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Haematological indicators may resent physiological variation by age. Vitamin B12 promotes haematopoiesis. The aims of this study were: 1) to compare the values of the haematological variables and the concentration of vitamin B12 in pre- or post-weaned veal calves and 2) to identify the possible association between the values of the haematological variables and the concentration of B12 in the blood of veal calves. Blood was collected on the same day from 31 pre-weaned and 31 weaned calves of the Limousine breed from the same farm. The complete blood count, including the blood cell morphology evaluation, was performed and the serum B12, total protein and albumin concentrations were determined. The serum concentration of vitamin B12, the haematocrit (HCT), the haemoglobin concentration (HGB), the platelet count and the lymphocyte count were significantly higher in the weaned calves. A very strong positive correlation was found between the concentration of the vitamin B12 and HCT and HGB before weaning, while these correlations were moderately positive following weaning and in the total population tested as well. The observed variation in the blood cell count and morphology, such as poikilocytosis and the presence of macrocytes and hypersegmented neutrophils, along with the age of the animal seem to be related to the vitamin B12 concentration.

The objectives of this prospective, randomized, blinded, crossover, experimental study were to detect the potential anaesthetic- and analgesic-sparing effects of classical music provided to dogs undergoing skin surgery, and to investigate the role of substance P as an intraoperative pain indicator. Twenty dogs were included, each subjected to three different treatments: Chopin music, Mozart music and no music. They were premedicated with acepromazine, butorphanol and meloxicam and anaesthetized with propofol and isoflurane. Fentanyl was used as rescue analgesia. The anaesthetic depth was monitored by using the bispectral index along with standard anaesthetic monitoring, and autonomic nervous system responses were used to monitor the adequacy of analgesia. Furthermore, measurements of substance P serum concentration were carried out. Dogs exposed to music required less isoflurane and fentanyl. Furthermore, a statistically significant effect of time on substance P concentration was observed regardless of exposure to music, and there was a significant interaction effect between different timepoints and the type of acoustic stimulus. Classical music seems to have an isoflurane and fentanyl sparing effect on dogs undergoing minor surgery. Following surgical stimulation, the serum substance P concentration increases rapidly, and thus appears to be a potentially useful pain indicator., (© 2024. The Author(s).)

Subacute rumen acidosis (SARA) is a current issue in intensive livestock farming, and it is particularly associated with feeding high-concentrate diets. This study investigated the prevalence and impact of SARA in forty-two Greek dairy sheep flocks by recording rumen pH, milk composition, and milk yield over a period of nine months. Moreover, it explored the use of computerized rumen colorimetry as a management and diagnostic tool for SARA in dairy sheep. In culled ewes, computerized rumen mucosal colorimetry was applied, and rumen wall samples taken for histological examination. SARA cases were identified in 19 farms (45%, n = 42). Farms with SARA cases had lower milk fat levels, while milk yield and milk protein levels did not differ based on the SARA status of the farms. In culled ewes, rumen color was significantly associated with the flock's SARA status, and affected ewes showed increased thickness in non-keratinized and total epithelial layers. It was concluded that computerized rumen mucosal colorimetry in aged, culled ewes shows promise as an indicator, post mortem , of SARA present in dairy sheep flocks whose impact can be minimized by making significant changes in dietary management.

Among the various zoonotic pathogens that infect horses, Anaplasma phagocytophilum, Borrelia spp. and Leishmania spp. have gained scientific interest, and relevant molecular and serological studies in horses have been conducted worldwide. Moreover, human and veterinary medicine have extensively applied alternatives to serum diagnostic samples-such as saliva-for detecting pathogens or antibodies. In this study, we investigated the exposure of horses in Greece to A. phagocytophilum , B. burgdorferi , and L. infantum , and we assessed the diagnostic accuracy of saliva compared to serum in detecting IgG antibodies against the abovementioned pathogens. Paired saliva and serum samples were collected from 317 horses from different regions in Greece. The paired samples were examined using the indirect fluorescent antibody test (IFAT) for detecting IgG antibodies against A. phagocytophilum, B. burgdorferi , and L. infantum. Sensitivity, specificity, positive likelihood ratio (PLR), and negative likelihood ratio (NLR) were determined to assess the validity of saliva as an alternative to serum. The receiver operating characteristic (ROC) curve revealed that the optimal cut-off value for detecting antibodies against all the examined pathogens in saliva was 1/10. Higher seropositivity rates were found for B. burgdorferi (15.14%) and A. phagocytophilum (14.19%) compared to L. infantum (1.26%). The detection of IgG antibodies using IFAT in saliva samples had a good test performance compared to serum. The two sample types had a substantial to almost perfect agreement. Although the sensitivity was moderate (70.83-75.56%) in all cases, the specificity was almost perfect to perfect (99.63-100%). This study provides the first evidence that horses in Greece are exposed to A. phagocytophilum and B. burgdorferi and confirms that the seroprevalence of L. infantum in horses in Greece remains low. Our findings suggest that saliva sampling coupled with IFAT could be successfully applied for detecting IgG antibodies against these important zoonotic pathogens in large-scale epidemiological studies in horses, at the population level, as an alternative to serum.

The objectives of the present study were to evaluate (a) the feasibility of using stromal vascular fraction (SVF) and nanocrystalline hydroxyapatite (nHA) paste in combination for the treatment of segmental bone defect, (b) the quality of the callus produced, (c) the potential improvement of the autograft technique, and (d) the direct comparison of the biomaterial to the use of autogenous cancellous bone. Unilateral, segmental mid-diaphyseal bone defect was created on the right metatarsus of skeletally mature sheep animals ( n = 24) under anesthesia (D0). Residual segments were stabilized by stainless-steel plates and appropriate screws. Defects were managed as follows: group A: use of nHA paste to filling, group B: use of autogenous bone graft mixed with nHA bone paste, placed in defect, group C: use of SVF mixed with nHA bone paste injected into defect, group D: use of bone graft and SVF with nHA paste before apposition in bone defect. SVF had been previously isolated from adipose tissue of the animals intra-operatively after digestion with collagenase solution and neutralization. Animals were evaluated clinically and by X-raying and ultrasonographic examination of the defect, at regular intervals, until D90. Ultrasonographic assessment performed along the length of the defect included calculation of the length of the bone defect and assessment of vascularization. SVF was successfully isolated from group C and D animals, with the average yield being 1.77 × 10 6 cells. The comparison of clinical scores (based on the 'Kaler scale') on each post-operative day indicated significant differences between the four groups on D1 to D30 ( p < 0.01); the median clinical score within group A was 2.5 for D1-D30 and 1 for the entire period; respective scores for other groups were 1.5 ( p = 0.07) and 0 ( p = 0.033). Differences in radiographic assessment scores were significant for scores obtained on D60 ( p = 0.049) and D90 ( p = 0.006). There was a significant difference between the four groups in the length of the bone defect, as assessed ultrasonographically, for the entire length of the study; median values were 8, 8.5, 6, and 8 mm for groups A, B, C, and D, respectively ( p = 0.008). There was a significance in the differences between median scores obtained during the histopathological examination: 2, 11, 13.5, and 12 for group A, B, C, and D ( p = 0.022). There was an inverse correlation between the overall scores of histopathological evaluations and the length of the bone defect (observed on D90) ( p < 0.0001) and a correlation between the overall scores and the radiographic assessment scores (obtained on D90) ( p < 0.0001). This is the first study in which the efficacy of fresh autologous Stromal Vascular Fraction (SVF) from adipose tissue in enhancing bone healing in a long, weight-bearing, diaphyseal bone was evaluated. It is concluded that the lumbosacral region was an attractive site for harvesting adipose tissue, the use of SVF contributed to faster rehabilitation post-operatively, and SVF significantly enhanced bone formation; in general, the results indicated an osteogenic potential of SVF comparable to the gold standard autologous bone graft.

