In Vitro Antagonistic Effect of Gut Bacteriota Isolated from Indigenous Honey Bees and Essential Oils against Paenibacillus Larvae.

The aim of study was to isolate and identify the gut bacteria of Apis mellifera and to evaluate antagonistic effect of the bacteriota against Paenibacillus larvae, which causes American foulbrood (AFB) in honeybees. The dilution plating method was used for the quantification of selected microbial groups from digestive tract of bees, with an emphasis on the bacteriota of the bees' intestines. Bacteria were identified using mass spectrometry (MALDI-TOF-MS Biotyper). Overall, five classes, 27 genera and 66 species of bacteria were identified. Genera Lactobacillus (10 species) and Bacillus (8 species) were the most abundant. Gram-negative bacteria were represented with 16 genera, whereas Gram-positive with 10 genera. Delftia acidovorans and Escherichia coli were the most abundant in the digestive tract of honey bee. Resistance to a selection of antimicrobials was assessed for the bacterial isolates from bee gut and confirmed against all antimicrobials included in the study, with the exception of cefepime. Lactobacillus spp., especially L. kunkeei, L. crispatus and L. acidophilus. showed the strongest antimicrobial activity against P. larvae, the causal pathogen of AFB. Antimicrobial activity of essential oils against isolated bacteria and two isolates of P. larvae were assessed. Application of a broad selection of plant essential oils indicated that Thymus vulgaris had the highest antimicrobial activity against P. larvae.

Properties of Ginkgo biloba L.: Antioxidant Characterization, Antimicrobial Activities, and Genomic MicroRNA Based Marker Fingerprints.

The aim of this study was to characterize extracts from the leaves of Ginkgo biloba L. from selected Slovakian localities in terms of the content of bioactive constituents, antioxidants and their antimicrobial properties. The results indicated that the content of antioxidants was sample-specific, and this specificity was statistically significant. Ginkgo biloba L. from the locality of Kosice had the best activity determined by the free radical scavenging activity (DPPH) (1.545 mg Trolox equivalent antioxidant capacity (TEAC)/g fresh matter (FM)) as well as the molybdenum-reducing antioxidant power (35.485 mg TEAC/g FM) methods. The highest content of total polyphenols (2.803 mg gallic acid equivalent (GAE)/g FM) and flavonoids (4.649 µg quercetin equivalent (QE)/g FM) was also detected in this sample. All samples of G. biloba leaf extracts showed significant antimicrobial activity against one or more of the examined bacterial species, and Staphylococcus aureus subsp. aureus CCM 2461 was found to be the most susceptible (minimal inhibition concentration MIC50 and MIC90 values of 64.2 and 72.2 µg/mL, respectively). Based on the results it was concluded that Ginkgo biloba L. extracts can be used as antimicrobial and antioxidant additives. Selected miRNA-based molecular markers were used to examine the environmental adaptability of Ginkgo biloba L. An almost-complete genotype clustering pattern based on locality was determined in the analysis that involved a species-specific gb-miR5261 marker. Morphologically specific exemplar, cv. Ohatsuki, was excluded.

Characterization of Rosa canina Fruits Collected in Urban Areas of Slovakia. Genome Size, iPBS Profiles and Antioxidant and Antimicrobial Activities.

The studies of plant bacterial endophytes, colonizing the plant tissues without any signs of diseases, are essential for understanding of ecological interactions. The aim of our study is to detect microbiological contamination and to assess the antimicrobial, antioxidant activity, total phenolic, carotenoid content, genome size, and ploidy of non-cultivated Rosa canina sampled from urban areas. Samples of Rosa canina fruits were collected in three locations in Slovakia. The highest total viable count and the Enterobacteriaceae count in fruits were 4.32 log CFU/g and 4.29 log CFU/g, respectively. Counts of the mesophilic anaerobic sporulating bacteria, Pseudomonas spp., and of the microscopic fungi and yeasts were 3.00, 2.15 log CFU/g, 3.65 log CFU/g, and 2.76 log CFU/g, respectively. Regarding the antimicrobial activity, Escherichia coli and Klebsiela oxytoca were the most sensitive species among the assayed microorganisms to the treatment with the ethanolic extracts of Rosa canina fruits. The fruits were rich in bioactive compounds, polyphenols, and carotenoids, that could be related to their antioxidant activity. Genome sizes of analyzed samples ranged from 2.3 to 2.96. DNA-based fingerprinting obtained by iPBS markers of the Rosa canina var. lapidicola Heinr. Braun., was characterized by some distinctive inserted loci. An interdisciplinary study was performed for the dog roses from different parts of Slovakia that resulted in deeper characterization of this species.

