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Somatic cells (SCs) in milk, which include epithelial cells from the gland and blood cells, are present in milk during the normal course of milking. Increase in SCs is found in mastitis-affected dairy cows and can be a useful indicator for estimating mammary health and milk quality worldwide. The aim of this study was to determine whether there was a pattern of somatic cell count (SCC) in mastitis-affected cows on three different farms. The study was conducted on three dairy farms of high milk-producing cattle breeds in Vojvodina during 2021. Samples were taken from 15 cows from each farm, all of the cows being diagnosed with clinical and subclinical mastitis. The SCC in milk samples was determined by the microscopic reference method according to the standard (SRPS EN ISO 13366-1:2010) of the Institute for Standardization of Serbia. The value of SCC was considered as high if >200.000 cells/mL, as this is the threshold indicating secretion disorder. In order to examine the differences between the observed three farms, one-factor analysis of variance (ANOVA) was applied, while a post-hoc LSD test was used for determination of statistically significant differences between the SCC in cows on three different farms. The mean values of the SCC on Farms 1, 2 and 3 were 7,055,266.67, 2,619,893.33 and 552,000 cells/mL, respectively. Based on the results, a statistically significant difference (p

Mastitis is one of the most common and costly diseases affecting dairy cows worldwide. Since antibiotic resistance has become a global threat to both animal and human health, it is becoming more urgent to continuously search for new therapeutical alternatives for the control and treatment of bovine mastitis. Hence, our research aimed to test the therapeutic use of two essential oils (EOs) based on their chemical composition, antibacterial and antioxidant potential. The present study was conducted by collecting milk samples from the cows diagnosed with clinical or subclinical mastitis with the aim of isolating and identifying bacterial strains. The antioxidant potential of essential oils of Menthae piperitae (MP) and Melissa officinalis (MO) was evaluated in several in vitro assays. In the MP EO, a total of 38 compounds were identified, with menthol as the dominant compound, whereas in MO EO 51 compounds were identified. Furthermore, the values of minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs) have been used to quantitatively measure the antibacterial activity of each essential oil. In accordance with which, MP EO samples exhibited a higher degree of antibacterial activity than MO EO. Thus, EOs have been shown to be promising alternatives to antibiotics because of their availability, biodegradability, and lower risk of side effects as compared with conventional, antimicrobial treatment. Nevertheless, further clinical studies are needed to test the potential role of EOs in treating mastitis in dairy cows.

Bovine mastitis is an important disease in the dairy industry responsi­ble for the welfare and significant economic losses in dairy cows. The treatment of choice for mastitis is the administration of antibiotics. However, this therapeutic choice has some disadvantages including presence of antibiotics residues in the milk, low cure rate as well as rapid increase in antibiotic-resistant pathogens. Therefore, new alternative approaches to antibiotics were investigated by different groups of researchers in order to find an effective approach for bovine mastitis therapy. This review was conducted in order to analyze different publications on usage of essential oils in relation to bovine mastitis. There are many in vitro studies for evaluating the antimicrobial efficacy of essential oils against many mastitis associated pathogens. In addition, numerous of tested essential oils have shown good efficacy with a wide range of minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs). On the other hand, only several in vivo studies have focused on therapeutic effects of essential oils. Moreover, recent studies indicate the possibility of using essential oils in the fight against biofilm which could be promising fight against bovine mastitis since unsuccessful antibiotic treatment can be associated with the presence of biofilms.

Ever since their discovery, antimicrobials have helped in controlling and treating infections in both humans and animals. The control of infectious diseases is endangered by the rise of microorganisms that are resistant to this group of drugs. Limited availability of authorized veterinary drugs leads to prescription of human approved drugs. The aim of our study was to describe metronidazole use patterns and its accordance with scientific literature in Serbia. Results have shown that majority of prescriptions were written to dogs, while 27.1% prescriptions were for cats. Most common general conditions were dental and digestive disorders. Our study shows that metronidazole is available in oral and injectable form, while cats were only treated with injectable formulation. Even though pre­scription of human approved drugs for companion animals is allowed by Law, there is no official record of data on the extent or nature of off-label use in Serbia. Such information is essential for guiding antimicrobial use policy in small animal veterinary practice as well as for assessing the risk of transmission of antimicrobial resistance to humans. Recognizing the importance of antimicrobial resistance and ensuring more judicious use of antibiotics are key elements to any strategy for maintaining the usefulness of antimicrobial drugs in animals and humans.