Modified live virus (MLV) vaccines for the control of porcine respiratory and reproductive syndrome virus (PRRSV) have been associated with the vertical and horizontal transmission of vaccine viruses. The present study aimed to describe pathological lung lesions in piglets born by gilts vaccinated with PRRSV-1 MLV. In total, 25 gilts were vaccinated at late gestation (100th day) and were divided into five groups according to the different vaccines (Vac) used: no vaccine-control group, Vac-1-strain DV, Vac-2-strain VP-046 BIS, Vac-3-strain 94881, Vac-4-strain 96V198. Within the first 0-9 h of the farrowing, blood samples were collected from all newborn piglets and lung samples were exanimated grossly, histopathologically and with scanning electron microscopy. PRRSV (RT-PCR-positive) and antibodies were detected in the serum of piglets from gilts vaccinated with Vac-2. In these piglets, moderate to severe interstitial pneumonia with thickened alveolar septa was noticed. Type II pneumocyte hyperplasia was also observed. The rest of the trial piglets showed unremarkable lung lesions. Phylogenetic analysis revealed the 98.7% similarity of the PRRSV field strain (GR 2019-1) to the PRRS MLV vaccine strain VP-046 BIS. In conclusion, the Vac-2 PRRSV vaccine strain can act as an infectious strain when vaccination is administrated at late gestation, causing lung lesions.

Flaxseed and lupin seed were offered as an alternative dietary approach in dairy cows, through the partial substitution of soybean meal. Milk production and fertility traits were investigated. A total of 330 animals were allocated into two groups, treated ( n = 176) and control ( n = 154). From each group, 30 animals were selected for hematological and cytological studies. The experimental feeding period lasted for 81 days (25 days prepartum and 56 days postpartum). The control ration (group C) contained corn, barley, soybean meal, rapeseed cake, corn silage and lucerne hay; whereas, in the treatment group (group T), 50% of the soybean meal was replaced by an equal mixture of flaxseed and lupins. The two rations were formulated to be isonitrogenous and isoenergetic. Milk samples were analyzed for chemical composition, somatic cell count (SCC) content and total colony forming units (CFU). Blood samples were collected, and serum was analyzed for non-esterified fatty acids (NEFA), acute phase proteins (haptoglobin and serum amyloid) and lipid oxidation indices, namely thiobarbituric-acid-reactive substances (TBARS) and catalase activity. To assess polymorphonuclear neutrophils (PMN) numbers, endometrial samples from each cow were collected on days 21 and 42. No difference was recorded between groups in milk yield ( p > 0.05). In multiparous cows, NEFA (mMol/L) concentrations were significantly lower in group T than in group C on day 14 ( p > 0.009) and on day 42 ( p = 0.05), while no difference was detected in the group of primiparous cows. At all time points, serum TBARS and catalase values were similar in both groups ( p > 0.05). Multiparous cows in group T expressed the first postpartum estrus and conceived earlier than cows in group C ( p ≤ 0.05). Between days 21 to 42 postpartum, the PMN reduction rate was higher in group T animals ( p ≤ 0.05). Acute phase protein levels were in general lower in group T animals, and at specific time points differed significantly from group C ( p ≤ 0.05). It was concluded that the partial replacement of soybean meal by flaxseed and lupins had no negative effect on milk yield or milk composition, and improved cow fertility; which, along with the lower cost of flaxseed and lupins mixture, may increase milk production profitability.

Urine test strips are commercially available and can be assessed with semi-automated analyzers or by visual assessment. This study aimed to compare the visual and automated evaluations of dipstick variables in canine urine samples. One hundred and nineteen urine samples were evaluated. Automated analysis was performed on a veterinary urine analyzer URIT-50Vet (URIT Medical Electronic) with UC VET13 Plus strips. Multistix 10 SG dipsticks (Siemens Healthcare GmbH, Erlangen, Germany) were used for visual evaluation, along with a refractometer (Clinical Refractometer Atago T2-Ne, Atago Co., Tokyo, Japan) for urine specific gravity measurements. A linear relationship was observed between the pH measurements ( p = 0.2) of the two methods; the Passing-Bablok procedure was valid since neither proportional nor systematic significant errors were observed. Comparing the two methods, the correlation for urine specific gravity was poor ( p = 0.01, CI 0.667-1.000). Moderate agreement was demonstrated for proteins (κ = 0.431), bilirubin (κ = 0.434) and glucose (κ = 0.450). Agreement was substantial for blood (κ = 0.620) and poor for leukocytes (κ = 0.100). Poor agreement was observed for ketones (κ = -0.006). Apart from the pH analysis, visual and automated dipstick urinalyses should not be used interchangeably. Multiple urine samples obtained from the same dog during the day should be evaluated using the same method to overcome erroneous results.

In endemic areas, dogs with leishmaniosis due to Leishmania infantum frequently have comorbidities, including mostly neoplastic, infectious, and parasitic diseases. The aim of this study was to compare the prevalence of comorbidities among dogs that are not infected by L. infantum, dogs that are infected but do not present leishmaniosis, and dogs with leishmaniosis, and to examine if certain comorbidities are independent risk factors for the infection by L. infantum and/or for the development of canine leishmaniosis (CanL). A total of 111 dogs, older than 1-year and non-vaccinated against CanL, were allocated into three groups: group A (n = 18) included dogs that were not infected by L. infantum, group B (n = 52) included dogs that were infected by L. infantum but did not present CanL, and group C (n = 41) included dogs with CanL. Signalment and historical data were obtained using a structured questionnaire. Laboratory examinations included complete blood count, serum biochemistry, urinalysis, fecal parasitology, modified Knott's test, microscopic examination of capillary blood, buffy coat, lymph node, bone marrow and conjunctival smears, qualitative serology for Dirofilaria immitis, Anaplasma phagocytophilum/A. platys, Borrelia burgdorferi and E. canis, IFAT for L. infantum, ELISA for Babesia spp. and Neospora caninum, and real-time PCR for L. infantum in bone marrow, skin biopsies and conjunctival swabs. A variety of comorbidities were found in all three groups. No independent risk factors for infection by L. infantum were found. On the contrary, among dogs infected by L. infantum, being a mongrel [odds ratio (OR): 11.2], not receiving prevention for dirofilariosis (OR: 26.5) and being seropositive to N. caninum (OR: 17.1) or to Babesia spp. (OR: 37.6), were independent risk factors for presenting CanL. Although no comorbidities influence the probability of canine infection by L. infantum, certain comorbidities may be precipitating factors for the transition from the subclinical infection by L. infantum to the overt CanL., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

(1) Background: Rabbit hepatic coccidiosis, caused by Eimeria stiedae , is a devastating disease with high morbidity and mortality rates. The disease is well described in rabbits, but little is known about E. stiedae infection in wild rabbits. In this study, we investigated the presence of E. stiedae infection in wild rabbits from the island of Lemnos, Greece, where this species is overpopulated, and the effects of infection on common hepatic biomarkers. (2) Methods: We used liver impression smears to detect the coccidian oocysts, and we defined the liver biochemical profile of the infected individuals. (3) Results: Overall, 13.3% of the liver imprints examined were positive for the presence of coccidial oocysts. The activities of liver enzymes, that is, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and glutamyltransferase (GGT), as well as globulins (GLOB), were increased while the concentrations of albumins (ALB), total proteins (TP) and the albumin to globulin (A/G) ratio were decreased in the infected individuals compared to the non-infected ones. (4) Conclusions: This study adds to the current knowledge on the pathogens affecting wild rabbits and those circulating in this population on the island of Lemnos, Greece. Moreover, we showed that E. stiedae infection exerts pathological effects on the hepatocyte integrity and liver function of wild rabbits, as reflected by the abnormal values of liver injury and dysfunction biomarkers.

