Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is an RNA virus responsible for coronavirus disease 2019 (COVID-19). While SARS-CoV-2 primarily targets the lungs and airways, it can also infect other organs, including the central nervous system (CNS). The aim of this study was to investigate whether the choroid plexus could serve as a potential entry site for SARS-CoV-2 into the brain. Tissue samples from 24 deceased COVID-19-positive individuals were analyzed. Reverse transcription real-time PCR (RT-qPCR) was performed on selected brain regions, including the choroid plexus, to detect SARS-CoV-2 viral RNA. Additionally, immunofluorescence staining and confocal microscopy were used to detect and localize two characteristic proteins of SARS-CoV-2: the spike protein S1 and the nucleocapsid protein. RT-qPCR analysis confirmed the presence of SARS-CoV-2 viral RNA in the choroid plexus. Immunohistochemical staining revealed viral particles localized in the epithelial cells of the choroid plexus, with the spike protein S1 detected in the late endosomes. Our findings suggest that the blood-cerebrospinal fluid (B-CSF) barrier in the choroid plexus serves as a route of entry for SARS-CoV-2 into the CNS. This study contributes to the understanding of the mechanisms underlying CNS involvement in COVID-19 and highlights the importance of further research to explore potential therapeutic strategies targeting this entry pathway.

• Klíčová slova
• COVID‐19, SARS‐CoV‐2, blood‐cerebrospinal fluid barrier, choroid plexus, neuroinvasion,
• MeSH
• COVID-19 * virologie   MeSH
• dospělí MeSH
• fosfoproteiny * metabolismus   MeSH
• glykoprotein S, koronavirus * genetika   metabolismus   MeSH
• hematoencefalická bariéra * virologie   MeSH
• internalizace viru MeSH
• koronavirové nukleokapsidové proteiny MeSH
• lidé středního věku MeSH
• lidé MeSH
• plexus chorioideus * virologie   MeSH
• RNA virová * genetika   MeSH
• SARS-CoV-2 * fyziologie   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fosfoproteiny * MeSH
• glykoprotein S, koronavirus * MeSH
• koronavirové nukleokapsidové proteiny MeSH
• nucleocapsid phosphoprotein, SARS-CoV-2 MeSH Prohlížeč
• RNA virová * MeSH
• spike protein, SARS-CoV-2 MeSH Prohlížeč

With the onset of the COVID-19 pandemic, a problem arose with classic body donation programmes for obtaining cadavers for anatomical dissections, science and research. The question has emerged whether bodies of individuals who died of COVID-19 or were infected by SARS-CoV-2 could be admitted to Departments of Anatomy. To determine the risk of SARS-CoV-2 transmission to employees or students, the presence and stability of SARS-CoV-2 RNA in cadavers after fixation agents' application and subsequent post-fixation baths over time were examined. The presence of viral RNA in swabs from selected tissues was assessed by the standardized routine RNA isolation protocol and subsequent real-time PCR analysis. To support the results obtained from the tissue swabs, samples of RNA were exposed in vitro to short and long-term exposure to the components of the injection and fixation solutions used for the bodies' conservation. Substantial removal of SARS-CoV-2 RNA was observed in post-mortem tissue following perfusion with 3.5% phenol, 2.2% formaldehyde, 11.8% glycerol and 55% ethanol, and subsequent post-fixation in an ethanol bath. In vitro experiments showed significant effects of formaldehyde on SARS-CoV-2 RNA, while phenol and ethanol showed only negligible effects. We conclude that cadavers subjected to fixation protocols as described here should not pose a considerable risk of SARS-CoV-2 infection while being handled by students and staff and are, therefore, suitable for routine anatomical dissections and teaching.

• Klíčová slova
• Coronavirus, Dissection, Embalming, Infections, Polymerase chain reaction,
• MeSH
• COVID-19 * MeSH
• ethanol MeSH
• fenoly MeSH
• formaldehyd MeSH
• lidé MeSH
• mrtvola MeSH
• pandemie MeSH
• RNA virová MeSH
• SARS-CoV-2 * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ethanol MeSH
• fenoly MeSH
• formaldehyd MeSH
• RNA virová MeSH