Biological Activity and Antibiofilm Molecular Profile of Citrus aurantium Essential Oil and Its Application in a Food Model.

The main aim of the study was to investigate the chemical composition, antioxidant, antimicrobial, and antibiofilm activity of Citrus aurantium essential oil (CAEO). The biofilm profile of Stenotrophonomonas maltophilia and Bacillus subtilis were assessed using the mass spectrometry MALDI-TOF MS Biotyper and the antibiofilm activity of Citrus aurantium (CAEO) was studied on wood and glass surfaces. A semi-quantitative composition using a modified version was applied for the CAEO characterization. The antioxidant activity of CAEO was determined using the DPPH method. The antimicrobial activity was analyzed by disc diffusion for two biofilm producing bacteria, while the vapor phase was used for three penicillia. The antibiofilm activity was observed with the agar microdilution method. The molecular differences of biofilm formation on different days were analyzed, and the genetic similarity was studied with dendrograms constructed from MSP spectra to illustrate the grouping profiles of S. maltophilia and B. subtilis. A differentiated branch was obtained for early growth variants of S. maltophilia for planktonic cells and all experimental groups. The time span can be reported for the grouping pattern of B. subtilis preferentially when comparing to the media matrix, but without clear differences among variants. Furthermore, the minimum inhibitory doses of the CAEO were investigated against microscopic fungi. The results showed that CAEO was most active against Penicillium crustosum, in the vapor phase, on bread and carrot in situ.

The evaluation of chemical, antioxidant, antimicrobial and sensory properties of kombucha tea beverage.

The aim of the present study was to determine the chemical composition (organic acids-acetic, tartaric, citric; sugars-sucrose, glucose, fructose; total acidity, alcohol content, pH-with FTIR instrument; content of selected mineral compounds-AAS instrument), antioxidant activity, antimicrobial activity and sensory profiles of prepared kombucha tea beverage. Black tea with white sugar as a substrate for kombucha beverage was used as a control sample. The dominant organic acid in kombucha tea beverage was acetic acid (1.55 g/L), followed by tartaric and citric acids. The sucrose (17.81 g/L) was the dominant sugar from detected sugars. Antioxidant activity of beverage tested by reducing power method (1318.56 mg TEAC/L) was significantly higher (p < 0.05) in comparison with black tea (345.59 mg TEAC/L). The same tendency was observed for total polyphenol content which was significantly higher (p < 0.05) in kombucha beverage (412.25 mg GAE/L) than in black tea (180.17 mg GAE/L). Among mineral compounds, the amount of manganese (1.57 mg/L) and zinc (0.53 mg/L) was the highest in kombucha tea beverage. Results of antimicrobial activity of kombucha tea beverage showed strong inhibition of Candida krusei CCM 8271 (15.81 mm), C. glabrata CCM 8270 (16 mm), C. albicans CCM 8186 (12 mm), C. tropicalis CCM 8223 (14 mm), Haemophilus influenzae CCM 4454 (10 mm) and Escherichia coli CCM 3954 (4 mm). Sensory properties of prepared beverage were evaluated overall as good with the best score in a taste (pleasant fruity-sour taste). The consumption of kombucha tea beverage as a part of drinking mode of consumers due to health benefits is recommended.

Comparison of MALDI-TOF MS Biotyper and 16S rDNA sequencing for the identification of Pseudomonas species isolated from fish.

Effective and reliable methods of identification of Pseudomonas species are important for the characterization of microorganisms. Freshwater ecosystems are an important source of Pseudomonas species, including those pathogenic to fish and humans. The aim of the present study was to compare the identification conducted with MALDI-TOF MS Biotyper and 16S rDNA sequencing of fish-borne Pseudomonas spp. Altogether, 13 different Pseudomonas spp. were isolated from freshwater fish. Phylogenetic analysis showed a clear taxonomic placement only for 13 out of 15 Pseudomonas isolates. Accordance of identification method was found only in 6 out of 15 isolates. The human pathogenic Pseudomonas spp. were not found in our study, indicating that the fish could be considered as safe for consumption. The present study revealed a high discriminatory power of the mass spectra investigation and 16S rDNA gene sequencing technology for the identification of Pseudomonas spp. associated with freshwater fish.

