State Veterinary Administration

BACKGROUNDSalmonellosis is the second most notified food-borne infection in Europe. In Czechia (CZ), the notification rate has consistently been above the European average, five times higher in 2013–2022.AIMWe aimed to describe the epidemiology of salmonellosis in CZ in 2012–2023 and identify areas for improving surveillance and public health actions.METHODSWe extracted data on notified salmonellosis cases from the national surveillance system and analysed the dataset for demographic characteristics, hospitalisation, transmission mode and suspected vehicle, descriptively and by chi-square tests and logistic regression.RESULTSIn 2012–2023, 130,990 cases (102.7/100,000 inhabitants) were notified. The overall annual notification rate decreased from 111.7 per 100,000 in 2012–2017 to 94.2 per 100,000 in 2018–2023. Most cases were children and adolescents aged < 15 years (n = 68,370; 52.2%) and most were females (n = 68,425; 52.2%) but males dominated among cases aged < 15 years (35,790/68,370; 52.3%). Few cases were imported (n = 2,627; 2.0%) or outbreak-related (n = 5,361; 4.1%); most were diagnosed in June–October (n = 83,057; 63.4%). Of all notified cases, 22.2% (n = 29,082) were hospitalised. Age of ≥ 40 years was associated with hospitalisation and Salmonella sepsis (p < 0.001). Transmission mode was identified for 59,729 (45.6%) cases.CONCLUSIONDespite a decline, the notification rates of salmonellosis remain high in CZ. The effectiveness of the surveillance system is reflected in relatively low hospitalisation rates compared with some other European countries, yet the source and transmission mode were unidentified in over half of the cases. We encourage enhancing epidemiological investigations, improving sample submission to the National Reference Laboratory and molecular surveillance.

Tracer tests are the key technique to characterize karst conduits. Hundreds of thousands tracer tests done worldwide showing typical flow velocities in conduits in km/day. In contrast slow flow in fractions of m/day is typical for porous aquifers. This study reports surprisingly slow flow velocity obtained from karst conduit in small metamorphosed carbonate occurrence in Czech Republic. Factors responsible for exceptionally slow flow are discussed.Ch&amp;#253;nov Karst is formed by several km long but few hundred meters wide carbonates strip inside gneiss in slightly undulated landscape. It hosts karst conduit traversed by small subterranean stream (5-13 L/s, observation since 2001), fed by diffuse recharge, no sinking streams occur in area. Karst conduit is accessible at two places: Ch&amp;#253;nov Cave and 1200 m distant spring Rutice. The carbonate strip is traversed perpendicularly by two small streams, which valleys contain Miocene Mydlovary Formation. The river network and karst conduit could be dated back at least to Miocene, as demonstrated by Miocene sediments below the recent streams. There are no signs of hypogenic origin. Cave is formed by deep phreatic loops indicating low density of fractures available for conduit forming. No transport of sediment was ever observed, the water remains limpid and flow in conduit is nearly constant.Tracer tests from 1960 indicated residence time in days, but individual results strongly differed, therefore new tracer test was performed using uranine in 2021. Uranine concentration has been monitored by field fluorometers GGUN-30 (Albilia, Switzerland) and laboratory fluorimeter Perkin Elmer LS55 for more than 3 years (continues). Breakthrough curve was analyzed by Qtracer2 code. New tracer test demonstrated residence time 1.4 year between cave and Rutice spring, which is equal to mean flow velocity 3.6 m/day. Volume of mobile water in conduit is 252000 m3 which is equal to average cross section of conduit 140 m2 . Tracer recovery is currently 25%. Tracer did not arrive to any other spring or stream in wider surrounding. Large conduit volume and low flow rate are responsible for extremely slow flow and long residence time. Large volumes are probably primarily caused by deep phreatic loops in cave. Phreatic loops disable under gentle landscape the transport of sediment from streams to cave.Therefore, the localized sinks can never develop in the area. As a consequence, the corrosion over more than 20 million years ever increases the conduit volume and no sediment is transported inside to infill it. Existence of this conduit indicates that low fracture density resulting in deep phreatic loops can in combination with gentle morphology over long time period result in evolution voluminous conduits with very slow flow. If Ch&amp;#253;nov cave is not known in the area the long residence time of water and low flow fluctuation will be considered as indication of non-existence of karst conduits.Funded by the GAUK No. 171624.

