Most human salmonellosis cases are linked to the consumption of poultry products contaminated with Salmonella enterica serovar Typhimurium. In poultry, infections cause intestinal inflammation, decreased weight gain, lower egg production, and potential economic losses for the poultry sector. This study examined the effects of a natural antimicrobial mixture (AuraShield, As) on Salmonella enterica serovar Typhimurium SE10/72 infection of chicken macrophage-like cells (HD11), focusing on infection inhibition, inflammatory response, cell viability, metabolic changes, and oxidative stress. The HD11 macrophage-like cell line was used in three infection scenarios: HD11 cell pretreatment, co-incubation with As, and co-incubation with As-pre-treated bacteria. The results showed that the natural antimicrobial mixture significantly reduced bacterial infection levels in macrophage-like cells without affecting cell viability. Low infectivity rates were also observed, along with a significant reduction in the expression of pro-inflammatory cytokines (IFN-β, IL-1β, IL-6, IL-8, IL-10). We have observed that itaconate increases, via Irg1 expression, in HD11 cells exposed to 0.25% As, alongside a significant decrease in the isocitrate lyase (AceA) gene expression in bacteria. The findings provide insights into the potential use of AuraShield as a feed additive to improve poultry health and resistance against bacterial pathogens, by minimizing Salmonella infection risk and the associated inflammatory response in macrophage-like cells. The observed modulation of metabolic and oxidative stress markers further suggests a multifaceted mode of action, presenting a promising pathway for alternative antimicrobial strategies in poultry health management.

This study investigates the effects of the organic acid mixture AuraAqua (Aq) on the pathogenicity of Aeromonas hydrophila in primary gut cells of shrimp (SGP) and tilapia (TGP). A. hydrophila, an important pathogen in aquaculture, damages gut barrier integrity and promotes systemic infection with a significant economic impact. Organic acids are proposed as antibiotic alternatives, and we aimed to investigate their impact as a mixture AuraAqua (Aq—5% maltodextrin, 1% sodium chloride, 42% citric acid, 18% sodium citrate, 10% silica, 12% malic acid, 9% citrus extract, and 3% olive extract) on the growth, virulence traits, and host cell protection during A. hydrophila infection SGP and TGP cells. Aq exhibited an MIC of 1% and an MBC of 3%. Sub-inhibitory concentrations significantly reduced adhesion and invasion in SGP and TGP cells, preserved TEER, and decreased LDH release, indicating reduced cytotoxicity and barrier disruption. Aq impaired motility and biofilm formation in a dose-dependent manner. Transcriptional analysis showed downregulation of hcp1, act, fla, hlyA and acg, whereas TonB, aerA and alt were largely unaffected. Aq also strongly reduces capsular polysaccharide production both in vitro and during infection, which likely weakens immune evasion and persistence. In host cells, Aq significantly decreases oxidative stress markers (H2O2, SOD, CAT) during infection, indicating a beneficial effect on host responses. Overall, Aq acts as an anti-virulence agent, weakening A. hydrophila’s ability to colonize and damage gut epithelia while reducing host oxidative stress, supporting its use as a non-antibiotic preventive strategy in aquaculture, pending further in vivo studies.

Environmental pollution with heavy metals (HMs) and microplastics (MPs) could enhance the global health challenge antimicrobial resistance (AMR). Herein, we explore the complicated mechanics of how HMs, MPs, and AMR are interlinked within microbial ecosystems, as well as the co-selection and cross-resistance mechanisms. Unlike antibiotics, HMs have influenced microbial evolution for billions of years, promoting resistance mechanisms that predate antibiotic resistance genes (ARGs). At the same time, this conundrum is further complicated by the pervasive spread of MPs in the aquatic and terrestrial environments, acting as substrates for bacterial pathogenic biofilms and accelerates the horizontal gene transfer (HGT) of ARGs and heavy metal resistance genes (MRGs). This review highlights that HMs such as lead (Pb), mercury (Hg), arsenic (As), chromium (Cr), cadmium (Cd), and nickel (Ni) have persistently selected for resistance traits through efflux systems and genetic co-regulation. Together, these interactions are amplified by MPs that create genetic exchange hotspots due to biofilm formation. These dynamics are modulated by organic matter, which serves both as a nutrient source and a mediator of HM bioavailability, directly influencing ARG abundance. Soil and water ecosystems, including riverine systems and landfill leachate, are reservoirs for ARGs and ARG–MRG combinations, with notable contributions originating from anthropogenic activities. This review also emphasizes the urgent need for integrated environmental and public health strategies to mitigate pollutant-driven AMR. This work seeks to approach HMs and MPs as synergistic drivers of AMR such that both HMs and MPs are upstream (causes) levers, a foundation from which future research on sustainable environmental management practices and health policy (One Health Approach), aimed at curbing the spread of resistance determinants can proceed.

