Ivo Pavlík | Mendel University in Brno (original) (raw)

Papers by Ivo Pavlík

Research paper thumbnail of Culture- and Quantitative IS 900 Real-Time PCR-Based Analysis of the Persistence of Mycobacterium avium subsp. paratuberculosis in a Controlled Dairy Cow Farm Environment

Applied and Environmental Microbiology, 2012

The aim of this study was to monitor the persistence of Mycobacterium avium subsp. paratuberculos... more The aim of this study was to monitor the persistence of Mycobacterium avium subsp. paratuberculosis in environmental samples taken from a Holstein farm with a long history of clinical paratuberculosis. A herd of 606 head was eradicated, and mechanical cleaning and disinfection with chloramine B with ammonium (4%) was carried out on the farm; in the surrounding areas (on the field and field midden) lime was applied. Environmental samples were collected before and over a period of 24 months after destocking. Only one sample out of 48 (2%) examined on the farm (originating from a waste pit and collected before destocking) was positive for M. avium subsp. paratuberculosis by cultivation on solid medium (Herrold's egg yolk medium). The results using real-time quantitative PCR (qPCR) showed that a total of 81% of environmental samples with an average mean M. avium subsp. paratuberculosis cell number of 3.09 × 10 3 were positive for M. avium subsp. paratuberculosis before destocking co...

Research paper thumbnail of Human-to-human and human-to-dog Mycobacterium tuberculosis transmission studied by IS6110 RFLP analysis: a case report

Veterinární medicína, 2011

This study reports on the transmission of Mycobacterium tuberculosis of the same IS6110 RFLP type... more This study reports on the transmission of Mycobacterium tuberculosis of the same IS6110 RFLP type between two acquaintances with open pulmonary tuberculosis and a five-year-old Doberman bitch. No clinical signs, gross lesions at necropsy or histopathological lesions were observed in the infected lungs and gastrointestinal tract of the dog, although M. tuberculosis was directly detected by IS6110 PCR and culture examinations in the respiratory and gastrointestinal tracts. IS6110 PCR positivity in the faeces and blood of the dog poses a risk of M. tuberculosis transmission between the dog and humans.

Research paper thumbnail of Systemic mixed infection in a brown caiman (Caiman crocodilus fuscus) caused by Mycobacterium szulgai and M. chelonae: a case report

Veterinární medicína, 2010

A five-year old female brown caiman (Caiman crocodilus fuscus) was admitted to a veterinary clini... more A five-year old female brown caiman (Caiman crocodilus fuscus) was admitted to a veterinary clinic because of anorexia and lethargy. Chronic deterioration of the patient’s condition together with the formation of slushy stools coloured from brown to red was observed during the previous eight weeks. Physical examination showed significant apathy and cachexia. Radiographic examinations of chest and abdomen revealed no pathological findings. Initial blood tests revealed decreased hematocrit and low levels of haemoglobin. Despite treatment with enrofloxacin and intensive supportive therapy with amino acids, vitamins and mineral matter, the animal died 14 days after admission to the clinic. Post mortem examination revealed splenomegaly with a total destruction of inner organ structure together with multiple granulomas in liver and lungs. Ziehl-Neelsen staining of tissue samples from liver, lungs and spleen revealed numerous acid-fast bacilli consistent with Mycobacterium spp. Identificat...

Research paper thumbnail of Peat as a feed supplement for animals: a literature review

Veterinární medicína, 2012

Peat is an easily available natural material and a source of biologically active substances widel... more Peat is an easily available natural material and a source of biologically active substances widely used, not only in agriculture but in human and animal medicine as well. In recent years, interest in the use of peat as a feed supplement has increased, particularly due to its capability to prevent enteric diseases and to stimulate growth in piglets and pigs. The purpose of this review was to compare the advantages and risks associated with the use of peat for animal nutrition based on the literature available. Beneficial effects of various peat preparations on digestion, growth and the immune systems of animals as well as the absorbent and detoxifying capabilities are associated with the high content of favourable humic substances. One disadvantage of using peat preparations is the considerable diversity of the various types of peat caused by different biological, chemical and geological conditions during formation. Biological activity of various peat preparations is associated not o...

Research paper thumbnail of Mycobacterium bovis in human population in four Central European countries during 1990-1999

Veterinární medicína, 2012

A survey on Mycobacterium bovis and M. tuberculosis in humans has been performed in four Central ... more A survey on Mycobacterium bovis and M. tuberculosis in humans has been performed in four Central European countries (Croatia, the Czech Republic, Slovak Republic and Slovenia) during the years 1990 to 1999. These countries cover an area of 204 688 km2 with 22 135 million population. During the period, new cases of tuberculosis were bacteriologically diagnosed in 47 516 patients. M. tuberculosis was detected in 47 461 (99.88%) cases, whereas M. bovis was found only in 55 (0.12%) patients. The rate of infection due to M. bovis in humans did not exceed 0.29% in the study countries. The annual incidence of bacteriological confirmed M. bovis cases did not exceed 0.1 per 100 000 inhabitants. In the Czech Republic out of 44 tuberculosis patients due to M. bovis, 32 (72.7%) were older than 61 years and originated from rural areas, where they lived during childhood and worked in agricultural occupations. These patients may have suffered a reactivation of persistent (long-standing) M. bovis i...

