•Analytical approaches to authentication of food of animal origin.•Determination of specific food ingredients.•Determination of geographical origin.•Substitution of ingredients.
Since adulteration ...can have serious consequences on human health, it affects market growth by destroying consumer confidence. Therefore, authentication of food is important for food processors, retailers and consumers, but also for regulatory authorities. However, a complex nature of food and an increase in types of adulterants make their detection difficult, so that food authentication often poses a challenge. This review focuses on analytical approaches to authentication of food of animal origin, with an emphasis put on determination of specific ingredients, geographical origin and adulteration by virtue of substitution. This review highlights a current overview of the application of target approaches in cases when the compound of interest is known and non-target approaches for screening issues. Papers cited herein mainly concern milk, cheese, meat and honey. Moreover, advantages, disadvantages as well as challenges regarding the use of both approaches in official food control but also in food industry are investigated.
To fulfill epidemiological data and investigate possible interspecies transmission, this study shall attempt to sequence representative HEV strains of human, swine and wild boar origin collected from ...2010 to 2017 in Croatia.
In total, 174 anti-HEV antibody positive human sera samples; 1419 blood or faeces samples of swine, as well as 720 tissue and/or blood samples of wild boar originating from different counties (18 in total) in Croatia were tested for the presence of HEV RNA.
HEV RNA was detected in 26 human sera samples (14.9%; 95% CI 10.4-21.0%). HEV RNA was detected in 216 tested swine (15.2%; 95% CI 13.5-17.1%), regardless of age, farm breeding system or geographical origin. Viral RNA was also detectable in faeces samples which prove that swine actively participate in shedding HEV into the environment. Of the total of 720 tested wild boar samples, 83 were HEV RNA positive (11.5, 95% CI 9.4-14.1%) originating from six counties. According to the sequence analysis all strains have shown to be members of Orthohepevirus A genotype HEV-3, regardless of host. The genotyping results confirm grouping of sequences into four subtypes of HEV strains of which subtypes 3a and 3c belong to the general cluster 3abchij, and were predominately detected during the study, while subtypes 3e and 3f fall within cluster 3efg. Strains within subtypes 3a and 3e were found in humans, swine and wild boars; subtype 3c strains were derived from humans and swine, whereas subtype 3f strains were found only in humans. Strains belonging to subtypes 3a and 3c were derived during the entire investigated period and may be considered endemic in Croatia, whereas strains within subtypes 3e and 3f were detected sporadically indicating the possibility of newly imported infections.
All detected strains show to be genetically highly related to strains found in humans and/or animals from other European Countries, indicating that trade of live animals or wild boar movement increases the risk of HEV infection spread. Furthermore, homologous strains found in different investigated species within this study indicate interspecies transmission of HEV and/or an existence of an accessible mutual source of infection.
Since the role of wild rodents/small mammals in hepatitis E virus (HEV) epidemiology has been a subject of considerable debate, this study was conducted to investigate the potential presence of HEV ...RNA in small rodents collected within their natural habitats and to detect if they can be potential reservoirs of the virus. A total of 483 small rodents were captured using snap traps placed at 11 regions in Croatia. Sampling was undertaken in 2008 and repeated from 2010 to 2014. Liver samples were tested for the presence of HEV RNA. HEV RNA was detected in only one liver sample (0.21%) originated from Apodemus flavicollis from the location Medvednica, nearby Zagreb collected in 2014. According to the sequence analysis, the isolate has shown to be a member of Orthohepevirus A species, genotype HEV-3. The genotyping results confirmed grouping into subtype 3a, general cluster 3abchij.The detected HEV strain showed to be genetically highly related to strains found in humans and/or domestic pigs and wild boars from Croatia. Our finding indicates that wild small mammals could play a role in the epidemiology of HEV-3 infection and therefore should be taken under consideration as potential reservoirs or/and transmitters of the disease. However, further investigation is needed to recognize their potential for maintaining the infection in natural conditions.
The canine distemper virus (CDV), a paramyxovirus that is closely related to the human measles virus and rinderpest virus of cattle, is a highly contagious viral disease in dogs and wild carnivores ...worldwide. CDV represents a serious threat to domestic and wild animals, especially to the conservation of endangered wild carnivores. Our study aims to investigate the occurrence of CDV in free-living wild canines in Croatia. For this purpose, 176 red foxes and 24 jackal brain samples collected in the frame of the active surveillance of rabies during winter 2021/2022 were tested. This study provided the first comprehensive overview of the prevalence and spatial distribution of CDV in the wildlife of Croatia, including the molecular phylogenetic analysis of the H gene sequence of field CDV strains circulating in red fox and jackal populations of Croatia. The molecular characterization of hemagglutinin gene genomic regions confirmed the phylogenetic clustering of obtained sequences into the Europa 1 genotype. The obtained CDV red fox sequences were mutually very similar (97.60%). This study indicates the high genetic similarity of Croatian CDV red fox sequences and CDV red fox sequences from Italy and Germany, badger sequences from Germany, polecat sequences from Hungary, and dog sequences from Hungary and Germany.