Campylobacter hepaticus is responsible for the emergence of Spotty Liver Disease (SLD) in commercial layer hens. Our experimental in vitro approach demonstrates that natural antimicrobial mixtures (AuraShield - As) can significantly inhibit (P < 0.0001) the growth and motility of C. hepaticus. The antimicrobial acted as a chemoattractant for the pathogen, an effect which is concentration-dependent and leads to reduced bacterial motility. The As reduced C. hepaticus invasion of chicken liver cells (LMH cells) by strengthening TEER. The As also modulates inflammatory responses (notably by lowering IL-8 expression via the NF-κB pathway) and enhances host cell defences. We demonstrate that the presence of As (0.5%) during the infection of LMH cells by C. hepaticus significantly mitigates oxidative stress by reducing the detectable levels of H2O2 released into the culture supernatants (P<0.0001). We also show that a reduced H2O2 level will not activate the NF-kB pathway, resulting in low levels of IL-8 orthologs expression. In vivo, As was able to significantly reduce (P=0.03) not only the number of C. hepaticus identified in the livers of the infected hens but also the number of liver lesions, while restoring the integrity of the intestinal barrier. These in vitro and in vivo findings present new opportunities for enhanced SLD management strategies that integrate dietary innovations with improved flock biosecurity. By adopting a holistic approach that combines dietary, environmental, and management interventions, producers may achieve more sustainable and effective outcomes in the ongoing battle against Campylobacter hepaticus and Spotty Liver Disease.

Campylobacteriosis is a major public health challenge with many human Campylobacter infections attributed to consuming contaminated poultry products. Despite a marked decrease in Campylobacter-positive chicken carcasses and improved control measures, the incidence of Campylobacteriosis remains high, a problem potentially exacerbated by the move away from routine antibiotic use in poultry production. This chapter looks at vectors of transmission for Campylobacter in the poultry supply chain. This chapter explores the range of strategies for enhancing poultry meat safety, including improved biosecurity measures, bacteriophages, probiotics, and natural antimicrobials, as well as chemical and physical treatments. Importantly, the delicate balance of these applications with environmental and sustainability The chapter discusses how these interventions, when applied optimally at various stages of the poultry production cycle, significantly reduce contamination, leading to safer poultry products.

The ever-growing global demand for animal protein forces the aquaculture industry to expand at a pace which imposes significant challenges in maintaining sustainable practices. This study aimed to investigate the efficacy of an organic acid mixture (Aq) in mitigating Lactococcus garvieae (L. garvieae) virulence through its effects on bacterial virulence (EPS production, biofilm, and haemolytic factors) and host pathogenicity, including its adherence to CHSE-214 cells, haemolysis, and proinflammatory responses. Our findings reveal that Aq significantly inhibits L. garvieae growth at a 0.125% concentration, suppresses EPS-related gene expression, reduces biofilm formation, and reduces cytotoxicity in fish epithelial cells (CHSE-214). Moreover, Aq decreased haemolysing gene expression (hly1, hly2 and hly3) and attenuated red blood cell haemolysis, a hallmark of L. garvieae pathogenicity. Lastly, Aq was demonstrated to induce modulation in the host immune responses, lowering IL-1β and IL-8 expression, which are critical mediators of inflammation and pathogen recruitment. Therefore, we conclude that the main mechanism of action of Aq involves inhibiting L. garvieae adhesion to epithelial cells, reducing EPS production, and downregulating key virulence-associated genes (e.g., hly1, hly2, and hly3). After preventing L. garvieae adherence and suppressing proinflammatory cytokine expression (IL-1β and IL-8), Aq disrupts the pathogen’s ability to breach epithelial barriers and induce red blood cell lysis, thereby mitigating its virulence and pathogenicity. Our results emphasised the potential of Aq as an alternative non-antibiotic intervention for controlling piscine lactococcosis, advancing our understanding of L. garvieae pathogenesis and providing the foundation for the future integration of environmentally friendly antimicrobials into aquaculture disease management.