Research paper thumbnail of Incidence of bovine tuberculosis in wild and domestic animals other than cattle in six Central European countries during 1990-1999

Veterinární medicína, 2012

The study was undertaken in Croatia, Czech Republic, Hungary, Poland, Slovakia and Slovenia layin... more The study was undertaken in Croatia, Czech Republic, Hungary, Poland, Slovakia and Slovenia laying between Baltic and Adriatic seas on 610 402 km2. Mycobacterium bovis infection was diagnosed in 70 animals belonging to 17 species other than cattle. The set of wild animals comprised 12 European bison (Bison bonasus), one red deer (Cervus elaphus), five wild boars (Sus scrofa), and one European wild goat (Capra aegagrus) bred in a game park. Further positive animals included two farmed red deer (Cervus elaphus) and one bactrian camel (Camelus ferus) owned by a circus. The infection was also demonstrated in 18 domestic animals belonging to 3 species living on farms where bovine tuberculosis was diagnosed in cattle. This set included 12 domestic pigs (Sus scrofa f. domestica), two domestic sheep (Ovis ammon f. aries), and four dogs (Canis lupus f. familiaris). The set of animals bred in zoological gardens consisted of 30 animals belonging to 9 species as follows: three bison (Bison biso...

Research paper thumbnail of The transmission and impact of paratuberculosis infection in domestic and wild ruminants

Veterinární medicína, 2001

Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) infects domestic cattle, sheep,... more Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) infects domestic cattle, sheep, goats, deer, camelids and wild ruminants leading to chronic enteritis known as paratuberculosis (Johne’s disease). The infection is chronic, progressive and unresponsive to treatment. Most infected animals do not develop clinical disease but may excrete the bacteria. Clinically sick animals suffer emaciation and in some species diarrhoea, followed by eventual death. During the course of the disease, excretion of M. paratuberculosis in faeces and milk occurs, and the organism spreads through the blood and lymph vessels of infected animals to multiple internal organs. The infection disseminates to both the female and male reproductive organs. Though M. paratuberculosis is not classified as a human pathogen, current opinions on the possible role of this mycobacteria in public health is discussed. This article attempts to review the ways and circumstances by which M. paratuberculosis is tra...

Research paper thumbnail of Peat as a feed supplement for animals: a review

Peat is an easily available natural material and a source of biologically active substances widel... more Peat is an easily available natural material and a source of biologically active substances widely used, not only in agriculture but in human and animal medicine as well. In recent years, interest in the use of peat as a feed supplement has increased, particularly due to its capability to prevent enteric diseases and to stimulate growth in piglets and pigs. The purpose of this review was to compare the advantages and risks associated with the use of peat for animal nutrition based on the literature available. Beneficial effects of various peat preparations on digestion, growth and the immune systems of animals as well as the absorbent and detoxifying capabilities are associated with the high content of favourable humic substances. One disadvantage of using peat preparations is the considerable diversity of the various types of peat caused by different biological, chemical and geological conditions during formation. Biological activity of various peat preparations is associated not o...

Research paper thumbnail of Molecular Fingerprinting ofMycobacterium bovissubsp.capraeIsolates from Central Europe

Journal of Clinical Microbiology, 2004

ABSTRACTTo study the dissemination ofMycobacterium bovissubsp.caprae, 79 European isolates from c... more ABSTRACTTo study the dissemination ofMycobacterium bovissubsp.caprae, 79 European isolates from cattle, humans, and other hosts were examined by spoligotyping and IS6110restriction fragment length polymorphism (RFLP) analysis. Among a total of 11 different spoligotypes identified, type C1 proved to be predominant (n= 62). Five of the spoligotypes are described for the first time. A total of 43 different RFLP types were identified, thus allowing further differentiation for epidemiological tracking. Isolates from a series of outbreaks in one village proved to be of the same spoligotype and of identical or closely related RFLP types.

Research paper thumbnail of Correlation of Mycobacterium avium subsp. paratuberculosis counts in gastrointestinal tract, muscles of the diaphragm and the masseter of dairy cattle and potential risk for consumers

International Journal of Food Microbiology, 2011

Tissues of cattle intended for human consumption can be contaminated by Mycobacterium avium subsp... more Tissues of cattle intended for human consumption can be contaminated by Mycobacterium avium subsp. paratuberculosis (MAP). Although different studies attribute varying roles of MAP in Crohn's disease progression it is thought that the exposure of humans to this bacterium should in any case be minimised. In this study, we have collected samples of intestine, mesenteric lymph nodes, muscles of diaphragm (musculus diaphragma) and masseter muscles (musculus masseter) from twenty-five cows in a slaughterhouse. The infectious status of all animals was confirmed by culture of faeces. MAP was found in almost all the intestines and mesenteric lymph nodes examined, including three faecal culture-negative animals indicating intermittent shedding. As intestine is used for the traditional production of sausages, it is alarming that 84.2% of intestine samples were positive for MAP. F57 and IS900 real time PCR revealed MAP in 40 to 68% of diaphragms and 11.1 to 38.9% of masseters. A noticeable dependence of the probability of MAP positivity of faeces versus gastrointestinal tract (GIT) and of GIT and muscles was observed. Due to the changing behaviour of consumers, both of these muscles have started to be widely used in cuisine. Therefore, the results of this paper imply that the processing of cows with paratuberculosis in abattoirs without any precautions (restrictions) and the usage of meat for human consumption should be rethought.

