Abstract The present study aimed to explore the connection between the acoustic parameters of dog sounds and the morphometric characteristics of the laryngeal cartilages in dogs with various body sizes. We compared the measurements of the laryngeal cartilages (n=15) of equal number of small, medium, and large size dog cadavers with the acoustic parameters recorded from other dogs of similar size and number. The morphometry was performed with a digital caliper. Animal sounds were analysed with Raven Pro 1.6 software. Waveform and spectrogram graphs were displayed for each sound with 512-point Hann window 50% overlap time. Sound selections were manually verified. Our findings confirmed that small-sized dogs generate the highest pitched barks in every measured frequency (5%-95% and peak). In addition, their vocal signals are of the lowest tonality expressed in sound-to-noise ratio (SNR). The Frequency parameters (F) 5%, 25%, and 95% showed significant correlations with the morphometric values of the laryngeal cartilages, which indicates their role in the sound formation. These acoustic parameters had strong to moderate negative correlations with the thyroid cartilage (TC) width and height; arytenoid cartilage (AC) – distance between proc. corniculatus and proc. cuneiformis (corn-cun) , depth of ventricle and distance between the two proc. cuneiformis (cun-cun) ; width, height and depth of cricoid cartilage (CC); width and height of the epiglottis (EP). Our finding suggested that F5%, F25% and F95% can be reliable parameters and can be used in the classification methods of dogs’ sounds as it varies predictably with laryngeal cartilages size.

Abstract Polycystic ovarian syndrome (PCOS) is the most predominant endocrine disorder responsible for female infertility. The clinical treatment strategies of PCOS only provide symptomatic relief but are often unsatisfactory. Asparagus racemosus and Vitex negundo have long been used as traditional herbal intervention in treating various metabolic and reproductive issues. Therefore, a pressing need for a better alternative approach is essential. The study aimed to assess the effect of A. racemosus (ARA) and V. negundo (VNA) aqueous extract on treating PCOS-like symptoms in female rats. Letrozole (1.0 mg/kg BW) was used to induce PCOS in rats which were then treated with ARA and VNA in a dose of 250 mg/kg BW orally for 21 consecutive days. These herbs improved the estrous cycle after being perturbed by letrozole. ARA and VNA significantly increased the level of estradiol and estradiol receptor ( ESR1 ) in PCOS rats, which further prevented uterine shrinkage. Post treatment of these herbs also revealed a notable decline in serum glucose and triglyceride levels in letrozole-induced PCOS rats. Letrozole caused reproductive and metabolic alterations without inducing oxidative stress, evidenced by higher activity of SOD and catalase in PCOS group. However, both supplemented groups showed baseline level of SOD and catalase similar to the vehicle-treated control. Moreover, ARA and VNA administration decreased the appearance of cystic follicles in histomorphological study by regulating ovarian folliculogenesis. Hence, this is the first time we reported that restoration of normal reproductive and metabolic function in letrozole induced PCOS by ARA and VNA were independent of oxidative stress.

Abstract This study aimed to assess the severity of anemia, associated parameters, and their impact on cardiac function in dogs naturally infected with Ehrlichia canis (E. canis), a zoonotic pathogen primarily transmitted by the brown dog tick. Forty-seven dogs, comprising 38 infected and 9 healthy controls, were classified into non-anemic (n=12), mildly mild (n=9), moderately (n=11), severely (n=4), and very severely anemic (n=2), groups. Hematological and echocardiographic assessments were performed, along with separate measurements of cardiac biomarkers (e.g., cardiac troponin I [cTnI]) and iron metabolism biomarkers (e.g., serum iron, total iron-binding capacity [TIBC], and transferrin saturation). Hematological analysis revealed a significant reduction in hematocrit (17.11±1.34%), hemoglobin (5.22±0.88 g/dL), and platelet counts (38.25±22.62×10⁹/L) in severely anemic dogs compared to other groups. Echocardiographic findings demonstrated increased interventricular septal thickness during diastole (1.16±0.60 cm) and systole (1.38±0.10 cm) in moderately anemic dogs, indicating compensatory adaptations. Dogs with severe anemia exhibited elevated left ventricular end-diastolic (3.56±0.54 cm) and end-systolic dimensions (1.71±0.22 cm) in comparison to control dogs (p<0.05). Fractional shortening (53.83±7.71%) and ejection fraction (77.58±8.19%) were diminished among the severely anemic group, denoting altered cardiac function. Elevated cardiac troponin I (cTnI) levels (up to 0.29±0.22 ng/mL) were noted in a limited number of cases, suggesting that myocardial injury was not widespread. This study demonstrated that anemia severity significantly influences myocardial and hematological parameters in E. canis-infected dogs, with evidence indicating compensatory cardiac remodeling in response to chronic anemia. Further research utilizing advanced diagnostic techniques is recommended to better understand disease progression and associated complications.