Prevalence and Antimicrobial Resistance of Escherichia coli, Enterococcus spp. and the Major Foodborne Pathogens in Calves in Latvia.

The aim of the present study was to detect the prevalence and antimicrobial resistance of fecal indicators and major foodborne pathogens in feces of calves and to identify the factors associated with increased prevalence of resistant bacteria on farms. Altogether, 180 rectal swabs were collected from 18 farms in Latvia. Samples were investigated to detect the prevalence and antimicrobial resistance of Escherichia coli, Enterococcus spp., Listeria monocytogenes, Yersinia enterocolitica, Salmonella spp., Staphylococcus aureus, and Campylobacter spp. Among all, 64% (74/110) of commensal E. coli, 100% (78/78) Enterococcus faecalis and 96% (22/23) Enterococcus faecium isolates were resistant at least to one antibiotic. The prevalence of extended-spectrum ß-lactamase (ESBL)/AmpC-producing E. coli were 11.1% (20/180) with blaCTX-M, blaTEM, and blaCMY genes identified. Campylobacter jejuni (12.8%, 23/180) and Campylobacter coli (2.8%, 5/180) were the most resistant to tetracycline (61%, 14/23; 100%, 5/5) and fluoroquinolones (61%, 14/23; 100%, 5/5). Prevalence of L. monocytogenes was 0.6% (1/180) and S. aureus 1.7% (3/180). All samples were Salmonella and Y. enterocolitica negative. Farm size, bought calves, contact with other calves, and antimicrobial treatment of cows were associated with increased prevalence of resistant E. coli and Enterococcus spp. Despite the low overall usage of antimicrobials in Latvia, the high rates of antimicrobial resistance in fecal indicators and Campylobacter, in addition to the high prevalence of ESBL-producing E. coli, highlights the necessity for the prudent use of antimicrobials in dairy farms in Latvia.

The antioxidant and antimicrobial activity of essential oils against Pseudomonas spp. isolated from fish.

Natural products of plant origin, which include essential oils (EO) could be used as a growth inhibitor of pathogenic and spoilage microflora in food. The objective of this study was to determine the antibacterial and antioxidant activity of 21 EO against 10 Pseudomonas species isolated from freshwater fish. The chemical composition of EO was determined by gas chromatography/mass spectrometry. The disc diffusion method and detection of minimum inhibitory concentration (MIC) were used for the determination of the antimicrobial activity. All the EO tested exhibited antimicrobial activity, however, Cinnamomum zeylanicum EO was the most effective against Pseudomonas spp. both according to the disc diffusion and MIC methods. The EOs of Cymbopogon nardus, Origanum vulgare, Foeniculum vulgare and Thymus serpyllum showed the highest antioxidant activity of 93.86 µg, 83.47 µg, 76.74 µg and 74.28 µg TEAC/mL. Application of EO could be an effective tool for inhibition of growth of Pseudomonas spp. on fish.

Identification of lactic acid bacteria isolated from wine using real-time PCR.

Different lactic acid bacteria strains have been shown to cause wine spoilage, including the generation of substances undesirable for the health of wine consumers. The aim of this study was to investigate the occurrence of selected species of heterofermentative lactobacilli, specifically Lactobacillus brevis, Lactobacillus hilgardii, and Lactobacillus plantarum in six different Slovak red wines following the fermentation process. In order to identify the dominant Lactobacillus strain using quantitative (real time) polymerized chain reaction (qPCR) method, pure lyophilized bacterial cultures from the Czech Collection of Microorganisms were used. Six different red wine samples following malolactic fermentation were obtained from selected wineries. After collection, the samples were subjected to a classic plate dilution method for enumeration of lactobacilli cells. Real-time PCR was performed after DNA extraction from pure bacterial strains and wine samples. We used SYBR® Green master mix reagents for measuring the fluorescence in qPCR. The number of lactobacilli ranged from 3.60 to 5.02 log CFU mL(-1). Specific lactobacilli strains were confirmed by qPCR in all wine samples. The number of lactobacilli ranged from 10(3) to 10(6) CFU mL(-1). A melting curve with different melting temperatures (T(m)) of DNA amplicons was obtained after PCR for the comparison of T(m) of control and experimental portions, revealing that the most common species in wine samples was Lactobacillus plantarum with a T(m) of 84.64°C.

