International Journal for Veterinary Sciences, Animal Sciences, and Biotechnology
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Prevalence of Salmonella spp. and Escherichia coli in Purple Dye Murex, Bolinus brandaris, Harvested from the Adriatic Sea off the Western Coast of Istria
The purple dye murex (Bolinus brandaris), a species of edible marine gastropod, is caught for human consumption in Mediterranean countries, including Croatia. When harvested for human consumption, marine gastropods are subject to European Food safety legislation. In this study, standard microbiological methods were used to determine the presence of Salmonella spp. and Escherichia coli in 48 samples of Bolinus brandaris caught in the Adriatic Sea off the western coast of Istria. Salmonella spp. were absent in all of the samples. E. coli was below the quantification limit of the method in 85.4% of samples. Four samples (8.3%) contained the most probable number (MPN) of E. coli between 18 and 230. Three samples (6.3%) contained E. coli in amounts between 230 and 700, and none of the samples exceeded 700 MPN/100g. While these marine gastropods contained low numbers of E. coli, and were safe regarding Salmonella, further studies are required to assess the presence of other microorganisms and contaminants.
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Urban Wild Boars – from Biodiversity to a Potential Public Health Concern
The growth of the human population, the expansion of settlements and the entry of people into the habitats of wild animals result in an increasingly frequent occurrence of wild animals in cities. Within urban areas, some wild species can find adequate conditions for survival, which are sometimes even more suitable than those in natural habitats. This has a positive impact on biodiversity, though such species can also have a negative impact on species already present in cities. Consequently, the impact of each species on the population of humans, domestic and wild animals should be observed separately. One species of particular importance is the wild boar (Sus scrofa), which adapts exceptionally well to the urban habitat. Green corridors, green areas in the city, abandoned orchards and vineyards, and inadequate waste disposal represent the basic prerequisites for their entry and survival in urban environments. Additionally, many studies have described the fundamental adaptations of wild boars to new, anthropogenic habitats. However, wild boars can cause problems in traffic, relationship with pets and people, and serve as a potential source of pathogens, though the available data suggest a lower prevalence of certain zoonotic pathogens (such as the hepatitis E virus, Campylobacter spp., Arcobacter spp., Streptococcus suis, Toxoplasma gondii, Trichinella spp., etc.) than from animals from rural areas. Nevertheless, the greater possibility of contact with wild boar excretions (primarily faeces) in cities emphasises the need to monitor the health status of wild boars and to conduct risk analysis for infection/invasion. Both activities are fundamental for the transfer of information and raising awareness as to the risks and protection of human health in urban areas.
Diversity and Antimicrobial Resistance Profile of Bacteria Isolated from Sheep Mastitis and Bulk-Tank Milk in the East-Centre Region of Portugal
The microbiological quality of milk reflects both udder health and the subsequent contamination that can occur through direct and indirect reservoirs. This study aimed to identify bacterial diversity in milk samples from ewes with mastitis and in bulk-tank milk, and to investigate their antibiotic susceptibility/resistance profiles. A total of 232 milk samples were analysed and 22 bacterial species were isolated, with Staphylococcus aureus (20.3%) and S. epidermidis (18.6%) as the dominant species. The diversity of bacterial species isolated from the bulk-tank was higher and included 44 species, dominated by E. coli (20.6%), S. aureus (7.8%) and Hafnia alvei (7.2%). Antimicrobial susceptibility testing was performed on 217 bacterial isolates. E. coli isolated from individual milk samples revealed high rates of resistance to penicillins and tetracyclines, whereas S. aureus revealed high rates of resistance to aminoglycosides, polymyxin, sulfonamides and tetracycline, maintaining high levels of susceptibility to third generation cephalosporins, lincosamides, some macrolides, penicillins, phenicols and pleuromutilins. The resistance profile of bacteria isolated from the bulk-tank milk was more concerning than that of the milk of ewe with mastitis. These results point to the need to implement structural improvements on farms and raise awareness about strengthening biosecurity, medical prophylaxis and control of access to veterinary medicines to improve milk quality and reduce the use of antibiotics.
Impact of Season on Fresh Semen Parameters and Post-Thawing Motility of Sperm of Arabian Stallions
The study examined the influence of season on several parameters of fresh semen ejaculates and the post-thawing motility of the sperm of Arabian stallions. Three Arabian stallions were used for semen collection using a Missouri-type artificial vagina. Semen was evaluated macroscopically for volume, colour and consistency, and microscopically for motility, concentration of sperm, and total sperm count in ejaculates. The semen was frozen using di-methyl formamide egg yolk based freezing extender and evaluated for post-thawing motility. Results revealed that the volume of semen, total number of sperm per ejaculate and the post-thawing motility in the breeding season was significantly higher than in the non-breeding season, while the opposite was true for sperm cell concentration. There was no difference in colour, consistency or motility between stallions in either the breeding or non-breeding season. In conclusion, season had a direct influence on semen volume, sperm cell concentration, total number of sperm, and post-thawing motility of sperm of Arabian stallions.