Research paper thumbnail of Mycobacterium avium subsp. paratuberculosis survival during fermentation of soured milk products detected by culture and quantitative real time PCR methods

International Journal of Food Microbiology, 2012

Mycobacterium avium paratuberculosis (MAP), etiological agent of paratuberculosis in ruminants, i... more Mycobacterium avium paratuberculosis (MAP), etiological agent of paratuberculosis in ruminants, is able to survive extreme conditions like very low pH (stomach), high temperature (pasteurization) or low temperature (refrigerated storage). Cheese, infant powder milk, cream and other milk and dairy products might thus be considered as possible sources of MAP for humans. The aim of this study was to investigate the survival of two MAP field isolates during fermentation of three different types of soured milk products (SMP; yogurt, acidophilus milk and kefir) under laboratory conditions. Pasteurized MAP-free milk was artificially contaminated with 10(6)MAPcells/mL and survival and absolute numbers of MAP were monitored during fermentation (4 or 16 h) and after six weeks of storage at 4°C by culture and quantitative real time PCR (qPCR). Viability of MAP was determined by culture using Herrold's egg yolk medium and Middlebrook 7H10 with antibiotics, supplemented with Mycobactin J and incubated at 37°C for up to 12 weeks. The absolute numbers of MAP were quantified by previously published qPCR assays targeting F57 and IS900 loci in MAP genome. We herein confirm that MAP can survive pH reduction, however, longer exposure to pH below 4 in SMP seems to be critical because it inhibits growth. Therefore, it is suggested that probiotic cultures that can decrease pH below 4 during fermentation could provide better inactivation of MAP in SMP.

Research paper thumbnail of Mycobacterium avium subsp. avium and Mycobacterium neoaurum detection in an immunocompromised patient

Epidemiology and Infection, 2013

SUMMARYNon-tuberculous mycobacteria are increasingly described as infectious agents in immunocomp... more SUMMARYNon-tuberculous mycobacteria are increasingly described as infectious agents in immunocompromised patients. A 17-year-old male patient suffering from secondary non-Hodgkin's lymphoma and treated with chemotherapeutic agents was admitted to hospital due to pleuropneumonia. Mycobacterium neoaurum was cultured repeatedly from his sputum and, Mycobacterium avium subsp. avium (M. a. avium) was detected by IS901 qPCR from detached fragments of his intestinal mucosa. We attempted to determine the possible sources of infection by analysing environmental samples from the closed oncology unit and conventional unit in the hospital, and from the patient's home residence and places which he frequented. The environment of the patient harboured mycobacteria (41 isolates in total); however, M. neoaurum was not recovered. M. a. avium was detected by qPCR in the environmental samples from a small flock of hens kept by his neighbour. Although it was not confirmed by DNA fingerprinting m...

Research paper thumbnail of Epidemiological Study of Paratuberculosis in Wild Rabbits in Scotland

Journal of Clinical Microbiology, 1999

A survey of 22 farms confirmed the presence of paratuberculosis in wild rabbits in Scotland. Regi... more A survey of 22 farms confirmed the presence of paratuberculosis in wild rabbits in Scotland. Regional differences were apparent in the prevalence of the disease in rabbits, with a significantly higher incidence occurring in the Tayside region. Statistical analysis showed a significant relationship between a previous history or current problem of paratuberculosis in cattle and the presence of paratuberculosis in rabbits on the farms. Molecular genetic typing techniques could not discriminate between selected rabbit and cattle isolates from the same or different farms, suggesting that the same strain may infect and cause disease in both species and that interspecies transmission may occur. The possibility of interspecies transmission and the involvement of wildlife in the epidemiology of paratuberculosis have important implications for the control of the disease.

Research paper thumbnail of Crohn's disease and related inflammatory diseases: from many single hypotheses to one "superhypothesis

Veterinární medicína, 2014

The aetiology of Crohn’s disease and paratuberculosis are the subjects of intensive study and als... more The aetiology of Crohn’s disease and paratuberculosis are the subjects of intensive study and also frequently, of dispute. However, a number of other nosological entities have a similar history, namely type 1 diabetes, multiple sclerosis, sarcoidosis, asthma, psoriasis, spondylarthritis, Blau syndrom etc. The zoonotic risk of Mycobacterium avium subsp. paratuberculosis (MAP) has been discussed for more than one hundred years. „The problem remains open, further research is needed“, is the sentence which seems to be obligatory in the conclusions of many scientific articles. A number of hypotheses have been suggested, all with a grain of truth in them. The infection hypothesis has many supporters and opponents, but it does not fit to all Crohn’s disease cases. The contribution of the genetic factor has been admitted a long time ago and has been experimentally confirmed by recent excellent studies. An environmental factor is expected and has been often mentioned, but has yet to be disco...

Research paper thumbnail of Bovine tuberculosis infection in animal and human populations in Ethiopia: a review

Veterinární medicína, 2008

Ethiopia is one among the nations that possesses the largest number of livestock population in th... more Ethiopia is one among the nations that possesses the largest number of livestock population in the African continent estimated to be 33 million cattle, 24 million sheep and 18 million goats. In contrast to the huge livestock resource, the livestock productivity is however, found to be very low. The major biological and socio-economical factors attributing to the low productivity includes: the low genetic potential and performance, poor nutrition (in quality and quantity terms), the prevailing of different diseases, traditional way of husbandry systems and inadequate skilled manpower, among others. Ethiopia is one of the African countries where tuberculosis is wide spread in both humans and cattle and the endemic nature of tuberculosis in humans and cattle has long been documented. The disease is considered as one of the major livestock diseases that results in high morbidity and mortality, although the current status on the actual prevalence rate of bovine tuberculosis (BTB) at a na...

Research paper thumbnail of Detection methods for Mycobacterium avium subsp paratuberculosis in milk and milk products: a review

Veterinární medicína, 2008

Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of paratuberculosis, a d... more Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of paratuberculosis, a disease with considerable economic impact, principally on dairy cattle herds. Animals with paratuberculosis shed viable MAP especially in their milk, faeces and semen. MAP may have a role in the development of Crohn’s disease in humans via the consumption of contaminated milk and milk products. The current methods of milk pasteurization are not sufficient to kill all MAP cells present in milk and MAP has been cultured from raw or pasteurized milk and isolated from cheese. The purpose of the present study was to review the different methods used for detection of MAP in milk and milk products. We analyze the current methods for direct or non direct identification of MAP and culture and molecular biology methods that can be applied to milk and milk products.