With the growing global pet population and increased spending on veterinary care, compounded medications offer customized, often more suitable and affordable treatment options compared to the limited available veterinary medications. This research aims to understand pet owners' attitudes towards compounded medications, focusing on their challenges and needs. A total of 300 respondents from the territory of Novi Sad, province of Vojvodina, Republic of Serbia completed the questionnaire, through face-to-face interviews at veterinary clinics. Pet owners stated that the majority of companion animals were dogs (66.7%), followed by cats (22.8%). A significant portion of them received medication in the past five years (70.7%), while nearly half of them were human-approved (47.9%) out of which more than half (55%) of the pets received antimicrobials for systemic use. Although the majority of pets did not receive compounded medication (79.8%), respondents believed that both they and their pets would benefit from it. Only small fraction of respondents knew that local pharmacies had the ability to compound medications for pets (14.5%). In the era of personalized medicine, compounded medication for specialized needs play an important role providing optimum therapy for veterinary patients. However, limitations persist, including variability in formulation quality, limited regulatory oversight, and challenges in ensuring consistency in potency and stability, as well as decreased efficacy for certain drugs and the challenges related to the lack of pharmacokinetic data for some administration routes. By customizing treatments for individual veterinary patients, we can optimize antimicrobial use, reduce the selective pressure that drives resistance, and enhance patient outcomes, while acknowledging the need for careful oversight and quality control in compounding practices to ensure safety and efficacy., Competing Interests: Declarations. Competing interests: The authors have no relevant financial or non-financial interests to disclose. Ethics approval: The study was approved by the Ethical Committee of the Faculty of Agriculture, University of Novi Sad (approval number 9000–104/1). Consent to participate: Informed consent was obtained from all individual participants included in the study., (© 2025. The Author(s), under exclusive licence to Springer Nature B.V.)

The increase in antimicrobial resistance (AMR) is a major global health problem with implications on human and veterinary medicine, as well as food production. In the poultry industry, the overuse and misuse of antimicrobials has led to the development of resistant or multi-drug resistant (MDR) strains of bacteria such as Salmonella spp., Escherichia coli and Campylobacter spp., which pose a serious risk to meat safety and public health. The genetic transfer of resistance elements between poultry MDR bacteria and human pathogens further exacerbates the AMR crisis and highlights the urgent need for action. Traditional methods of preserving poultry meat, often based on synthetic chemicals, are increasingly being questioned due to their potential impact on human health and the environment. This situation has led to a shift towards natural, sustainable alternatives, such as plant-derived compounds, for meat preservation. Essential oils (EOs) have emerged as promising natural preservatives in the poultry meat industry offering a potential solution to the growing AMR problem by possessing inherent antimicrobial properties making them effective against a broad spectrum of pathogens. Their use in the preservation of poultry meat not only extends shelf life, but also reduces reliance on synthetic preservatives and antibiotics, which contribute significantly to AMR. The unique chemical composition of EOs, that contains a large number of different active compounds, minimizes the risk of bacteria developing resistance. Recent advances in nano-encapsulation technology have further improved the stability, bioavailability and efficacy of EOs, making them more suitable for commercial use. Hence, in this manuscript, the recent literature on the mechanisms of AMR in the most important Gram-negative poultry pathogens and antimicrobial properties of EOs on these meat isolates was reviewed. Additionally, chemical composition, extraction methods of EOs were discussed, as well as future directions of EOs as natural food preservatives. In conclusion, by integrating EOs into poultry meat preservation strategies, the industry can adopt more sustainable and health-conscious practices and ultimately contribute to global efforts to combat AMR.