Increasing litter size may lead to low-birth-weight piglets (LBW) and further negative long-term effects. This study aimed to evaluate the effects of intramuscular administration (IM) of dexamethasone (Dexa) alone or in combination with vitamin E/Se on LBW piglets during the early postnatal period. The study included a total of 100 LBW piglets that were divided into 5 groups (20 LBW piglets per group) and treated with IM Dexa alone or in combination with vitamin E/Se (Vit E/Se) after birth as follows: (a) Group A-Cont: Control group, (b) Group B-Dexa1: Dexa on D1 (1st day of life), (c) Group C-Dexa3: Dexa on D1, D2, D3 (D2: 2nd day of life, D3: 3rd day of life), (d) Group D-Dexa + VitE/S1: Dexa + Vit E/Se on D1, and (e) Group E-Dexa + VitE/S3: Dexa + Vit E/Se (IM) on D1, D2, D3. Body weight (BW) and the Average Daily Weight Gain (ADWG) were recorded for all piglets on days 1, 7, 14, and 25, and vitality score (VS) was recorded on days 1, 2, 3, 4, and 14. A significant increase in BW and ADWG in Group E-Dexa + VitE/S3 and a significant reduction in Group C-Dexa3 were noticed in comparison to other groups. VS in groups Group B-Dexa1 and Group C-Dexa3 were significantly lower in comparison to other groups. Furthermore, piglets of Group C-Dexa3 had a significantly higher frequency of clinical findings compared to other groups. In conclusion, the administration of Dexa and vitamin E/Se combined after the birth of LBW piglets for 1-3 days has beneficial effects on their growth and survival scores.

The occurrence of co-infected hosts and questing ticks with more than one tick-borne pathogen-as in the case of Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato-is expected in endemic regions. Their synergy-in terms of pathogenesis and disease severity-has been suggested previously in humans. Limited data exist on the clinicopathological alterations in co-infected sheep. In this study, we investigated the impact of A. phagocytophilum and B. burgdorferi s.l. seropositivity, alone and in combination, on the hematological parameters of naturally infected sheep. A complete blood count was performed, and indirect immunofluorescence assays were used to detect IgG antibodies against A. phagocytophilum and IgG and IgM antibodies against B. burgdorferi s.l. Single natural exposure to B. burgdorferi s.l. was characterized by low Packed Cell Volume (PCV) values and platelet (PLT) counts, while single exposure to A. phagocytophilum was characterized by low PCV values, low white blood cell (WBC) counts, and an increased risk for leukopenia and neutropenia. Co-exposure resulted in the most severe blood abnormalities; all the blood parameters decreased, and the sheep presented an increased risk for anemia. Our study showed that natural co-exposure to A. phagocytophilum and B. burgdorferi s.l. in sheep leads to more severe blood abnormalities and enhances the pathogenic processes. More studies are needed to clarify the possible background mechanisms.

This study aimed to report, for the first time, histopathological lesions caused by an outbreak of acute Actinobacillus pleuropneumoniae serotype 8 infections in two farms in Cyprus. Lung tissue samples were collected from two different affected farms (a total of eight samples) for bacterial culture, multiplex polymerase chain reaction (PCR)-based serotyping and histopathological evaluation. Severe respiratory clinical signs, vomiting, anorexia, sudden deaths, a morbidity rate of around 25.00% and a mortality rate of over 60.00% in the fattening stage were reported. Macroscopic lesions included acute to subacute fibrotic, hemorrhagic and necrotizing pneumonia with occasionally encapsulated nodule-like abscesses and fibrous pleuritis. Histopathological evaluation revealed fibrous exudate between alveolar spaces and connective tissue, areas of necrosis mixed with alveolar macrophages, lymphocytes, plasma cells and necrotic leukocytes surrounding colonies of cocci. The bronchial and bronchiolar epithelia were degenerated and replaced by eosinophilic cell debris mixed with inflammatory cells. Several arteries and capillaries were clotted and/or infiltrated by inflammatory cells. In conclusion, these A. pleuropneumoniae serotype 8 cases were accompanied by acute illness, death and more pronounced bronchitis and bronchiolitis., Competing Interests: There is no conflict of interest., (© 2023 Urmia University. All rights reserved.)

The objective of the present study was to measure the concentration of Paraoxonase-1 (PON-1) and N-terminal-prohormone-B-type natriuretic peptide (NT-proBNP), in the serum of dogs with degenerative Mitral Valve Disease (MVD), in order to identify their association with the clinical stage and specific clinico-pathologic and echocardiographic findings.Eighty dogs diagnosed with MVD and staged according to the ACVIM (American College of Veterinary Internal Medicine) consensus statement (B1, B2, C and D), based on their clinical, radiographic, and echocardiographic findings, were included in the study. NT-proBNP was measured only in stage B1 and B2 dogs. Clinical stage did not have a significant effect on PON-1 concentrations ( p = 0.149), but NT-proBNP levels were lower in B1 dogs ( p = 0.001). A significant correlation between PON-1 and total plasma proteins ( p = 0.001), albumin ( p = 0.003) and white blood cell count ( p = 0.041) was detected, whereas there was no significant correlation ( p = 0.847) between PON-1 and NT-proBNP concentrations. PON-1 showed a significant but weak negative correlation with normalized left ventricular internal diameter at diastole (LVIDdn) ( p = 0.022) and systole (LVIDsn) ( p = 0.012), as well as mitral valve E to A wave velocity ratio (MV E/A) ( p = 0.015), but not with Left Atrial to Aortic root ratio (LA/Ao) ( p = 0.892) or fractional shortening (FS%) ( p = 0.944). PON-1 seems to be an insensitive marker of clinical stage and disease severity in MVD, but can be indicative of some clinico-pathological and echocardiographic changes. NT-proBNP changes are independent of oxidative stress.

Porcine reproductive and respiratory syndrome virus (PRRSV) causes respiratory disease in weaning and growing pigs. A vaccination against PRRSV is one of the most important control measures. This trial aimed to evaluate the effect of the intradermal (ID) administration of a PRRSV-1 modified live virus (MLV) vaccine in comparison to the intramuscular (IM) administration on the piglets’ health and performance. A total of 187 suckling piglets of a PRRSV-positive commercial farrow-to-finish farm were assigned to four groups: group A—PRRSV ID, group B—PRRSV IM, group C—control ID, and group D—control IM. At 2 weeks of age, all the study piglets were either vaccinated with a PRRSV-1 MLV vaccine or injected with the vaccine adjuvant (controls). The collected blood serum samples were tested by ELISA and qRT-PCR. The side effects, body weight (BW), average daily gain (ADG), mortality rate, and lung and pleurisy lesions scores (LLS, PLS) were also recorded. The ELISA results indicated that the vaccination induced an important seroconversion at 4 and 7 weeks. Significant differences in the qRT-PCR results were noticed only at 10 weeks in group A vs. group C (p < 0.01) and group B vs. group C (p < 0.05). High viral loads, as evidenced by the qRT-PCR Ct values, were noticed in animals of both non-vaccinated groups at 7, 10, and 13 weeks. An ID vaccination has a positive impact on the BW at the piglets’ slaughter, while both an ID and IM vaccination had a positive impact on the ADG. The mortality rate was lower in vaccinated groups at the finishing stage. The LLS and PLS were significantly lower in the vaccinated groups. In conclusion, our study demonstrated that the ID vaccination of suckling piglets with a PRRSV-1 MLV vaccine has a positive effect on the piglets’ health and performance, including an improved BW and a lower LLS and PLS index at their slaughter, as well as a decreased mortality rate at the growing/finishing stage.

Background: Urine dipstick and Heller's reaction are easy first-line screening tests for the detection of proteinuria; however, the performance of these methods in alkaline ovine urine is largely unknown., Objective: The aim of this study was to evaluate the diagnostic performance of Heller's reaction alone or in combination with dipstick for the detection of proteinuria in sheep, using the urine protein to creatinine ratio (UP/C) with two cut-off values as the reference method., Methods: Ninety-eight urine samples were collected from sheep using the transient apnea method. Heller's reaction, the dipstick method, and the UP/C ratio were used to assess proteinuria. The results were statistically analyzed twice, based on two different UP/C cut-off values of 0.2 and 0.5. Cohen's kappa value was used to determine the agreement between the UP/C ratios and Heller's reaction, the dipstick method, or the combination of methods. Sensitivity, specificity, and positive and negative likelihood ratios were calculated. ROC curves were also generated, and the areas under the curve (AUC) were evaluated to determine the optimal threshold for the numerical values of the two methods., Results: Heller's reaction is more specific (96.67% and 96.00% when the cut-off value is 0.2 and 0.5, respectively) than the dipstick method, while the dipstick method was more sensitive (91.18% and 91.30%, when the cut-off value was 0.2 and 0.5, respectively) than Heller's reaction for the detection of proteinuria. Both tests were accurate when any grade >0 was considered positive., Conclusions: Proteinuria can almost be excluded in ovine urine samples with negative Heller's reaction and dipstick test., (© 2022 American Society for Veterinary Clinical Pathology.)

The Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) induces apoptosis in different organs. Angiotensin II (Ang II) is the main effector of the renin-angiotensin system and participates in apoptosis. Thus, this study aimed to investigate changes in piglet serum Ang II levels following intradermal (ID) and intramuscular (IM) vaccination with a commercial PRRS modified live virus (MLV) vaccine. The trial was conducted in a commercial pig farm, including 104 piglets which were randomly allocated to four groups: Group A-Porcilis PRRS ID, Group B-Porcilis PRRS IM, Group C-Diluvac ID and Group D-Diluvac IM. The study piglets were either vaccinated or injected at 2 weeks of age and they were tested by qRT-PCR for PRRSV and by ELISA for Ang II. The results indicated differences in viremia of tested piglets at 7 weeks of age, while piglets at 10 weeks of age were all found qRT-PCR positive for PRRSV. In addition, significant differences were noticed in Ang II in 7-week-old piglets. In conclusion, the present study provides evidence that ID vaccination induces less tissue damage, based on the lower measurements of Ang II in the serum of ID vaccinated piglets.

Background: The optimal microscopic magnification and number of optical fields of adhesive tape strip cytological slides that should be examined when searching for Malassezia yeasts on canine skin are unknown., Objectives: To determine the optimal magnification and the minimum number of optical fields that should be examined to maximise intraobserver repeatability and interobserver reproducibility., Materials and Methods: Seven experienced examiners counted, twice, the number of yeasts in 10, 20, 30, 40 and 50 optical fields of 40 slides at ×400 and ×1000 magnification., Results: The number of yeasts per unit surface area was significantly higher at ×1000 compared to ×400 magnification. Repeatability and reproducibility for counting the yeasts was very poor., Conclusions and Clinical Relevance: Adhesive tape strip cytological slides should be examined microscopically for Malassezia spp. at ×1000 magnification. The repeatability of this examination for counting the yeasts is poor., (© 2022 ESVD and ACVD.)

The redox status shortly after the vaccination of pregnant ewes is rather unexploited. Thus, the present study was designed to evaluate the fluctuation of redox status after the administration of the annual booster dose of a polyvalent clostridial vaccine in pregnant ewes, 3 to 4 weeks before lambing, with or without a simultaneous injection of Vit E/Se. In total, 24 pregnant Lacaune ewes 3-4 weeks before lambing were randomly allocated into four equal groups: the V (vaccinated with a polyvalent clostridial vaccine), VE (vaccinated and injected IM with Vit E/Se), E (injected IM with Vit E and Se), and C (neither vaccinated nor injected with Vit E/Se). The study period lasted for 21 days, starting on the day of administration. Four redox biomarkers, the antioxidant capacity (TAC), the thiobarbituric acid reactive substances (TBARS), the reduced glutathione (GSH), and the catalase (CAT) were evaluated in blood samples collected from all ewes before the injections (0 h) and then at 12 (12 h), 24 (D1), and 48 h (D2), and thereafter on days 4 (D4), 6 (D6), 10 (D10), 14 (D14), and 21 (D21). The results reveal that the TAC was the only biomarker evaluated that was significantly affected by group and significantly lower in vaccinated animals (V and VE groups) compared to non-vaccinated (E and C groups). The absence of an increase in the TBARS values after vaccination in groups V and VE indicates the absence of significant oxidative stress. Overall, it can be assumed that annual booster immunizations against clostridial diseases do not impose acute oxidative stress on pregnant ewes in the last month of pregnancy.

The aim of this study was to assess the agreement between anti- Toxoplasma gondii IgG antibody detection in serum and filter paper (FP) blood spots using the indirect immunofluorescence antibody assay (IFA) and to evaluate the potential impact of the packed cell volume (PCV) on antibody detection in FPs. A pair of a serum and an FP sample was collected from 96 sows at various farms in Greece, with previously identified high seropositivity and/or risk factors associated with high seropositivity against T. gondii . The PCV value was determined using the microhematocrit method. IFA was used for the detection of antibodies against T. gondii . T. gondii -specific antibodies were detected in 45.8% serum samples and 41.6% FP samples showing almost perfect agreement. Detection in FP samples presented high sensitivity (87.1-92.8%) and excellent specificity (100%) when compared with detection in serum, regardless of the PCV values. The findings of this study support the reliability of FPs for the evaluation of the serological status of swine against T. gondii . FPs could be a good alternative sample type compared with serum for large-scale epidemiological studies.

Colibacillosis is the most common bacterial disease in poultry and it is caused by avian pathogenic Escherichia coli (APEC), which is assigned to various O-serogroups. Previous studies have shown that APEC strains are more often related to certain O-serogroups such asO78, O2 and O1. E. coli has been reported to act either as a primary or secondary agent in complicating other infections. The aim of this study was to investigate the occurrence of and characterize the O-serogroups of E. coli strains isolated from commercial layer and layer breeder flocks showing macroscopic lesions of colibacillosis and increased or normal mortality in Greece. Furthermore, we attempted to assess the interaction between infectious agents such as Mycoplasma gallisepticum (MG), Mycoplasma synoviae (MS), infectious bronchitis (IBV) and infectious laryngotracheitis (ILT) with E. coli infections in layer flocks with increased mortality. Our study revealed that in addition to the common serogroups (O78, O2), many other, and less common serogroups were identified, including O111. The O78, O111 and O2 serogroups were frequently detected in flocks with lesions of colibacillosis and increased mortality whereas O2, O88 and O8 were reported more commonly in birds with colibacillosis lesions but normal mortality rates. These data provide important information for colibacillosis monitoring and define preventative measures, especially by using effective vaccination programs because E. coli vaccines are reported to mainly offer homologous protection. Finally, concerning the association of the four tested infectious agents with E. coli mortality, our study did not reveal a statistically significant effect of the above infectious agents tested with E. coli infection mortality.

The aim of this study was to investigate the effects of two commercial phenolic phytogenic feed additives (PFAs) on sows under heat stress conditions of high summer temperatures for seven days before and seven days after the farrowing. The PFA-1 product was a mixture based on the plants Emblica officinalis , Foeniculum vulgare , Citrus sinensis and nut fiber, while the PFA-2 product was a mixture based on plants Andrographis paniculata , Glycyrrhizia glabra , Tinospora cordifolia and nut fiber. A total of 48 primiparous sows were divided into three groups: T1-control group: regular gestation (GF) and lactation feed (LF); T2 group: regular GF and LF supplemented with PFA-1; T3 group: regular GF and LF supplemented with PFA-2. Each sow in the T2 and T3 groups received 5 g daily of the PFA-1 and PFA-2 product, respectively, for seven days before and seven days after the farrowing. Blood samples were collected from all groups 24 h after farrowing. Thiobarbituric acid--reactive substances (TBARS) and protein carbonyl (CARB) concentrations were determined in the sow plasma. The body condition scoring (BCS) and the backfat of sows on the farrowing and weaning days along with reproductive parameters and litter characteristics were recorded. The highest number of stillborn piglets and the largest interval from weaning to estrus were observed in the T1 group. The lowest number of alive 24 h after birth and weaning piglets and the lowest BCS and backfat at weaning were also recorded in the T1 group. TBARS and CARB concentrations were significant higher in the T1 group compared to all other groups. In conclusion, the use of phenolic PFAs seems to reduce oxidative damage caused by heat stress and ameliorate performance in primiparous sows.