Antimicrobial Resistance in Enterococcus spp. Isolates from Red Foxes (Vulpes vulpes) in Latvia.

Antimicrobial resistance (AMR) is an emerging public health threat and is one of the One Health priorities for humans, animals, and environmental health. Red foxes (Vulpes vulpes) are a widespread predator species with great ecological significance, and they may serve as a sentinel of antimicrobial resistance in the general environment. The present study was carried out to detect antimicrobial resistance, antimicrobial resistance genes, and genetic diversity in faecal isolates of red foxes (Vulpes vulpes). In total, 34 Enterococcus isolates, including E. faecium (n = 17), E. faecalis (n = 12), E. durans (n = 3), and E. hirae (n = 2), were isolated. Antimicrobial resistance to 12 antimicrobial agents was detected with EUVENC panels using the minimum inhibitory concentration (MIC). The presence of antimicrobial resistance genes (ARGs) was determined using whole-genome sequencing (WGS). Resistance to tetracycline (6/34), erythromycin (3/34), ciprofloxacin (2/34), tigecycline (2/34), and daptomycin (2/34) was identified in 44% (15/34) of Enterococcus isolates, while all the isolates were found to be susceptible to ampicillin, chloramphenicol, gentamicin, linezolid, teicoplanin, and vancomycin. No multi-resistant Enterococcus spp. were detected. A total of 12 ARGs were identified in Enterococcus spp., with the presence of at least 1 ARG in every isolate. The identified ARGs encoded resistance to aminoglycosides (aac(6')-I, ant(6)-Ia, aac(6')-Iih and spw), tetracyclines (tet(M), tet(L) and tet(S)), and macrolide-lincosamide-streptogramin AB (lnu(B,G), lsa(A,E), and msr(C)), and their presence was associated with phenotypical resistance. Core genome multilocus sequence typing (cgMLST) revealed the high diversity of E. faecalis and E. faecium isolates, even within the same geographical area. The distribution of resistant Enterococcus spp. in wild foxes in Latvia highlights the importance of a One Health approach in tackling AMR.

Giardia duodenalis Styles, 1902 Prevalence in Cattle (Bos taurus Linnaeus, 1758) in Europe: A Systematic Review.

Giardia duodenalis Styles, 1902 is an infectious agent which can cause enteritic disease in cattle (Bos taurus Linnaeus, 1758) worldwide. As a zoonotic protozoan, it is important to acknowledge Giardia prevalence and assemblages found in cattle and risk factors associated with the Giardia infection in herds. This systematic review aims to estimate the prevalence of G. duodenalis and its assemblages in cattle and to identify the risk factors associated with Giardia infection in cattle in Europe. A systematic review was performed according to Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines to collect information from published studies in Europe. A total of 1414 studies were identified and 17 relevant studies were included in this review. Mean Giardia prevalence in cattle in Europe was 35.1%, with the highest prevalence found in neonatal animals (39.6%), but mean heard prevalence was 67.0%. Mixed infections of Giardia assemblages A and E were found most frequently (55.6%), while assemblages A and B were found more frequently in animals under 24 months old. Risk factors, such as deep litter with run-out, prolonged calf contact with the dam, and seasonality, such as winter and spring, were found to be potential risk factors for Giardia presence in the herds.

Giardia duodenalis (Styles, 1902) in Cattle: Isolation of Calves with Diarrhoea and Manure Treatment in the Lagoon Presented as Risk Factors in Latvian Herds.