Bovine mastitis, as a significant and prevalent health problem in dairy herds, is primarily attributed to bacterial infections. Yeasts, although considered relatively rare causative agents, have also been associated with mastitis in dairy cattle. Current mastitis treatment predominantly relies on antibiotics, with limited emphasis on antifungal treatment. However, mycotic mastitis is challenging to treat, since these fungi are often resistant to antibiotics and may even utilize them for energy. In the current research, the in vivo antimicrobial activity of the essential oil-based formulation (Phyto-Bomat), as a possible alternative mastitis treatment associated with yeasts, was studied. This study involved a total of 68 animals from two dairy farms with diagnosed mastitis, and three treatment groups were established: conventional antibiotic treatment, Phyto-Bomat treatment, and the combination of both. The findings suggest significant variations in the presence of Candida samples based on the treatment administered, and the most significant difference was noted in cows treated with the combination (Phyto-Bomat and antibiotics). Yet, it is important to note that the results reveal that, regardless of the treatment type, there are statistically significant differences in the presence of Candida samples across the examined time points. These results aim to provide valuable insights into the potential of EOs as an alternative therapy in bovine mastitis, specifically targeting yeasts. Such findings could offer new strategies in the mycotic mastitis control and reducing the occurrence of secondary infections following antibiotic treatment.

The dairy sector is essential for the world's food systems, playing a vital role in agricultural production. Yet, mastitis, an inflammation of the mammary gland, affects a considerable portion of the dairy cow population annually, leading to reduced productivity and sustainability. Insight into therapeutic protocols is essential for a better understanding of the situation on farms with the aim of developing new or harmonizing existing protocols in the mastitis treatment. This study conducted on dairy farm in Serbia aimed to explore the most commonly used antibiotics during bovine mastitis treatment and their association with daily milk yield. Data from 100 lactating cows, including therapeutic protocols, lactation numbers, mastitis episodes, milk yield, and antibiotic usage, were obtained from a computerized database and statistically analysed. The results demonstrated a high prevalence of narrow-spectrum antibiotics usage, with the majority of cows treated with combination therapy. The route of antibiotic application did not significantly impact daily milk yield, while treatment duration had no discernible effect. Nonetheless, specific antibiotics were associated with milk yield variations, suggesting the need for careful antibiotic selection and management in mastitis therapy. These findings emphasize the importance of prudent antibiotic use to safeguard animal health and long-term milk production sustainability., (© 2023 Wiley-VCH GmbH. Published by John Wiley & Sons Ltd.)

Mastitis is one of the most important diseases in dairy cows, leading to substantial economic losses associated with decreased milk production and quality. Early detection of changes in metabolic and milk parameters is crucial for maintaining animal welfare and milk quality. This study aimed to detect patterns in metabolic and milk composition parameters in Serbian dairy cows affected by mastitis. It also examined the relationship between these factors in cows with clinical and subclinical mastitis, as well as in healthy cows. This study included 60 Holstein-Friesian cows with the same body score condition that were in the same lactation phase. They were divided into three groups of 20: clinical and subclinical mastitis and a control group of healthy cows. The categorization was based on clinical udder health and the California mastitis test. Blood serum metabolic profiles were measured using a Rayto spectrophotometer (Shenzhen, China), and milk composition was determined using MilcoScan TM (Foss, Hilleroed, Denmark) and Fossomatic TM (Foss, Hilleroed, Denmark) instruments. Significant increases in non-esterified fatty acids (NEFAs), beta-hydroxybutyrate (BHB), total protein, globulin, urea, total bilirubin, magnesium, and enzyme activity were noted in mastitis-affected cows compared to healthy ones. Additionally, mastitis-affected cows had higher total protein and globulin levels and increased somatic cell counts (SCCs), while albumin concentrations were decreased. Furthermore, a negative correlation between total protein and lactose suggested inflammation leading to reduced lactose levels due to cell damage, infection, and lactose use by mastitis pathogens. Hence, indicators of the energy and protein status of the metabolic profile, together with the chemical composition of milk, may be significant diagnostic tools for detecting, monitoring, and predicting the outcome of mastitis in cows.