•Porcine astroviruses (PAstV) are prevalent in the faeces of domestic pigs in Croatia, regardless of the age, the type of holding or its geographical location.•There is wide genetic heterogeneity of ...PAstV sequences in Croatia, with all five known lineages being detected.•This is the first report of the PAstV-3 lineage outside North America.
Porcine astroviruses (PAstVs) are common and genetically diverse viruses of domestic pigs. In the present study, the prevalence of PAstV in healthy domestic pigs in Croatia was 89%. Phylogenetic analysis revealed genetic heterogeneity among PAstV sequences; five lineages were detected, with PAstV-2 being predominant, while PAstV-3 was detected for the first time outside North America.
Hepatitis E virus (HEV) can infect a wide range of domestic and wild animals, and the identification of new host species is reported successively worldwide. Nevertheless, its zoonotic potential and ...natural transmission, especially in wildlife remains unclear, primarily due to the discrete nature of HEV infections. Since the red fox (
) is the most widespread carnivore worldwide, and has been recognized as a potential HEV reservoir, its role as a potent host species is of increasing interest. Another wild canine species, the jackal （
）, is becoming more important within the same habitat as that of the red fox since its number and geographical distribution have been rapidly growing. Therefore, we have chosen these wild species to determine their potential role in the epidemiology and persistence of HEV in the wilderness. The main reason for this is the finding of HEV and a rather high HEV seroprevalence in wild boars sharing the same ecological niche as the wild canine species, as well as the risk of the spread of HEV through red foxes into the outskirts of cities, where possible indirect and even direct contact with people are not excluded. Therefore, our study aimed to investigate the possibility of natural HEV infection of free-living wild canines, by testing samples for the presence of HEV RNA and anti-HEV antibodies to gain better epidemiological knowledge of the disease. For this purpose, 692 red fox and 171 jackal muscle extracts and feces samples were tested. Neither HEV RNA nor anti-HEV antibodies were detected. Although HEV circulation was not detected in the tested samples, to our knowledge, these are the first results that include jackals as a growing and important omnivore wildlife species for the presence of HEV infection in Europe.
The circulation of SARS-CoV-2 in the environment has been confirmed numerous times, whilst research on the bioaccumulation in bivalve molluscan shellfish (BMS) has been rather scarce. The present ...study aimed to fulfil the knowledge gap on SARS-CoV-2 circulation in wastewaters and surface waters in this region and to extend the current knowledge on potential presence of SARS-CoV-2 contamination in BMS. The study included 13 archive wastewater and surface water samples from the start of epidemic and 17 influents and effluents from nine wastewater treatment plants (WWTP) of different capacity and treatment stage, sampled during the second epidemic wave. From that period are the most of 77 collected BMS samples, represented by mussels, oysters and warty venus clams harvested along the Dalmatian coast. All samples were processed according to EN ISO 15216-1 2017 using Mengovirus as a whole process control. SARS-CoV-2 detection was performed by real-time and conventional RT-PCR assays targeting E, N and nsp14 protein genes complemented with nsp14 partial sequencing. Rotavirus A (RVA) real-time RT-PCR assay was implemented as an additional evaluation criterion of virus concentration techniques. The results revealed the circulation of SARS-CoV-2 in nine influents and two secondary treatment effluents from eight WWTPs, while all samples from the start of epidemic (wastewaters, surface waters) were negative which was influenced by sampling strategy. All tertiary effluents and BMS were SARS-CoV-2 negative. The results of RVA amplification were beneficial in evaluating virus concentration techniques and provided insights into RVA dynamics within the environment and community. In conclusion, the results of the present study confirm SARS-CoV-2 circulation in Croatian wastewaters during the second epidemic wave while extending the knowledge on wastewater treatment potential in SARS-CoV-2 removal. Our findings represent a significant contribution to the current state of knowledge that considers BMS of a very low food safety risk regarding SARS-CoV-2.