Porcine reproductive and respiratory syndrome virus (PRRSV) induces apoptosis through the activation of death receptors, including cell-surface Fas receptor. The aim of this study was to investigate the impact of intradermal (ID) and intramuscular (IM) vaccination with a commercial PRRSV-modified live vaccine in piglets on Fas-related apoptosis. The study included 104 suckling piglets from a commercial farrow-to-finish pig farm, suffering from positive unstable PRRSV status. Animals were assigned in four groups: group A-Porcilis PRRS ID-vaccinated pigs, group B-Porcilis PRRS IM-vaccinated pigs, group C-Diluvac ID adjuvant-administered pigs, and group D-Diluvac IM adjuvant-administered pigs. Vaccines were administered at 2 weeks of age. Blood samples were collected from the same pigs at 4, 7, and 10 weeks of age. Sera were examined by quantitative real-time reverse transcription-PCR (qRT-PCR) for PRRSV and by ELISA for soluble Fas (sFas). At 4 weeks of age, all groups were negative qRT-PCR for PRRSV; at 7 weeks only group A was negative; and at 10 weeks all groups were positive. sFas was significantly increased in groups C (4 vs. 7, 4 vs. 10, and 7 vs. 10 weeks) and D (7 vs. 10 weeks). Significant differences among groups were noticed only at 10 weeks (A vs. C, A vs. D, B vs. C, B vs. D). A significant positive and moderate correlation between PRRSV viral load and Fas level was observed. In unvaccinated piglets, increased serum sFas levels reveal apoptotic suppression compared with vaccinated piglets. In the latter, vaccine-derived antibodies limit the infection and may attribute to the reduced Fas expression, suggesting a weak induction of lymphocyte-mediated cytotoxicity.

Diagnosis of anaplasmosis is challenging considering the great variation in clinical signs and the limitations of the available diagnostic assays, while the detection of carrier animals that play a significant role in disease epidemiology as reservoirs is of great significance. In this study, we evaluated the diagnostic accuracy of a newly developed indirect immunofluorescent assay (Ag-IFAT) for the detection of A. phagocytophilum antigens in buffy coat specimens, alone and in combination with cytology, using PCR as a reference. Blood samples were collected from 138 sheep of the Chios breed from six farms in Greece. A buffy coat was extruded from the centrifuged blood. Buffy coat smears were used for cytological examination and the Ag-IFAT assay. The Ag-IFAT assay presented excellent specificity (100%) and high sensitivity (85.4%) for the detection of A. phagocytophilum antigens in buffy coats, and it has an almost perfect agreement with PCR and cytology (κ value = 0.88 and 0.85, respectively). A. phagocytophilum antigens are likely to be detected using Ag-IFAT in a PCR-positive animal, as indicated by the good performance of the assay. Overall, this assay presents high diagnostic accuracy, and it could be used for the detection of animals during the early stage of infection.

Despite being important, there are no equations for prediction of ionized calcium (iCa) in sheep and cattle. The objectives of this study were i) to create equations for the calculation of serum iCa concentration based on the serum concentrations of total calcium (tCa), albumin (Alb) and total proteins (TP) and ii) to investigate whether predicted serum iCa values are beneficial in clinical practice. Serum samples from 30 sheep and 30 dairy cattle were used. Serum tCa was determined colorimetrically, while serum iCa was determined with an ion selective electrode method. Serum Alb and TP concentration were determined using bromo-cresol green and biuret methods, respectively. Ionized calcium was also calculated based on serum tCa, using regression analysis, and with two equations based on Alb and TP concentration. Bland-Altman plots were plotted to evaluate the agreement between measured and predicted iCa; Passing and Bablok (P - B) regression analysis was used to assess their agreement. The initial equations were corrected using the P - B generated equation and Bland - Altman plots were run to evaluate the level of agreement between measured and predicted iCa using the final equations. Six equations were finally created for cattle and 6 for sheep. The total bias exceeded 10.00% in all of them indicating that they are clinically unacceptable for iCa prediction especially when the predicted result is very close to the cut-off point of < 1.00 mmol L -1 . So, it could be suggested that, when necessary, iCa concentration should be directly determined., Competing Interests: The authors declare no conflict of interests.

Wild rabbits ( Oryctolagus cuniculus ) can be important sentinel species for the presence of zoonotic pathogens. Therefore, we collected blood samples from wild rabbits harvested by hunters during the hunting season 2019-2020 on the island of Lemnos, to determine exposure of wild rabbits to the zoonotic pathogens Leishmania infantum , Toxoplasma gondii , Anaplasma phagocytophilum and Babesia caballi , as well as aqueous humor to assess its diagnostic performance in terms of sensitivity, specificity, positive and negative likelihood ratios. Antibodies against these pathogens were detected by Indirect Immunofluorescence Antibody (IFA) assay. Out of the 72 wild rabbits included in the study, 4.2%, 5.5%, 18% and 9.7% were seropositive to L. infantum , T. gondii , A. phagocytophilum and B. caballi , respectively. Although less frequently, antibodies were also detected in aqueous humor of wild rabbits. The antibody detection in aqueous humor presented 100% specificity but decreased sensitivity compared to serum suggesting that aqueous humor could be successfully used in epidemiological studies to confirm exposure at the population level but has little diagnostic value at the individual level. This is the first report on the seropositivity of wild rabbits to A. phagocytophilum and B. caballi and the detection of antibodies against A. phagocytopylum , L. infantum , T. gondii and B. caballi in the aqueous humor.

Toxoplasmosis, due to Toxoplasma gondii , is a parasitic disease with global importance. Among livestock, chronic T. gondii infection has been reported in higher rates in pigs and small ruminants, but with subclinical infections in case commonly encountered in pigs. Seroprevalence in the global pig population ranges according to the age or species of pigs, geographical distribution, production programs, and systems. Generally, T. gondii infections are noticed in low prevalence rates in conventional pig farms with high hygiene standards. In contrast, higher prevalence is common on free-ranging farms, outdoor or backyard small pig fams, as well as in farmed or hunted wild boars. The T. gondii average worldwide seroprevalence in pigs is reported to be 13% in Europe, 21% in Africa, 25% in North America, 21% in Asia, and 23% in South America. Human toxoplasmosis outbreaks have been correlated with the consumption of raw or undercooked meat, especially from infected pigs or wild boars, as well as of contaminated drinking water. The risk of infection in processed pork products is lower compared with fresh pork, as meat processing can reduce or inactivate T. gondii tissue cysts. Hence, the prevalence of T. gondii in the pig population may be a useful indicator of the risk of human toxoplasmosis associated with the consumption of pork products. The lack of obligatory screening methods at farm level for the detection of antibodies in farmed animals or the viable T. gondii in carcasses at slaughterhouse level increases the risk of contaminated pork or meat products. For this reason, the application of biosecurity and surveillance programs at farm level is very important to prevent a T. gondii infection.

The urine protein:creatinine (UPC) ratio is considered the reference method to assess proteinuria. Its diagnostic value in ovine medicine needs further elucidation. In population monitoring and/or for research purposes, it is convenient to collect many samples simultaneously and store them for later analysis. However, analyte stability data are required to ensure reliable results. We used 15 of 90 urine samples collected from sheep to assess the effect of storage time on the UPC ratio. After centrifugation, the supernatant of each sample was divided into 6 aliquots. Urine protein and creatinine concentrations were determined immediately in one aliquot using the pyrogallol red and a modified Jaffè method, respectively. The other aliquots were stored at -18°C. Based on the absence of active sediment, alkaline urine pH, and UPC ratio ≥0.2, we included 15 samples in our study. The UPC ratio was determined in the stored aliquots 2, 7, 14, 21, and 60 d after collection. The data were analyzed with univariate ANOVA. No significant difference was observed in the urinary concentrations of protein, creatinine, and the UPC ratio (0.8 ± 0.84 in conventional units and 0.09 ± 0.095 in SI units) among different times ( p  > 0.05). The UPC ratio remained stable for 2 mo in ovine urine samples stored at -18°C.

Neospora caninum and Toxoplasma gondii affect both humans and animals worldwide. To investigate their seroprevalence and differences in seropositivity between pigs vaccinated and unvaccinated against porcine circovirus 2 (PCV2), as well as differences in muscle enzyme activity between seropositive and seronegative pigs, blood samples were collected from 380 sows. Antibodies against T. gondii and N. caninum were detected by an indirect immunofluorescence antibody (IFA) assay, while the activities of creatine kinase (CK) and aspartate aminotransferase (AST) were biochemically assessed. Out of the 364 sows finally included in the study, 4.4%, 3.5%, and 0.5% were seropositive to T. gondii , N. caninum , or both. A significantly higher percentage of seropositivity against T. gondii and/or N. caninum in PCV2 unvaccinated pigs compared with vaccinated pigs was observed. Increased serum activities of CK and AST were detected in 71.43% and 100% of only against T. gondii (T+) and 63.64% and 90.91% of only against N. caninum (N+) seropositive sows, respectively, and were significantly higher compared to seronegative animals. T. gondii and N. caninum seropositivity, especially in presumed immunocompromised pigs, and the evidence of muscle damage highlight their importance as a zoonotic pathogen and animal model of human infection, respectively.