Research paper thumbnail of Fish: a potential source of bacterial pathogens for human beings

Veterinární medicína, 2012

Human infections caused by pathogens transmitted from fish or the aquatic environment are quite c... more Human infections caused by pathogens transmitted from fish or the aquatic environment are quite common and depend on the season, patients’ contact with fish and related environment, dietary habits and the immune system status of the exposed individual. They are often bacterial species facultatively pathogenic for both fish and human beings and may be isolated from fish without apparent symptoms of the disease. The infection source may be fish kept for both for food and as a hobby. Human infections and intoxications with the following bacteria have been recorded: Mycobacterium spp., Streptococcus iniae, Photobacterium damselae, Vibrio alginolyticus, V. vulnificus, V. parahaemolyticus, V. cholerae, Erysipelothrix rhusiopathiae, Escherichia coli, Aeromonas spp., Salmonella spp., Staphylococcus aureus, Listeria monocytogenes, Clostridium botulinum, C. perfringens, Campylobacter jejuni, Delftia acidovorans, Edwardsiella tarda, Legionella pneumophila, and Plesiomonas shigelloides. Fish ti...

Research paper thumbnail of Characterization of IS900 loci in Mycobacterium avium subsp. paratuberculosis and development of multiplex PCR typing

Microbiology, 2000

Mycobacterium avium subsp. paratuberculosis is a pathogen that causes chronic inflammation of the... more Mycobacterium avium subsp. paratuberculosis is a pathogen that causes chronic inflammation of the intestine in many animals, including primates, and is implicated in Crohn's disease in humans. It differs from other members of the M. avium complex in having 14-18 copies of IS900 inserted into conserved loci in its genome. In the present study, genomic DNA flanking 14 of these insertions was characterized and homologues in the Mycobacterium tuberculosis and M. avium subsp. avium genomes were identified. These included regions encoding a sigma factor (sigJ) at locus 3, a nitrate reductase (nirA) at locus 4, a transcription regulator (tetR) and polyketide synthase at locus 6, and a 6-O-methylguanine methyltransferase at locus 9. In addition, locus numbers were assigned to 9 of 15 RFLP bands previously described. IS900 insertion at 7 of the 14 characterized loci was into the RBS of a gene substituting an RBS encoded by IS900 sited two bases closer to the initiation codon. IS900 insertion at five loci interrupted an ORF at the target site, one of which encoded a homologue of the immunodominant mycobacterial DesA1 protein. Eleven of eighty-one M. avium subsp. paratuberculosis isolates lacked the insertion site at locus 6 together with flanking genomic DNA. This region was also absent from seven reference strains of M. avium subsp. avium, from one M. avium subsp. silvaticum and from six other mycobacterial species. A multiplex PCR of IS900 loci (MPIL) typing method was developed which was able to discriminate 10 different types of M. avium subsp. paratuberculosis from the panel of 81 isolates with consistent differences between those of bovine and ovine origin. Nine MPIL types corresponded with a single PstI/BstEII RFLP type, suggesting that this method may be applicable to typing of M. avium subsp. paratuberculosis directly from a sample without the need for culture. The remaining MPIL type corresponded with seven PstI/BstEII RFLP types. Further resolution of these may come from sequencing the remaining four uncharacterized IS900 loci.

Research paper thumbnail of IS900 Restriction Fragment Length Polymorphism (RFLP) Analysis of Mycobacterium avium subsp. paratuberculosis Isolates from Goats and Cattle in Norway

Acta veterinaria Scandinavica

In Norway, paratuberculosis has been frequently diagnosed in goats, while cattle have been almost... more In Norway, paratuberculosis has been frequently diagnosed in goats, while cattle have been almost free of the infection. This difference in prevalence between goats and cattle has led to speculations about the existence of a Mycobacterium avium subsp. paratuberculosis (M. a. paratuberculosis) isolate that is non-pathogenic for cattle. There is little information available on genotypic variation of M. a. paratuberculosis isolated from animals in Norway. In the present study, genotypic information on 51 isolates from goats and four isolates from cattle in Norway was obtained by use of IS900 restriction fragment length polymorphism (RFLP) analysis. All isolates from cattle and 84% of the isolates from goats had the same RFLP pattern (B-C1). Five RFLP patterns not previously detected were found. No genotypic variation that could explain a difference in host origin was found between the isolates from cattle and the majority of the Norwegian goat isolates. This lack of difference indicate...

Research paper thumbnail of Bovine tuberculosis: an old disease but a new threat to Africa

The international journal of tuberculosis and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease, 2004

Bovine tuberculosis (TB) is a disease characterised by progressive development of specific granul... more Bovine tuberculosis (TB) is a disease characterised by progressive development of specific granulomatous lesions or tubercles in lung tissue, lymph nodes or other organs. Mycobacterium bovis is the causative agent of the disease. Bovine species, including bison and buffaloes, are susceptible to the disease, but nearly all warm-blooded animals can be affected. All species are not equally susceptible to the disease; some are spill-over (end) hosts and others maintenance hosts. In Africa, bovine TB primarily affects cattle; however, infection in other farm and domestic animals, such as sheep, goats, pigs, dogs and cats, is not uncommon. Wild ruminants and carnivores are also affected and are the natural reservoirs of the infectious agent in the wild. Man is also susceptible to the disease, the highest risk groups being individuals with concomitant HIV/AIDS infection. In Africa, human TB is widely known to be caused by M. tuberculosis; however, an unknown proportion of cases are due to ...

