Utilisation of Actiphage in combination with IS qPCR as a diagnostic tool for rapid determination of paratuberculosis infection status in small ruminant herds.

Introduction: subsp. (MAP) is the causative agent of paratuberculosis, a chronic infectious intestinal disease occurring in domestic and wild ruminants. Early diagnosis of infected herds enabling timely adoption of control measures is tremendously important in view of the fact that the disease has a significant economic impact on farmers.

Identification of and discrimination between the complex and complex directly from sputum using quadruplex real-time PCR.

. 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 complex (MAC) and complex (MABSC) being the most frequently identified.

First Evidence of the Presence of the Causative Agent of Caseous Lymphadenitis- in Dairy Products Produced from the Milk of Small Ruminants.

This study focused on the detection and quantification of selected bacteria and on the presence of enterotoxin genes in milk and dairy products from sheep and goat farms in the Czech Republic using quantitative real-time PCR (qPCR) and multiplex PCR (PCR). The presence of (CP), subsp. (MAP), , , enterotoxin genes and methicillin-resistant (MRSA) was determined in 18 milk samples, 28 fresh cheeses, 20 ripened cheeses and 14 yoghurts.

A Novel Approach to the Viability Determination of subsp. Using Platinum Compounds in Combination With Quantitative PCR.

Mycobacterium avium subsp. (MAP) represents a slow-growing bacterium causing paratuberculosis, especially in domestic and wild ruminants. Until recently, the assessment of MAP viability relied mainly on cultivation, which is very time consuming and is unable to detect viable but non-culturable cells.

Application of the Actiphage® Assay to Detect Viable subsp. Cells in Fresh Sheep and Goat Milk and Previously Frozen Milk and In-Line Milk Filters.

subsp. (MAP) is a well-known causative agent of paratuberculosis, a chronic infectious granulomatous enteritis of ruminants contributing to significant economic losses worldwide. Current conventional diagnostic tools are far from being sufficient to manage and control this disease.

Development of a reference standard for the detection and quantification of Mycobacterium avium subsp. paratuberculosis by quantitative PCR.

Quantitative PCR (qPCR) has become a frequently employed direct method for the detection and quantification of Mycobacterium avium subsp. paratuberculosis (MAP). The quantity of MAP determined by qPCR, however, may be affected by the type of qPCR quantification standard used (PCR product, plasmid, genomic DNA) and the way in which standard DNA quantity is determined (absorbance, fluorescence).

Phage Amplification Assay for Detection of Mycobacterial Infection: A Review.

An important prerequisite for the effective control, timely diagnosis, and successful treatment of mycobacterial infections in both humans and animals is a rapid, specific, and sensitive detection technique. Culture is still considered the gold standard in the detection of viable mycobacteria; however, mycobacteria are extremely fastidious and slow-growing microorganisms, and therefore cultivation requires a very long incubation period to obtain results. Polymerase Chain Reaction (PCR) methods are also frequently used in the diagnosis of mycobacterial infections, providing faster and more accurate results, but are unable to distinguish between a viable and non-viable microorganism, which results in an inability to determine the success of tuberculosis patient treatment or to differentiate between an active and passive infection of animals.

Molecular Methods for the Detection of Oocysts in Fresh Produce: An Extensive Review.

Human infection with the important zoonotic foodborne pathogen has been associated with unwashed raw fresh produce consumption. The lack of a standardised detection method limits the estimation of fresh produce as an infection source. To support method development and standardisation, an extensive literature review and a multi-attribute assessment were performed to analyse the key aspects of published methods for the detection of oocyst contamination in fresh produce.

Efficiency of DNA Isolation Methods Based on Silica Columns and Magnetic Separation Tested for the Detection of Subsp. in Milk and Faeces.

Timely and reliable detection of animals shedding subsp. (MAP) should help to effectively identify infected animals and limit infection transmission at early stages to ensure effective control of paratuberculosis. The aim of the study was to compare DNA extraction methods and evaluate isolation efficiency using milk and faecal samples artificially contaminated by MAP with a focus on modern instrumental automatic DNA isolation procedures based on magnetic separation.

Toxoplasma gondii in vegetables from fields and farm storage facilities in the Czech Republic.

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.

Control of paratuberculosis: who, why and how. A review of 48 countries.

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.

[Mycobacterium marinum as a cause of human and animal infections].

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.