Canine parvovirus type 2 (CPV-2) primarily infects dogs, which are the main host reservoir, causing severe gastrointestinal disease associated with immunosuppression. The present study was conducted in Thessaly, Greece and aimed to identify risk and environmental factors associated with CPV-2 infection in diarrheic dogs. Fecal samples were collected from 116 dogs presenting diarrhea and were tested by polymerase chain reaction (PCR) for the presence of CPV-2 DNA. Supplementary data regarding clinical symptoms, individual features, management factors and medical history were also gathered for each animal during clinical evaluation. Sixty-eight diarrheic dogs were found to be positive for the virus DNA in their feces. Statistical analysis revealed that CPV-2 DNA was less likely to be detected in senior dogs, while working dogs, namely hounds and shepherds, had higher odds to be positive for the virus. Livestock density and land uses, specifically the categories of discontinuous urban fabric and of human population density, were identified as significant environmental parameters associated with CPV-2 infection by using Geographical Information System (GIS) together with the Ecological Niche Model (ENM). This is the first description of the environmental variables associated with the presence of CPV-2 DNA in dogs' feces in Greece.

Objectives: The aim of this study was to investigate the antimicrobial resistance (AMR) patterns of Escherichia coli strains isolated from poultry flocks suffering from colibacillosis in Greece and to detect the presence of the mcr-1 gene in isolates being phenotypically resistant to colistin. Results: A total of 150 E. coli strains were isolated from commercial layers and layer breeder flocks in Greece and tested for antimicrobial susceptibility. A high level of susceptibility was revealed for cephalosporins, neomycin, and colistin. Susceptibility varied for other antimicrobials (tetracycline, doxycycline, lincospectin, trimethoprim/sulfamethoxazole, enrofloxacin, amoxicillin), whereas no susceptibility was reported for macrolides, tiamulin, lincomycin, oxacillin. Concerning colistin resistance, 20 E. coli strains were found to be phenotypically resistant (13 strains showed intermediate resistance pattern and 7 strains fully resistance trait). Further investigation was performed by PCR, which has revealed the presence of the mcr-1 gene in one phenotypically colistin-resistant isolate. Conclusion: AMR is prevalent in layer poultry production, including resistance against colistin confirmed by the presence of the mcr-1 gene.

Anaplasma phagocytophilum is the causative agent of a disease known as tick borne fever in sheep, although fever is not always present. Due to inconclusive clinical signs, diagnosis is based on the cytological or molecular detection of the microorganism in blood and/or the determination of antibodies against A. phagocytophilum. The aim of the study was to determine the alterations caused by the presence of antibodies and/or the antigen of A. phagocytophilum in the blood cell count and morphology in sheep. Cytology and indirect immunofluorescence assay were performed for detection of antibodies and the antigen of A. phagocytophilum , respectively. The samples were divided into four groups depending on the result of the antigen and antibody detection. The samples that were only positive for antigen detection had mild anemia, leukopenia (lymphopenia), and thrombocytopenia. The samples that were positive in both assays had anemia, leukopenia (neutropenia and lymphopenia), and thrombocytopenia. Samples that were positive only for antibody detection had mild leukopenia. Morphological findings in infected sheep included band neutrophils, toxic neutrophils, reactive lymphocytes, and activated monocytes. The hematological findings along with cytological and serological tests can contribute to the assessment of the stage of the disease. A combination of leukopenia and thrombocytopenia raises a strong suspicion of the disease. When the microorganism and antibodies are simultaneously present, sheep are more susceptible to secondary complications. The first reported morphological findings and the quantitative hematological alterations are indicative of an inflammatory reaction, antigenic stimulation, and stress.

The aim of this study was to evaluate the diagnostic performance of a multiplex bead assay for the simultaneous detection of antibodies against Mycobacterium bovis , Brucella suis , and Trichinella spiralis . Sera from Eurasian wild boar of known serological status for TB (64 seropositive, 106 seronegative), Brucella (30 seropositive, 39 seronegative), and Trichinella (21 seropositive, 97 seronegative) were used for the development and evaluation of the assay. Magnetic beads coated with recombinant MPB83 antigen (TB), a whole-cell B. suis 1330 antigen, and an E/S T. spiralis antigen were used for the detection of specific antibodies using Bio-Rad Bio-Plex technology. The sensitivities (Se) and specificities (Sp) of the multiplex assay were, for M. bovis , 0.98 and 0.86; for B. suis , 1.00 and 0.97; and for T. spiralis , 0.90 and 0.99 (Se and Sp, respectively). The results show the diagnostic potential of this assay for the simultaneous detection of antibodies against M. bovis , B. suis , and T. spiralis in wild boar.

Exposure of sheep to Borrelia burgdorferi sensu lato ( s.I. ) complex, the causative agent of Lyme borreliosis (LB), has been reported in tick-abundant areas worldwide, while no data have been reported in Greece. The aim of the study was to identify the hematological alterations in sheep with seropositivity against Borrelia burgdorferi ( s.I. ). Blood samples were obtained from 318 tick infested sheep for blood analysis and serological determination of IgG and IgM antibodies against B. burgdorferi by indirect immunofluorescence antibody (IFA) assay after exclusion of endo-ectoparasites and other tick-borne infections. A total number of 162 sheep met the inclusion criteria, allocated in four groups based on the presence or absence of IgG and/or IgM; sheep found negative for IgM and IgG (Group A), positive for IgM (Group B), positive for both IgM and IgG (Group C) and positive for IgG (Group D). Anemia, thrombocytopenia and normal or decreased leukocyte count, mainly due to lymphopenia were the main hematological features observed in seropositive sheep. The presence of these features raises the suspicion of Borrelia infection in tick infested sheep. The seropositivity of 23.58% in sheep raises concerns of Borrelia circulation, especially in rural areas and potential risk of transmission to humans.

Bluetongue is a vector-borne disease with epidemic potential. Recently, outbreaks of Bluetongue were reported across Greece, caused by the Bluetongue virus (BTV) serotype 4. Regarding its pathogenesis, BTV infection involves various target organs with limited data referring to the kidneys. The objective of this study was to identify the possible impact of BTV infection on kidneys using common renal biomarkers. Urine and blood samples collected from 30 sheep with clinical signs of bluetongue (BTV sheep) and 30 clinically healthy sheep (normal sheep) from the same farms were finally selected and included in the study from an initial population of 47 sheep per group, based on the absence of active urine sediment. Complete urinalysis was performed and urine protein to creatinine ratio (UPC) and urine gamma-glutamyl transferase to creatinine (UGGTC) ratio were determined. Blood urea nitrogen (BUN), creatinine, total proteins, albumin (ALB), and inorganic phosphate (P) were determined in serum samples. UPC and UGGTC were significantly higher ( p < 0.05) in BTV sheep compared to normal, whereas urine specific gravity (USG) was significantly lower ( p < 0.05). Cylindruria was also detected in BTV sheep, and absence of azotemia in BTV and normal sheep. All these findings are indicative of renal tubular injury and/or dysfunction and suggestive of an association between BTV infection and acute damage of renal tissue.