The present paper comprises a systematic survey of helminths (trematodes, an acanthocephalan and nematodes) found in nine species of freshwater fishes in Ecuador collected in March 1999 and those (a trematode and acanthocephalans) collected from an amphibian and two species of freshwater fishes in Venezuela in 1992, 1996 and 2001. The following 17 helminth species were recorded: Trematoda: Prosthenhystera ornamentosa sp. n., P. obesa (Diesing, 1850), Crassicutis intermedius (Szidat, 1954), C. cichlasomae Manter, 1936 and Glypthelmins eleutherodactyli sp. n. Acanthocephala: Quadrigyrus torquatus Van Cleave, 1920, Gracilisentis variabilis (Diesing, 1851) and Neoechinorhynchus (Neoechinorhynchus) ecuadoris sp. n. Nematoda: Cosmoxynema vianai Travassos, 1949, Travnema travnema Pereira, 1938, Touzeta ecuadoris Petter, 1987, Sprentascaris hypostomi Petter et Cassone, 1984, Sprentascaris sp., Contracaecum sp. Type 1 larvae, Contracaecum sp. Type 2 larvae, Procamallanus (Procamallanus) peraccuratus Pinto, Noronha et Rolas, 1976 and Procamallanus (Spirocamallanus) sp. juv. Nearly all of these parasites are reported from Ecuador or Venezuela for the first time and many of these findings represent new host records. The new species P. ornamentosa sp. n. was collected from the gall-bladder of an unidentified anostomid (Anostomidae, Characiformes) in Ecuador, G. eleutherodactyli sp. n. from the digestive tract of the frog Eleutherodactylus sp. (Eleutherodactylidae, Anura) in Venezuela and N. (N.) ecuadoris sp. n. from the intestine of Lebiasina sp. (Lebiasinidae, Characiformes) in Ecuador. Most parasites are briefly described and illustrated and problems concerning their morphology, taxonomy, hosts and geographical distribution are discussed.

In this study, our primary objective was to develop an effective analytical method for studying trypsin-digested peptides of two proteins commonly found in cow's milk: β-casein (βCN) and β-lactoglobulin (βLG). To achieve this, we employed two distinct approaches: traditional in-gel protein digestion and protein digestion using immobilized enzyme microreactors (μ-IMER). Both methods utilized ZipTip pipette tips filled with C18 reverse phase media for sample concentration. The μ-IMER was fabricated through a multi-step process that included preconditioning the capillary, modifying its surface, synthesizing a monolithic support, and further surface modification. Its performance was evaluated under HPLC chromatography conditions using a small-molecule trypsin substrate (BAEE). Hydrolysates from both digestion methods were analyzed using MALDI-TOF MS. Our findings indicate that the μ-IMER method demonstrated superior sequence coverage for oxidized molecules in βCN (33 ± 1.5%) and βLG (65 ± 3%) compared to classical in-gel digestion (20 ± 2% for βCN; 49 ± 2% for βLG). The use of ZipTips further improved sequence coverage in both classical in-gel digestion (26 ± 1% for βCN; 60 ± 4% for βLG) and μ-IMER (41 ± 3% for βCN; 80 ± 5% for βLG). Additionally, phosphorylations were identified. For βCN, no phosphorylation was detected using classical digestion, but the use of ZipTips showed a value of 27 ± 4%. With μ-IMER and μ-IMER–ZipTip, the values increased to 30 ± 2% and 33 ± 1%, respectively. For βLG, the use of ZipTip enabled the detection of a higher percentage of modified peptides in both classical (79 ± 2%) and μ-IMER (79 ± 4%) digestions. By providing a comprehensive comparison of traditional in-gel digestion and μ-IMER methods, this study offers valuable insights into the advantages and limitations of each approach, particularly in the context of complex biological samples. The findings set a new benchmark in protein digestion and analysis, highlighting the potential of μ-IMER systems for enhanced sequence coverage and post-translational modification detection.

We present the first data on nation-wide surveillance of plasmid-mediated colistin resistance in the Czech Republic. High occurrence of plasmid-mediated colistin resistance was found in meat samples, especially in poultry from both domestic production and import, while the presence of mcr genes was lower in the gut of slaughter animals. In contrast to culture-based approach, testing of whole-community DNA showed higher prevalence of mcr and presence of various mcr variants. Our results support the importance of combining cultivation methods with direct culture-independent techniques and highlight the need for harmonized surveillance of plasmid-mediated colistin resistance. Our study confirmed the importance of livestock as a major reservoir of plasmid-mediated colistin resistance and pointed out the risks of poultry meat for the transmission of mcr genes toward humans. We identified several mcr-associated prevalent STs, especially ST1011, which should be monitored further as they represent zoonotic bacteria circulating between different environments.

