion: These findings indicate that the thymus atrophied after lipopolysaccharide induction in weaned Sprague-Dawley rats, and excessive production of intrathymic IL-1β, IL-6 and TNF-α was probably involved in the atrophic process.

The study was undertaken to determine the effect of continuation or changes of the diet on the morphometry and histomorphometry of bone in male and female Wistar rats with experimen- tally induced obesity by high energetic diet. Sixty-four 90-day-old Wistar rats obtained from obese parents (16 male, 16 female) and control parents (16 male, 16 female) were used in this study. After 21 days of the baby period, rats were divided into four groups: obese rats fed with high energy feed (F/F), control rats fed with a standard diet (C/C), obese rats with changed diet from high energy diet to control diet (F/C) and control rats with changed diet from control diet to high energy diet (C/F). After 90 days of experimental feeding, the rats were sacrificed. Thereafter, body weight and the isolated humerus were measured and next, the histological stainings and counts were done. Our results revealed that change in the parent’s diet from F to C in the female leads to increased bone growth length and reduction of body weight in female and male. Reverse diet changes (from C to F) lead to decreased bone length only in the female. Moreover, the con- tinuation by offspring of both sexes with a high-energy diet contributes to a reduction in osteo- cytes, reduction in bone marrow cavity and cortical expansion, but a change in nutrition from parents’ standard diet to high-energy diet leads to increase in osteocytes dimensions. The contin- uation of feeding with F diet promotes the accumulation of adipocytes in the bone marrow in female and male, and correction of nutrition from F to standard diet leads to a reduction in their number in the bone marrow compared to groups continuing feeding with high-energy diet.

Culture gas atmosphere is one of the most important factors affecting embryo development in vitro. The main objective of this study was to compare the effects of CO concentration on the subsequent pre-implantation developmental capacity of pig embryos in vitro, including embryos obtained via parthenogenesis, in vitro fertilization (IVF), and intracytoplasmic sperm injection (ICSI). Pig embryos were developed in four different CO2 concentrations in air: 3%, 5%, 10%, or 15%. The cleavage rate of pig parthenogenetic, IVF, or ICSI embryos developed in CO2 concen- trations under 5% was the highest. There were no significant differences in the oocyte cleavage rate in ICSI embryos in CO2 concentrations under 3% and 5% (p>0.05). However, as CO2 levels increased (up to 15%) the blastocyst output on day 7, from parthenogenetic, IVF, and ICSI em- bryos, decreased to 0%. These findings demonstrate that CO2 positively affects the developmen- tal capacity of pig embryos. However, high or low CO2 levels do not significantly improve the developmental capacity of pig embryos. The best results were obtained for all of the pig embryos at a 5% CO2 concentration.

To evaluate the role of preovulatory progesterone on canine sexual behavior and the course of proestrus and estrus, seven bitches in spontaneous cycles were treated with aglepristone for temporary elimination of progesterone action. Aglepristone was administered at the dose 10 mg/kg b.m., two times 24 hours apart, beginning in early proestrus when progesterone concen- tration was <0.5 ng/ml. Seven untreated bitches served as a control group. Reproductive sexual behavior (standing behavior, display vulva, tail deviation) was evaluated according to behavioral score. Cytologic, clinical and vaginoscopic examinations and progesterone measurements were used for the determination of proestrus and estrus and estimation of ovulation time.

Although, a similar pattern and magnitude of sexual behavior were observed in both groups, the duration of a total reproductive behavior was significantly extended (28.71 ± 2.06 vs 17.00 ± 2.45, p <0.05) in experimental group; similarly, the length of cytologic estrus (23.86 ± 3.02 vs 11.14 ± 2.41, p <0.05) was prolonged in treated bitches. In contrast, ovulation rate, duration of proestrus did not differ between the groups (p >0.05). We conclude, that during the canine estrus cycle the preovulatory progesterone terminates the duration of reproductive behavior and cytologic estrus.