Introduction. Cystic fibrosis (CF) is a serious disease with multisystemic clinical signs that is easily and frequently complicated by bacterial infection. Recently, the prevalence of nontuberculous mycobacteria as secondary contaminants of CF has increased, with the Mycobacterium avium complex (MAC) and Mycobacterium abscessus complex (MABSC) being the most frequently identified. The MABSC includes subspecies of significant clinical importance, mainly due to their resistance to antibiotics.Gap statement. Sensitive method for early detection and differentiation of MABSC members and MAC complex for use in routine clinical laboratories is lacking. A method based on direct DNA isolation from sputum, using standard equipment in clinical laboratories and allowing uncovering of possible sample inhibition (false negative results) would be required. The availability of such a method would allow accurate and accelerated time detection of MABSC members and their timely and targeted treatment.Aim. To develop a real time multiplex assay for rapid and sensitive identification and discrimination of MABSC members and MAC complex.Methodology. The method of DNA isolation directly from the sputum of patients followed by quadruplex real-time quantitative PCR (qPCR) detection was developed and optimised. The sensitivity and limit of detection (LOD) of the qPCR was determined using human sputum samples artificially spiked with a known amount of M. abscessus subsp. massiliense (MAM).Results. The method can distinguish between MAC and MABSC members and, at the same time, to differentiate between M. abscessus subsp. abscessus/subsp. bolletii (MAAb/MAB) and MAM. The system was verified using 61 culture isolates and sputum samples from CF and non-CF patients showing 29.5 % MAAb/MAB, 14.7 % MAM and 26.2 % MAC. The LOD was determined to be 1 490 MAM cells in the sputum sample with the efficiency of DNA isolation being 95.4 %. Verification of the qPCR results with sequencing showed 100 % homology.Conclusions. The developed quadruplex qPCR assay, which is preceded by DNA extraction directly from patients' sputum without the need for culturing, significantly improves and speeds up the entire process of diagnosing CF patients and is therefore particularly suitable for use in routine laboratories.

• Klíčová slova
• cystic fibrosis, mycobacteria, quadruplex quantitative PCR, sputum,
• MeSH
• atypické mykobakteriální infekce * diagnóza   farmakoterapie   MeSH
• cystická fibróza * komplikace   mikrobiologie   MeSH
• DNA terapeutické užití   MeSH
• intracelulární infekce bakterií Mycobacterium avium * epidemiologie   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé MeSH
• Mycobacterium abscessus * genetika   MeSH
• Mycobacterium avium komplex genetika   MeSH
• netuberkulózní mykobakterie MeSH
• sputum mikrobiologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DNA MeSH

The risk of Alzheimer's disease (AD) has a strong genetic component, also in the case of late-onset AD (LOAD). Attempts to sequence whole genome in large populations of subjects have identified only a few mutations common to most of the patients with AD. Targeting smaller well-characterized groups of subjects where specific genetic variations in selected genes could be related to precisely defined psychological traits typical of dementia is needed to better understand the heritability of AD. More than one thousand participants, categorized according to cognitive deficits, were assessed using 14 psychometric tests evaluating performance in five cognitive domains (attention/working memory, memory, language, executive functions, visuospatial functions). CD36 was selected as a gene previously shown to be implicated in the etiology of AD. A total of 174 polymorphisms were tested for associations with cognition-related traits and other AD-relevant data using the next generation sequencing. Several associations between single nucleotide polymorphisms (SNP's) and the cognitive deficits have been found (rs12667404 with language performance, rs3211827 and rs41272372 with executive functions, rs137984792 with visuospatial performance). The most prominent association was found between a group of genotypes in six genetically linked and the age at which the AD patients presented with, or developed, a full-blown dementia. The identified alleles appear to be associated with a delay in the onset of LOAD. In silico studies suggested that the SNP's alter the expression of CD36 thus potentially affecting CD36-related neuroinflammation and other molecular and cellular mechanisms known to be involved in the neuronal loss leading to AD. The main outcome of the study is an identification of a set of six new mutations apparently conferring a distinct protection against AD and delaying the onset by about 8 years. Additional mutations in CD36 associated with certain traits characteristic of the cognitive decline in AD have also been found.

• MeSH
• Alzheimerova nemoc * genetika   psychologie   MeSH
• antigeny CD36 * genetika   MeSH
• exekutivní funkce fyziologie   MeSH
• jednonukleotidový polymorfismus MeSH
• lidé MeSH
• mutace MeSH
• neuropsychologické testy MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD36 * MeSH
• CD36 protein, human MeSH Prohlížeč

At the time of sampling (2020/2021), the number of new cases of SARS-CoV-2-positive individuals in the Czech Republic significantly exceeded the numbers in neighboring countries and in the EU. In terms of the number of deaths, the country ranked near the top of the list. Legislative orders required wearing masks indoors, disinfecting surfaces in public places, and limiting the number of people per sales area in commercial spaces. Due to an situation, most schools and shops were closed. The entire country anticipated a total lockdown. To assess the risk to public health regarding SARS-CoV-2 transmission, air and surfaces were sampled in two public places: a post office and a shopping center. Samples were also collected at the COVID-19 unit at the local hospital. Neither air nor surface samples were positive for SARS-CoV-2 virus particles in the post office or shopping center. Positive results were found in the hospital ward, with floors being the most and highest contaminated surface. Based on our results, we believe that public places do not pose a risk in relation to SARS-CoV-2 transmission, especially when epidemiological measures to reduce transmission are followed, such as wearing masks, using disinfectant or limiting the number of customers per retail establishment.

