The current state of Alex’s line in the Bohemian Shepherd Dog breed

The Bohemian Shepherd Dog is a Czech working dog breed of medium size with a long black and tan coat and characteristic ear shape. The dogs of this breed are friendly, trainable, and with no sign of nervousness. The population of this historic breed of dogs, known since the 13th century, was almost destroyed until 1984, when Jan Findejs and Dr. Vilém Kurz partnered to take the first steps to regenerate the breed. One of the sires used in the project was a sire named Alex. This study aimed to determine the state of Alex’s line (IV. line), and the number of descendant dogs born from 2021 until the first quarter of 2024. Used 13 years after the start of the breed regeneration project, and with only two breeding first-generation descendants, Alex’s line accounted for 2.7% of the population from the beginning in 1996 until 2008. In 2009 an expansion started, caused by three of his male descendants who sired more than 100 puppies each, resulting in Alex’s progeny accounting for 41.25% of all registered puppies from 2021 to 2024. Therefore, genetic testing before mating is now an essential measure to prevent genetic diseases in purebred dogs

Quantification of biosecurity measures in Mediterranean European seabass and gilthead sea bream farms

In the Mediterranean region, aquaculture drives economic growth and food supply through the production of seabass and sea bream. However, the region faces disease risks attributable to aquaculture intensification. Biosecurity is crucial for maintaining fish health and sustaining the aquaculture industry. This study adopted a novel risk-based scoring tool, originally developed for terrestrial animals, to benchmark biosecurity levels in seabass and sea bream. The tool considers all relevant aspects of aquaculture biosecurity, subdivided into external and internal biosecurity, with an accompanying questionnaire subjected to expert scoring. Land-based hatcheries and pre-growing facilities exhibited higher preventative measures, achieving overall biosecurity scores of 73.0 % and 75.0 %, respectively. These scores stem from stringent external and internal preventative measures, contrasting with lower overall biosecurity scores in land-based on-growing (50.0 %) and sea-based on-growing (59.0 %) facilities. Maintaining biosecurity during on-growing phases seems demanding due to uncontrolled interactions with the seawater environment and potential water quality issues. This benchmarking tool allows farmers to audit their biosecurity programs and pinpoint areas needing improvement. One such area is vaccination, which not only received the lowest scores but also exhibited high disparities across farm types. Overall, the study emphasizes the need for tailored measures to create synergy between external and internal components for robust biosecurity in Mediterranean aquaculture

Morphological and Molecular Identification of Obligatory Myiasis-Causing Species in Wild Cervids in Croatia

The infestation of ruminants by fly larvae of the Oestridae family can lead to nasopharyngeal or subcutaneous myiasis, which can harm animal welfare and productivity on sheep and cattle farms. The infestation of wild ruminants allows them to serve as reservoirs for subsequent transfer to domestic animals. The fly species most commonly responsible for myiasis have been well studied in many parts of the world, but less so in southeastern Europe. Here, we identified three botfly species among 197 subcutaneous larvae and 138 nasopharyngeal larvae from 36 wild roe deer (Capreolus capreolus) and 9 wild red deer (Cervus elaphus) at 30 locations across Croatia. Instars were stored in 75% ethanol and their species and developmental stage were identified based on morphological and morphometric keys. Subcutaneous larvae in both cervid species were identified as Hypoderma diana, while nasopharyngeal larvae were identified as Cephenemyia simulator in roe deer and Pharyngomyia picta in red deer. Partial sequencing of the gene-encoding mitochondrial cytochrome oxidase subunit I confirmed the taxonomic classification of C. stimulator and P. picta but was unable to confirm that of H. diana for lack of previously published sequences. This study appears to be the first to combine the morphological and molecular characterization of myiasis-causing botflies in wild cervids in southeastern Europe

Non-zoonotic tick-borne pathogens in Western Balkan

Ixodid ticks are present throughout the Western Balkan countries, including Albania, Bosnia and Herzegovina, Croatia, Montenegro, North Macedonia and Serbia, with many species serving as vectors for pathogens of both veterinary and medical importance. A thorough examination of existing literature, encompassing historical documents, has been conducted to gather information on all documented non-zoonotic tick-borne pathogens found in ticks, pets, farm animals, and wild animals across the Western Balkan region. This comprehensive review was necessary due to the scarcity and scattered nature of available data from this area. The tick fauna in the Western Balkans consists of 32 species across five genera: Ixodes, Haemaphysalis, Dermacentor, Rhipicephalus and Hyalomma. Various pathogens responsible for diseases in animals, including bacteria and parasites, have also been documented, many of which can cause important diseases and significant reductions in animal productivity. Initial efforts were directed towards pathogen surveillance and the characterization of non-zoonotic tick-borne pathogens, resulting in the identification of Theileria orientalis, Anaplasma bovis and Anaplasma marginale in cattle, although significant gaps remain in the current research. Studies on small ruminants have been sparse, with confirmed cases of Anaplasma ovis and Babesia ovis in sheep, but no comprehensive and systematic research on pathogens in goats. In contrast, research on canine piroplasms has identified several species, including Babesia canis and Babesia vulpes. Studies on wild animals, however, have predominantly focused on wild canines and carnivores, with limited attention given to non-zoonotic pathogens. Notably, only one study has reported non-zoonotic tick-borne pathogens in artiodactyl species and wild felids. The aim of this review is to provide an overview of the existing research on non-zoonotic tick-borne diseases in the Western Balkans, including the historical context, current data, and research gaps. Given the significant impact of these diseases on animal health and productivity, as well as their potential biodiversity, further comprehensive studies and the establishment of national surveillance systems for tick-borne diseases are essential for a better understanding and mitigation of their impact