Antimicrobial resistance (AMR) poses a significant threat in veterinary medicine due to the excessive and inappropriate use of antimicrobial agents, compromising the effectiveness of these drugs. To combat AMR, the collection of data on the consumption of antibiotics is paramount, as there is a well-established connection between antibiotic use and AMR in both humans and food-producing animals. Hence, the current study aimed to generate measurable data concerning the sales patterns of antimicrobial drugs used in animal treatment in Montenegro over a five-year period (from 2017 to 2021). Furthermore, the study aimed to compare these sales figures with the overall sales of antimicrobial veterinary medicine products (AMVMPs) during the same period, with particular emphasis on the utilization of critically important antimicrobials (CIAs) for human use. Data on AMVMPs consumption from 2017 to 2021 were expressed in euros and were sourced from the Institute for Medicines and Medical Devices of Montenegro (CInMED) annual reports, complying with the regulatory framework of the Law on Medicines in this country. Research results indicate that the sales of AMVMPs increased from 2017 to peak in 2019, followed by a stable decline of 21.79% in 2021. However, the portion of selected CIAs AMVMPs in total sales experienced a minor decline from 2017 to 2019, followed by a noticeable 6.11% increase from 2019 to 2021. In order to address AMR challenges, these findings emphasize the importance of enhancing surveillance and monitoring of veterinary antimicrobial use, as well as CIAs for human use.

Despite preventive and therapeutic measures, mastitis continues to be the most prevalent health problem in dairy herds. Considering the risks associated with antibiotic therapy, such as compromised effectiveness due to the emergence of resistant bacteria, food safety issues, and environmental impact, an increasing number of scientific studies have referred to the new therapeutic procedures that could serve as alternatives to conventional therapy. Therefore, the aim of this review was to provide insight into the currently available literature data in the investigation of non-antibiotic alternative approaches. In general, a vast number of in vitro and in vivo available data offer the comprehension of novel, effective, and safe agents with the potential to reduce the current use of antibiotics and increase animal productivity and environmental protection. Constant progress in this field could overcome treatment difficulties associated with bovine mastitis and considerable global pressure being applied on reducing antimicrobial therapy in animals.

Increasing pressure on animal and human health as well as food security, due to antimicrobial resistance, is leading to increased demand for natural-product-derived antibiotics worldwide. Considering the importance of bovine mastitis as well as the huge challenge posed by mastitis therapy in cattle production, the main goal of this research was to evaluate the effectiveness of the essential oil-based pharmaceutical (Phyto-Bomat), as an alternative to the existing treatment with antibiotics. The therapeutic response of the cows was monitored using clinical and bacteriological cure of the proposed formulation. Among 550 dairy cows, 75 were diagnosed with a clinical or subclinical form of mastitis and divided into three experimental groups: treated with antibiotic; Phyto-Bomat; antibiotic and Phyto-Bomat. Indicators of bacteriological cure were somatic cell count (SCC), total bacterial count, and causative agent isolation, while the clinical cure was accessed by observing physical udder conditions. All mentioned indicators were assessed before and after the treatment (1st and 7th day post-treatment), with all of the three therapeutic protocols. After Phyto-Bomat treatment, no visible signs of irritation were noticed, while no significant effect in reducing SCC and total bacterial count was observed. Phyto-Bomat has shown promising results in the treatment of mastitis, without the risk of udder irritation, alone or in combination with antibiotics as part of a mastitis control program during lactation.

Mastitis in dairy cows is responsible for major economic losses on dairy farms worldwide as the most expensive and prevalent disease in dairy cattle. In spite of the fact that antibiotic therapy still remains the main treatment strategy for bovine mastitis, concerns about the shortcomings of this treatment approach are continuously raised. Hence, research on alternative treatments with increased effectiveness and reduced costs is needed. Therefore, we conducted a pharmacoeconomic analysis of conventional antibiotic vs. a proposed Phyto-Bomat treatment based on essential oils in bovine mastitis therapy. Treatments were compared from the farmer's perspective in the domain of costs (expressed in total, direct and indirect, cost differences) and effectiveness (expressed in daily milk yield differences). Economic calculations were based on data from a dairy farm in Serbia. The average cost of conventional antibiotic treatment was estimated at EUR 80.32 consisting of therapy costs, veterinary services and milk rejection costs at EUR 16.54, EUR 17.68 and EUR 46.10, respectively. The average cost of Phyto-Bomat treatment was estimated at EUR 76.34 with therapy costs of EUR 34.34, veterinary costs of EUR 32.00 and rejection of milk costs of only EUR 10.00. Therefore, Phyto-Bomat results in cost savings approximating EUR 4 per each mastitis episode with the highest cost reductions obtained in milk rejection costs. This estimation of Phyto-Bomat's economic benefits could be used as a starting point for the inclusion of this formulation as an alternative treatment approach with a focus on subclinical mastitis since it contributes to most of the financial losses.