The absence of efficient on-farm interventions against white spot syndrome viral (WSSV) infections can cause significant economic losses to shrimp farmers. With this exploratory study we aimed to test, both in vitro and in vivo, the efficacy of an organic acid mixture (Aq) against WSSV infections in shrimp. In vitro, using shrimp gut primary cells (SGP), 2% Aq significantly reduced WSSV infection and the amounts of H2O2 released but had no impact on CAT and SOD expression. In vivo, in a shrimp challenge test, 2% Aq significantly downregulated the expression of proteins involved in WSSV virulence, such as the lipopolysaccharide-β-1,3-glucan-binding protein (LGBP) and the TLR signalling pathway (LvECSIT), and increased the expression of HO-1 oxygenase. Additionally, at 2% Aq, the expression of the digestive-related enzyme carboxypeptidase B was upregulated in the gut, alongside a significant decrease in IL-22 expression, a cytokine usually increased during WSSV infection in shrimp. A low mortality rate (7.33%) was recorded in infected shrimp treated with 2% Aq compared to the 96.66% mortality in the absence of Aq. The peritrophic membrane (PM) was proven essential to ensure Aq efficacy, as the infected and treated PM deficient shrimp (PM−) had a mortality rate of 27.8%, compared to only 9.34% mortality in the infected shrimp at 2% Aq and in the presence of PM (PM+). Aq significantly increased the expression of mucin-1, mucin-2, mucin-5AC, mucin-5B, and mucin-19 in both PM+ and PM− shrimp. Conclusively, organic acid in mixtures can protect farmed shrimp against WSSV infection and increase their survivability through a mediated gut health effect which includes resistance to oxidative stress and improved immunity.

This study investigated the effects of prepared innovative feed (PIF) on the saturated and unsaturated fatty acid profiles of dairy cow milk. PIF was composed of rape seed meal, sunflower meal, protected fat, source of conjugated linoleic acid (CLA), calcium carbonate, monocalcium phosphate (MCP), sodium bicarbonate, and vitamin mineral premix. Dairy cows were divided into two groups (10 cows/group) including a control group (CG) fed basal diet without any nutritional supplements and experimental group (EG) fed basal diet + 1.5 kg PIF/cow/day of the new products. The PIF resulted to change the overall fatty acid content of the milk from the experimental cows and raised the levels of UFA from 64.164% to 73.229%. The SFA content slightly decreased from 35.607% to 26.910%. Furthermore, the PUFA content has elevated from 37.824% (CG) to 45.415% (EG). Not significant difference was recorded for n3, n6 and the n6/n3 ratio (p≥0.05). In conclusion during the experimental period the content of SFA tended to decrease, while the amount of UFA has increased notably (p≤0.05). Similar observations were also noticed for PUFA and MUFA. The inclusion of PIF as dietary supplement in dairy cows modified the fatty acid profile of milk, with major impact on consumer health.

Background Starting primarily as an inflammation of the mammary gland, mastitis is frequently driven by infectious agents such as Staphylococcus aureus . Mastitis has a large economic impact globally, which includes diagnostic, treatment, and the production costs not to mention the potential milk contamination with antimicrobial residues. Currently, mastitis prevention and cure depends on intramammary infusion of antimicrobials, yet, their overuse risks engendering resistant pathogens, posing further threats to livestock. Methods In our study we aimed to investigate, in vitro, using bovine mammary epithelial cells (MAC-T), the efficacy of the AuraShield an antimicrobial mixture (As) in preventing S. aureus attachment, internalisation, and inflammation. The antimicrobial mixture (As) included: 5% maltodextrin, 1% sodium chloride, 42% citric acid, 18% sodium citrate, 10% silica, 12% malic acid, 9% citrus extract and 3% olive extract (w/w). Results and discussion Herein we show that As can significantly reduce both adherence and invasion of MAC-T cells by S. aureus , with no impact on cell viability at all concentrations tested (0.1, 0.2, 0.5, 1%) compared with untreated controls. The anti-apoptotic effect of As was achieved by significantly reducing cellular caspase 1, 3 and 8 activities in the infected MAC-T cells. All As concentrations were proven to be subinhibitory, suggesting that Ac can reduce S. aureus virulence without bacterial killing and that the effect could be dual including a host modulation effect. In this context, we show that As can reduce the expression of S. aureus clumping factor (ClfB) and block its interaction with the host Annexin A2 (AnxA2), resulting in decreased bacterial adherence in infection of MAC-T cells. Moreover, the ability of As to block AnxA2 had a significant decreasing effect on the levels of pro inflammatory cytokine released upon S. aureus interaction with MAC-T cells. Conclusion The results presented in this study indicate that mixtures of natural antimicrobials could potentially be considered an efficient alternative to antibiotics in treating S. aureus induced mastitis.