Urinary Bladder Cancer in Dogs: A Promising Model for Advancing Human Urothelial Carcinoma Research?
Dogs are widely recognised as valuable models in comparative oncology because they develop cancer spontaneously, have shorter lifespans, and share significant biological and clinical similarities with human tumours. The objective of this review was to assess the relevance of canine urothelial carcinoma (UC) as a translational model for human bladder cancer by comparing epidemiological patterns, molecular alterations, and immunological features across species. Urothelial carcinoma is of particular interest due to its aggressive nature and high mortality rates. In dogs, invasive forms of UC are more common, underscoring the relevance of this species for modelling human disease. While advanced age and exposure to environmental carcinogens are shared risk factors, notable epidemiological differences exist, including a higher prevalence in female dogs than in male humans. Molecularly, canine and human UCs share tumour subtypes and dysregulation of key oncogenic pathways, such as p53/p63, RTK/RAS/MAPK, PI3K/AKT/mTOR, and CDKN2A/CDK4, despite distinct initiating genetic events. These alterations converge on pathways controlling proliferation, survival, and immune evasion. Shared overexpression of therapeutic targets and similar immune checkpoint mechanisms further support the translational value of the canine model. Collectively, these features reinforce the utility of dogs in advancing biological understanding and therapeutic development for urothelial carcinoma in both veterinary and human medicine.
Prediction of Frozen Chicken Meat Storage Time through Artificial Neural Network Analysis of Colour Parameters
Poultry meat colour deteriorates during prolonged frozen storage, affecting consumer perception and product value. This study investigated changes in colour parameters (L*, a*, b*) of frozen chicken breast meat, and developed an artificial neural network to predict storage time non-destructively. Ten chicken breast samples were analysed at different freezing storage times (0, 3, 6, 9, and 12 weeks) at -18°C, with colour measurements performed directly on each sample. Lightness (L*) increased from 51.82 ± 1.1 to 58.21 ± 1.0 units, redness (a*) decreased from 3.25 ± 0.2 to 2.12 ± 0.2 units, and yellowness (b*) increased from 8.12 ± 0.5 to 9.91 ± 0.6 units, resulting in a total colour difference of ΔE = 6.87, and all changes were statistically significant. The multilayer perceptron artificial neural network showed good agreement between predicted and actual storage times (R² ≈ 0.98), demonstrating the potential of colour-based modelling as a rapid, non-destructive tool for monitoring frozen chicken meat quality.
Implementation and Comparison of Real-Time Molecular Serotyping and Conventional PCR for Listeria monocytogenes Strains in Food
Listeria monocytogenes is an important foodborne pathogen responsible for listeriosis, a severe human infection with high morbidity and mortality. Molecular serotyping is a universally accepted subtyping method for L. monocytogenes. Identification of strain serotype permits differentiation between significant foodborne strains (1/2a, 1/2b, and 4b) and allows a better understanding of their distribution and epidemiological significance. This study evaluated the implementation of real-time molecular serotyping of L. monocytogenes and compared its performance with conventional PCR methods. Eighty L. monocytogenes strains isolated from food and the food chain were analysed. Conventional multiplex PCR and qualitative real-time PCR (ReT-PCR) were used to identify molecular serogroups based on specific genetic markers to define the presence or absence of a particular serogroup of L. monocytogenes in strains. The results showed that ReT-PCR offers faster processing times than the conventional method. These strains belonged predominantly to serogroup IIa, followed by IIc, IIb, and IVb. ReT-PCR is becoming a standard tool for the detection and expression profiling of selected strains. This research highlights the importance of providing a rapid method for the qualitative detection of L. monocytogenes in food and integrating ReT-PCR into routine diagnostic applications.
First Molecular Detection of Ehrlichia canis in Croatia Reveals its Presence in Rhipicephalus bursa Ticks from Ruminants
Ehrlichia canis, the causative agent of canine monocytic ehrlichiosis, is considered endemic in Mediterranean Europe and is primarily associated with the brown dog tick, Rhipicephalus sanguineus sensu lato. In Croatia, previous molecular investigations in dogs failed to detect the pathogen, despite sporadic serological findings and the confirmed presence of competent tick vectors. The aim of this study was to investigate the presence of E. canis DNA in ticks collected in Croatia. A total of 583 ticks from five genera were collected from the environment, domestic animals, wildlife, and humans across approximately 130 locations. All ticks were screened by PCR targeting the 16S rRNA gene, and positive samples were further characterised by sequencing of the full-length 16S rRNA gene. Six ticks tested positive for Ehrlichia sp. DNA, and sequencing confirmed all isolates as E. canis. All positive ticks were identified as Rhipicephalus bursa and originated from coastal Mediterranean regions of Croatia. Positive specimens were collected from goats, sheep, and chamois (Rupicapra rupicapra). These findings represent the first molecular evidence of E. canis in Croatia and indicate its circulation in ruminant-associated tick populations within the Mediterranean part of the country. Further studies are needed to clarify transmission dynamics and the potential epidemiological relevance for domestic animals.
