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Antimicrobial resistance (AMR) is an increasing global concern in small animal veterinary practices. The overuse and misuse of antimicrobials in companion animals can contribute to the spread of AMR between animals and humans. This study aimed to assess small animal veterinarians’ knowledge, attitudes, and behaviors towards antimicrobial use (AMU) and AMR in North Macedonia. A web-based questionnaire was developed, including questions addressing demographics, owner influence on antibiotic prescription and prescribing practices. The response rate of the small animal veterinarians was 47.50% (57/120), with most of them (84.20%) having 1-15 years of work experience. Pearson’s chi-square test of independence was used for contingency tables that met the chi-square assumption, and Fisher’s exact test was conducted for contingency tables that did not. Most veterinarians responded that they had perceived pressure and direct demands from pet owners to prescribe antimicrobials (35.09% frequently, 45.61% occasionally). In the survey, 50.77% of the veterinarians reported frequently administering antibiotics without conducting bacteriological or antimicrobial susceptibility testing. They typically resorted to these tests in cases with recurrent infections, such as skin infections (19.63%), or when empirical therapy is ineffective (14.72%). Based on the findings, it can be concluded that veterinarians play a crucial role in tackling the AMR in North Macedonia. The lack of stewardship programs or guidelines for responsible antimicrobial use in companion animals is a critical issue that requires urgent attention, emphasizing the vital role of veterinarians in combating AMR.

Infectious hematopoietic necrosis (IHN) is a common disease in the intensive production of salmonids. The IHN virus (IHNV) was isolated for the first time in North Macedonia in 2018. This study aimed to investigate the distribution and genotype of IHN in farmed rainbow trout and autochthonous salmonid fish in North Macedonia following the first detection. The samples were collected from 47 trout farms. Trout fry with or without clinical signs of IHN were selected as individual samples. Kidney, spleen, and heart were taken from each fish during the dissection. Three pooled samples were collected from each farm. A total of 141 pooled samples were collected: Rainbow trout (Oncorhynchus mykiss) n=127, Macedonian trout (Salmo macedonicus) n=11, and Ohrid trout (Salmo letnica) n=3. The virus was detected in 43 samples (30.50%): rainbow trout (n=40), Macedonian trout (n=2), and Ohrid trout (n=1). There were 18 (38.30%) positive fish farms. The MAKIHNV1 isolate from 2018 (MN641902) and the newly isolated virus shared a similarity of >99 and were placed in clade E-1 of European genogroup E. The IHN has spread throughout the country and is also present in the autochthonous salmonids.

The present study aimed to determinate the effect of external progesterone (P4) supplementation on luteolysis in cows under heat stress. Forty-eight (n=48) dairy cows in the period from July September 2018 were part of and at day 35±3 postpartum scored for BSC, synchronized using PG-3-G + Ovsynch protocol and randomly allocated into two treatments: PRID group (n=27) treated with external P4 device between G1 and PGF2α and CON group (n=21) left without treatment. Collection of blood samples to assess P4 concentrations was done at Pre-PG, at G1, at PGF2α, at 72 h after PGF2α (at timed artificial insemination TAI) and at d 21 after TAI. The pregnancy diagnosis was done at d 21 and d 30 after TAI by ultrasound. The average temperature humidity index (THI) was 79.5±0.6. At G1, the P4 was significantly lower in the PRID group (1.84±0.99 ng/mL) in comparison to the CON group (2.97±1.82 ng/mL). In contrast, at PGF2α, there was a tendency (p=0.09) of increased P4 concentration in PRID group compared with the CON group (4.26±1.68 and 3.74±2.39 ng/mL), respectively. At TAI, more PRID cows (p=0.0001) had a lower P4 (0.06±0.03 ng/mL), in comparison to CON (1.28±2.41 ng/mL). At d 21 and d 30 after TAI, more PRID cows were predicted and diagnosed pregnant (16/27 or 59.25% and 13/27 or 48.14%) compared with the CON group (11/21 or 52.38% and 8/21 or 38.08%) respectively, but without any significant differences. Supplementation of the P4 during the Ovsynch protocol increases the P4 before TAI and reduces the incomplete luteolysis in heat stressed dairy cows.