This experimental trial explored the impact of incorporating specific nutritional feed into the diet of dairy cows and assessed their influence on the quality of products derived from the processing of raw milk. Results showed that nutritional feed changed several parameters of dairy products, such as dry matter, crude protein, crude fat, ash, and fatty acid profiles between the two groups for produced dairy products. The resulting two types of cheeses, namely Caș and Telemea, made from the milk of Experimental Group (EG) cows had a decreased content of saturated fatty acids (SFA) and an elevated content of unsaturated fatty acids (UFA) compared to the Control Group (CG), whereas the situation was the opposite. The associations of the n6:n3 ratio were lower in the Caș cheese from the EG compared to CG, while in the Telemea cheese, the observations also revealed inverted results.

In this paper were studied the possibilities of some flavonoides (quercetin and rutin) identification from the propolis ethanolic extract (EEP), depending on the harvesting region from the west of Romania (Timis county), the obtained results were compared to those obtained for similar products in literature (Hungary, Czechia, China, Slovenia). Representative samples were collected from three different regions of Timis county (Lugoj 1, Lugoj 2 and Faget). The identification of quercitin and rutin from propolis ethanolic extracts (EEP) was realized with high performance liquid chromatography (HPLC). For separation and quantification the ethanolic extracts of propolis (EEP), I used the high performance liquid chromatography. Quantification was performed on the basis of the calibration curve obtained for pure quercitine. I used the HPLC Hewelett Packard apparatus Agilent 1100 type with UV detection, with the following characteristics: column: Nucleosil C18; 5µm dimension for stationary phase particles; column dimension: 150 x 4,6mm x mm; eluent acetonytrile:water = 1:1, eluent flow: 1 mL/min; wave length: 365 nm; temperature 30°C; injected volume: μL.

The metals contents of plants are variable, due to the factors like differences between the plants species, geographical area, conditions of drying process. Metals contents in soil are a great importance for their effect of animals and humans, through the biologic chain: soil – plant – feed and food. Analysis of metals content was made with ContrAA-300, Analytik-Jena device, by flame atomic absorption spectrometry (FASS) in air/acetylene flame. It were analyzed the macro elements content for 33 medicinal plants. The main macro elements (Ca, Mg K and Na) were quantified for each sample. The contents in macroelements for analysed samples were in range: 3.763 % (Plantago major) – 0.442 % (Hippophae rhamnoides), for Ca; 0.718% (Urtica dioica) – 0.107% (Hippophae rhamnoides and Pinus), for Mg; 1.417% (Chelidonium majus) – 0.319% (Rhamnus frangula), for K and 1.945% (Cynara scolymus) – 0.021% (Pinus), for Na.

The purpose of the paper was to assess the rheological properties of some kinds of sausages from the local market. Three different kinds of sausages were chosen: Cabanos (CS), Peasant (PS) and Grill (GS). In order to analyze the viscoelastic characteristics stress relaxation tests were run on the above food at the room temperature. With the "Origin" program, the graphic part was made, obtaining for each three compression curves. From these curves were calculated the compression module (or elasticity module or Young) as well as the energy required for compression. Comparing the compression curves of all three types of sausages, it is noticed that the highest values of the compression tension, corresponding to a specific deformation ? = 0.2, were noticed in the case of Cabanos sausages (162.0-174.2 kPa), followed by Grill sausages (58.3-89.7 kPa) and the Peasent sausages (60.78-78.31 kPa). The value for the compression stress at the end of compression ranged between: 10-13 kPa for PS; 28-34 kPa for CS and 7.8-14.3 kPa for GS. The energy required for the 20% compression of the Cabanos sausages, 2.24�0.31 KJ�kg-1 was more than two times higher than the same energy obtained for the Peasent sausages (0.87�0.13 KJ�kg-1) respectively the Grill sausages (0.69�0.33 KJ�kg- 1). The relaxation tests were performed in continuation of the compression tests by measuring the variation in time of the force necessary to maintain the compressed sample at a deformation of ? = 0.2 (compression of 20%). It was found substantial differences between the relaxation times distribution curves of the investigated sausages.