Hock Arthritis in Pet Chickens (Gallus gallus domesticus): Physical, Cytological and Bacteriological Study of Synovial Fluid
Hock arthritis is a pathology with significant repercussions on the health of pet chickens, and on the economics of the poultry sector. The objective of this study was to detail the physical, cytological, and bacteriological characteristics of synovial fluid from hocks of 52 pet chickens affected by arthritis. The appearance of synovial fluid was assessed in terms of volume, colour, turbidity and viscosity. The mucin clot test was performed. Synovial fluid smears were stained with May-Grunwald-Giemsa to perform differential cell counts. Isolated bacteria were identified by standard microbiological and biochemical tests. Analysis of the synovial fluid samples revealed variations in their physical characteristics. The majority of samples exhibited a volume of 200 µL, slight turbidity, a bloody colour and decreased viscosity. The number of erythrocytes, platelets and white blood cells varied depending on the sample for cytological parameters. One sample showed an absence of leukocytes. Synovial fluids infected with Staphylococci showed a higher number of leukocytes per field compared to those infected with Enterobacteriaceae. Bacteriological analysis revealed that 65.38% of synovial fluid samples were negative for bacteria, while 34.61% were positive, of which Staphylococcus aureus (S. aureus) (35%), Staphylococcus epidermidis (S. epidermidis) (40%), Escherichia coli (E. coli) (5%) and Salmonella enterica (S. enterica) (20%). Synovial fluid in hock arthritis shows variable physical and cytological characteristics, reflecting changes in cell types and fluid properties. In pet chickens, the main bacterial agents identified are S. aureus, S. epidermidis, E. coli and S. enterica. These findings provide essential information for veterinarians and poultry practitioners, aiding in joint health assessment, early diagnosis and timely treatment.
Antibiotic-Free Broiler Meat Production: Feasibility and Economic Viability
The excessive use of vaccines, antibiotics, and other preventive therapeutic agents in conventional broiler production has resulted in the frequent occurrence of antibiotic residues in edible tissues, posing risks to both animal and human health. These residues contribute to the global problem of antimicrobial resistance, highlighting the need for alternative approaches to poultry disease prevention. This study aimed to evaluate whether a comprehensive preventive programme encompassing biosecurity, hygiene, and an antibiotic-free prophylaxis regimen could eliminate the need for prophylactic antibiotic therapy, thereby enabling the production of broiler meat free of antibiotic residues. This approach was contrasted with the conventional production system, in which less rigorous preventive practices necessitate the prophylactic use of broad-spectrum antibiotics, ultimately resulting in detectable residues within the muscle and liver tissues of conventionally reared chickens. Ross 308 broilers were reared under field conditions. in. The control group was reared using conventional prophylactic protocols, including broad-spectrum antibiotics, vitamins, bio-stimulants, mineral supplements, and acidifiers via drinking water. The experimental group received no antibiotics; and instead were treated with hydro-soluble probiotic preparations containing vitamin C, lactose, fructose, and baker’s yeast, combined with continuous water disinfection throughout the fattening period. Microbiological analyses showed the presence of antibiotic residues in the soft tissues of conventionally reared broilers, while no residues were detected in the experimental group. Production performance indicators—mortality rate, vitality, final body weight, and overall cost—were similar to or superior to those in the antibiotic-free group, confirming that broiler meat production without antibiotics is both feasible and economically viable.
Faunal Composition and Seasonal Dynamics of Stable Flies (Diptera: Muscidae and Calliphoridae) in Pasture Ecosystems of the Samarkand Region, Uzbekistan
Stable flies (Diptera: Muscidae) and other zoophilic dipterans represent a major veterinary and economic challenge in livestock production due to their haematophagous feeding behaviour and their capacity to transmit pathogens. This study aimed to characterise the species composition and seasonal dynamics of synanthropic flies associated with cattle pastures in the Samarkand region of Uzbekistan. Systematic sampling was conducted from April 2022 to May 2023 in the Payariq and Nurobod districts using active net collection from cattle and their immediate surroundings. A total of 3,512 specimens were collected, representing 14 species across six genera and two families. Muscidae were overwhelmingly dominant (95.4%), whereas Calliphoridae accounted for 4.6% of the fauna. Musca domestica (13.1%) and M. autumnalis (12.6%) were the most abundant species, followed by Stomoxys calcitrans (8.1%), Musca simplex (8.5%), and Lyperosia spp. (7.7%). Pronounced seasonal patterns were observed, with peak fly abundance during summer months, when 62.8% of all specimens were collected, and a complete absence of adult flies from December to February. Blood-feeding species with piercing–sucking mouthparts (S. calcitrans, Lyperosia irritans, L. titillans, Haematobia dentipes) comprised nearly one quarter of all collected specimens and reached their highest densities in summer, indicating a critical period for pathogen transmission risk. In contrast, lapping-type flies were present from spring through autumn and, despite lacking direct haematophagy, may act as important mechanical vectors. These findings provide the first comprehensive overview of pasture-associated stable fly fauna in Uzbekistan, highlighting seasonal windows of increased veterinary risk, and underscoring the need for integrated, region-specific vector management strategies.