Hyperadrenocorticism (HAC) in dogs is routinely treated with trilostane single-dose (CTG) which is reported to cause adverse reactions. The current retrospective study of several dogs with terminal stage of HAC aimed to compare the clinical, hematological, and biochemical effects of trilostane titration-dose treatment (TTG) with the single-dose treatment (CTG). All clinical cases (n=7) were confirmed on HAC by anamnestic, clinical, hematology, biochemistry, and low-dose dexamethasone suppression test findings, indicative for Cushing’s disease. Two cases were treated with CTG (2.2-6.7 mg/kg, single dose daily) and their treatment was discontinued on the second week due to adverse reactions. The TTG cases were treated for up to 12 weeks (0.5 mg/kg once daily for 7 days, and then with 0.5 mg/kg twice daily for 7 days). Blood samples and clinical checks were performed on 0., 4., and 12. weeks of the treatment. Hemoglobin was non-significantly higher in TTG at 12 weeks. Alanine transaminase was significantly lower in the TTG cases on the 12. week of the treatment (78.04±15.37 U/L) compared to the 0-week (137.81±24.03 U/L), and 4-week samples (131.92±23.36 U/L). No significant differences were observed with the CTG cases. Alkaline phosphatase was significantly lower on 12-week samples in TTG (251.02±93.06) compared to the 4-week (567.94±283.93 U/L), and 0-week samples (1,341.84 U/L). In conclusion, TTG has indicated to have significantly higher tendency to decrease alanine transaminase and alkaline phosphatase, alleviating the negative effects on the liver. The clinical findings were more adverse for the CTG.

Brilliant cresyl blue (BCB) staining is a method used for selection of developmentally competent porcine oocytes, in addition to the standard morphological classification. The aim of the current study was to assess the relationship between BCB staining in porcine oocytes with high and low morphological characteristics and its maturation rate. In the first part of the study, cumulus-oocytes complexes (COCs) (n=434) were aspirated from 60 ovaries. According to the morphological classification and BCB staining, they were divided in 4 groups: COC+/BCB+, COC-/BCB+, COC+/BCB-, and COC-/BCB-. In the second experiment, COCs (n=445) were categorized into 3 groups: control, BCB+, and BCB-. Significant differences in the maturation rate were observed between COC+/BCB+ and COC-/BCB- (66% vs. 23%), and between COC+/BCB+ and COC-/BCB+ (66% vs. 38%) (p

The incidence of clinical mastitis (CM) in small and large-scale dairy cow farms can be highly variable and can be affected by age, parity, post-calving status, and atmospheric conditions. The current study aimed to assess the CM-incidence and its association with the post-calving days, number of lactations, season, and number of affected udder quarters in dairy cows in small-scale dairy farms. The study was conducted within one calendar year in 177 small-scale farms with 864 dairy cows. Clinically confirmed CM cows (n=72) were sampled from each udder quarter and processed for bacteriology examination. The CM-positive samples were grouped according to the season (spring, summer, autumn, and winter), the number of days after calving (180), the number of lactations (1-st, 2-5-th, and >6-th), and the number of affected udder quarters (one, two, three, and four). The CM-positive samples (n=72, 8.3%) were confirmed on bacteriology examination in significantly lower count (n=56, 6.5%). The 2-5-th lactations cows (n=35, 68.6%) were significantly more compared to the first lactation (n=6, 11.8%), and >6-th lactation cows (n=10, 19.6%). CM cows with infection of one udder quarter (n=40, 78.4%) were significantly more than the cows with two (n=6, 11.8%), and four infected quarters (n=5, 9.8%). The CM-incidence in small-scale dairy cow farms in North Macedonia was 8.3% and 6.5% by clinical and bacteriology examination, respectively. The highest CM susceptibility was observed in the cows between the second and fifth lactations. One udder quarter was most frequently affected in CM-positive cases.