Giardia duodenalis is a waterborne zoonotic protozoan that causes gastrointestinal tract inflammation in humans, cattle, and other animals. The aim of the present study was to estimate the prevalence and potential risk factors for Giardia infection in cattle in Latvia. During 2020-2021, a total of 973 individual faecal samples from cattle aged from 1 day to 12 years old, from 32 cattle herds, were tested for Giardia cyst presence with immunofluorescence staining followed by Giardia assemblage differentiation targeting beta-giardin gene. Using a questionnaire, information was collected to estimate the potential risk factors for G. duodenalis infection in cattle herds. Giardia was found in 8.4% of the examined cattle with a mean intensity of 5756 cysts per gram of faeces. The highest prevalence was observed in the 0 to 3-month-old calves (16.4%). At least one Giardia shedding animal was found in 27 herds with an overall prevalence of 84.4%. Significantly higher prevalence was found for cattle infected with G. duodenalis assemblage E compared to that infected with assemblage A: 88.7% and 11.3%, respectively. Protective factors such as age and rodent control and change of shoes were found to be significant for Giardia infection, while isolating calves for diarrhoea and water bodies (ponds/lakes) in pasture were potential risk factors in Latvian cattle.

Evaluation of Genetic Diversity and Virulence Potential of Legionella pneumophila Isolated from Water Supply Systems of Residential Buildings in Latvia.

Legionella is an opportunistic pathogen with a biphasic life cycle that occasionally infects humans. The aim of the study was to assess the distribution of virulence genes and genetic diversity among L. pneumophila isolated from water supply systems of residential buildings in Latvia. In total, 492 water samples from 200 residential buildings were collected. Identification of Legionella spp. was performed according to ISO 11731, and 58 isolates were subjected to whole-genome sequencing. At least one Legionella-positive sample was found in 112 out of 200 apartment buildings (56.0%). The study revealed extensive sequence-type diversity, where 58 L. pneumophila isolates fell into 36 different sequence types. A total of 420 virulence genes were identified, of which 260 genes were found in all sequenced L. pneumophila isolates. The virulence genes enhC, htpB, omp28, and mip were detected in all isolates, suggesting that adhesion, attachment, and entry into host cells are enabled for all isolates. The relative frequency of virulence genes among L. pneumophila isolates was high. The high prevalence, extensive genetic diversity, and the wide range of virulence genes indicated that the virulence potential of environmental Legionella is high, and proper risk management is of key importance to public health.

Genetic diversity and known virulence genes in Listeria innocua strains isolated from cattle abortions and farm environment.

Listeria innocua is considered as non-pathogenic bacteria living in an environment although several cases of immunocompromised humans and ruminant listeriosis infections have been reported. Previously, L. innocua was identified as a potential pathogen and virulence in association with L. monocytogenes PrfA dependent virulence (LIPI-1) gene cluster was demonstrated in hemolytic L. innocua. L. innocua usually considered non-pathogenic versus pathogenic L. monocytogenes and L. ivanovii because of the main virulence gene loss. There are limited studies and reports available about L. innocua-caused illness in cattle. A total of 18 STs were identified in cattle abortions while 17 STs in the farm environment with majority of STs were present in both abortions and environmental samples. Genome sequencing showed that in one farm identical L. innocua clones were represented in water, feed, soil, and faeces sample groups, suggesting that animals most likely through the faecal shedding may remain as the main source of L. innocua in a farm environment. Out of all L. innocua isolates PrfA-dependent virulence genes were not found in aborted foetuses isolates and environmental L. innocua isolate groups; however, in 20% of isolates a complete LIPI-3 pathogenicity island encoding listeriolysin S was identified. In this study, we demonstrated that genetically diverse L. innocua clones were widely distributed in cattle farm environment and certain isolates had a significant pathogenicity potential for cattle, thus causing adverse health effects, including abortions.

Prevalence, virulence determinants, and genetic diversity in Yersinia enterocolitica isolated from slaughtered pigs and pig carcasses.