•The first report of SARS-CoV-2 circulation in wastewaters in Croatia.•Wastewater treatment contributes to the SARS-CoV-2 reduction and removal.•The first SARS-CoV-2 study in bivalves during autumn-winter and 2nd epidemic wave.•All bivalves were SARS-CoV-2 negative indicating very low food safety risk.
• Porcine astrovirus viremia in apparently healthy domestic pigs. • Possible high fecal prevalence and persistence in domestic pigs in Croatia. • The vast genetic diversity of PoAstV strains on a ...relatively confined area. • The first report of PoAstV lineage 5 presence in Europe.
Astroviruses are emerging viral agents, primarily enteropathogenic in mammals, but recently have been acknowledged to have extra-intestinal implications in humans and mink. Porcine astrovirus is thought to be widely distributed and highly prevalent among pigs, nevertheless its clinical significance remains doubtful as it can be detected in diarrheic as well as in healthy pigs. Recent reports imply the immense genetic variability among porcine astrovirus strains with five distinct lineages being characterized so far. Herein, we report porcine astrovirus circulation in the blood of healthy pigs in different age categories bred on two large industrial holdings in Croatia, with viral RNA seroprevalence of 3.89%. These are the first extra-intestinal findings of astrovirus in pigs, indicating a more complex pathogenesis than previously thought. Partial polymerase sequences of serum-derived strains provisionally clustered into porcine astrovirus lineages 2 and 4, sharing high genetic identity with previously described porcine astrovirus strains. The results were supported by detecting porcine astrovirus strains in composite fecal samples, regardless of pig category or holding tested. Phylogenetic analysis of derived strains suggested the presence of porcine astrovirus lineages previously detected in pig sera with an additional highly genetically divergent lineage 5, reported for the first time in Europe. Moreover, the existence of possible sub lineages should not be excluded. The results obtained in the present study, contribute to knowledge of porcine astrovirus pathogenesis; even though it’s possible clinical significance remains unclear. High fecal prevalence accompanied with vast genetic diversity on a relatively confined area, underscores the importance of pigs as porcine astrovirus reservoirs with eventual recombination events as a possible outcome.
Aujeszky disease is a viral disease, primarily of suids, caused by Suid-alphaherpesvirus 1, also known as the Aujeszky disease virus. The potential transmission of virus from wild boars to domestic ...pigs underlines the necessity of studying the epidemiology of Aujeszky disease in wild boars. The aim of this study was to evaluate the potential of demographic, environmental and population management predictors in predicting that ELISA tested animals will be seropositive. We used 222 wild boar blood samples, collected from 10 different hunting grounds and Medvednica Nature Park. In total, 12 predictors were used in this study, three of which were demographic (age category, sex, population density), two represented population management (hunting method, relative hunting bag) and seven environmental characteristics (altitude, watercourses, meadows, scrublands, forests, agricultural area and infrastructure area). The model selection was based on the Akaike Information Criterion, corrected AIC and Akaike weight. The model with the lowest value (ΔAIC <2 units) was selected as representing the better model fit. Predictors were selected using logistic forward stepwise regression. The age of the wild boars proved to be positively correlated with prediction of seropositive animals, while altitude and population density proved to be negatively correlated. The selected models were shown to be able to predict ADV seropositive animals with an average accuracy between 77.12 and 78.67%, and seronegative animals between 82.46 % and 85.09. The odds ratio values ranged between 12.53 and 14.52. Our results are in accordance with previous studies, and the impact of population density in the case of Aujeszky disease in wild boars remains unclear.
In this study we investigated the presence of Bovine Viral Diarrhoea Virus (BVDV) antibodies and antigens in a dairy cow herd that had reported clinical signs related to BVDV infection. The aim of ...the study was to confirm BVDV infection on the dairy farm with 1500 Holstein-Friesian cattles, and genetically analyse the BVDV type/s circulating in the herd. Serum samples (n=233) were collected from clinically infected cattles and tested for the presence of BVDV antibodies and antigens. The virus neutralization test showed that 194 out of 233 sera of the tested cows were positive (83.26%), and by Ab-ELISA 203 sera samples of the tested cows were shown to be positive (87.12%). The presence of antigens was proven with the Ag-ELISA test in 2 out of 233 samples (0.86%). After RT-PCR and phylogenetic analysis of two BVD virus isolates, it was found that both isolates belonged to genotype 1, subtype BVDV-1d. This is the first time that the BVDV-1d subtype has been confirmed in Croatia, although the BVDV-1b and BVDV-1f subtypes have been detected previously on Croatian dairy farms.
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