First documented case of canine distemper virus in a leopard cub (Panthera pardus) in Tamil Nadu, India
A four-month female leopard cub (Panthera pardus), rescued from a residential area in Kaliyal, Kanyakumari, Tamil Nadu, India with exhibited sign of dullness, convulsions, seizures, and profuse salivation was screened for infectious disease conditions and confirmed as canine distemper (CD) by conventional PCR method. To our best knowledge this was the first documented case of CD in a leopard in Tamil Nadu, India and this report focused on the integrative treatment with allopathic and siddha medicine for the disease condition.
Prevalence and antimicrobial susceptibility profiles of Staphylococcus spp. isolated from broilers in Croatia
Staphylococcal infections can be a serious public health problem for farm workers and other persons in contact with poultry, due to their ability to cause a wide range of infections in other animal species and humans. In addition, the presence of pathogenic bacteria in the intestines of poultry can lead to contamination of poultry products. Coagulase-positive (CoPS) and coagulase-negative (CoNS) staphylococci, which are potentially involved in staphylococcal infections in humans, have been associated with poultry. The best-known CoPS is Staphylococcus aureus (S. aureus). The aim of this study was to evaluate the prevalence of Staphylococcus spp. in broilers on commercial farms in Croatia, and to determine their antimicrobial susceptibility profiles. Staphylococcus spp. strains isolated from cloacal swabs of apparently healthy broiler flocks in Croatia were analysed, and a relatively high Staphylococcus infection rate was detected, with a total percentage of isolated Staphylococcus strains of 38%. Among these, 3/100 (3%), 5/100 (5%), 3/100 (3%), 2/100 (2%) were S. aureus, and three CoNS: S. sciuri, S. simiae and S. lentus, respectively. The majority of isolated strains were resistant to penicillin (88.8%), clindamycin (85%), cefoxitin (78%), trimethoprim/sulfamethoxazole (65%), ciprofloxacin (53%), erythromycin (40%) and tetracycline (25%). The highest susceptibility of isolated strains was to chloramphenicol (95%) and gentamicin (89%). All S. aureus, S. lentus and S. simiae isolates were multi-drug-resistant strains that exhibited resistance to three or more antimicrobials, while S. sciuri isolates were resistant to two of the tested antimicrobials.
Ergot alkaloids in cereal-based systems: Pathways of food and feed contamination, toxic effects and control measures
Ergot alkaloids (EAs) are a class of mycotoxins synthesised by various fungi, mainly by Claviceps purpurea and other members of the genus Claviceps. EAs are found in many economically important cereals, causing production losses in agriculture around the world. They are mycotoxins with well-documented toxicological effects on humans and animals, including vasoconstriction, neurotoxicity, hormonal disorders and impairment of reproductive function. Their presence in cereal crops, especially in rye, wheat and barley, poses a persistent risk to food and feed safety. Despite advances in analytical detection methods, the heterogeneous distribution of these alkaloids and the variability of their toxic profiles make effective risk assessment difficult. Chronic dietary exposure, even at subclinical levels, can lead to adverse health effects, emphasising the importance of continuous monitoring. Therefore, the establishment and enforcement of harmonised legal limits is essential, accompanied by comprehensive monitoring and the application of good agricultural and storage practises. A systematic and science-based control strategy is necessary to mitigate the risks associated with EAs and to ensure the protection of human and animal health within the food production chain. The aim of this review is to characterise EAs, their pathways of contamination, toxic effects in the body and control measures which should be applied for food and feed.
Fusarium moulds, mycotoxins and their metabolites
Moulds of the genus Fusarium are present in a range of aquatic and soil environments as pathogens, endophytes and/or saprophytes. They have ability to easily adapt to different environmental conditions, enabling the ease of their spread worldwide. However, their growth and development occur more often in the areas with a humid and warm climate. The majority of Fusarium species are plant pathogens, but certain strains can cause diseases in humans and animals either as direct pathogens or by indirect action of their mycotoxin metabolites. They can cause acute and chronic intoxication and disrupt the function of various organ systems. The most important groups of Fusarium mycotoxins are the trichothecenes, fumonisins and zearalenones. Modern technologies, instruments and methods have enabled discovery of a large number of Fusarium moulds and their metabolites, though toxigenic functions have been described only for a small number of species. For most newly discovered metabolites, there is no detailed information on their structural and chemical characteristics, mode of biosynthesis or evidence of their potential (un)desirable bioactive effects. Considering the unavoidable common presence of Fusarium moulds and/or their metabolites, as well as their potential harmful effects, further research on newly discovered species and metabolites is necessary. The aim of this review paper is to provide an overview of the current knowledge on Fusarium species and their metabolites, with a focus on their growth characteristics and metabolite biosynthesis.