The objectives of this experimental work were the evaluation of the potential role of Salmonella enterica subsp. diarizonae in diarrhoeic syndrome in lambs and the investigation of facets of the pathogenesis of the infection. In total, 12 lambs were challenged orally on the first day of life, with a S. enterica subsp. diarizonae isolate from a clinical case of diarrhoeic syndrome. Sequential blood, faecal and buccal samples were collected from lambs and faecal and milk samples were taken from their dams. Lambs were euthanised 1, 2, 4, 7, 10, 14 and 21 days after challenge. Samples were processed for recovery of the challenge organism; they were also subjected to examination by PCR for detection of the invA gene. Tissue samples from lambs were also examined as above and histopathologically. S. enterica subsp. diarizonae was recovered from faecal samples of all lambs, in total, from 45/77 samples (median duration: 2.4 days post-inoculation). It was also recovered from buccal samples (10/77) from seven lambs (median duration: 0.8 days), and from tissue samples (small intestine, abomasum, liver, gallbladder) of nine lambs. It was recovered from two consecutive milk samples from the same ewe, but not from any faecal sample from ewes. The invA gene was detected in samples from all lambs (median duration: 5.5 days in faecal and 1.3 days in buccal samples), as well as in milk samples from three ewes. Histopathological findings included abomasitis with subepithelial presence of eosinophils, lymphocytes and plasma cells, consistently observed in all lambs. In the small intestine, salient lesions initially included distension and oedema of intestinal villi, leucocytic infiltration and hyperplasia of lymphoid nodules with apparent germinal centres; this was followed at later stages by atrophy and/or degeneration of the lymphoid tissue of the intestine with marked subepithelial infiltration of lymphocytes, plasma cells and eosinophils.

This prospective study included four dog groups (group A: healthy dogs, groups B: dogs with idiopathic epilepsy under antiepileptic medication (AEM), C: idiopathic epilepsy dogs without AEM administration, D: dogs with structural epilepsy). The purpose of the study was to compare the proteomic profile among the four groups. Samples were analyzed by a quantitative Tandem Mass Tags approach using a Q-Exactive-Plus mass-spectrometer. Identification and relative quantification were performed using Proteome Discoverer, and data were analyzed using R. Gene ontology terms were analyzed based on Canis lupus familiaris database. Data are available via ProteomeXchange with identifier PXD018893. Eighteen proteins were statistically significant among the four groups (P < 0.05). MMP2 and EFEMP2 appeared down-regulated whereas HP and APO-A1 were up-regulated (groups B, D). CLEC3B and PEBP4 were up-regulated whereas APO-A1 was down-regulated (group C). IGLL1 was down-regulated (groups B, C) and up-regulated (group D). EFEMP2 was the only protein detected among the four groups and PEBP4 was significantly different among the epileptic dogs. Western blot and SPARCL immunoassay were used to quantify HP abundance change, validating proteomic analysis. Both, showed good correlation with HP levels identified through proteomic analysis (r = 0.712 and r = 0.703, respectively). SIGNIFICANCE: The proteomic analysis from CSF of dogs with epileptic seizures could reflect that MMP2, HP and APO-A1 may contribute to a blood-brain barrier disruption through the seizure-induced inflammatory process in the brain. MMP2 change may indicate the activation of protective mechanisms within the brain tissue. Antiepileptic medication could influence several cellular responses and alter the CSF proteome composition., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Introduction: Ruminants (cattle and sheep) with Mycobacterium avium (MAP)-induced paratuberculosis (ptb), the ruminant model of Crohn's disease (CD), exhibit pancreatic specific autoantibodies (PAB) against GP2 but not against CUZD1. Since anti-Saccharomyces cerevisiae antibodies (ASCAs) is a CD marker, we tested MAP-infected ptb ruminants for ASCA, and compared them with ruminants lacking evidence of anti-MAP serology or with ruminants, which were positive for anti-GP2 antibodies., Material and Methods: A total of 98 samples from ruminants (48 cattle and 50 sheep) were studied. IgG anti-MAP antibodies, and CD-related ASCA and anti-GP2 antibodies were tested by modified ELISAs., Results: Nine cattle (18.75%) and 20 sheep (40%) were suffered from ptb. ASCA antibodies were present in 21/48 (43.7%) cattle and 10/50 (20%) sheep while anti-GP2 antibodies were present in 14/48 (29.2%) cattle, and 8/50 (16%) sheep. ASCA antibodies were more prevalent in anti-MAP antibody positive (14/29, 48.3%) than in anti-MAP negative ruminants (17/69, 24.6%, P=0.022) and also in anti-GP2 antibody positive (13/23, 56.5%) than in anti-GP2 negative ruminants (18/75, 24%, P=0.003). No association between ASCA and anti-MAP antibody concentrations were found (r=0.159, P=0.117). A significant association between ASCA and anti-GP2 antibody concentration were observed (r=0.211 and P=0.037)., Conclusion: ASCA are present in a significant proportion of ruminants with ptb and correlate with anti-GP2 antibody positivity, a finding further supporting the notion that Crohn's disease and ptb share common immunological mechanisms of antigen-driven loss of self-tolerance., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Introduction: The objective of this study was to determine the renal dimensions in adult cows and to investigate whether they are affected by the breed, the age of the animals and the presence of subclinical renal histopathological lesions., Materials and Methods: The kidneys of 57 adult female dairy (44 Holstein) and beef (13 Limousine-Cross) cattle aged≥2years obtained at an abattoir were used for the purpose of the study. The greatest pole-to-pole length and the maximum width were measured with a digital caliper. A renal sample obtained from a central pole of each kidney underwent histopathologic examination., Results: Renal dimensions were significantly affected by the breed and the age of the animals without however been affected by the presence of mild histopathologically detected lesions. The renal size was significantly higher in Holstein cows compared to Limousine-Cross ones and significantly lower in cows aged more than 6years than the youngest ones. Left kidney was shorter and wider in the majority of animals., Conclusions: These results indicate that breed and age of animals are significant factors that affect renal size in cows. They further prove that mild histologically detected renal lesions do not significantly alter renal dimensions. Although no significant difference was detected, the obtained results tend to confirm the well documented morphological difference (asymmetry) in cattle between right and left kidney., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

European brown hare (Lepus europaeus, EBH) is probably the most important game animal in Europe throughout its historical distribution. The decline in its populations across its geographic range in Europe have been attributed to factors such as reproductive rate and the ability for adaptation, climate, feed availability, predators, anthropogenic factors and diseases. Apart from common diseases of hares with a high impact on their mortality such as European Brown hare Syndrome, EBH has been involved in the epidemiology of pathogens with zoonotic potential. In this work, the role of EBH as a source of Crimean-Congo haemorrhagic fever virus (CCHFV), Hepatitis E virus (HEV), Yersinia spp., Brucella spp., Francisella tularensis, Toxoplasma gondii and Leishmania infantum is discussed. Hares may significantly contribute to the epidemiology of important emerging zoonotic pathogens through maintenance of high endemicity levels as in the case of CCHFV, as a reservoir of important pathogens such as Yersinia spp., B. suis, F. tularensis and L. infantum and as a potential source of T. gondii for other animals, especially for carnivores but also for humans. However, EBH may also be a host of minor importance as in the case of HEV. The continuous surveillance of hare populations will enable the collection of information on the population health status and the pathogens currently circulating in the area posing risk for wildlife, domestic animals and humans. The possible live animal translocations of infected hares, the fact that this species acts as a host of vectors (fleas, ticks, mosquitoes and sandflies) and the prey of carnivores and omnivores that travel in great distances getting into contact with domestic animals and humans, further highlights the need to be included in surveillance studies. Besides, the hunter-harvested EBH population is an excellent indicator for recent pathogen transmission due to its short lifespan., (© 2020 The Authors. Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