Yersinia enterocolitica is an important zoonotic foodborne pathogen that could be transferred from infected pigs to their carcasses at slaughter, with subsequent introduction of the pathogen into the food chain. The aim of the present study was to study the prevalence, virulence characteristics, and genetic diversity of Y. enterocolitica isolates present in slaughtered pig tonsils and carcasses by using the WGS approach. A total of 200 slaughtered pig tonsils from 11 pig farms were collected in 2020-2021 at six slaughterhouses located in Latvia. Out of these samples, n = 190 were obtained from slaughtered pigs raised on Latvian farms while n = 10 were of Lithuanian origin, with the number of farms sampled being 10 and 1, respectively. Additionally, 30 pig carcasses were sampled at five slaughterhouses from pigs originating from five farms in 2021. Samples were investigated microbiologically, Y. enterocolitica isolates were biotyped and serotyped. Y. enterocolitica 4/O:3 was screened for antimicrobial resistance with the EUVSEC test panels. Whole genome sequence analysis (WGS) was performed in order to detect virulence genes and to assess the genetic diversity of Y. enterocolitica isolates. A total of 139 isolates, including one to three isolates from 84 Y. enterocolitica positive slaughtered pig tonsils and 13 pig carcass samples, were subjected to WGS analysis. The prevalence of Y. enterocolitica 4/O:3 in slaughtered pig tonsils and carcasses was 35% (70/200) and 13% (4/30), respectively. Antimicrobial resistance to ampicillin and tetracycline was detected in 97% (72/74) and 1% (1/74) of Y. enterocolitica 4/O:3 isolates. Y. enterocolitica 4/O:3 was represented only by ST18, while Y. enterocolitica 1A by ST3, ST147, ST304, ST307, and ST473. The ST18 isolates harbored the same main chromosomal (ail, inv, myfA, ystA) and majority shared plasmid-borne virulence genes (virF, yadA, yop virulon). The main virulence genes were not identified within the STs of Y. enterocolitica 1A and only minor differences were found between ST3, ST147, ST304, ST307, and ST473. Among ST18 isolates, cgMLST analysis revealed 43 cgMLST genotypes while 16 cgMLST genotypes were found among Y. enterocolitica 1A STs. The present study has shown the distribution of genetically distant cgMLST genotypes in slaughtered pigs from pig farms located in different geographical regions of Latvia, with one to 11 cgMLSTs identified within each sampled farm. The presence of undistinguishable cgMLST genotypes in slaughtered pig tonsils and the respective carcasses supported the link between the slaughter of Y. enterocolitica - positive pigs and carcass contamination with Y. enterocolitica 4/O:3.

Bacteriota and Antibiotic Resistance in Spiders.

Arthropods are reported to serve as vectors of transmission of pathogenic microorganisms to humans, animals, and the environment. The aims of our study were (i) to identify the external bacteriota of spiders inhabiting a chicken farm and slaughterhouse and (ii) to detect antimicrobial resistance of the isolates. In total, 102 spiders of 14 species were collected from a chicken farm, slaughterhouse, and buildings located in west Slovakia in 2017. Samples were diluted in peptone buffered water, and Tryptone Soya Agar (TSA), Triple Sugar Agar (TSI), Blood Agar (BA), and Anaerobic Agar (AA) were used for inoculation. A total of 28 genera and 56 microbial species were isolated from the samples. The most abundant species were Bacillus pumilus (28 isolates) and B. thuringensis (28 isolates). The least isolated species were Rhodotorula mucilaginosa (one isolate), Kocuria rhizophila (two isolates), Paenibacillus polymyxa (two isolates), and Staphylococcus equorum (two isolates). There were differences in microbial composition between the samples originating from the slaughterhouse, chicken farm, and buildings. The majority of the bacterial isolates resistant to antibiotics were isolated from the chicken farm. The isolation of potentially pathogenic bacteria such as Salmonella, Escherichia, and Salmonella spp., which possess multiple drug resistance, is of public health concern.

Antimicrobial Activity and Chemical Composition of Essential Oils against Pathogenic Microorganisms of Freshwater Fish.

Antimicrobials are widely applied in aquaculture for treatment of infectious diseases in fish. The increased antimicrobial resistance of fish pathogens to conventional antimicrobial treatment highlights the need for research on the antibacterial properties of natural products-in this case, essential oils (EOs). The aim of the present study was to detect the antimicrobial activity of the essential oils on pathogenic microorganisms found in freshwater fish. Freshwater fish isolates of Aerococcus spp., Aeromonas spp., Enterococcus spp., Escherichia spp., Pseudomonas spp., Shewanella spp., Yersinia spp., and Vagococcus spp. were tested for antimicrobial resistance and antimicrobial activity against 14 commercially available essential oils. Antimicrobial resistance was identified in Pseudomonas spp. isolates against cefepime and ciprofloxacin; while all Aeromonas, Enterococcus, and Yersinia isolates were fully susceptible. All tested EOs revealed antimicrobial activity against the tested freshwater fish isolates at different extents. Cinnamomum camphora exhibited strong antimicrobial activity against Aeromonas spp. (3.12 µL/mL), Enterococcus spp. (0.78-1.56 µL/mL), and Pseudomonas spp. with the MIC method. EOs of Gaultheria procumbens and Litsea cubeba showed strong antibacterial activity against Yersinia spp. and Vacococcus spp. (6.25 µL/mL). The study shows the antimicrobial activity of EOs against the most relevant freshwater fish pathogens and indicates the application opportunities in aquaculture.