Research paper thumbnail of Culture- and Quantitative IS 900 Real-Time PCR-Based Analysis of the Persistence of Mycobacterium avium subsp. paratuberculosis in a Controlled Dairy Cow Farm Environment

Applied and Environmental Microbiology, 2012

The aim of this study was to monitor the persistence of Mycobacterium avium subsp. paratuberculos... more The aim of this study was to monitor the persistence of Mycobacterium avium subsp. paratuberculosis in environmental samples taken from a Holstein farm with a long history of clinical paratuberculosis. A herd of 606 head was eradicated, and mechanical cleaning and disinfection with chloramine B with ammonium (4%) was carried out on the farm; in the surrounding areas (on the field and field midden) lime was applied. Environmental samples were collected before and over a period of 24 months after destocking. Only one sample out of 48 (2%) examined on the farm (originating from a waste pit and collected before destocking) was positive for M. avium subsp. paratuberculosis by cultivation on solid medium (Herrold's egg yolk medium). The results using real-time quantitative PCR (qPCR) showed that a total of 81% of environmental samples with an average mean M. avium subsp. paratuberculosis cell number of 3.09 × 10 3 were positive for M. avium subsp. paratuberculosis before destocking co...

Research paper thumbnail of Human-to-human and human-to-dog Mycobacterium tuberculosis transmission studied by IS6110 RFLP analysis: a case report

Veterinární medicína, 2011

This study reports on the transmission of Mycobacterium tuberculosis of the same IS6110 RFLP type... more This study reports on the transmission of Mycobacterium tuberculosis of the same IS6110 RFLP type between two acquaintances with open pulmonary tuberculosis and a five-year-old Doberman bitch. No clinical signs, gross lesions at necropsy or histopathological lesions were observed in the infected lungs and gastrointestinal tract of the dog, although M. tuberculosis was directly detected by IS6110 PCR and culture examinations in the respiratory and gastrointestinal tracts. IS6110 PCR positivity in the faeces and blood of the dog poses a risk of M. tuberculosis transmission between the dog and humans.

Research paper thumbnail of Systemic mixed infection in a brown caiman (Caiman crocodilus fuscus) caused by Mycobacterium szulgai and M. chelonae: a case report

Veterinární medicína, 2010

A five-year old female brown caiman (Caiman crocodilus fuscus) was admitted to a veterinary clini... more A five-year old female brown caiman (Caiman crocodilus fuscus) was admitted to a veterinary clinic because of anorexia and lethargy. Chronic deterioration of the patient’s condition together with the formation of slushy stools coloured from brown to red was observed during the previous eight weeks. Physical examination showed significant apathy and cachexia. Radiographic examinations of chest and abdomen revealed no pathological findings. Initial blood tests revealed decreased hematocrit and low levels of haemoglobin. Despite treatment with enrofloxacin and intensive supportive therapy with amino acids, vitamins and mineral matter, the animal died 14 days after admission to the clinic. Post mortem examination revealed splenomegaly with a total destruction of inner organ structure together with multiple granulomas in liver and lungs. Ziehl-Neelsen staining of tissue samples from liver, lungs and spleen revealed numerous acid-fast bacilli consistent with Mycobacterium spp. Identificat...

Research paper thumbnail of Peat as a feed supplement for animals: a literature review

Veterinární medicína, 2012

Peat is an easily available natural material and a source of biologically active substances widel... more Peat is an easily available natural material and a source of biologically active substances widely used, not only in agriculture but in human and animal medicine as well. In recent years, interest in the use of peat as a feed supplement has increased, particularly due to its capability to prevent enteric diseases and to stimulate growth in piglets and pigs. The purpose of this review was to compare the advantages and risks associated with the use of peat for animal nutrition based on the literature available. Beneficial effects of various peat preparations on digestion, growth and the immune systems of animals as well as the absorbent and detoxifying capabilities are associated with the high content of favourable humic substances. One disadvantage of using peat preparations is the considerable diversity of the various types of peat caused by different biological, chemical and geological conditions during formation. Biological activity of various peat preparations is associated not o...

Research paper thumbnail of Mycobacterium bovis in human population in four Central European countries during 1990-1999

Veterinární medicína, 2012

A survey on Mycobacterium bovis and M. tuberculosis in humans has been performed in four Central ... more A survey on Mycobacterium bovis and M. tuberculosis in humans has been performed in four Central European countries (Croatia, the Czech Republic, Slovak Republic and Slovenia) during the years 1990 to 1999. These countries cover an area of 204 688 km2 with 22 135 million population. During the period, new cases of tuberculosis were bacteriologically diagnosed in 47 516 patients. M. tuberculosis was detected in 47 461 (99.88%) cases, whereas M. bovis was found only in 55 (0.12%) patients. The rate of infection due to M. bovis in humans did not exceed 0.29% in the study countries. The annual incidence of bacteriological confirmed M. bovis cases did not exceed 0.1 per 100 000 inhabitants. In the Czech Republic out of 44 tuberculosis patients due to M. bovis, 32 (72.7%) were older than 61 years and originated from rural areas, where they lived during childhood and worked in agricultural occupations. These patients may have suffered a reactivation of persistent (long-standing) M. bovis i...