Prevalence of arrhythmias in dogs examined at the Clinic for Internal Diseases, Faculty of Veterinary Medicine, University of Zagreb, Croatia (2018-2020)
The aim of this retrospective study was to determine the prevalence of arrhythmias in dogs admitted to the Clinic for Internal Diseases, Faculty of Veterinary Medicine, University of Zagreb, Croatia. Medical records of 307 dogs that underwent electrocardiographic examination between October 2018 and October 2020 were analysed. The study population (65 breeds, median age 9 years, median body weight 15 kg) showed arrhythmias in 36.2% of cases. The most common arrhythmia was atrial fibrillation (18% of the total population; 26.7% of dogs with heart disease), followed by premature ventricular and atrial contractions. Arrhythmias were also detected in 18.6% of dogs without primary cardiac disease, most often associated with systemic disorders such as tumours, epilepsy, hypoadrenocorticism and arterial hypertension. The results confirm that arrhythmias are frequent in dogs, occurring both in the presence of heart disease and as a consequence of non-cardiac conditions, emphasising the importance of routine electrocardiographic screening in veterinary clinical practice.
Histomonas meleagridis infections in turkeys
This report describes an infection of Histomonas meleagridis in a turkey flock kept together with pheasants and geese on a farm. The owner observed a number of turkey deaths and two turkey carcasses were sent to the Pathology Laboratory of the Poultry Centre of the Croatian Veterinary Institute for parasitological diagnosis. Histomonosis was confirmed through histopathological and parasitological methods. In total, 150 of 500 turkeys in the flock perished. The owner was advised to enhance biosecurity measures on the farm.
Metabolomics in sepsis-associated acute kidney injury research
Metabolomics is a scientific discipline focused on the analysis of metabolites in biological samples with the aim of understanding physiological and pathological processes. The development of analytical technologies, such as mass spectrometry and nuclear magnetic resonance, has enabled advances in untargeted and targeted metabolomic analysis. In untargeted analysis, a broad spectrum of metabolites is analysed without prior assumptions, whereas targeted analysis focuses on the quantification of known molecules. Biological samples such as serum, plasma and urine are most commonly used, with the choice depending on the clinical context and the nature of the disease. Despite its potential, metabolomics faces challenges such as technical complexity, biological variability, lack of standardisation, and the need for advanced statistical methods. However, in the last decade, it has been used extensively in the study of sepsis and associated acute kidney injury, a condition with high mortality and limited therapeutic options. In this review, we present metabolomics, the challenges it faces, and the current state of research concerning sepsis-associated acute kidney injury (SA-AKI). Metabolomic studies of SA-AKI have identified potential key biological changes, including enhanced inflammatory signalling, oxidative stress, osmolyte disturbances, and cellular metabolic dysfunction. Although individual metabolites have been recognised as potential diagnostic markers, none can fully capture the complex metabolic changes characteristic of SA-AKI. In the future, the integration of metabolomics with other "omics" and advanced analytical approaches could advance early disease detection, the understanding of pathophysiology, and the development of personalised therapies.
Prevalence of individual bacterial uropathogens isolated from urine samples of dogs and cats and the relationship between a positive urine culture and the presence of organic, inorganic elements and proteins in the urine
Clinical signs of lower urinary tract inflammation are common in domestic carnivores. In dogs, they are usually the result of a bacterial infection, while in cats, they are more often caused by sterile inflammation or behavioural disorders. The prevalence of individual bacterial pathogens isolated from urine samples of dogs and cats was determined, and the relationship between a positive urine culture and the presence of organic and inorganic elements, and proteins was investigated. From January 2022 to June 2024, a total of 563 dog urine samples and 86 cat urine samples were analysed in the Split branch of the Laboratory for Diagnostics, Croatian Veterinary Institute. A clinically significant number of bacteria were isolated from 196 of 563 (34.81%) dog urine samples and 21 of 86 (24.41%) cat urine samples. Of the total number of bacterial isolates, 141 (64.4%) were gram-negative and 78 (35.6%) were gram-positive. Escherichia coli was the most common isolate (45.9% in dogs, 33.3% in cats). In dogs, the second most common isolates were Staphylococcus spp. (20.9%), Streptococcus spp. (9.2%), Proteus spp. (7.7%), Enterococcus spp. (4.6%), Klebsiella spp. (4.08%), and Pseudomonas spp. (3.6%), while other bacterial isolates were <2%. In cats, the most frequently isolated bacteria were Proteus spp. (23.8%), Pseudomonas spp. (14.3%), Streptococcus and Enterococcus spp. (9.5% each), and Staphylococcus spp. and Pasteurella spp. (4.8% each). In dogs, a significant and positive correlation was found between a positive bacteriological result and pyuria, haematuria, proteinuria, and age. In cats, no significant correlations were found between a positive bacteriological culture and variables such as pyuria, haematuria, proteinuria, crystals, or animal age or sex.