The purpose of this experiment was to determine the effect of essential oils individually administered in broiler feed on the bioproductive performance in each growing period. The broilers were raised according to the ROSS 2014 Growth Guide in three experimental periods, namely: hatching - 10 days, 11 days-21 days, 22-35 days. The chickens were divided into 9 experimental groups, 3 replicates each and 10 chickens per replication. Oils were incorporated into compound feed on a support of sunflower oil. The offspring in the experimental T0 group received unleavened compound feed. Chickens from T1 and T2 were fed with a combined feed in which the incorporated essential oil was oregano 125 or 250 ppm in combined feed, those in T3 and T4 consumed aniseed feed at the same doses, T5 and T6 a feed combined with fennel, and those in T7 and T8 were treated with a feed mixed with the non-aryl in the same amount. In the first growth period it was found that the highest body weight was recorded at T2 (259.9 g) and also the smallest feed conversion index was recorded in this group. In the 11-21 day period T2 recorded both the highest body weight (914.80g) and the lowest feed conversion index (1.293). In the period 22-35 days, the highest body weight was recorded at T2 (2159.80) and the lowest conversion index of food at T2 (1.599). In the last growth period, namely 35-42 days, the highest body weight was recorded at T1 (2759.20) and the lowest FCR at T2 (1.948). It can be concluded that the individual administration of essential oils (Aetheroleum Origanum vulgare L.) influences the bioproductive performances of the broilers causing an improvement in them.

The aim of this study was to quantify the effect that the Optima 100 nutritional supplements (protected fats) and CLA (conjugated linoleic acid) administered in the ration of mother sheep during lactation had on blood indices in lambs during lactation. The experimental period began immediately after lambing and lasted for 60 days. 40 sheeps were grouped as follows: The control group (CG) received only the basic ration (BR); In experimental group 1 (EG1) the basic ration was supplemented with 12 g/head/day with the product Optima 100 which represents protected fats; In experimental group 2 (EG2), BR was supplemented with CLA in an amount of 12 g/head/day. The two nutritional supplements were added to BR, each in an amount of 12 g/head/day, to experimental group 3 (EG3). The values obtained in EG1 lambs were significantly (p <0.05) lower for TP, BUN and TBLI, significantly higher (p <0.05) for TCHO, TG, HDL, LDL, GPT, GOT and LIP, and insignificant (p> 0.05) for GLU. For the lambs in group EG 2 the parameters were significantly (p <0.05) lower for GLU and BUN, significantly higher (p <0.05) for TCHO, TG, HDL, LDL, GPT, GOT and LIP, and insignificant (p> 0.05) for TP and TBIL. By combining the two supplements for group EG3, the blood parameters were significantly (p <0.05) lower for LDL, GLU, BUN and TBLI, respectively significantly higher (p <0.05) for TCHO, TG, HDL, TP, GPT, GOT and LIP.

The purpose of this experiment was to determine the effect of fennel and sage essential oils administered in a dose of 125, respectively 250 mg/kg CF in the feeding of broiler chickens on the bioproductive indices. The biological material was represented by sexed (male) broiler chickens belonging to the ROSS 308 hybrid on which the phasial breeding (1-10d, 11-21d, 22-35d) was practiced, while observing the recommendations from the ROSS 308 Broiler Management Guide, 2014. The chickens were divided into 5 experimental variants (T0, T1, T2, T3, T4), with 3 replications and 10 chickens/replication. The chickens from T0 (control) have received only the basic diet, for T1 and T2 the basic diet+fennel essential oil (125 and 250 mg/kg CF) were administered, and for T3 and T4, the basic diet+sage essential oil (125 and 250 mg/kg CF). During the first breeding period (0-10d), the highest value for the body weight was recorded at T2 (258.93 g), which also had the lowest feed conversion ratio (1.03 kg CF/kg WG). In the period 11-21d, also in T2, the highest body weight (913.80 g) and the lowest feed conversion ratio (1.29 kg CF/kg WG) were recorded. In the finisher phase (22-35d), T2 maintained differences compared to T0, for body weight +8.01% and -10.56%, for FCR. During the entire breeding period 1-35d, the lowest FCR was recorded in T2 (1.45 kg CF/kg WG). The difference between T2 and T0 was statistically ensured, which highlighted the positive effect of the fennel essential oil (250 mg/kg CF) on the bioproductive performance of broiler chickens.