Artificial insemination (AI) as a part of assisted reproductive technologies represents the oldest and most widespread method used to accelerate genetic progress in all domestic animals. After its first implementation in ovine reproduction and almost 80 years afterward, AI is continuously used for improving the genetic merit, utilizing either fresh or shorttime chilled semen. Nevertheless, regardless of the semen used for insemination, the conception rate (CR) is still lower in comparison to natural service. At least two factors are commonly thought to limit the success of the AI and reduce the CR: (1) failure of placing the semen directly into the uterus due to the specific anatomic structure of the ewe’s cervix; (2) lower viability of ram spermatozoa during cryopreservation (

Cystic echinococcosis is a cosmopolitan zoonotic infection, commonly seen in areas where livestock and dogs are kept together. The objective of this study was to investigate the prevalence of echinococcosis in slaughtered cattle and sheep in North Macedonia and to evaluate the location and fertility/sterility rates of the cysts. A total of 302 slaughtered animals were examined with visual inspection, palpation, and incision of the internal organs in order to detect cysts. Microscopic examination was carried out to determine the presence of protoscoleces. The overall prevalence of echinococcosis in slaughtered cattle and sheep was 60%. The presence of cysts increased with age in cattle. In both cattle and sheep, the most common affected organs were the liver and lungs infected with more than one cyst. The findings showed higher fertility rate in sheep which confirmed their role as the most important intermediate host. This study shows that North Macedonia is an endemic region, and serious control strategies should be implemented, with special emphasis on safety disposal of infected organs and anthelmintic treatment of the dogs.

The present study aimed to evaluate whether the induction and the formation of an accessory corpus luteum (CL) after AI might increase the pregnancy per AI (P/AI) in heat stressed dairy cows. Starting at d 50±3 post-partum, 113 lactating Holstein cows from one commercial herd during summer were scored for body condition, blood sampled and examined by ultrasound. Those bearing a CL>25mm and progesterone (P4) level>2ng/mL were synchronized using a double PGF2α injection given 12 h apart and AI-ed at detected estrus. In total 18 cows, there were not any signs of estrus (n=10) nor a P4 level 0.05). In conclusion, the induction of an accessory CL at d 5 after AI might increase P/AI in heat stressed dairy cows.

The bovine kappa-casein (κ-CN) is a phospho-protein with 169 amino acids encoded by the CSN3 gene. The two most common gene variants in the HF breed are CSN3*A and CSN3*B while CSN3*E has been found with lower frequency. The aim of this study was to optimize a laboratory method for genotyping of these three alleles as well as to determine their genotype and allele frequencies in the HF cattle population in the Republic of North Macedonia. Genomic DNA was extracted from full blood from 250 cows. The target DNA sequence was amplified with newly designed pair of primers and the products were subjected to enzymatic restriction with HindIII and HaeIII endonucleases. Genotype determination was achieved in all animals. The primers successfully amplified a fragment of 458 bp and the digestion of this fragment with both endonucleases enabled differentiation of five different genotypes with the following observed frequencies: AA (0.39), AB (0.29), BB (0.16), AE (0.10), and BE (0.06). The estimated allele frequencies were: CSN3*A (0.584), CSN3*B (0.336) and CSN3*E (0.08). The observed genotype frequencies differed significantly (P

