The clinical correlation of findings obtained by fine needle fenestration biopsy of the canine spleen with visible ultrasound changes

The medical records databases (March 2016 to March 2021) of the Faculty of Veterinary Medicine, University of Zagreb, were examined to determine the frequency and clinical relevance of cytological diagnoses from fine-needle fenestration biopsy (FNFB) of the spleen in dogs with visible ultrasound changes. Seventy-five dogs were divided into clinically relevant and irrelevant groups, according to the clinical relevance of the diagnosis. The incidence of clinically relevant diagnoses was 28/75 (37.3%). Malignant diagnoses were over-represented (23/28; 82.1%), followed by hemorrhages/hematomas (3/28; 10.7%) and suppurative inflammation (2/28; 7.1%). The most common malignancy was lymphoma (12/28; 42.9%). There was no correlation between the ultrasound lesions examined and the relevant cytological diagnoses, except in cases of patchy echo texture (P = 0.010). Lesion size greater than 1.74 cm had the highest sensitivity/specificity values (91.3%; 42.1%) with a significant area under the curve (AUC) of 0.68 (P = 0.029) for predicting clinically relevant findings. The complication rate due to bleeding was 2/130 (1.5%). In conclusion, splenic FNFB can be safely performed in dogs as it carries a low risk of development of complications. Specific ultrasonographic findings, such as patchy echo texture pattern, may increase the suspicion of the presence of neoplastic disease in the form of lymphoma. According to ROC analysis, lesion size greater than 1.74 increases the possibility of predicting clinically relevant findings.

Association of genetic variants in STAT5B gene with milk performance and mastitis related traits in dairy cattle

Udder performance and health are important traits in dairy cattle worldwide. The present study aimed to explore the association of single nucleotide polymorphisms (SNPs) in the STAT5B gene in dairy cattle with milk performance and mastitis related traits. The study included 201 cows of three pure breeds (i.e. Holstein Friesian, Jersey and Achai) and two crossbred cattle at four established dairy farms in Khyber Pakhtunkhwa, Pakistan. The milk samples were analysed for somatic cell count (SCC) and milk composition (i.e. fat , protein and lactose percentages). The generalized linear model was deployed for association analysis using SAS. The pool DNA sequencing showed four (three synonymous and a 3/ UTR) SNPs in STAT5B. These SNPs were further validated in all DNA samples using SNaPshot assay. The breed-wise analysis showed that most of the SNPs were consistent with Hardy-Weinberg equilibrium (P>0.05). The association analysis revealed a significantly higher protein percentage in TT genotype and lower SCC in CC genotypes of SNP 1 (exon 2, C>T), whereas in SNP 2 (exon 16, T>C) the TT genotypes revealed significantly lower SCC and SCS compared with other genotypes (P<0.05). SNP 4 (3/ UTR, C>T) showed significantly lower SCC and frequency of clinical mastitis in the heterozygous (CT) genotype compared to the homozygous genotypes. The in silico predictions revealed changes in the RNA secondary structure for SNP 2 and SNP 3. The study suggests that STAT5B should be considered as a candidate gene, and the variants identified as useful genetic markers for improved milk composition and udder health.

Differential expression of antioxidant genes during clinical mastitis of cow caused by Staphylococcus aureus and Escherichia coli

There is considerable interest in the hypothesis that oxidative stress is enhanced in the pathophysiology of clinical mastitis. The main goal of this research was to establish profiles of antioxidant gene expression in the milk of cows with clinical mastitis. Standard bacteriology was conducted on pretreatment milk specimens from 77 cows with clinical mastitis between 15 and 70 days in milk (DIM). Examinations were performed on mRNA expression of antioxidant genes, such as catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD). Additionally, levels of lipid peroxidation were measured in milk samples from healthy cows and those with clinical mastitis. The isolated bacteria consisted of Staphylococcus aureus (S. aureus, 10.48%), Streptococcus agalactiae (7.69%), Streptococcus dysagalactiae (6.29%), and Escherichia coli (E. coli, 29.37%). E. coli was the most prevalent pathogen found in the milk of cows with clinical mastitis in early lactation. The mean level of malondialdehyde (MDA) in clinical mastitis samples was significantly higher (P<0.05) than that of healthy cows. The results revealed that the expression profiles of SOD in mastitis milk induced by S. aureus were significantly (P<0.0001) up-regulated compared with E.coli. In addition, the mRNA levels of GPx in mastitis milk due to E.coli were significantly (P<0.0001) over expressed compared to S. aureus. CAT gene expression had a tendency to be enhanced in mastitis milk induced by S. aureus compared with mastitis in cows due to E.coli. These results showed that the mRNA levels of antioxidant genes may differ depending on the type of bacteria, and diminished expression of antioxidant genes might increase susceptibility to mastitis.