Research paper thumbnail of Incidence of bovine tuberculosis in wild and domestic animals other than cattle in six Central European countries during 1990-1999

Veterinární medicína, 2012

The study was undertaken in Croatia, Czech Republic, Hungary, Poland, Slovakia and Slovenia layin... more The study was undertaken in Croatia, Czech Republic, Hungary, Poland, Slovakia and Slovenia laying between Baltic and Adriatic seas on 610 402 km2. Mycobacterium bovis infection was diagnosed in 70 animals belonging to 17 species other than cattle. The set of wild animals comprised 12 European bison (Bison bonasus), one red deer (Cervus elaphus), five wild boars (Sus scrofa), and one European wild goat (Capra aegagrus) bred in a game park. Further positive animals included two farmed red deer (Cervus elaphus) and one bactrian camel (Camelus ferus) owned by a circus. The infection was also demonstrated in 18 domestic animals belonging to 3 species living on farms where bovine tuberculosis was diagnosed in cattle. This set included 12 domestic pigs (Sus scrofa f. domestica), two domestic sheep (Ovis ammon f. aries), and four dogs (Canis lupus f. familiaris). The set of animals bred in zoological gardens consisted of 30 animals belonging to 9 species as follows: three bison (Bison biso...

Research paper thumbnail of The transmission and impact of paratuberculosis infection in domestic and wild ruminants

Veterinární medicína, 2001

Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) infects domestic cattle, sheep,... more Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) infects domestic cattle, sheep, goats, deer, camelids and wild ruminants leading to chronic enteritis known as paratuberculosis (Johne’s disease). The infection is chronic, progressive and unresponsive to treatment. Most infected animals do not develop clinical disease but may excrete the bacteria. Clinically sick animals suffer emaciation and in some species diarrhoea, followed by eventual death. During the course of the disease, excretion of M. paratuberculosis in faeces and milk occurs, and the organism spreads through the blood and lymph vessels of infected animals to multiple internal organs. The infection disseminates to both the female and male reproductive organs. Though M. paratuberculosis is not classified as a human pathogen, current opinions on the possible role of this mycobacteria in public health is discussed. This article attempts to review the ways and circumstances by which M. paratuberculosis is tra...

Research paper thumbnail of Peat as a feed supplement for animals: a review

Peat is an easily available natural material and a source of biologically active substances widel... more Peat is an easily available natural material and a source of biologically active substances widely used, not only in agriculture but in human and animal medicine as well. In recent years, interest in the use of peat as a feed supplement has increased, particularly due to its capability to prevent enteric diseases and to stimulate growth in piglets and pigs. The purpose of this review was to compare the advantages and risks associated with the use of peat for animal nutrition based on the literature available. Beneficial effects of various peat preparations on digestion, growth and the immune systems of animals as well as the absorbent and detoxifying capabilities are associated with the high content of favourable humic substances. One disadvantage of using peat preparations is the considerable diversity of the various types of peat caused by different biological, chemical and geological conditions during formation. Biological activity of various peat preparations is associated not o...

Research paper thumbnail of Molecular Fingerprinting ofMycobacterium bovissubsp.capraeIsolates from Central Europe

Journal of Clinical Microbiology, 2004

ABSTRACTTo study the dissemination ofMycobacterium bovissubsp.caprae, 79 European isolates from c... more ABSTRACTTo study the dissemination ofMycobacterium bovissubsp.caprae, 79 European isolates from cattle, humans, and other hosts were examined by spoligotyping and IS6110restriction fragment length polymorphism (RFLP) analysis. Among a total of 11 different spoligotypes identified, type C1 proved to be predominant (n= 62). Five of the spoligotypes are described for the first time. A total of 43 different RFLP types were identified, thus allowing further differentiation for epidemiological tracking. Isolates from a series of outbreaks in one village proved to be of the same spoligotype and of identical or closely related RFLP types.

Research paper thumbnail of Correlation of Mycobacterium avium subsp. paratuberculosis counts in gastrointestinal tract, muscles of the diaphragm and the masseter of dairy cattle and potential risk for consumers

International Journal of Food Microbiology, 2011

Tissues of cattle intended for human consumption can be contaminated by Mycobacterium avium subsp... more Tissues of cattle intended for human consumption can be contaminated by Mycobacterium avium subsp. paratuberculosis (MAP). Although different studies attribute varying roles of MAP in Crohn's disease progression it is thought that the exposure of humans to this bacterium should in any case be minimised. In this study, we have collected samples of intestine, mesenteric lymph nodes, muscles of diaphragm (musculus diaphragma) and masseter muscles (musculus masseter) from twenty-five cows in a slaughterhouse. The infectious status of all animals was confirmed by culture of faeces. MAP was found in almost all the intestines and mesenteric lymph nodes examined, including three faecal culture-negative animals indicating intermittent shedding. As intestine is used for the traditional production of sausages, it is alarming that 84.2% of intestine samples were positive for MAP. F57 and IS900 real time PCR revealed MAP in 40 to 68% of diaphragms and 11.1 to 38.9% of masseters. A noticeable dependence of the probability of MAP positivity of faeces versus gastrointestinal tract (GIT) and of GIT and muscles was observed. Due to the changing behaviour of consumers, both of these muscles have started to be widely used in cuisine. Therefore, the results of this paper imply that the processing of cows with paratuberculosis in abattoirs without any precautions (restrictions) and the usage of meat for human consumption should be rethought.