• Klíčová slova
• RT-qPCR, SARS-CoV-2, air, fomites, indoor, surface contamination,
• MeSH
• COVID-19 * MeSH
• kontrola infekčních nemocí MeSH
• lidé MeSH
• masky MeSH
• nemocnice MeSH
• SARS-CoV-2 * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Hepatitis E virus (HEV) is the etiological agent behind hepatitis E infection. Domestic pigs and wild boars are the main animal reservoirs of HEV. Very few papers describe HEV infection in goats and sheep. As the data pertaining to the presence of HEV virus in the milk of small ruminants in Europe are lacking, the aim of this paper was to examine a representative number of milk samples from these animals. The detection of HEV genome (HEV RNA) was performed using reverse transcriptase real-time polymerase chain reaction (RT-qPCR). HEV RNA was found in 2.8% of the examined samples. Positivity ranged from 101 to 103 genome equivalents/mL (GE/mL) with a median of 9.99 × 102 GE/mL. On the basis of these results, the milk of small ruminants could represent a source of HEV infection to consumers.

• Klíčová slova
• HEV, RT-qPCR, milk, small ruminant,
• MeSH
• hepatitida E veterinární   MeSH
• kozy virologie   MeSH
• mléko virologie   MeSH
• nemoci zvířat diagnóza   virologie   MeSH
• ovce virologie   MeSH
• RNA virová MeSH
• virus hepatitidy E genetika   izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• RNA virová MeSH

Infection with Toxoplasma gondii has usually been connected with consumption of improperly treated meat. However, contaminated water and products of plant origin have emerged as new sources of infection in the last few years. Here, 292 vegetable samples-carrot, cucumber and lettuce-obtained from nine farms in the Czech Republic were examined using triplex real time PCR targeting two specific T. gondii sequences. Irrigation water and water used for washing of vegetables were also included. Overall, a positivity rate of 9.6% was found in vegetables. The concentration varied between 1.31 × 100 and 9.00 × 102 oocysts/g of sample. A significant difference was found between the positivity of vegetables collected directly from fields and that of vegetables collected from farm storage rooms (4.4-8.6% vs 10-24.1%, respectively). All samples of irrigation water and water used to rinse vegetables were negative. Genotyping based on restriction fragment length polymorphism (RFLP) analysis using seven markers revealed the exclusive presence of genotype II.

• Klíčová slova
• Toxoplasma gondii, farm, food safety, genotyping, qPCR, vegetable,
• MeSH
• bezpečnost potravin MeSH
• farmy * MeSH
• genotyp MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• potravinářská parazitologie * MeSH
• Toxoplasma * klasifikace   genetika   izolace a purifikace   MeSH
• zelenina parazitologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

Paratuberculosis, a chronic disease affecting ruminant livestock, is caused by Mycobacterium avium subsp. paratuberculosis (MAP). It has direct and indirect economic costs, impacts animal welfare and arouses public health concerns. In a survey of 48 countries we found paratuberculosis to be very common in livestock. In about half the countries more than 20% of herds and flocks were infected with MAP. Most countries had large ruminant populations (millions), several types of farmed ruminants, multiple husbandry systems and tens of thousands of individual farms, creating challenges for disease control. In addition, numerous species of free-living wildlife were infected. Paratuberculosis was notifiable in most countries, but formal control programs were present in only 22 countries. Generally, these were the more highly developed countries with advanced veterinary services. Of the countries without a formal control program for paratuberculosis, 76% were in South and Central America, Asia and Africa while 20% were in Europe. Control programs were justified most commonly on animal health grounds, but protecting market access and public health were other factors. Prevalence reduction was the major objective in most countries, but Norway and Sweden aimed to eradicate the disease, so surveillance and response were their major objectives. Government funding was involved in about two thirds of countries, but operations tended to be funded by farmers and their organizations and not by government alone. The majority of countries (60%) had voluntary control programs. Generally, programs were supported by incentives for joining, financial compensation and/or penalties for non-participation. Performance indicators, structure, leadership, practices and tools used in control programs are also presented. Securing funding for long-term control activities was a widespread problem. Control programs were reported to be successful in 16 (73%) of the 22 countries. Recommendations are made for future control programs, including a primary goal of establishing an international code for paratuberculosis, leading to universal acknowledgment of the principles and methods of control in relation to endemic and transboundary disease. An holistic approach across all ruminant livestock industries and long-term commitment is required for control of paratuberculosis.