The effects of Pinus brutia bark extract on pure and mixed continuous cultures of rumen bacteria and archaea, and fermentation characteristics in vitro

The aim of the study was to investigate the effects of Pinus brutia bark extract, which is rich in polyphenolic compounds of tannins, on both pure and mixed continuous cultures of rumen bacteria and archaea, as well as on rumen fermentation characteristics in vitro. Antimicrobial susceptibility assay with pure cultures was carried out in an anaerobic chamber. Pinus brutia bark extract exhibited a potential inhibitor activity (P<0.05) against pure cultures of Ruminococcus flavefaciens, Eubacterium ruminantium, and Methanobacterium formicicum while a growth stimulatory effect (P<0.05) was observed for Ruminoccocus albus, Butyrivibrio fibrisolvens, and Streptococcus bovis. Pinus brutia bark extract only had a potential inhibitor effect (P<0.05) on R. albus at the highest dose (1200 µg/mL). Pinus brutia bark extract also stimulated (P<0.05) the growth of pure cultures of Fibrobacter succinogenes, while it did not affect Megasphaera elsdenii, except at the highest dose. The effects of two doses (75 and 375 mg/L) of P. brutia bark extract on in vitro mixed cultures and rumen fermentation parameters were determined by the rumen simulation technique (Rusitec). Supplementation with P. brutia bark extract led to a quadratic decrease (P<0.05) in the cell numbers of R. flavefaciens. Production of total and individual short chain fatty acids (SCFA), acetate to propionate ratio (C2/C3), total protozoa, ruminal pH, and dry matter digestibility (DMD) did not change in the presence of P. brutia bark extract. Supplementation with both doses of P. brutia bark extract decreased (P<0.05) the ammonia-N concentrations. Ammonia-N concentration was lowest in the high-supplemented group (P<0.05). As a conclusion, inhibitory effects of P. brutia bark extract on some species in the pure cultures were in the same direction as with mixed ruminal cultures, while stimulatory effects disappeared. The lack of inhibitory effects on protozoa and on a large number of Gram-positive rumen bacteria in the mixed cultures suggests that its mechanism of action is not exactly similar to antibiotics. Although P. brutia bark extract did not alter ruminal SCFA, it could have potential to improve ruminal protein utilization without depressing rumen microbial fermentation.

The effect of the red blood cell system disorders on the further development and productivity of Holstein calves that had had bronchopneumonia

The aim of this research was to study the effect of red blood cell system disorders on the further development and productivity of calves that had had bronchopneumonia. The study included 170 Holstein heifers at the age of 180-195 days: healthy heifers (n = 92) and heifers with moderate bronchopneumonia (n = 78). All animals had been under constant clinical supervision for 27 months. Blood sampling was performed in 10 animals from each group at the beginning of the experiment and on day 22, and also when they reached a body weight of 380 kg (the first artificial insemination), 30 days before the planned calving date, and on days 7, 90, 180 of lactation. The blood samples obtained were examined by a hematological analyzer, and the content of fetal hemoglobin was determined by Singer’s method. It was shown that after completion of the course of treatment the clinical symptoms of bronchopneumonia disappeared in calves, but microcytic hypochromic anemia appeared. Further, in these animals an increase was registered in the age of fruitful fertilization by 10.8% and of the insemination index by - 36.7%. After calving and during lactation, the severity of hypochromia decreased, but there was a tendency for macrocytosis, which, combined with increased anisocytosis, indicated hypoplastic anemia. This form of anemia occurred against the background of pathological residual changes after pneumonia due to the depletion of the compensatory potential of the bone marrow and its functional overload during lactation. The operational anemia revealed in lactating cows caused a decrease in milk productivity by 23.2-26.7% (P<0.01).

Combination of photodynamic therapy and phototherapy for the treatment of cutaneous open wounds in dogs - case reports

The evolution of cutaneous open wounds was evaluated in five client-owned dogs treated with photodynamic therapy (PDT), mediated by methylene blue and phototherapy with light-emitting diodes (LED) as the light source. On the first day of treatment, the wound was irrigated with sterile 0.9% sodium chloride solution, the surface was dried, and 3% solution of hydrogen peroxide soaked for 1 minute. The hydrogen peroxide was mechanically removed and, immediately after, 0.01% methylene blue was used in the wound for five minutes, after which a cluster of red LEDs (650 - 670 nm wavelength, 350 mW power, and irradiance of 200 mW/cm2) was used. Subsequently, single phototherapy sessions, every 48 hours or more, were applied until wound healing. In general, 24J was used for PDT and 12J for phototherapy. In all cases, the cutaneous open wounds treated with PDT associated with phototherapy healed without complications, and it may be considered as a treatment option, especially for extensive lesions.