Seafood Labeling in Croatia: Molecular Evidence and Regulatory Insights

Fisheries and aquaculture play a crucial role in global food security, yet species mislabeling remains a persistent challenge, undermining consumer trust and market transparency. Proper food labeling is essential for protecting public health due to the presence of unknown toxic or allergenic substances and preventing illegally sourced products from entering the market. Despite extensive research across Europe, seafood mislabeling in Croatia has remained unexplored. This study aims to provide the first comprehensive assessment of seafood labeling accuracy in Croatia, where fisheries are integral to the coastal economies and tourism. Using DNA barcoding of the COI gene, 109 seafood samples were collected over two years from various sources, including restaurants, markets, and fishing vessels, and analyzed for potential mislabeling. Results revealed a mislabeling rate of 3% among fish samples and 20% among cephalopods, with notable substitutions, such as the yellowfin tuna mislabeled as bigeye tuna and Bluefin tuna and the European squid mislabeled as Patagonian squid. Additionally, 38.5% of samples were partially labeled, while 32% lacked clear country-of-origin information, complicating traceability. While the findings align with the mislabeling rates in other European countries, this study underscores the ongoing challenges in seafood labeling compliance. Establishing standardized monitoring protocols will be essential for improving comparability and effectively addressing seafood fraud

Prevalence of Salmonella spp., Listeria monocytogenes, Campylobacter spp. and Enterobacteriaceae in raw pet food

Raw feeding of dogs and cats is becoming increasingly popular worldwide. The raw material for such feed is muscles, offal and bones from mammals, fish or poultry. The production of such pet food is regulated by the European Commission legislation and should be carried out in a facility approved by the competent authority. Although proponents of such a diet point to the positive effects on the health and condition of the animals, studies have shown that raw pet food often contains pathogenic microorganisms. In the analysis of 28 samples of frozen raw pet food, we detected Salmonella spp. in 21.4% of the samples, Enterobacteriaceae more than 5000 cfu/g in 85.7% of the samples, Listeria monocytogenes more than 10 cfu/g in 39.3% of the samples, while Campylobacter spp. was below the limit of quantification in all samples. Of the eight samples containing poultrymeat, Salmonella spp. was detected in five of them (62.5%), indicating that poultry is a high-risk raw material for Salmonella contamination. Meanwhile, meat from domestic ungulates was identified as a high-risk source for Listeria monocytogenes, with that bacteria detected in 9 out of 20 (45%) of the samples containing this type of meat. The results indicate that hygienein the production of raw pet food is unsatisfactory in most cases, that such food poses a risk to animal health and can be an indirect source of infection for humans via animal excretions and contaminated surfaces