The artisanally produced cheese feels a major comeback. Shops specialising in artisanal cheeses are increasingly selling artisanal cheeses made by local producers. In this work we made an assortment of matured cheese, from cow's milk, according to a traditional recipe from Harghita County, Romania. The technological process of obtaining ripened cheeses from cow's milk, studied by us, complies with the traditional technology used for many years. The products obtained were physicochemical evaluated in terms of moisture content, dry matter content, protein content, ash content and fat content in compliance with the methods of the quality standards regarding dairy products. All the results obtained from the physicochemical analysis were compared with the results from the specialized literature. At the same time, a sensory analysis was performed for the matured cow cheese produced by its own production versus other cheese samples from the same category from different areas of Romania. The sensory evaluation was carried out by the method of comparison with unitary score scales, the characteristics pursued being: the appearance and shape of the product, the consistency of the product, the color of the product, the aroma of the product and the taste of the product.

The objective of this experiment was to assess and fight against the NSP’s anti-nutritional effect from meat chickens’ alimentation. The experiment lasted for six weeks and was performed on a number of 120 chickens distributed in four experimental variants (LE1, LE2 and LE3, LE4). The hybrid used was Ross 308. We formed four experimental groups, as follows: the experimental group LE1 fed with a CF containing triticale in proportion of 40 % in its structure, the experimental group LE2 fed with a combined forage in which we had incorporated triticale in proportion of 60%, the experimental group LE3 fed with forage including triticale 60 % with supplementation of xylanase 100 ppm, and the experimental group LE4 fed with forage including triticale 60% and xylanase in a quantity of 250 ppm. We could observe that the NSPs, NSPi and NSP total contents increased with the increase of triticale participation; so, the combined forage including 60 % triticale during the growth period from eclosion to 6 weeks presented up to 1.08 percentage points the NSPs content, up to 0.28 p% the NSPi content and up to 1.36 p% the NSPt content. Xylanase incorporation in a quantity of 250 ppm determines body weight increase with up to approx. 12% in the case of 6-week old chickens and feed conversion ratio reduction with up to 10 %. The utilization of triticale in proportion of 40-60 % in combined forage structure, for broiler chickens, is possible only in association with enzyme incorporation; these determine the reduction of intestinal viscosity at duodenum and jejunum level, with up to 15 %.

The problems approached by our team are represented by the involvement of different sources and levels of calcium, namely calcium carbonate, fructoborate and alfalfa, in the mineralization of bone tissue. So, we made fix histological preparations of tissue fragments taken from tibia bone, from 9 pigs belonging to three batches: the control batch, where calcium was provided in a proportion of 1% through calcium carbonate, experimental batch 1, where calcium was provided in a proportion of 1.04% through fructoborate, on a calcium carbonate support, and experimental batch 2, where calcium was provided in a proportion of 1.13% through fructoborate + alfalfa, on a calcium carbonate support. The histomorphometric parameters assessed were represented by the volume of bone trabeculae (BV/TV, %) or the percentage of bone tissue in a given volume and the mean width of bone trabeculae. At the same time, in order to establish fructoborate and alfalfa implication in bone mineralization, we supervised the presence and activity of osteoblasts respectively osteoclasts. In the case of the experimental batch 1, the histomorphometric study shows an increase of bone trabeculae dimension, with a mean width of about 142,5μ μ, and also an increase of their mean volume, which is about 37,11%. The trabecular system is dense and present mineralised and ossified territory where are formed by osteoclasts with osteocytes. Peritrabeculary are presented active osteoblasts which are involved in plurilamellar stratification by deposition of young collagen. In the case of individuals from experimental batch 2, trabeculae mean width is about 140,7μ, while their volume is 35,62%.

Romanian Raven Shepherd Dog is the fourth breed of shepherd dog of national interest, formed in the area of the southern Carpathians and the related sub-Carpathian area. The breed is nationally recognized and approved in Romania, with a national breed standard. In order to be internationally approved, breed morphometric studies are required. The purpose of this study is to evaluate body measurements in certain target population of Romanian Raven Shepherd Dog in relation to the national breed standard.