Abstract Swine influenza caused by the influenza A virus significantly affects pig production and pig health due to acute respiratory disease and huge economic losses. Pigs play an essential role in the epidemiology of influenza because they act as a mixing vessel for the formation of potentially pandemic zoonotic strains. The objective of our study was to assess the seroprevalence of Swine influenza A viruses (swIAV) in commercial pig farms in Macedonia. A total of 373 blood samples were collected from piglets aged 1 to 4 weeks of sows with different parities from 19 different commercial farrow-to-finish pig farms. For the detection of anti-IAV antibodies, sera samples were analyzed using a competitive ELISA. All farms were seropositive to swIAV. Seropositivity was detected in 258 (69.2%) samples, ranging between 10 and 100% at farm level. The highest seroprevalence was found in piglets from sows in 5-6th parity. In contrast, the lowest seropositivity was found in samples from the youngest sows (1-2nd parity), which indicates that the virus has circulated for a longer period in these farms. Furthermore, large farms with more than 120 sows had a significantly greater percentage of seropositive animals than small farms with less than 120 breeding sows (83% vs. 54%, respectively). In conclusion, our results demonstrated that swIAV circulates endemically in Macedonian commercial farrow-to-finish pig farms, underscoring the need of immunization in preventing infection on these farms.

Abstract The objective of the current research was to determine expression, function and regulation of bone morphogenetic protein 15 (BMP15) during follicular development in laying hens. A trial was conducted with 40 layers from Lohman Klassik Brown breed (40 weeks old). At the end of the study fifteen layers were humanely killed and their ovaries were then dissected. Ribonucleic acid (RNA) expression of BMP15 was analyzed in the ooplasm and in granulosa cells. It was significantly higher in the ooplasm (p<0.01). BPM15 expression was not found in the granulosa cells from 6-8 mm and >9 mm follicles. The expression for bone morphogenetic protein 15 receptors (BMPR1B and BMPR2) in the granulosa cells was in significant positive correlation with the follicle size (p<0.05). The results obtained in this study demonstrate the possible role of BMP15 in developing oocytes. BMP15 expression is important for the growth regulation and signaling in the follicular cells in the preovulatory phase.

Abstract Back pain is one of the most common triggers of performance failure in athletic and riding horses. Diagnosis of equine back pain has been very challenging for equine practitioners, particularly in chronic cases. Therefore, the identification of blood biomarkers would facilitate the clinical differentiation of chronic back pain. This study aimed to investigate serum biomarkers of glial cell activation, axonal damage, and inflammation for the diagnosis of equine chronic back pain. Serum samples from forty horses comprising chronic back pain (CBP), back pain concurrent with lameness (BPL), lameness (LN), and healthy control (HC) (n=10 per group) were screened for ionized calcium-binding adaptor molecule 1 (Iba-1), glial fibrillary acidic protein (GFAP), phosphorylated neurofilament-H (pNF-H) by ELISA, and proinflammatory cytokines (IL-1β, IL-6, and TNF-α) by multiplex assay. Serum concentrations of GFAP (3.81±1.72 ng/mL) and pNF-H (0.76±0.18 ng/mL) were significantly (p<0.05) higher in horses with CBP when compared with other groups. Iba-1 was not significantly higher in CBP horses. There was no significant difference between the pro-inflammatory cytokines among the groups. The levels of IL-1β, IL-6, and TNF-α were also increased in the CBP than the HC control horses but lower in relation to BPL and LN horses. In addition, serum Iba-1, GFAP, and pNF-H showed a high discriminatory capacity for horses with CBP with high sensitivity (50-100%) and specificity (70-100%). This study provides evidence that serum levels of the GFAP and pNF-H may be useful in the clinical differentiation of horses with chronic back pain.

Abstract The study aimed to investigate the Staphylococcus species from dogs with chronic otitis externa in Istanbul and to determine their antibiotic susceptibility patterns. Ear swab samples were collected from 100 dogs suspected of otitis externa admitted at the clinics of Istanbul University - Cerrahpaşa, Faculty of Veterinary Medicine. The dogs were of different age, gender, and breed. The bacterial isolation was performed by conventional methods. BD Phoenix Automated Microbiology System was used to confirm bacterial identification by conventional methods and to test antimicrobial susceptibility. Staphylococcus spp. were isolated from 36% of the samples collected from the dogs. S. pseudintermedius, S. aureus, S. epidermidis, S. hyicus and S. chromogenes were identified in 41.6%, 22.2%, 11.1%, 5.5%, and 5.5%. In Staphylococcus spp. isolates, enrofloxacin, penicillin, and ampicillin-sulbactam resistance was 8.3%, marbofloxacin resistance was 11.1%, doxycycline resistance was 16.6%, amoxicillin-clavulanic acid, erythromycin, and gentamicin resistance was 19.4%, tetracycline, clindamycin, and sulphonamide resistance was 25%. Methicillin resistance was not observed in any of the isolates. However, multiple drug resistance (MDR) was detected in 11 (30.5%) of 36 isolates. In conclusion, the early detection and antimicrobial sensitivity testing of Staphylococcus spp in dog otitis externa cases that do not respond to empiric therapy could be beneficial for appropriate antibiotic selection and treatment thus preventing MDR.