Analysis of residues of veterinary drugs – nitroimidazoles in rare honey species

Honey is a natural product producedby honeybees valued for its nutritional andhealth benefits. However, contaminationwith veterinary drug residues, such as ni-troimidazoles (NMZ), poses a health risk toconsumers. Nitroimidazoles, banned in theEuropean Union due to their carcinogenicand genotoxic properties, have been mis-used in beekeeping to treat diseases suchas nosemosis. The aim of this study was todetect nitroimidazole residues in rare uni-floral honey species from Croatia, Ailanthusaltissima (Mill) Swingle and Mentha spp.,using a validated ultra-performance liquidchromatography-tandem mass spectrometrymethod. Although the applied method wasoriginally developed for the determinationof NMZs in poultry muscle and eggs, the re-sults of the validation study show that it isalso suitable for determination in honey withsome modifications. For all analytes, the de-cision limit and detection capability valueswere between 0.33 and 0.71 µg/kg, which isbelow the minimum method performancerequirement for NMZs in honey of 1 µg/kgset by the European Union Reference Labo-ratories. Ten nitroimidazoles were analysedin 11 honey samples and no residues weredetected, confirming compliance with Eu-ropean regulations. These results are in linewith broader studies showing rare NMZ con-tamination in the global honey market. How-ever, they emphasise the need for continuousHoney is a natural product producedby honeybees valued for its nutritional andhealth benefits. However, contaminationwith veterinary drug residues, such as ni-troimidazoles (NMZ), poses a health risk toconsumers. Nitroimidazoles, banned in theEuropean Union due to their carcinogenicand genotoxic properties, have been mis-used in beekeeping to treat diseases suchas nosemosis. The aim of this study was todetect nitroimidazole residues in rare uni-floral honey species from Croatia, Ailanthusaltissima (Mill) Swingle and Mentha spp.,using a validated ultra-performance liquidchromatography-tandem mass spectrometrymethod. Although the applied method wasoriginally developed for the determinationof NMZs in poultry muscle and eggs, the re-sults of the validation study show that it isalso suitable for determination in honey withsome modifications. For all analytes, the de-cision limit and detection capability valueswere between 0.33 and 0.71 µg/kg, which isbelow the minimum method performancerequirement for NMZs in honey of 1 µg/kgset by the European Union Reference Labo-ratories. Ten nitroimidazoles were analysedin 11 honey samples and no residues weredetected, confirming compliance with Eu-ropean regulations. These results are in linewith broader studies showing rare NMZ con-tamination in the global honey market. How-ever, they emphasise the need for continuousmonitoring, especially as the possible use ofbanned NMZs in honey production cannot becompletely ruled out. Future studies shouldalso investigate the transfer of NMZs frombeeswax to honey, emphasising the impor-tance of good beekeeping practises to avoidcontamination.monitoring, especially as the possible use ofbanned NMZs in honey production cannot becompletely ruled out. Future studies shouldalso investigate the transfer of NMZs frombeeswax to honey, emphasising the impor-tance of good beekeeping practises to avoidcontamination

Is golden jackal (Canis aureus) the most important indicator species of Trichinella infection in Croatia?

Epidemiologic monitoring of wild animals is always an important step in defining potential zoonoses that can threaten humans. Particular emphasis should be given to those zoonotic agents permanently cycling within wild animal populations and represent a permanent reservoir for other wild or domesticated animals that can be direct sources of disease for humans. In Croatia, there are two European jackal populations: the Dalmatian population (DP) that has been inhabiting the islands and coastal area along the Adriatic Sea since medieval times, and the South East European population (SEEP) that is found in continental Croatia. Research on Trichinella infections in jackal populations in Croatia was conducted from 2008 to 2022. During this 15-year period, we tested 186 jackal samples and confirmed infection in 47 individuals (25.3%). The dominant species was T. spiralis, identified in 28 samples (60%), T. britovi was found in 13 samples (28%), while for six samples (12%) the PCR test was unsuccessful. In both populations, the Trichinella species of the domestic cycle (T. spiralis) was found, though in varying ratios: in DP the ratio of identified species was 10:6 in favour of T. britovi, as opposed to 22:3 in favour of T. spiralis in SEEP. The frequency of infection with parasites from the genus Trichinella was significantly different in DP (22.9%) than in SEEP (26.7%) (p<0.001), while the larval count in analysed tissue did not differ by type of Trichinella species (p=0.1028). Infected jackals were found in nine of ten tested counties. The results were analysed statistically and the origin of tested and positive samples shown on a map of Croatia. Based on these findings, both jackal populations can be considered to represent an exceptionally important indicators of parasites from the genus Trichinella in Croatia, both for the sylvatic and domestic cycles. There is an evident need for epidemiological monitoring for members of both populations

Efficient Management of Sample Reception and Documentation at the Croatian Veterinary Institute

The reception office at the Croatian Veterinary Institute plays a crucial role in ensuring the accurate and efficient handling of samples and accompanying documentation, and serves as the first point of contact for veterinarians and inspectors regarding samples, or for clients who call by phone. This paper outlines the best practices for managing sample reception, emphasising the importance of proper documentation, the use of technology to streamline processes, and the implementation of quality control measures. Proper documentation is essential for maintaining sample traceability and integrity, which is critical for accurate testing and analysis. The use of technology, such as electronic documentation systems and barcode scanning, can significantly enhance the efficiency and accuracy of the sample reception process. Additionally, implementing quality control measures, including regular audits and staff training, ensures the reliability and consistency of the reception process. By adopting these best practices, the Croatian Veterinary Institute can improve its operational efficiency and reduce the risk of errors, thereby contributing to public health and safety

Control of glyphosate residues in food in the European union in the period 2007-2022

Most agricultural crops are treated several times with different types of pesticides during cultivation and it is therefore very likely that pesticide residues remain on the treated agricultural crops. Pesticides are therefore a very important group of chemical compounds that need to be controlled, mainly because of their high toxicity and widespread use in the production of agriculturalproducts. The most commonly used herbicide is glyphosate. To ensure a high level of consumer protection, legal limits or maximum residue levels have been set in Regulation (EC) No. 396/2005. The European Food Safety Authority (EFSA) publishes anannual report that provides an overview of the official monitoring of pesticide residues in the European Union Member States, Iceland and Norway. This paper summarises the data from EFSA's annual reports from 2007 to 2022, referring to data related to the best-selling herbicide glyphosate