Analysis of chromium distribution in tissues of bottlenose, striped and Risso’s dolphins from the Croatian Adriatic over a 20-year period
This study aimed to quantify chromium (Cr) concentrations in tissues of 190 individuals of three odontocete species—159 bottlenose dolphin (Tursiops truncatus), 25 striped dolphin (Stenella coeruleoalba), and six Risso’s dolphin (Grampus griseus)—stranded along the Croatian coast of the Adriatic Sea between 1995 and 2013. Chromium concentrations were measured in liver, muscle, kidney, skin, lung, spleen, and fat using inductively coupled plasma optical emission spectrometry. Significant interspecific differences in Cr concentrations were observed in the kidney, liver, and spleen, suggesting dietary habits as a major factor in chromium accumulation. Risso’s dolphins exhibited the highest Cr levels, likely due to the higher Cr concentrations in cephalopods that make up the basis of their diet. Chromium concentrations exceeding 1 mg/kg wet weight were found in only 16 tissue samples—15 from bottlenose dolphin and one from striped dolphin—indicating a low overall prevalence of elevated Cr levels. Regression analysis revealed no significant correlation between Cr concentrations and body mass, age, sex, or geographic location. However, a significant decline in Cr levels over the study period was observed across all tissues.
Cortisol as a biomarker for stress: A review of sampling methods in small ruminants
Cortisol is a key biomarker of stress and hypothalamic-pituitary-adrenal (HPA) axis activity in an organism. Different biological samples are used for measuring cortisol levels, such as blood, saliva, milk, urine, faeces, and hair. Blood plasma or serum cortisol measurements are widely applied to quantify acute stress response in farm animals, but stress sampling and rapid hormonal changes can undermine accuracy in results. Saliva has proven to be a reliable alternative sample for assessing free cortisol, offering a less stressful approach with minimal triggering of the stress response. Cortisol determination in milk is a non-invasive method of limited applicability for lactating ruminants, though physiological parameters such as milk yield and stage of lactation can influence its interpretation. Concentrations of cortisol in urine reflect the cumulative cortisol secretion over time, making it a valuable indicator of chronic stress, though research on small ruminants remains limited. Similarly, faecal cortisol metabolites offer a non-invasive means of assessing adrenal activity with delayed but integrated hormonal fluctuations. Hair and wool analysis provide insight into long-term stress exposure, though factors such as seasonal variation and local synthesis complicate its application. This review highlights the strengths and challenges associated with different cortisol sampling methods, emphasising the importance of multiple biological sample approaches for accurate welfare assessments in ruminants.
Variance components for weight and type traits in a multiracial population of meat sheep in eastern Antioquia
The objective of genetic evaluation is to provide a tool to identify and select genetically superior animals that transmit desirable traits to their offspring, improving flock productivity over generations. This study aimed to evaluate genetic and phenotypic parameters for weight and type traits in a multiracial meat sheep population in eastern Antioquia, using Bayesian inference analysis. The study was conducted at two farms: “Ovinos de la Sierra” in La Ceja del Tambo, and “El Charrascal” in Copacabana, Antioquia, Colombia. Animals from various racial groups were evaluated, focusing on productive traits, namely birth weight (BW) and adjusted weaning weight at 90 days (AW90), together with twelve phenotypic traits. A single-trait animal model was applied, incorporating the fixed effects of breed type, calving type, sex, and live weight. Variance components and genetic parameters and expected breeding values were estimated. The mean (± standard deviation) for BW and AW90 were 3.655 ± 0.806 kg and 19.610 ± 5.181 kg, respectively. Direct and maternal heritability estimates for BW were 0.6734 and 0.2963, and for AW90 were 0.9323 and 0.0670. Among phenotypic traits, thoracic perimeter showed the highest heritability, followed by rump length, while the lowest heritability estimates were for withers width and rump width. All estimates were convergent, except maternal and phenotypic estimates for BW. Proper data recording is essential for evaluating genetic and phenotypic parameters, and Bayesian analysis provides reliable variability through posterior distributions, supporting the grouping of animals by age and production group for uniform data.
Gonadotropin regimen and dose predict superovulatory ovarian response in Ouled Djellal ewes: a stepwise regression study
This study aimed to evaluate the effects of gonadotropin regimen (PMSG alone vs. PMSG + GnRH or PMSG + hCG) and dose on superovulatory responses in 35 Ouled Djellal ewes during the breeding season. Intravaginal sponges containing 40 mg fluorogestone acetate were inserted for 14 days in all ewes. Two days before sponge removal, ewes were randomly assigned to seven treatment groups differing by regimen and dose. Outcomes were corpora lutea (CL) count, anovulatory follicles (AF), ovulation rate (OR), and serum progesterone (P4). Associations were assessed using stepwise regression analysis. Ewes receiving PMSG + GnRH or PMSG + hCG exhibited stronger ovarian responses than those given PMSG alone. Final stepwise models explained 41.3% of variance in CL, 20.5% in AF, 27.10% in P4, and 23.60% in OR. In each model, molecule type entered first and contributed more to model fit than dose, indicating that regimen exerted a greater influence than dose escalation. In conclusion, combining PMSG with either hCG or GnRH improves ovarian response parameters and overall model predictability compared with PMSG alone. These findings support evidence-based optimisation of superovulation protocols in Ouled Djellal ewes to enhance prolificacy, increase embryo yield, and accelerate dissemination of elite genetics in advanced breeding programmes.