Antimicrobials have had an important impact on animal health and production performance. However, non-prudent antimicrobial use (AMU) in food producing animals is considered to contribute to the emergence of antimicrobial resistance (AMR), with a potential impact on both animal and public health. Considering the global importance of AMR, and the threats and challenges posed by mastitis and mastitis therapy in livestock production, the main objective of this study was to quantify AMU on three dairy farms in Serbia and to examine whether there is an association between AMU and the emergence of antimicrobial resistance of mastitis-associated pathogens. Antimicrobial susceptibility testing was performed by the disk diffusion method using causative agents isolated from the milk samples of 247 dairy cows. AMU data were obtained for a one-year period (May 2021 to May 2022) based on antibiotic prescriptions listed in electronic databases kept by farm veterinarians. To estimate antimicrobial drug exposure at the farm level, the veterinary drug Defined Daily Dose was calculated by multiplying the total amount of antibiotic used on the farms during the study period by the quantity of antibiotic in the administered drug and number of original drug packages used. The results on the association between the use of common antibiotics in mastitis treatment and AMR of isolated mastitis-associated pathogens confirm a pattern that could raise awareness of the importance of this aspect of good veterinary and clinical practice to combat the global threat of AMR.

Due to the emergence of antibiotic-resistant bacteria, the risk it represents to public health, and the possible consequences for animal health and welfare, there is an increasing focus on reducing antimicrobial usage (AMU) in animal husbandry. Therefore, a great interest in developing alternatives to AMU in livestock production is present worldwide. Recently, essential oils (EOs) have gained great attention as promising possibilities for the replacement of antibiotics. The current study aimed to test the potential of using a novel EO-based pharmaceutical formulation (Phyto-Bomat) in bovine mastitis treatment. The antibacterial activity was performed using the microdilution technique. Lactating dairy cows were treated with 15 mL of Phyto-Bomat in the inflamed quarter for 5 consecutive days in order to analyze blood and milk samples for thymol and carvacrol residues using gas chromatography and mass spectrometry (GC-MS). Antimicrobial activity expressed as the minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) indicates that this formulation has the highest activity against Gram-positive strains. The dominant compounds in Phyto-Bomat were thymol and carvacrol, at 12.58 ± 1.23 mg/mL and 23.11 ± 2.31 mg/mL, respectively. The quantification of these two compounds in evaluated biological samples showed that 24 h after administration the concentration of thymol and carvacrol in milk samples was at the same level as before application. On the other hand, thymol and carvacrol were detectable in plasma samples even after 24 h post-treatment, with values ranging from 0.15-0.38 and 0.21-0.66 µg/mL, respectively. The tested formulation showed encouraging results of antibacterial activity against bovine mastitis pathogens, as well as the withdrawal period of dominant compounds, which implies that further testing regarding the bacteriological and clinical cure rates in clinical settings is needed.

Mastitis is considered to be one of the most important diseases of dairy cows in terms of health, production, and economy. Being the most common cause of antibiotic consumption in dairy cows, treatment of this disease is one of the biggest challenges in the veterinary profession as an increasing number of pathogens develop resistance to antibiotics used in the treatment. Therefore, new alternative approaches for limiting the use of antibiotics in livestock are required. For this reason, our study aimed to investigate prevalence of environmental mastitis associated bacterial strains, as well as the sensitivity of isolated strains to different antibiotics. Additionally, the therapeutic potential of three essential oils (EOs) was tested against bovine Serratia spp. and Proteus spp. mastitis pathogens, based on their chemical composition, as well as antibacterial potential. The study was carried out on 81 milk samples collected from dairy cows with mastitis. In order to determine prevalence of S. marcescens and P. mirabilis , microbiological isolation and identification were performed. Antimicrobial susceptibility testing was performed by disk diffusion method and the microdilution method was used to determine the antibacterial activity of selected EOs. In the oregano EO, a total of 23 compounds were detected, with carvacrol as a dominant component (78.94%). A total of 26 components were present in the EO of common thyme, where thymol was the most abundant compound (46.37%). Thymol also dominated (55.11%) the wild thyme EO. All tested EOs displayed antibacterial activity against all strains to different extents, while wild and common thyme EOs were the most effective. It could be concluded that the tested EOs represent promising therapeutic candidates for effective non-antibiotic treatment of mastitis.