Reduced glutathione (GSH) and homologous ram seminal plasma (HSP), used as additives in cryopreserving (CP) media prior to freezing, showed conflicting results in retaining structural integrity and progressive motility in post-thawed ram spermatozoa. The aims of this research were: (1) to assess the effect of GSH and/or HSP supplementation via soybean-lecithin CP extender on cryopreserved ram spermatozoa viability, morphology and motility pattern; and (2) to assess the effect of incubation in the context of the previous aim. Quantitatively and qualitatively, homogenized and pooled ram ejaculates (N=10) were extended with one of the following extenders: control (C) – tris-based, GSH and HSP-free, experimental-1 (E1) – C + GSH 5 mM, experimental-2 (E2) – C + HSP 20 % and experimental-3 (E3) - GSH 5 mM + HSP 20 %. Following thawing, samples were taken at 0- and 3-hours from each group (n=10) and were assessed for spermatozoa viability, morphology, and motility pattern. C-0h samples yielded a spermatozoa population with low viability, altered head morphology and highly deviated motility pattern. E3-3h samples yielded spermatozoa with unaffected viability, head morphology and high progressive motility. In conclusion, E3 extender added to cryopreserved-thawed ram spermatozoa is most efficient in obtaining high viability, unaltered head morphology, and progressive motility.

Several reports indicated that a large proportion of dairy cows have not resumed cyclicity until day 60 after calving. These cows are traditionally classified as non-cycling (anoestrous or anovular cows). Static ovaries (SO, lack of luteal tissue and follicles >8 mm, and progesterone < 0.5 ng/mL) could be a possible underlying reason that contributes to a non-cycling status. Although SO affects both primiparous (PP) and multiparous (MP) cows, PP cows are more prone to be non-cycling than MP. Therefore, this study aims to compare the metabolic profiles and hormonal status between non-cycling PP and MP cows diagnosed with SO. One hundred and twenty one animals that did not express signs of oestrus until day 60 postpartum were grouped by parity (PP, n=58 and MP, n=63), then blood sampled and examined using transrectal ultrasonography. Blood samples were collected before the ultrasonographic examination. Out of those, 42 PP (72.4%) and 28 MP (44.4%) were diagnosed as non-cycling (bearing SO). Serum concentrations of triglycerides, cholesterol, total protein and albumin did not differ between parity groups. The glucose concentrations in PP cows (1.43 ± 0.59 mmol/L) and MP cows (1.69 ± 0.71 mmol/L) did not differ, however, they were less than the normal physiological concentration. In addition, no differences were detected between parity groups for concentrations of NEFA, β-HBA, progesterone and estradiol. In summary, we concluded that non-cycling PP and MP cows bearing SO have similar hormonal status and metabolic profiles.

Postpartum anestrus is a physiological phenomenon in high-producing dairy cows. Static ovaries have been related as major contributors for its occurrence causing a significant reproductive problem to the dairy industry. Different treatment methods have been employed with inconsistent rate of success in initiation of cyclicity, requiring further investigations in order to achieve satisfactory results. The aim of the present study was to compare the ovarian response in cows diagnosed with static ovaries, more than 60 days postpartum using two different hormonal treatment (GnRH and eCG) methods. A total of 58 acyclic cows (no CL, follicles0.05). Cows in Group 2 had significantly higher incidence of multiple ovulations than cows in Group 1 (p

Motility patterns of spermatozoa are an indicator of their fertilizing capacity. Reduced glutathione (GSH) has been reported to induce positive effects on biological quality of the frozen-thawed spermatozoa. The aim of this study was to investigate the effects of GSH addition to semen extender on the following kinetic parameters: continuous line velocity (VSL), average path velocity (VAP), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), linearity (LIN), straightness (STR), beat cross frequency (BCF), total motility (tMOT) and progressive motility (pMOT), and to appoint them as indicators of its presence. A soybean-based extender was used for dilution of the semen samples, fractioned in two parts, one containing GSH (5 mMol/ml) and second without GSH. The ejaculates (n=48) were collected from two rams (January – May, 2013) which were classified in two groups according to the used extender: Group 1 (with GSH, n=24) and Group 2 (without GSH, n=24), and then frozen in liquid nitrogen on -196°C degrees in a programmable freezer. Assessment of the samples has been performed postthawing (30 sec. at 37°C) on CASA equipment, acquiring kinetic parameters. Results showed that only VSL and BCF have a statistically significant difference between group1 and group 2 (102.98±15.13 vs. 88.47±20.63, t=2.77, p