Canine distemper virus (CDV) infects wild and domestic Canidae worldwide. The hemag- glutinin (H) gene has the highest genetic variation in the genome of this virus. Thus, the H gene is commonly used for lineage identification and genetic analyses. In order to study the genetic characteristics and pathogenicity of CDV strains prevalent in China, 132 samples were collected from domestic dogs with suspected CDV infection, 58 samples were confirmed to be positive, and the H gene was successfully amplified from 15 samples. The epidemic strain was identified as type Asia-1 and the novel mutations, A51T, V58I, R179K and D262N, were detected in this strain. Isolated strains, BJ16B53, BJ16B14, and BJ17B8, were used for an animal infection experiment in raccoon dogs. BJ16B53 and BJ16B14 were found to cause clinical symptoms, death, and exten- sive lesions in various organs. These results are expected to facilitate the development of effective strategies to monitor and control CDV infection in China.

The aim of this study was to determine the influence of feed on the pharmacokinetics of flumequine (FLU) administered to broiler chickens as follows: directly into the crop (10 mg/kg of BW) of fasted (group I/control) and non-fasted chickens (group II), or administered continu- ously with drinking water (1 g/L for 72 h) and with unlimited access to feed (group III). Plasma concentration of FLU was determined by high-performance liquid chromatography with fluo- rescence detection. In group II, a significant decrease in the maximum concentration (Cmax = 2.13±0.7 μg/mL) and the area under the concentration curve from zero to infinity (AUC0→∞ = 7.47±2.41 μg·h/mL) was noted as compared to the control group (Cmax = 4.11±1.68 μg/mL and AUC0→∞ = 18.17±6.85 μg·h/mL, respectively). In group III, the decrease in AUC was signifi- cant only in the first 3 hours (AUC0→3 = 5.02±1.34 μg·h/mL) as compared to the control group (AUC0→3 = 7.79±3.29 μg·h/mL). The results indicate that feed reduced the bioavailability of FLU from the gastrointestinal tract by at least 50% after the administration of a single oral dose. However, continuous administration of FLU with drinking water could compensate for the feed-induced decrease in absorption after single oral dose.

Oxygen is used for medical treatment and general anesthesia. However, high concentrations of oxygen can have toxic effects on cells. In veterinary medicine, 100% oxygen is usually used during general anesthesia and it can be toxic to animals. However, there is little concern about its harmful effects in humans. The objective of this study was to demonstrate that using a high con- centration of oxygen increases the partial pressure of oxygen in arterial blood (PaO2) more so than a lower concentration, by comparing PaO2 at three different oxygen concentrations (100%, 60%, and 40%) in six dogs under general anesthesia for 3 hours. The mean PaO2 and standard error values at the 100%, 60%, and 40% oxygen concentrations were 535.8 ± 24.01, 374 ± 17.19, and 239 ± 8.78 mmHg, respectively (p<0.05). These results show that 100% and 60% oxygen concentrations could increase oxidative stress. Further studies are needed to examine the oxygen concentration that causes toxicity.

In the present study on Bubalus bubalis of the Campania Region (Italy) the serum levels of derivatives of reactive oxygen metabolites (d-ROMs), anti-ROM and oxidative stress index (Osi) were evaluated. These data were then related to the seropositive status of the animals against alpha-herpesviruses, precisely Bubaline herpesvirus 1 (BuHV-1) and Bovine herpesvirus 1 (BoHV-1). Clinically healthy Mediterranean buffaloes were selected for this study. The serum samples of these animals were taken, and d-ROMs, anti-ROM and Osi were measured using commercially available tests. The preliminary data demonstrated that animals seropositive to both BuHV-1 and BoHV-1 present more oxidative stress than seronegative animals, as revealed by a significant increase in d-ROMs. Our results provide, for the first time, insight into the reac- tive oxygen species (ROS) modulation induced by the herpesvirus in Bubalus bubalis.

Vaccination is a common routine for prevention and control of human and animal diseases by inducing antibody responses and cell-mediated immunity in the body. Through vaccinations, smallpox and some other diseases have been eradicated in the past few years. The use of a patho- gen itself or a subunit domain of a protein antigen as immunogens lays the basis for traditional vaccine development. But there are more and more newly emerged pathogens which have expe- rienced antigenic drift or shift under antibody selective pressures, rendering vaccine-induced im- munity ineffective. In addition, vaccine development has been hampered due to problems includ- ing difficulties in isolation and culture of certain pathogens and the antibody-dependent enhancement of viral infection (ADE). How to induce strong antibody responses, especially neu- tralizing antibody responses, and robust cell-mediated immune responses is tricky. Here we re- view the progress in vaccine development from traditional vaccine design to reverse vaccinology and structural vaccinology and present with some helpful perspectives on developing novel vac- cines.