The aim of this 6-month, randomized, blinded, controlled clinical trial was to compare the efficacy and safety of aminosidine-allopurinol combination with that of meglumine antimoniate-allopurinol combination for the treatment of leishmaniosis in dogs without stage III or IV chronic kidney disease. Forty client-owned dogs were randomly assigned to group A [n = 20; aminosidine (15 mg/kg, subcutaneously, once daily, for 28 days) and allopurinol (10 mg/kg, per os, twice daily, for 6 months)] or group B [(n = 20; meglumine antimoniate (100 mg/kg SC, once daily, for 28 days) and allopurinol (10 mg/kg, per os, twice daily, for 6 months)]. Clinical and clinicopathological evaluations, parasitic load measurement (lymph node and bone marrow microscopy, bone marrow real-time PCR), specific serology and leishmanin skin test (LST) were performed at baseline (time 1) and after 14 (time 2), 28 (time 3), 60 (time 4) and 180 (time 5) days. Both treatments were safe and resulted in significant clinical and clinicopathological improvement, reduction of parasitic load and of indirect immunofluorescence antibody test (IFAT) titer and induction of positive LST. There was no significant difference between groups with regards to the primary outcome measures of the trial that included the proportion of dogs that presented severe treatment-related side effects, were cured and were parasitologically negative at time 5. However, some (proportion of dogs that presented no clinical signs, no hyperglobulinemia and negative serology at time 5) secondary outcome measures showed significant differences in favor of the meglumine antimoniate-allopurinol treatment arm. Treatment-related death occurred in one dog in each group, while injection site reactions appeared at a similar frequency in both groups. Due to the differences in some secondary outcome measures in association with the low power of this trial, it cannot be definitively concluded that the two treatments are equally effective. Therefore, the aminisodine-allopurinol combination cannot be proposed as a first-line treatment of CanL but rather as a second-line treatment that may be particularly useful to avoid repeated administration of meglumine antimoniate and in countries where the latter is not available or registered., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to declare., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: Manual evaluation of blood cell counts on stained blood films is a common procedure in resource-limited laboratories of farm animal clinics. Moreover, settings for sheep blood cell counts are not provided on most veterinary hematology analyzers., Objectives: We aimed to (a) compare the results of white blood cell (WBC) counts evaluated microscopically on ovine blood smears with those obtained by the ADVIA 120 hematology analyzer and validate appropriate correction factors for the manual technique; and (b) assess the two suggested factors to calculate platelet counts on blood smears in sheep., Methods: The blood samples of 57 sheep were used to generate a regression equation between the average WBC count per field and the WBC count determined using the ADVIA 120 analyzer. Thirty-one new ovine samples were used to assess the agreement between the calculated WBC counts based on a generated equation and those obtained by the analyzer using the Passing-Bablok test and Bland-Altman plots. Similarly, agreements between platelet counts using two different factors for platelet calculation were assessed using the Bland-Altman plot., Results: The average bias of calculated WBC counts was 0.4%, with precision and accuracy being over 95%. Regarding calculated platelet counts, Bland-Altman plots revealed a bias of 26.4% and 1.4% when the average number of platelets per field was multiplied by 15 000 and 20 000, respectively., Conclusions: Microscopic WBC counting in ovine blood is a reliable alternative to automated analyses using the generated equation. A better agreement between the two methods was observed when a factor of 20 000 was used to calculate platelet counts in ovine blood smears., (© 2020 American Society for Veterinary Clinical Pathology.)

Trichinellosis is an important zoonosis and the most common source of human infection is meat from pigs and wild boars. The European Union (EU) supports alternative pig production systems (organic and free-ranging) as sustainable farming systems. However, these systems that allow outdoor access for farm animals, may create new or reintroduce old risks to public health. During the last years, alternative pig production systems (free-ranging or organic pig) are growing in popularity in Greece due to the increasing interest of consumers for organic products. The majority of the trichinellosis outbreaks in the EU were associated with pork and meat products including wild boars. In Greece, from 2009 to 2012, 37 Trichinella spp. positive free-ranging pigs were reported in free-ranging pig farms of Northern-Eastern Greece (31 were identified as T. britovi ). The recent re-emerging present of the Trichinella spp . infections in free-ranging pigs and wild boars are a high risk for the consumers and should alarm the Public Health Authorities in Greece and the EU. During the last years, the organic or free-ranging pig production systems are growing in popularity in the EU. However, these systems increase the risk of Trichinella spp. infections, since pigs are possible to be infected by feeding on carcasses or the offal of hunted or dead wild animals. For this reason, it is important for Public Health Authorities to be focused on the training of hunters and farmers in order to avoid the transmission among free-ranging pigs and prevent the cases of human infection., (© 2020 Urmia University. All rights reserved.)

The aim of this study was to investigate the occurrence of Bartonella spp, Brucella spp, Coxiella burnetii , and Francisella tularensis in European Brown hares ( Lepus europaeus ) hunter harvested during 2-year hunting periods in northern and central Greece. Serum samples were examined for the presence of IgG antibodies by using an immune fluorescence test and/or an enzyme-linked immunosorbent assay. PCR was used to detect Bartonella spp DNA in blood samples and Brucella spp, C. burnetii , and F. tularensis DNA in liver samples. Antibodies against Bartonella spp were detected in 12 hares (12/105); whereas none of the hares examined was seropositive for Brucella spp , C. burnetii , and F. tularensis. The presence of Bartonella spp, Brucella spp, C. burnetii , and F. tularensis DNA was not detected in the samples examined. This study did not provide any evidence that the European Brown hare is involved in the epidemiology of Brucella spp , C. burnetii , and F. tularensis in Greece. However, our results suggest that this species is exposed to Bartonella spp, which gives the impetus for further investigation of its role as another host of this bacterium.

An incident of sudden deaths in the breeding stock was reported from a farrow-to-finish commercial pig farm in Greece. The 8·4% of sows during lactation and gestation period presented anorexia, fever, haematuria, return-to-oestrus and sudden deaths (mortality rate: 2·3%). Blood and urine samples were collected from four diseased sows. Furthermore, swabs from urine bladders were collected from two dead sows and four culled sows at the slaughterhouse. Blood testing demonstrated mild leucocytosis and absence of azotaemia. Urinalysis revealed haematuria, proteinuria, bilirubinuria and active urine sediment with bacilli, epithelial cells and leucocytes, crystals and granular casts. Histopathological evaluation of the bladder demonstrated chronic active polypoid cystitis. The bacterial culture revealed the presence of Serratia liquefaciens. The antibiotic susceptibility testing showed high resistance to the most common antibiotics, with the highest sensitivity of the isolate towards quinolones. After the administration of a single dose of 7·5 mg kg -1 body weight enrofloxacin intramuscularly, the mortality rate decreased to less than 0·5% along with a remarkable reduction in the severity of clinical signs. Based on our findings, S. liquefaciens induced severe clinical signs and deaths in sows, mainly due to urinary infection. Inadequate water sanitation might have been responsible for increased exposure to S. liquefaciens. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, the isolation of Serratia liquefaciens from the urinary tract of pigs associated with clinical signs and increased mortality was described for the first time. Serratia liquefaciens is an important cause of hospital-acquired human infections. The isolate in this study was resistant to the most common antibiotics. Therefore, the use of quinolones which are drugs of last resort for treatment of infections was the only therapeutic option. The presence of the resistant bacterium in the urinary tract raises concerns for its zoonotic potential., (© 2019 The Society for Applied Microbiology.)

Background: Urinalysis is not routinely used in bovine medicine, and there is no evidence as to whether urine protein-to-creatinine ratio (UPC) could be used for the diagnosis of renal diseases in cattle., Objective: The goal of the study was to determine alterations in UPCs observed with different subclinical renal diseases in clinically healthy cattle and to investigate whether UPC can efficiently differentiate cattle with and without subclinical renal pathology., Methods: Kidney and urine samples from 57 clinically healthy adult dairy (44) and beef (13) cattle were collected after slaughter. Urinary protein and creatinine concentrations were measured in an automatic analyzer, and urinary-specific gravity (USG) was measured using a temperature compensated refractometer. Kidney samples underwent histopathologic examination, and the cattle were classified as NL (no renal lesion) and L (lesions detected even in one kidney). Based on USG, the cattle were divided into the Normal USG (≥1.020) and Low USG (<1.020) groups. The cattle with either histopathologic lesions or low specific gravities were considered to have renal disease., Results: Renal lesions were detected in 37 cattle. UPC values were significantly affected (P < .05) by USG values, and not by the type of lesion detected, breed, or age, and their interactions (P > .05). The analysis revealed that a UPC of ≥0.19 provided an optimal cut-off point for the differentiation between normal animals and those with renal disease with 66.0% sensitivity and 90% specificity., Conclusions: The UPC calculation is a useful tool for the differentiation of normal cattle and those with renal disease. A UPC of less than 0.19 is associated with the absence of renal damage, whereas higher values raise suspicion for renal disease., (© 2020 American Society for Veterinary Clinical Pathology.)