Abstract This article, part 2 of a 2-part series, describes the next two steps in the application of the Modified Calgary-Cambridge Guides (MCCG) to consultations in bovine medicine, ‘explanation and planning’, and ‘closing the consultation’, and introduces concepts that are associated with all the components of the guide, ‘building the relationship with the client’ and ‘providing structure to the consultation’. Part 1 introduced the aim and framework of the MCCG which enables the practitioner to gain an insight into the client’s understanding of the problem, including understanding aetiology, epidemiology and pathophysiology. Part 2 introduces the framework that provides the opportunity to understand the client’s expectations regarding the outcome, their motivation and willingness to adhere to recommendations. It also describes how to engage and acknowledge the client as an important part of the decision-making process, how to establish responsibilities of both the client and practitioner, and how to reach out to the client at the conclusion of the consultation to make certain that the client’s expectations were met.

Abstract The objective of this study was to examine the effect of ketoprofen with or without combination with xylazine on the level of cyclooxygenase-2 in mice. The intraperitoneal (i.p.) dose of ketoprofen and xylazine that caused an analgesic response in half of the mouse population was 1.26 mg/kg and 6.63 mg/kg, respectively. Serum cyclooxygenase-2 concentration (activity) in the control mice was 16.94 ng/ml. The ketoprofen-treated group (2.52 mg/kg, i.p.) decreased the cyclooxygenase-2 concentration by 58% (7.16 ng/ml). The combined ketoprofen and xylazine treatment (13.26 mg/kg, i.p.) decreased the cyclooxygenase-2 by 94% (0.98 ng/ml). The ketoprofen plasma concentration in the combined treatment group was significantly higher compared to the ketoprofen treatment group. Ketoprofen plasma concentrations measured at 0.25, 0.5, 1, 2, 4, and 24 hours were 19.07, 18.94, 14.66, 6.53, 5.44, and 5.54 µg/ml, respectively. Plasma concentrations of ketoprofen and xylazine were raised to 28.74, 29.74, 15.32, 13.04, 14.64, and 11.95 µg/ml or by 51%, 56%, 5%, 100%, 169%, and 116%, respectively. Ketoprofen pharmacokinetic variables were increased (AUC0-∞ (515%), AUMC0-∞ (2389%), MRT (305%), t1/2β (375%), Tmax (100%), and Cmax (55%)), while other values were decreased (Kel (79%), Vss (25%), and Cl (88%)). Our findings suggested a synergistic interaction between ketoprofen and xylazine on the level of cyclooxygenase-2 (pharmacodynamic interaction) which was exerted by modification of the ketoprofen pharmacokinetic properties in mice.

Abstract The objective of our research was to estimate the therapeutic index and assess the interaction of alfaxalone (IP) with ketamine or xylazine (IM) in chicks by using isobolographic analysis. The up-and-down technique was involved to calculate the median effective anesthetic dosages (ED50) of alfaxalone, xylazine, and ketamine given separately or at the same time in young chicks. Then the up-and-down technique was involved to estimate the median lethal dosage (LD50) of alfaxalone (IP) to determine the safety profile. The ED50 of all anesthetics was evaluated isobolographically to assess the type of interaction between alfaxalone and xylazine or alfaxalone and ketamine. The alfaxalone ED50 was 32.88 mg/kg (IP), whereas the LD50 was 102.40 mg/kg (IP). The ED50 values for alfaxalone, ketamine, and xylazine were 32.88, 12.24, and 2.45 mg/kg, respectively. The ED50 values of alfaxalone with ketamine or xylazine (25:25 ED50 values) were: 7.39+2.35, and 8.61+0.63 mg/kg, respectively. ED50 values were decreased when the combinations of alfaxalone/ketamine or alfaxalone/xylazine were administered by 22-21% and 26-25%, respectively. The anesthesia of chicks with alfaxalone is safe, produces a surgical stage of anesthesia, and can be used for minor surgical procedures. The use of alfaxalone with ketamine or xylazine has been shown to have a synergistic effect and these findings may be of clinical relevance in poultry or may be extended to mammals following further clinical trials.