Agriculture is inextricably linked with the production of not only the main types of food products, but also a significant amount of waste. Therefore, the main task of modern agroengineering today is to develop or test new, more efficient and environmentally friendly agricultural technologies based on the intelligent use of natural resources. Problems with livestock manure are well known to experts, but the rest of the population is practically unaware of them and considers them insignificant. Since its production is constantly growing and negatively affects the state of the environment, it is important to set up effective measures that will be sustainable and financially acceptable in the long term. Recycling livestock manure with subsequent use as bedding is the right ecological way to use livestock manure.

In 2021/2022, the re-emergence of highly pathogenic avian influenza (HPAI) occurred in Europe. The outbreak was seeded from two sources: resident and reintroduced viruses, which is unprecedented in the recorded history of avian influenza. The dominant subtype was H5N1, which replaced the H5N8 subtype that had predominated in previous seasons. In this study, we present a whole genome sequence and a phylogenetic analysis of 57 H5N1 HPAI and two low pathogenic avian influenza (LPAI) H5N1 strains collected in the Czech Republic during 2021/2022. Phylogenetic analysis revealed close relationships between H5N1 genomes from poultry and wild birds and secondary transmission in commercial geese. The genotyping showed considerable genetic heterogeneity among Czech H5N1 viruses, with six different HPAI genotypes, three of which were apparently unique. In addition, second-order reassortment relationships were observed with the direct involvement of co-circulating H5N1 LPAI strains. The genetic distance between Czech H5N1 HPAI and the closest LPAI segments available in the database illustrates the profound gaps in our knowledge of circulating LPAI strains. The changing dynamics of HPAI in the wild may increase the likelihood of future HPAI outbreaks and present new challenges in poultry management, biosecurity, and surveillance.

Simple SummaryLaboratory diagnostics of the presence of Melissococcus plutonius is necessary for the confirmation of European foulbrood (EFB) in honey bee colonies. Seeking EFB positive colonies is based on inspections performed by beekeepers or authorized inspectors. Bee brood, adult bees, or honey are usually sampled and confirmed by PCR. Here, we tested a new concept of searching suspected colonies by simple and effective collecting hive debris. Conventional PCR detection of M. plutonius in hive debris gives comparable sensitivity and specificity as for adults or honey. The main advantage of testing hive debris combines its effectiveness and costs compared to the sampling of adult bees or honey. Easy and fast collection of hive debris samples in areas with EFB outbreaks could fasten disease early detection and elimination.European foulbrood (EFB) is an infectious disease of honey bees caused by the bacterium Melissococcus plutonius. A method for DNA isolation and conventional PCR diagnosis was developed using hive debris, which was non-invasively collected on paper sheets placed on the bottom boards of hives. Field trials utilized 23 honey bee colonies with clinically positive symptoms and 21 colonies without symptoms. Bayes statistics were applied to calculate the comparable parameters for EFB diagnostics when using honey, hive debris, or samples of adult bees. The reliability of the conventional PCR was 100% at 6.7 × 103 Colony Forming Unit of M. plutonius in 1 g of debris. The sensitivity of the method for the sampled honey, hive debris, and adult bees was 0.867, 0.714, and 1.000, respectively. The specificity for the tested matrices was 0.842, 0.800, and 0.833. The predictive values for the positive tests from selected populations with 52% prevalence were 0.813, 0.833, and 0.842, and the real accuracies were 0.853, 0.750, and 0.912, for the honey, hive debris, and adult bees, respectively. It was concluded that hive debris can effectively be utilized to non-invasively monitor EFB in honey bee colonies.

Early detection of biohazardous bacteria that can be misused as biological weapons is one of the most important measures to prevent the spread and outbreak of biological warfare. For this reason, many instrument platforms need to be introduced into operation in the field of biological warfare detection. Therefore the purpose of this study is to establish a new detection panel for biothreat bacteria (Bacillus anthracis, Yersinia pestis, Francisella tularensis, and Brucella spp.) and confirm it by collaborative validation by using a multiplex oligonucleotide ligation followed by polymerase chain reaction and hybridization to microspheres by MagPix detection platform (MOL-PCR). Appropriate specific sequences in bacterial DNA were selected and tested to assemble the detection panel, and MOLigo probes (short specific oligonucleotides) were designed to show no cross-reactivity when tested between bacteria and to decrease the background signal measurement on the MagPix platform. During testing, sensitivity was assessed for all target bacteria using serially diluted DNA and was determined to be at least 0.5 ng/µL. For use as a diagnostic kit and easier handling, the storage stability of ligation premixes (MOLigo probe mixes) was tested. This highly multiplex method can be used for rapid screening to prevent outbreaks arising from the use of bacterial strains for bioterrorism, because time of analysis take under 4 h.

The designations and denominations employed