Research paper thumbnail of Mycobacterium avium subsp. paratuberculosis survival during fermentation of soured milk products detected by culture and quantitative real time PCR methods

International Journal of Food Microbiology, 2012

Mycobacterium avium paratuberculosis (MAP), etiological agent of paratuberculosis in ruminants, i... more Mycobacterium avium paratuberculosis (MAP), etiological agent of paratuberculosis in ruminants, is able to survive extreme conditions like very low pH (stomach), high temperature (pasteurization) or low temperature (refrigerated storage). Cheese, infant powder milk, cream and other milk and dairy products might thus be considered as possible sources of MAP for humans. The aim of this study was to investigate the survival of two MAP field isolates during fermentation of three different types of soured milk products (SMP; yogurt, acidophilus milk and kefir) under laboratory conditions. Pasteurized MAP-free milk was artificially contaminated with 10(6)MAPcells/mL and survival and absolute numbers of MAP were monitored during fermentation (4 or 16 h) and after six weeks of storage at 4°C by culture and quantitative real time PCR (qPCR). Viability of MAP was determined by culture using Herrold's egg yolk medium and Middlebrook 7H10 with antibiotics, supplemented with Mycobactin J and incubated at 37°C for up to 12 weeks. The absolute numbers of MAP were quantified by previously published qPCR assays targeting F57 and IS900 loci in MAP genome. We herein confirm that MAP can survive pH reduction, however, longer exposure to pH below 4 in SMP seems to be critical because it inhibits growth. Therefore, it is suggested that probiotic cultures that can decrease pH below 4 during fermentation could provide better inactivation of MAP in SMP.

Research paper thumbnail of Mycobacterium avium subsp. avium and Mycobacterium neoaurum detection in an immunocompromised patient

Epidemiology and Infection, 2013

SUMMARYNon-tuberculous mycobacteria are increasingly described as infectious agents in immunocomp... more SUMMARYNon-tuberculous mycobacteria are increasingly described as infectious agents in immunocompromised patients. A 17-year-old male patient suffering from secondary non-Hodgkin's lymphoma and treated with chemotherapeutic agents was admitted to hospital due to pleuropneumonia. Mycobacterium neoaurum was cultured repeatedly from his sputum and, Mycobacterium avium subsp. avium (M. a. avium) was detected by IS901 qPCR from detached fragments of his intestinal mucosa. We attempted to determine the possible sources of infection by analysing environmental samples from the closed oncology unit and conventional unit in the hospital, and from the patient's home residence and places which he frequented. The environment of the patient harboured mycobacteria (41 isolates in total); however, M. neoaurum was not recovered. M. a. avium was detected by qPCR in the environmental samples from a small flock of hens kept by his neighbour. Although it was not confirmed by DNA fingerprinting m...

Research paper thumbnail of Epidemiological Study of Paratuberculosis in Wild Rabbits in Scotland

Journal of Clinical Microbiology, 1999

A survey of 22 farms confirmed the presence of paratuberculosis in wild rabbits in Scotland. Regi... more A survey of 22 farms confirmed the presence of paratuberculosis in wild rabbits in Scotland. Regional differences were apparent in the prevalence of the disease in rabbits, with a significantly higher incidence occurring in the Tayside region. Statistical analysis showed a significant relationship between a previous history or current problem of paratuberculosis in cattle and the presence of paratuberculosis in rabbits on the farms. Molecular genetic typing techniques could not discriminate between selected rabbit and cattle isolates from the same or different farms, suggesting that the same strain may infect and cause disease in both species and that interspecies transmission may occur. The possibility of interspecies transmission and the involvement of wildlife in the epidemiology of paratuberculosis have important implications for the control of the disease.

Research paper thumbnail of Crohn's disease and related inflammatory diseases: from many single hypotheses to one "superhypothesis

Veterinární medicína, 2014

The aetiology of Crohn’s disease and paratuberculosis are the subjects of intensive study and als... more The aetiology of Crohn’s disease and paratuberculosis are the subjects of intensive study and also frequently, of dispute. However, a number of other nosological entities have a similar history, namely type 1 diabetes, multiple sclerosis, sarcoidosis, asthma, psoriasis, spondylarthritis, Blau syndrom etc. The zoonotic risk of Mycobacterium avium subsp. paratuberculosis (MAP) has been discussed for more than one hundred years. „The problem remains open, further research is needed“, is the sentence which seems to be obligatory in the conclusions of many scientific articles. A number of hypotheses have been suggested, all with a grain of truth in them. The infection hypothesis has many supporters and opponents, but it does not fit to all Crohn’s disease cases. The contribution of the genetic factor has been admitted a long time ago and has been experimentally confirmed by recent excellent studies. An environmental factor is expected and has been often mentioned, but has yet to be disco...

Research paper thumbnail of Bovine tuberculosis infection in animal and human populations in Ethiopia: a review

Veterinární medicína, 2008

Ethiopia is one among the nations that possesses the largest number of livestock population in th... more Ethiopia is one among the nations that possesses the largest number of livestock population in the African continent estimated to be 33 million cattle, 24 million sheep and 18 million goats. In contrast to the huge livestock resource, the livestock productivity is however, found to be very low. The major biological and socio-economical factors attributing to the low productivity includes: the low genetic potential and performance, poor nutrition (in quality and quantity terms), the prevailing of different diseases, traditional way of husbandry systems and inadequate skilled manpower, among others. Ethiopia is one of the African countries where tuberculosis is wide spread in both humans and cattle and the endemic nature of tuberculosis in humans and cattle has long been documented. The disease is considered as one of the major livestock diseases that results in high morbidity and mortality, although the current status on the actual prevalence rate of bovine tuberculosis (BTB) at a na...

Research paper thumbnail of Detection methods for Mycobacterium avium subsp paratuberculosis in milk and milk products: a review

Veterinární medicína, 2008

Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of paratuberculosis, a d... more Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of paratuberculosis, a disease with considerable economic impact, principally on dairy cattle herds. Animals with paratuberculosis shed viable MAP especially in their milk, faeces and semen. MAP may have a role in the development of Crohn’s disease in humans via the consumption of contaminated milk and milk products. The current methods of milk pasteurization are not sufficient to kill all MAP cells present in milk and MAP has been cultured from raw or pasteurized milk and isolated from cheese. The purpose of the present study was to review the different methods used for detection of MAP in milk and milk products. We analyze the current methods for direct or non direct identification of MAP and culture and molecular biology methods that can be applied to milk and milk products.