Pelvimetry of the creole goat of Santa Elena: reference parameters and values
This study evaluated the pelvic indices and diameters, pelvic area and circumference of the Santa Elena Creole goat in Ecuador. A total of 340 Creole goats between one and three years of age, with one to three parturitions, the last of which was eutocic. The indices and dimensions of the pelvis were determined and compared by means of a simple analysis of variance. Reference values for the screening range were calculated as  ± 1.65 SD. The pelvic indices classify the investigated goats as concavilinear. Pelvic diameters, area, and pelvic circumference in first and second kidding oats were smaller (P<0.05) than in third kidding goats. The lower limit of the reference values for each parameter is its appropriate value before kidding. Prior to first kidding, this is 8.97, 5.77, 6.07 and 7.37 cm, for the true conjugate and the upper, middle, and lower transverse diameters, respectively. For the pelvic area and circumference, the appropriate values are 47.3 cm2 and 15.44 cm, respectively. Prior to the second and third kidding, the ideal values of each parameter were higher than for the first kidding. It is concluded that, according to their pelvic indices, the goats investigated are concavilinear and their pelvic dimensions in the first and second parturition were smaller (P<0.05) than in those of the third parturition.
Immunohistochemical detection and localisation of prion protein in brain tissue of cattle, sheep and goats
The misfolding of the cellular prion protein into its pathogenic isoform prion—a proteinaceous infectious particle-causes fatal neurodegenerative diseases. These prion diseases are classified as transmissible spongiform encephalopathies, which may arise from genetic mutations, infectious transmission or sporadic occurrence. They are characterised by protein aggregation and progressive neurodegeneration. In humans, prion diseases include Kuru, Creutzfeldt-Jakob disease, variant Creutzfeldt-Jakob disease, Gerstmann-Sträussler-Scheinker-Syndrome, and fatal familial insomnia. In animals, they include atypical and classical scrapie in sheep and goats, atypical and classical bovine spongiform encephalopathy, also known as “mad cow disease”, and chronic wasting disease in cervids. Although the mechanisms underlying neurodegeneration and the role of neuroinflammation in these disorders remain insufficiently understood, characteristic features include spongiform changes, neuronal loss, gliosis, and accumulation of the pathogenic isoform of the prion. To date, there is no reliable ante mortem diagnostic test for these encephalopathies. The diagnosis is confirmed postmortem, and immunohistochemistry is one of the key diagnostic techniques. This review article summarises the available research data about prion diseases, focusing on the detection and localisation of the pathogenic isoform of the prion protein in brain tissue of cattle, sheep, and goats using immunohistochemical analysis.
Determination of metals and aflatoxin M1 in the milk of Croatian Cold-Blooded mares in different stages of lactation
In this study, the concentrations of metals (arsenic (As), cadmium (Cd), copper (Cu), mercury (Hg) and lead (Pb)), and aflatoxin M1 were analysed in mare's milk samples taken from eight mares over a period of six months during lactation. The results showed that the concentrations of metals in milk occurred in the following order: Cu > Pb > Hg > Cd > As. Key findings of the study included the determined content of metals, depending on lactation stages and animal husbandry practices. Copper had the highest concentration among the metals analysed, while Cd and As were not detected. No statistically significant differences were found in the levels of heavy metals or aflatoxin M1 across different lactation stages. However, statistically significant differences in aflatoxin M1 concentrations were observed depending on the type of animal husbandry, illustrating the influence of environmental and husbandry conditions on milk contamination. The study concludes that there is no toxicological risk for consumers of Croatian Cold-Blooded mare's milk, which is an important finding for food safety and public health. The significance of this study lies in the provision of valuable data on the safety of mare’s milk, especially with regard to its possible contamination with toxic heavy metals and aflatoxin M1. The main objective was to determine the presence and concentrations of the listed contaminants, to evaluate the influence of lactation stage and husbandry, and to assess compliance with established safety standards.