Sequence Analysis of Changes in Microbial Composition in Different Milk Products During Fermentation and Storage.

The objective of this study was to analyze the changes in the microbiota of milk products during fermentation and storage. Two kinds of Yoghurt, one Kefir, and one Acidophilus milk were observed during the fermentation process and storage using 16S rDNA amplicon sequencing. Cow's, goat's, raw and pasteurized milk were also examined.

Magnetic Separation Methods for the Detection of subsp. in Various Types of Matrices: A Review.

The main reasons to improve the detection of subsp. (MAP) are animal health and monitoring of MAP entering the food chain via meat, milk, and/or dairy products. Different approaches can be used for the detection of MAP, but the use of magnetic separation especially in conjunction with PCR as an end-point detection method has risen in past years.

Avian Mycobacteriosis: Still Existing Threat to Humans.

The nontuberculous mycobacteria are typically environmental organisms residing in soil and water. These microorganisms can cause a wide range of clinical diseases; pulmonary disease is most frequent, followed by lymphadenitis in children, skin and soft tissue disease, and rare extra pulmonary or disseminated infections. Mycobacterium avium complex is the second most common cause of pulmonary mycobacterioses after M.

Faecal bacterial composition in dairy cows shedding Mycobacterium avium subsp. paratuberculosis in faeces in comparison with nonshedding cows.

The aim of this study was to determine possible differences in the faecal microbiota of dairy cows infected with Mycobacterium avium subsp. paratuberculosis (Johne's disease) in comparison with noninfected cows from the same herds. Faecal samples from cows in 4 herds were tested for M.

Seasonal changes in microbial community composition in river water studied using 454-pyrosequencing.

The aims of this study were to determine the microbial community in five rivers in the proximity of a city in the Czech Republic using 454-pyrosequencing, as well as to assess seasonal variability over the course of 1 year and to identify the factors influencing the structure of bacterial communities. Samples from five rivers around the city of Brno were obtained during four seasons and analysed using 454 pyrosequencing of the 16S rRNA gene. The core composition of bacterial communities consisted of Actinobacteria, Bacteroidetes, Proteobacteria, Firmicutes, Fusobacteria, TM7 and others.

Detection of viable Mycobacterium avium subspecies paratuberculosis in powdered infant formula by phage-PCR and confirmed by culture.

Surveys from different parts of the world have reported that viable Mycobacterium avium subsp. paratuberculosis (MAP) can be cultured from approximately 2% of samples of retail pasteurised milk samples. Pasteurised milk is used for the production of powdered infant formula (PIF) and therefore there is a concern that MAP may also be present in these products.

Comparison of filtering methods, filter processing and DNA extraction kits for detection of mycobacteria in water.

Introduction And Objective: Mycobacteria have been isolated from almost all types of natural waters, as well as from man-made water distribution systems. Detection of mycobacteria using PCR has been described in different types of water; however, currently, there is no standardised protocol for the processing of large volumes of water.

Material And Methods: In the present study, different filtering methods are tested and optimised for tap or river water filtration up to 10 L, as well as filter processing and DNA isolation using four commercially available kits.

Comparison of methods for the isolation of mycobacteria from water treatment plant sludge.

Nontuberculous mycobacteria (NTM) are ubiquitous organisms in all natural ecosystems, including water environments. Several of these species are potential pathogens which affect human health. NTM most commonly cause pulmonary, skin or soft tissue infections.

The water environment as a source of potentially pathogenic mycobacteria.

Nontuberculous mycobacteria (NTM) are ubiquitous organisms of a wide variety of environmental reservoirs, including natural and municipal water, soil, aerosols, protozoans, animals and humans. Several of these species are potential pathogens which affect human health. The aim of this study was to determine the occurrence of NTM in the water environment.

Evidence of passive faecal shedding of Mycobacterium avium subsp. paratuberculosis in a Limousin cattle herd.

It has been suggested that passive shedding of Mycobacterium avium subsp. paratuberculosis (MAP) in faeces may occur, but reliable data are missing. Passive shedding assumes the ingestion of MAP in contaminated feed and passive passage through the gastrointestinal tract without causing infection.

Presence of Mycobacterium avium subspecies and hepatitis E virus in raw meat products.

Meat and meat products may be the source of various pathogenic and potentially pathogenic agents for humans. We ascertained the occurrence of Mycobacterium avium subsp. paratuberculosis, subsp.