Research paper thumbnail of Fish: a potential source of bacterial pathogens for human beings

Veterinární medicína, 2012

Human infections caused by pathogens transmitted from fish or the aquatic environment are quite c... more Human infections caused by pathogens transmitted from fish or the aquatic environment are quite common and depend on the season, patients’ contact with fish and related environment, dietary habits and the immune system status of the exposed individual. They are often bacterial species facultatively pathogenic for both fish and human beings and may be isolated from fish without apparent symptoms of the disease. The infection source may be fish kept for both for food and as a hobby. Human infections and intoxications with the following bacteria have been recorded: Mycobacterium spp., Streptococcus iniae, Photobacterium damselae, Vibrio alginolyticus, V. vulnificus, V. parahaemolyticus, V. cholerae, Erysipelothrix rhusiopathiae, Escherichia coli, Aeromonas spp., Salmonella spp., Staphylococcus aureus, Listeria monocytogenes, Clostridium botulinum, C. perfringens, Campylobacter jejuni, Delftia acidovorans, Edwardsiella tarda, Legionella pneumophila, and Plesiomonas shigelloides. Fish ti...

Research paper thumbnail of Characterization of IS900 loci in Mycobacterium avium subsp. paratuberculosis and development of multiplex PCR typing

Microbiology, 2000

Mycobacterium avium subsp. paratuberculosis is a pathogen that causes chronic inflammation of the... more Mycobacterium avium subsp. paratuberculosis is a pathogen that causes chronic inflammation of the intestine in many animals, including primates, and is implicated in Crohn's disease in humans. It differs from other members of the M. avium complex in having 14-18 copies of IS900 inserted into conserved loci in its genome. In the present study, genomic DNA flanking 14 of these insertions was characterized and homologues in the Mycobacterium tuberculosis and M. avium subsp. avium genomes were identified. These included regions encoding a sigma factor (sigJ) at locus 3, a nitrate reductase (nirA) at locus 4, a transcription regulator (tetR) and polyketide synthase at locus 6, and a 6-O-methylguanine methyltransferase at locus 9. In addition, locus numbers were assigned to 9 of 15 RFLP bands previously described. IS900 insertion at 7 of the 14 characterized loci was into the RBS of a gene substituting an RBS encoded by IS900 sited two bases closer to the initiation codon. IS900 insertion at five loci interrupted an ORF at the target site, one of which encoded a homologue of the immunodominant mycobacterial DesA1 protein. Eleven of eighty-one M. avium subsp. paratuberculosis isolates lacked the insertion site at locus 6 together with flanking genomic DNA. This region was also absent from seven reference strains of M. avium subsp. avium, from one M. avium subsp. silvaticum and from six other mycobacterial species. A multiplex PCR of IS900 loci (MPIL) typing method was developed which was able to discriminate 10 different types of M. avium subsp. paratuberculosis from the panel of 81 isolates with consistent differences between those of bovine and ovine origin. Nine MPIL types corresponded with a single PstI/BstEII RFLP type, suggesting that this method may be applicable to typing of M. avium subsp. paratuberculosis directly from a sample without the need for culture. The remaining MPIL type corresponded with seven PstI/BstEII RFLP types. Further resolution of these may come from sequencing the remaining four uncharacterized IS900 loci.

Research paper thumbnail of IS900 Restriction Fragment Length Polymorphism (RFLP) Analysis of Mycobacterium avium subsp. paratuberculosis Isolates from Goats and Cattle in Norway

Acta veterinaria Scandinavica

In Norway, paratuberculosis has been frequently diagnosed in goats, while cattle have been almost... more In Norway, paratuberculosis has been frequently diagnosed in goats, while cattle have been almost free of the infection. This difference in prevalence between goats and cattle has led to speculations about the existence of a Mycobacterium avium subsp. paratuberculosis (M. a. paratuberculosis) isolate that is non-pathogenic for cattle. There is little information available on genotypic variation of M. a. paratuberculosis isolated from animals in Norway. In the present study, genotypic information on 51 isolates from goats and four isolates from cattle in Norway was obtained by use of IS900 restriction fragment length polymorphism (RFLP) analysis. All isolates from cattle and 84% of the isolates from goats had the same RFLP pattern (B-C1). Five RFLP patterns not previously detected were found. No genotypic variation that could explain a difference in host origin was found between the isolates from cattle and the majority of the Norwegian goat isolates. This lack of difference indicate...

Research paper thumbnail of Bovine tuberculosis: an old disease but a new threat to Africa

The international journal of tuberculosis and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease, 2004

Bovine tuberculosis (TB) is a disease characterised by progressive development of specific granul... more Bovine tuberculosis (TB) is a disease characterised by progressive development of specific granulomatous lesions or tubercles in lung tissue, lymph nodes or other organs. Mycobacterium bovis is the causative agent of the disease. Bovine species, including bison and buffaloes, are susceptible to the disease, but nearly all warm-blooded animals can be affected. All species are not equally susceptible to the disease; some are spill-over (end) hosts and others maintenance hosts. In Africa, bovine TB primarily affects cattle; however, infection in other farm and domestic animals, such as sheep, goats, pigs, dogs and cats, is not uncommon. Wild ruminants and carnivores are also affected and are the natural reservoirs of the infectious agent in the wild. Man is also susceptible to the disease, the highest risk groups being individuals with concomitant HIV/AIDS infection. In Africa, human TB is widely known to be caused by M. tuberculosis; however, an unknown proportion of cases are due to ...