Diversity of microbiota in Slovak summer ewes' cheese "Bryndza".

"Bryndza" cheese is an important Slovak traditional regional product. New knowledge on the role of microorganisms involved the "Bryndza" ripening process may provide valuable data on its quality and safety. In our study, the "Bryndza" made from pasteurized ewes milk was studied towards total count of bacteria, coliforms bacteria, enterococci, lactic acid bacteria, and microscopic filamentous fungi. All those groups of microbiota were detected using classical microbiological methods and identified using mass spectrometry. A total of 3,758 isolates were identified with score higher than 2.00. Altogether, 13 families, 24 genus, and 44 species of microbiota were identified in Slovak cheese "Bryndza." The most often isolated species were yeasts Yarrowia lipolitica and Dipodascus geotrichum and the lactic acid bacteria Lactobacillus paracasei subsp. paracasei.

Characterization and Genetic Diversity of Listeria monocytogenes Isolated from Cattle Abortions in Latvia, 2013-2018.

Listeria monocytogenes can cause disease in humans and in a wide range of animal species, especially in farm ruminants. The aim of the study was to determine the prevalence and genetic diversity of L. monocytogenes related to 1185 cattle abortion cases in Latvia during 2013-2018. The prevalence of L. monocytogenes among cattle abortions was 16.1% (191/1185). The seasonality of L. monocytogenes abortions was observed with significantly higher occurrence (p < 0.01) in spring (March-May). In 61.0% of the cases, the affected cattle were under four years of age. L. monocytogenes abortions were observed during the third (64.6%) and second (33.3%) trimesters of gestation. Overall, 27 different sequence types (ST) were detected, and four of them, ST29 (clonal complex, CC29), ST37 (CC37), ST451 (CC11) and ST7 (CC7), covered more than half of the L. monocytogenes isolates. Key virulence factors like the prfA-dependent virulence cluster and inlA, inlB were observed in all the analyzed isolates, but lntA, inlF, inlJ, vip were associated with individual sequence types. Our results confirmed that L. monocytogenes is the most important causative agent of cattle abortions in Latvia and more than 20 different STs were observed in L. monocytogenes abortions in cattle.

Virulence Determinants and Genetic Diversity of Yersinia Species Isolated from Retail Meat.

Yersinia enterocolitica is an important foodborne pathogen, and the determination of its virulence factors and genetic diversity within the food chain could help understand the epidemiology of yersiniosis. The aim of the present study was to detect the prevalence, and characterize the virulence determinants and genetic diversity, of Yersinia species isolated from meat. A total of 330 samples of retailed beef (n = 150) and pork (n = 180) in Latvia were investigated with culture and molecular methods. Whole genome sequencing (WGS) was applied for the detection of virulence and genetic diversity. The antimicrobial resistance of pathogenic Y. enterocolitica isolates was detected in accordance with EUCAST. Yersinia species were isolated from 24% (79/330) of meats, and the prevalence of Y. enterocolitica in pork (24%, 44/180) was significantly higher (p < 0.05) than in beef (13%, 19/150). Y. enterocolitica pathogenic bioserovars 2/O:9 and 4/O:3 were isolated from pork samples (3%, 6/180). Only resistance to ampicillin was confirmed in Y. enterocolitica 4/O:3 and 2/O:9 isolates, but not in other antimicrobials. Major virulence determinants, including ail, inv, virF, ystA and myfA, were confirmed with WGS in Y. enterocolitica 2/O:9 and 4/O:3. MLST typing revealed 15 STs (sequence types) of Y. enterocolitica with ST12 and ST18, which were associated with pathogenic bioserovars. For Y. enterocolitica 1A, Y. kristensenii, Y. intermedia and Y. frederiksenii, novel STs were registered (ST680-688). The presence of virulence genes and genetic characteristics of certain Y. enterocolitica STs confirm the common knowledge that pork could be an important source of pathogenic Yersinia.