The most common toxinotypes of the bacterium Clostridium perfringens in small ruminants and their resistance to antibiotics
Infection with the bacterium C. perfringens in sheep and goat herds leads to considerable losses. The most common diseases in small ruminants caused by this bacterium are pulpy kidney disease, enterotoxaemia, yellow lamb disease, and necrotic enteritis. C. perfringens is widely distributed in the environment and is part of the intestinal microbiota of humans and animals. When physiological conditions in the body change, it begins to produce toxins that lead to various pathological changes. Depending on the toxins it produces, it is classified into seven types (A–G), defined based on the presence of genes encoding the CPA toxin (all types), CPB toxin (type B and type C), ETX toxin (type B and D), ITX toxin (type E), CPE toxin (type F) and NetB toxin (type G). Strains A and D are most commonly associated with enterotoxaemia in small ruminants. According to the available literature, C. perfringens toxinotype A is most frequently isolated in small ruminant herds. In addition to toxin genes, antibiotic resistance genes also play an important role in bacterial survival. Of the genetic determinants of antimicrobial resistance discovered to date, most are associated with the following antimicrobial drugs: bacitracin (bcrR gene), tetracyclines (tet genes), macrolides (erm genes), lincosamides (erm and lnu genes), streptogramins (erm genes) and chloramphenicol (cat genes). Due to sudden death after the onset of symptoms and the economic unprofitability, very few results are available on antibiotic resistance of C. perfringens in small ruminants. Therefore, it has proven to be more economical and effective to work on prevention than on treatment. Since there is no vaccine against enterotoxaemia in Croatia, prevention of the disease is difficult.
Evaluation of microbiological safety and mycotoxin contamination of household produced meat products originating from Croatian indigenous pig breeds
This study aimed to assess the microbiological safety and mycotoxin contamination of traditional meat products made from Croatian indigenous pig breeds, produced in family households under uncontrolled conditions. Different factors, including natural, seasonal, and uncontrolled production conditions, pose challenges in maintaining consistent product microbiological and mycotoxin contamination. No bacteria from the genera Salmonella or Clostridium, or the species Staphylococcus aureus, were detected in any of the investigated samples. However, Listeria monocytogenes was found in a cured sausage sample from Black Slavonian pigs, rendering it unsafe for consumption. Additionally, Listeria innocua was identified in a dried sausage sample from Turopolje pigs. Yeast and mould contamination levels ranged as follows: cured sausages: 10² – 1.4 × 10⁴ cfu/g; whole ham: 1.4 × 10² – 2.7 × 10⁴ cfu/g; bacon: 2.6 × 10² – 3.5 × 10³ cfu/g. The dominant mould genus colonising the dry meat products was Penicillium (30 isolates), followed by Aspergillus (20 isolates), with Cladosporium and Mucor species present in lower numbers. The most frequently isolated Penicillium species were P. brevicompactum, P. commune, and P. solitum (85.7%), while the most common Aspergillus species were A. proliferans and A. tubingensis (57.1%). Regarding product type, bacon and ham met the respective safety standards, but sausages were contaminated with L. monocytogenes and L. innocua, making them unsafe for consumption. All products were safe in terms of mycotoxin contamination.
High-mortality outbreak of Yersinia pseudotuberculosis infection in a guinea pig herd: a case report
Yersinia pseudotuberculosis caused a fulminant outbreak in a Croatian breeding colony of 84 guinea pigs, in which 80 animals (95%) died within 72 hours after abrupt food refusal and marked lethargy, without diarrhoea. Seven freshly dead animals were examined. At necropsy, 2–5 mm, white-yellow nodules were found distributed in the liver, spleen, lungs, mesenteric lymph nodes and partly also in the small intestine. Aerobic culture of these organs on Columbia blood agar (28°C, 24–48 h) revealed pure growth of Y. pseudotuberculosis. Histology showed well demarcated necro-pyogranulomatous lesions with central bacterial colonies, margins of neutrophils and a peripheral coat of macrophages and few lymphocytes. Immunohistochemistry confirmed abundant lysozyme-positive macrophages and few CD3-positive T and CD79α-positive B cells, indicating a predominantly innate response. This case report emphasises the extreme lethality and rapid progression of guinea pig yersiniosis and highlights its zoonotic potential.
Preliminary analysis of cattle deaths suspected to be an adverse reaction of vaccination against lumpy skin disease in Croatia
Vaccination against lumpy skin disease was carried out in Croatia during 2016 and 2017 to prevent this emerging viral disease from causing significant damage to the cattle population. Two unauthorised live vaccines were used for immunoprophylaxis, and adverse reactions in cattle were expected. Adverse events were monitored and 14 selected reported cases of deaths were analysed using the available necropsy results. The cases analysed were classified as probable, possible, inconclusive, or unlikely to be caused by vaccination, primarily based on results of post-mortem examination, taking into consideration factors that contributed to or resulted in death or euthanasia. Most cases were classified as possibly caused by vaccination due to subcutaneous oedema of varying consistency (at the injection site) and intramuscular haemorrhage, in some cases leading to purulent and necrotic processes. These processes eventually led to fatal purulent bronchopneumonia, pulmonary emphysema, septic conditions, or euthanasia. In the remaining cases, necropsy results were unable to be linked to vaccination or the findings were inconclusive due to the presence of other factors, such as Klebsiella infection. It can be assumed that neither vaccine had a direct connection with animal deaths, but they may have contributed to the fatal outcomes due to the complications of severe local reactions and general impact on the immune system.