The effect of follicle size and cryoprotectants on nuclear maturation and early embryonic development of vitrified - thawed Awassi sheep oocytes (Ovis aries)

In this study, two experiments were conducted to study the effect of both the follicle size and the cryoprotectants dimethyl sulfoxide (DMSO) and ethylene glycol (EG) on the main phases of nuclear maturation (Experiment I), cleavage stages and embryo quality (Experiment II) of Awassi sheep oocytes. Follicles were classified into two groups: small follicles (SF) (1-2 mm) and large follicles (LF) (> 2 mm). Oocytes were vitrified in three solutions: A (30% DMSO), B (30% EG) and C (15% DMSO and 15% EG). In Experiment I, the resulting vitrified-thawed oocytes in solution C achieved the best rates after the control group (fresh), respectively as the rates of maturation, germinal vesicle (GV), metaphase II(M-II), arrest, and lyses were 85.71% (P = 0.04), 8.33% (P = 0.02), 72.92% (P = 0.04); LF group, 15.25% (P = 0.04), and 5.08% (P = 0.04); SF group, respectively. In Experiment II, the same group of oocytes achieved the best rates after the control group, as the rates of fertilization, cleavage, 2-16 cell, Type3, blastocyst, and Type1 embryos were 63.28% (P = 0.001), 57.46% (P = 0.001), 40.38% (P = 0.04), 38.46% (P = 0.04); LF group, 30.00% (P = 0.01), and SF group 36.67% (P = 0.001), respectively, while the vitrified-thawed oocytes in A solution (SF group) reached the highest rate of Type 2 embryo quality (58.06%; P = 0.01). No significant differences were noticed in the germinal vesicle breakdown (GVBD), metaphase I (M-I) and morula stage. Vitrification of oocytes obtained from follicles with a diameter of more than 2 mm in a cocktail solution of DMSO (15%) and EG (15%) led to a significant increase in the yield and quality of the resulting sheep embryos.

Establishment of a sandwich ELISA for bovine plasma PON-1 and its predictive capabilities for dairy cows with fatty liver

The aim of this study to improve the clinical diagnosis of fatty livers (FL) in dairy cows by using the paraoxonase-1 (PON-1) enzyme as a detection index. Prokaryotic expression technology was used to generate recombinant bovine PON-1 protein. Mice were immunized with this protein to generate hybridoma cells, stably secreting anti-PON-1. Cells were injected into the peritoneal cavity of mice, and ascites were purified to generate bovine PON-1 monoclonal antibody. Rabbits were then immunized with this antigen, and a polyclonal antibody against bovine PON-1 was obtained. Using monoclonal and polyclonal antibodies, a double-antibody sandwich ELISA for plasma PON-1 was constructed. Plasma samples were collected from healthy (n = 13) and FL (n = 13) cows, and plasma PON-1 levels were detected using the PON-1 ELISA. Receiver operating characteristic curve (ROC) analysis was used to analyze correlations between PON-1 levels and FL. Results showed that the ideal working concentration of the monoclonal antibody was 0.8 mg/mL, and the quantitative detection limit was 90 ng/mL. Plasma PON-1 levels were significantly lower in FL cows, when compared with healthy animals. It is concluded that PON-1 ELISA predicts risk factors for dairy cows with FL. PON-1 levels in plasma can be used as an early warning indicator for FL and concentration of 61.87 nmol/L was identified as warning index.

Antibodies to Bluetongue, Akabane and Schmallenberg viruses in native dromedary camels in Turkey

Sera from 86 Turkish native camels from seven provinces in Turkey were collected and tested for specific antibodies to Bluetongue virus (BTV), Akabane virus (AKAV) and Schmallenberg virus (SBV) using ELISA. The BTV, AKAV and SBV antibodies were found in 53.5%, 51.2% and 15.1%, respectively. Furthermore, the seropositivity for multiple infection was the highest for dual infection with AKAV and BTV (25.6%), followed by triple seropositivity (9.3%). These findings indicated that BTV, AKAV and SBV circulate in camels in Turkey at a relatively high rate, and that an active surveillance program is needed for the management and tracing the dynamics of these infections in the Turkish camel population.

Determining the effect of natural inhibitors on sesame meal degradability using in vitro three step method

The aim of this experiment was to investigate the beneficial effect of monensin, tannic acid and cinnamon essential oil addition on sesame meal degradability by the three-step in vitro method. The effect of experimental additives on the degradability of sesame meal in the rumen, after rumen and in the whole gastrointestinal tract was significant (P<0.05). The in vitro ruminal and intestinal digestibility of sesame meal crude protein with experimental additives was in the range of 76 to 84% and 49 to 60%, respectively. The intestinal degradability of crude protein increased with the addition of cinnamon essential oil (about 10%). Addition of monensin, tannic acid, and cinnamon essential oil significantly increased the degradability of Neutral Detergent Fiber (NDF) and Acid Detergent Fiber (ADF) in the rumen, intestines and the whole gastrointestinal tract. The results showed that cinnamon essential oil (125 mg/L) increased the degradability of dry matter (DM), organic matter (OM), crude protein (CP), ADF and NDF in the rumen, after rumen and the whole digestive tract compared to all treatments (P<0.05). The results showed that addition of tannic acid (100 mg/L) decreased the disappearance of crude protein in the rumen, while it increased crude protein’s disappearance in the after rumen (P<0.05).