• Klíčová slova
• Camelid, Cattle, Control, Deer, Goat, Paratuberculosis, Prevalence, Review, Sheep, Wildlife,
• MeSH
• chov zvířat MeSH
• divoká zvířata mikrobiologie   MeSH
• hlášení nemocí normy   MeSH
• incidence MeSH
• Mycobacterium avium subsp. paratuberculosis izolace a purifikace   MeSH
• paratuberkulóza ekonomika   epidemiologie   prevence a kontrola   MeSH
• přežvýkavci mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Field-grown strawberries, the environment of strawberry farms and fresh strawberries from marketplaces were examined for bacterial, viral, and protozoan pathogens. The presence of bacteria was determined using culture and real-time PCR (qPCR), presence of protozoa and viruses using qPCR and reverse transcription qPCR, respectively. The highest proportion of positivity was observed for Escherichia coli both in field and purchased strawberries (up to 48.6%). Finding of Cronobacter ranged from 0.6% to 9% both for field and market strawberries. The prevalence of other pathogens (Listeria monocytogenes, Giardia intestinalis, Cryptosporidium sp., and Norovirus) in strawberries was below 4.5%; HAV was not detected at all. Positivity of the environment was determined to be lower than 2.1% for all microorganisms, except for E. coli. The concentration of pathogens in most samples did not exceed 100 CFU/g using culture and 1.8 × 102 GE/g of strawberries or swabbing area 6.1 × 102 GE/mL or swabbing area of environmental samples using qPCR. All studied farms applied preventive measures such as drip irrigation, avoidance of organic fertilizers, and use of mulch foils or gloves for workers to decrease contamination of strawberries. Despite this, certain pathogens were found in fresh strawberries. Even at low concentrations, these pathogens can be a source of infection for consumers. Thus, their presence in strawberries is of particular significance as these are mostly consumed fresh and without any thermal processing. PRACTICAL APPLICATION: Nonlegislatively monitored pathogens of bacterial, viral and parasitic origin were found in strawberries. Monitoring the presence of these pathogens in ready-to-eat food is therefore meaningful and important in terms of food safety, especially in relation to pathogens with low infectious dose (for example, viruses, parasites).

• Klíčová slova
• Cronobacter, Norovirus, berry, environment, protozoa, ready-to-eat,
• MeSH
• bezpečnost potravin MeSH
• Cryptosporidium izolace a purifikace   MeSH
• Escherichia coli izolace a purifikace   MeSH
• farmy MeSH
• Giardia lamblia izolace a purifikace   MeSH
• jahodník mikrobiologie   parazitologie   virologie   MeSH
• kontaminace potravin analýza   MeSH
• lidé MeSH
• Listeria monocytogenes izolace a purifikace   MeSH
• Norovirus izolace a purifikace   MeSH
• počet mikrobiálních kolonií MeSH
• potravinářská mikrobiologie * MeSH
• potravinářská parazitologie MeSH
• průmyslově zpracované potraviny mikrobiologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Mycobacterium marinum is a slowly growing non-tuberculous (environmental, atypical) mycobacterium with zoonotic potential. It occurs in the aquatic environment and causes diseases in fish and other aquatic animals known as mycobacterioses. In humans, it primarily causes skin infections, which are most commonly located in the upper limbs. The disease commonly appears in connection with the aquarium environment and is thus referred to as fish tank granuloma. As with all mycobacterial diseases, treatment is complicated and lengthy. For a definitive determination of the pathogen, biological materials should always be examined in a laboratory specializing in diagnosing mycobacteria. Critical for the right diagnosis is proper sample collection and assessment of the patient's history. To detect mycobacteria, culture and microscopy are generally used. Species are identified using modern biological methods such as mass spectrometry (MALDI), polymerase chain reaction, hybridization probes or sequencing.

• MeSH
• atypické mykobakteriální infekce * diagnóza   farmakoterapie   mikrobiologie   MeSH
• lidé MeSH
• Mycobacterium marinum * MeSH
• nemoci ryb * diagnóza   farmakoterapie   mikrobiologie   prevence a kontrola   MeSH
• netuberkulózní mykobakterie MeSH
• polymerázová řetězová reakce MeSH
• ryby MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH