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Control and Eradication of Bovine Tuberculosis in Central Europe
... The test and slaughter policy may be of significance in the developed countries because of the effective and compulsory reporting of M. bovis infection of all animals, quarantine of infected herds, testing of animals in-contact with suspected or confirmed cases of bovine TB, movement restrictions on cattle herds in TB endemic areas and also of cattle herds not yet tested for TB [12,13], However, the World Health Organization (W.H.O) has proposed that the test-and-segregation program, a modified form of the test-andslaughter policy, will be more significant in developing countries, where the test-and-slaughter policy is usually impracticable because of the endemicity of TB in the cattle population. Measures are taken to segregate positive reactors slaughter them [14]. ...
... Pigs are susceptible to all the three types of tubercle bacilli [1,3]. It has been suggested that there is a correlation between the occurrence of TB in pigs and a direct or indirect contact of pigs with tuberculous humans, cattle or birds [4,5]. The presence of TB in pigs in virtually all countries, in which pigs are farmed, has long been reported [4]. ...
Background
Tuberculosis (TB) is an infectious, granulomatous disease caused by acid-fast bacilli of the genus Mycobacterium. The disease affects practically all species of vertebrates. Although mammalian tuberculosis has been nearly controlled in many developed countries, it is still a serious problem in humans and domestic animals including pigs in developing countries. In Ethiopia, the prevalence of TB in pigs is not known. Therefore, this study was designed to estimate the prevalence of TB in pigs in central Ethiopia and to characterize the causative agents using molecular techniques.
Results
The estimated prevalence of TB was 5.8% (49/841). Age and origin of pigs were significantly associated (P<0.001) with the prevalence. In contrast, an association of sex, floor type and water source with the prevalence could not be shown. Culture positivity was confirmed in 30.6% (15/49) of the tuberculous-like lesions. Of the 15 isolates, 12 were acid fast positive while five of the latter were confirmed by multiplex PCR as members of the M. tuberculosis complex. Speciation of the five isolates further confirmed that they were M. tuberculosis, belonging to SIT1088 (two isolates) and SIT1195 (one isolate). The remaining two isolates belong to an identical spoligotype, the pattern of which was not found in the spoligotype database (SpolDB4).
Conclusions
The isolation of M. tuberculosis from pigs suggests a possible risk of transmission between humans and pigs. Hence, establishing feasible control methods is required.
... Meat inspection was an integral part of both quality assurance and quality control systems, and gross inspection of carcasses was effectively carried out to provide the quality demanded and protection of consumers [50]. Rigorous meat inspections and tracing of TB lesions back to the animal farms [51][52][53][54] are complementary to eradication or continuous reduction of zoonotic bovine TB [51][52][53][55][56][57]. However, bovine TB and zoonotic TB due to M. bovis are poorly investigated and controlled in most of Africa including Cameroon where bovine TB is widespread in cattle. ...
Bovine tuberculosis (BTB) is widespread but poorly controlled in Africa and M. bovis is posing threats to human health. The risk of cattle handlers to M. bovis prevalence and public health significance of BTB in Cameroon were assessed. Slaughter inspection records from major cities revealed that BTB detection rates in cattle from 0.18% to 4.25% and BTB lesions were most common. Analyses of tissues and sera confirmed BTB in 31% (Ziehl-Neelsen), 51% (culture), and 60% (antibody detection) of test cattle. Among cattle handlers, 81.9% were aware of BTB, 67.9% knew that BTB is zoonotic, and 53.8% knew one mode of transmission but over 27% consumed raw meat and/or drank unpasteurized milk. Respondents who had encountered tuberculosis cases were more informed about zoonotic BTB (P < .05). Tuberculosis is prevalent in cattle destined for human consumption in Cameroon with serious public health implications. Targeted monitoring of infected animal populations and concerted veterinary/medical efforts are essential for control.
... Moreover, the susceptibility of human immunodeficiency virus (HIV)‐infected individuals to M. tuberculosis complex is of major concern to public health officials in developing countries where acquired immune deficiency syndrome (AIDS) is rampant (254). M. bovis accounts for only a small percentage of the cases of tuberculosis reported in humans; however, it is a pathogen of significant economic importance in wild and domestic animals around the globe, especially in countries where little information is available on the incidence of M. bovis infection in humans (44, 52, 57, 71, 196, 211, 249, 261, 282, 285). The genus Mycobacterium includes several species that cause disease in humans and other animals. ...
Tuberculosis continues to be an important disease both in humans and animals. It causes morbidity, mortality and economic loss worldwide. The occurrence of Mycobacterium bovis disease in humans, domesticated and wild animals confirms the relevance of this zoonosis. M. bovis in humans continues to be reported in industrialised countries and in immigrants from regions of the world where tuberculosis in cattle is endemic. The real incidence of M. bovis in humans in developing countries continues to be roughly under-estimated due to the scarcity of appropriate laboratory facilities to isolate and to differentiate M. bovis strains. In Latin America, less than 1% of tuberculosis cases are reported as being due to M. bovis. However, the economic relevance that meat and dairy industries play in these countries stimulates the promotion of bovine tuberculosis eradication programmes. Human-to-human airborne transmission of M. bovis does occur and it may be important where human immunodeficiency virus (HIV) infection in humans is prevalent, M. bovis infection in cattle is enzootic and pasteurisation of dairy products is not routinely practised. Eradication of M. bovis in cattle and pasteurisation of dairy products are the cornerstones of prevention of human disease. Measures should be developed to identify and control M. bovis infection in wild animals as these may be important reservoirs of infection for domesticated food-producing animals. There is a need for medical and veterinary professionals to cooperate on disease outbreaks. The information presented herein strongly supports the 'One World/One Health/One Medicine' concept.
This chapter presents an analysis of data on the occurrence of bovine tuberculosis in countries of the Central Europe and countries of the former Soviet Union before 2010, available from OIE official reports and published papers. Between 2000 and 2010, bovine tuberculosis was not diagnosed in cattle either in the Czech Republic or in Slovakia. In the other five countries, namely, Bosnia and Herzegovina, Croatia, Hungary, Poland, and Slovenia, bovine tuberculosis was diagnosed sporadically. The occurrence of bovine tuberculosis in Central Europe, notably in feral animals, can be considered a health hazard, being most serious in the wild boar in Croatia and Hungary and in the European bison in Poland. Observations provide evidence for long-term bovine tuberculosis infection in red deer population in western Austria. The chapter also discusses, in detail, bovine tuberculosis in cattle in Russia and the former states of the Soviet Union including the Baltic countries.
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 infection as they got older. Bovine tuberculosis in cattle was eliminated from these countries during the second half of the 1960s (Croatia in 1966, Czech Republic and Slovak Republic - former Czechoslovakia in 1968, Slovenia in 1973) and the incidence of outbreaks of bovine tuberculosis in cattle were very low, thus the disease in humans was unexpected.
Sources of mycobacterial infections in 50 pig herds in the Czech Republic were investigated during the years 1996 to 2002. A total of 2 412 samples from the external environment (feeds, bedding materials, drinking water, biofilms on drinkers, scrapings from the walls, floors and pen barriers, dust, spider webs, peat, kaolin, faeces, organs of rodents, and birds, etc.) were examined. After staining by the Ziehl-Neelsen technique, acid-fast rods were detected in 95 (3.9%) samples by direct microscopic examination and mycobacteria were cultured from 575 (23.8%) samples. From Mycobacterium avium complex (MAC), M. avium subsp. hominissuis (genotype IS901-, IS1245+) of serotypes 4, 6, 8, and 9 (272; 47.0% isolates), M. a. avium (genotype IS901+, IS1245+) of serotype 2 (13; 2.2% isolates) and M. intracellulare (genotype IS901-, IS1245-) of different serotypes (2; 0.3% isolates) were detected most frequently. Other isolates from among 14 other mycobacterial species ranked as follows: 64 M. gordonae, 47 M. fortuitum, 17 M. chelonae, 14 M. flavescens, 11 M. terrae, seven M. phlei, seven M. scrofulaceum, three M. diernhoferi, three M triviale, three M. smegmatis, two M. xenopi, one M. szulgai, one M. gastri, and one M. ulcerans. The remaining 111 isolates of unidentified species did not contain specific sequences IS901 and IS1245 characteristic for the pathogenic members of MAC (M. a. avium and M. a. hominissuis). Peat, drinking water, biofilms on drinkers, bedding materials, feeds, free living birds, kaolin and charcoal were identified as potential sources of mycobacterial infections for pigs. Peat given to piglets as a feed supplement was the most important source of mycobacteria (65.1% positive of 327 examined samples); 81.2% of them were positive for M. a. hominissuis of serotypes 4, 6, 8, and 9. By contrast, mycobacteria of other species (M. gordonae, M. fortuitum, M. chelonae, M. flavescens, etc.) were the main isolates obtained from drinking water and biofilms on drinkers for pigs. By culture examination, the detection rate was higher in the biofilm samples (36.4%) than in the samples of drinking water (29.6%). The third group of sites with detected high levels (26.4%) of mycobacterial contamination were various types of beddings of woody material. M. a. hominissuis of serotypes 6, 8, and 9 were the most frequent isolates from sawdust; M. a. avium serotype 2 was sporadically detected. Mycobacterial findings in other samples from the external environment (wall and floor scrapings, dust, soil from the runs, and invertebrates) gave an indication of the pressure of infection in the herds. High contamination levels in faecal samples (15.6%) and in scrapings (18.4%) from respective parts of pens and stables indicated exposure of pigs to mycobacteria. In those materials, isolation of M. a. hominissuis of serotypes 4, 6, 8, and 9 prevailed. Mycobacteria were also detected in 7.9% of 430 samples of various invertebrate species. Various mycobacterial species were identified in the larvae and puparia of Eristalis tenax and Musca spp. and in imagoes of Drosophila spp., Musca spp., family Scatophagidae, Stomoxys calcitrans, E. tenax, and in earthworms. All of the constituents of the external environment that are potential sources of mycobacterial infections should be considered during implementation of preventative measures and the control of mycobacterial infections in pig herds.
An increased incidence of tuberculous lesions in head and mesenteric lymph nodes from slaughtered pigs weighing about 115 kg was recorded in a herd of pigs kept in two farms A and B in the Czech Republic. Tuberculous lesions were more frequently (P < 0.01) diagnosed in pigs from Farm A (10.4%) than from Farm B (1.1%). The follow-up investigation of potential sources of infection on Farm A revealed that the piglets were fed kaolin from a nearby mine as a supplement. Among 20 samples from the pigs' environment, atypical conditionally pathogenic mycobacteria (ACPM) were detected in four samples as follows: dust (n = 2), pig faeces (n = 1) and kaolin fed as a supplement (n = 1). Mycobacterium avium subsp. hominissuis of genotype IS901- and IS1245+ and serotype 8 was isolated from kaolin and pig lymph nodes, M. fortuitum from stable dust and pig faeces and other ACPM from stable dust. When kaolin feeding to piglets ceased, the incidence of tuberculous lesions in these pigs at slaughter 5-6 months later decreased from 16.1% to 3.4%. No ACPM was detected on Farm B in 27 samples from the environment. An investigation of surface kaolin mines did not detect mycobacteria in non-extracted kaolin. However, surface water (three isolates among 13 samples) from the pond used for kaolin levigation and 23 batches of the final product of kaolin (10 samples from each batch, i.e. a total of 230 samples were examined) were contaminated with ACPM. Among the latter, ACPM were isolated from three samples originating from three different batches. ACPM were likely to survive during transport of the kaolin, as a suspension through the pipeline, and during its further processing to the final product (sedimentation, addition of colloid substances, drying and other procedures).
In the decade monitored a total of 45 873 318 pigs were slaughtered and examined according to veterinary hygiene standards. Apart from 1991, when results of tuberculous findings were not obtained, tuberculous lesions were found in 134 088 (0.32%) of the 41 458 565 pigs examined in the remaining nine years. During a detailed analysis of the pathological anatomical examination of 190 940 pigs slaughtered in one district, tuberculous lesions in lymph nodes were found in 4 107 (2.2%) pigs: mesenteric (65.3% pigs), submandibular (18.6% pigs), inguinal (0.1% pigs) and simultaneously intestinal and head lymph nodes (15.9% pigs). Miliary tuberculosis was found only in the parenchymatous organs of four (0.1%) pigs. The following financial losses resulted: 6% for confiscating the head, intestines and stomach, and from 22 to 24% for assessing meat as conditionally edible after processing, i.e. intended only for heat-processed products. Mycobacteria were isolated from 7 246 (41.8%) pigs through the cultivation of tissue samples from 17 326 pigs. Mycobacterium bovis was detected in only five (0.07%) animals which originated from the last outbreak of bovine tuberculosis in cattle in the Czech Republic in 1995. M. avium complex (MAC) isolates came from 6 870 (94.8%) animals: 55.7% M. a. avium isolates were mainly of serotypes 2 and 3 and genotype IS901+ and IS1245+ and 39.2% M. a. hominissuis isolates were mainly of serotypes 4, 8 and 9 and genotype IS901- and IS1245+. Conditionally pathogenic mycobacteria (M. chelonae, M. terrae, M. phlei and M. fortuitum) were isolated from 371 (5.1%) pigs. In the whole period monitored, two marked increases in the findings of tuberculous lesions were recorded: In the mid-1990s as a result of using deep bedding with wood shavings and at the end of the 1990s as a result of supplementing the pigs' feed with peat. The predominant occurrence of M. a. avium isolates of genotype IS901+ and IS1245+ in the first half of the 1990s was replaced above all by M. a. hominissuis isolates of genotype IS901- and IS1245+. The reason for this was probably a change in the sources of infection for pigs. While at the beginning of the 1990s the most frequent source of infection were wild and domestic birds, various parts of the external environment became the source of the infection for pigs from the mid-1990s. In the years 1996 to 1999, Rhodococcus equi was isolated from 203 (11.6%) of the 1 745 animals examined. It was solely isolated from 154 (8.8%) animals and from 49 (2.8%) animals together with mycobacteria.
Infected animals in the wild, which can act as a reservoir and/or vector for the origin of bovine tuberculosis, are a great problem for national programmes seeking to free herds of cattle from the infection. The circulation of Mycobacterium bovis in the wild animal population might cause a slow-down in the progress of control programmes through the reinfection of herds of livestock. The Eurasian badger (Meles meles) and red deer (Cervus elaphus) living in the wild in Great Britain and Ireland, brushtail possum (Trichosurus vulpecula), ferrets (Mustela putorius f. furo) in New Zealand and wild buffalo (Bubalus arnee) in Australia are among already known reservoirs and vectors of bovine tuberculosis. In 7 countries of Central Europe (Bosnia and Herzegovina, Croatia, Czech Republic, Hungary, Poland, Slovakia and Slovenia) bovine tuberculosis in cattle was controlled as part of national control programmes more than 20 years ago. In the last decade M. bovis has been diagnosed extremely sporadically in cattle and other domestic animals as well as in wild animals held in captivity or living in the wild. This favourable situation could be threatened by the mycobacteria spreading via the wild boar (Sus scrofa) which is susceptible to mycobacterial infection and very abundant in Central Europe. According to available literary data, mycobacteria were detected in 361 wild boar originating from countries other than those of Central Europe, such as Australia, Bulgaria, Germany, the Hawaiian island of Molokai, Italy and Spain. M. tuberculosis complex (33.9%) and M. bovis complex (39.8%) isolates were most frequently detected in the faeces and/or parenchymatous organs of wild boar. Of other mycobacterial species, M. intracellulare (3.8%), M. avium subsp. avium (3.8%), M. terrae (2.4%), M. fortuitum (2.2%), M. scrofulaceum (2.2%), M. gordonae (0.8%), M. simiae (0.5%), M. szulgai (0.5%), M. xenopi (0.5%), M. smegmatis (0.2%), M. vaccae (0.2%), fast-growing, further unspecified species (0.2%) and unidentified mycobacteria (8.8%) were isolated. Following the analysis of literary data and our own results, it was found that, in the area covered by the above-mentioned 7 countries of Central Europe, a total of 431 wild boar were examined for mycobacterial infections in the years 1983-2001. Tuberculous lesions in parenchymatous organs were found in 43 (10.0%) animals. M. bovis was identified in 22 (5.1%) animals, M. a. avium in 2 (0.4%), M. a. paratuberculosis in 1 (0.2%) animal and atypical mycobacteria in 27 (6.3%) animals. The wild boar may therefore represent, under certain unfavourable epizootiological conditions, a vector of some mycobacterial infections in not only animals, but also humans.
Results of Mycobacterium tuberculosis detection in animals from six Central European countries (Croatia, the Czech Republic, Hungary, Poland, Slovakia and Slovenia) spreading over 610 402 km2 with a population of 11.8 million heads of cattle were analysed. In the monitoring period, 1990 to 1999, M. tuberculosis from animals was isolated only in two countries (Poland and Slovak Republic) from 16 animals with tuberculous lesions. These comprise nine heads of cattle (Bos taurus), four domestic pigs (Sus scrofa f. domestica) and three wild animals, an African elephant (Loxodonta africana), agouti (Dasyprocta aguti) and terrestrial tapir (Tapirus terrestris), originated form a zoological garden Gdansk in Poland. Steady decrease in the incidence of tuberculosis in humans was recorded during the monitoring period in all countries. The human population of the study countries was 68.03 million. In the period monitored, infection caused by M. tuberculosis was identified in a total of 241 040 patients with a decreasing incidence of tuberculosis found in all countries. The lowest relative bacteriologically confirmed disease was found in the Czech Republic, Slovak Republic and Slovenia. Given the low number of infected domestic and wild animals, the epidemiological and epizootiological situation may be considered auspicious.
In the nineties, cattle stocks gradually decreased from 3 506 222 head of cattle in 1990 to 1 657 337 head in 1999. Skin testing of cattle was carried out annually using bovine tuberculin. Animals for sale were also simultaneously tested with avian tuberculin. In records from 1991 to 1999 a total of 14 611 393 bovine tuberculin skin tests and 611 405 simultaneous avian tuberculin skin tests are registered. A total of 1 457 (0.01%) animals reacted positively with bovine tuberculin and 1 790 (0.29%) with avian tuberculin. In the period monitored a total of 7 268 274 head of cattle were slaughtered and given veterinary hygienic examinations. Statistical data on the post-mortem detection of tuberculous lesions have been available for nine years since 1992 when tuberculous lesions were found in 1 186 (0.019%) out of 6 273 441 slaughtered animals. Mycobacteria were isolated from the organs of only 561 (17.5%) out of 3 202 culturally examined animals. M. bovis only was isolated from 48 (8.6%) animals originating from seven herds (two infected herds in 1991, 1992 and 1994 and one infected herd in 1995): four outbreaks were detected by annual skin testing, one outbreak by movement tuberculin skin testing and two outbreaks by the detection of tuberculous lesions at slaughter. M. avium complex strains of serotypes 1, 2 and 3 and of genotypes IS901+ and IS1245+ were isolated from 331 (59.0%) animals and strains of serotypes 4 to 6, 8 to 11 and 21 and of genotypes IS901- and IS1245+ were isolated from 132 (23.5%) animals. Potentially pathogenic bacteria of the M. chelonae, M. terrae, M. phlei and M. fortuitum species were isolated from 50 (8.9%) animals. Neither miliary nor generalised tuberculosis was found in any of the animals. Between 1996 and 1999, the proportion of cattle in which tuberculous lesions were recorded decreased.
e last outbreak of bovine tuberculosis in cattle in the Czech Republic was detected in 1995. Signs of diarrhoea, weight loss and occasional coughing appeared in one 14-year-old cow after giving birth for the thirteenth time. Two months after these symptoms had been observed, it had to be slaughtered and numerous tuberculous lesions were found in its lung tissue, including the pleura. Within three months after the confirmation of the infection and consecutive intra-vitam and post-mortem diagnostics, all 28 remaining head of cattle from the herd (nine cows, seven bulls, six heifers and six calves) and five pigs were slaughtered. Patho-anatomical lesions were detected in all animals indicative of tuberculosis, from which Mycobacterium bovis was cultured and identified on the basis of biochemical tests and virulence test in a guinea-pig. e culture of 33 samples of other biological material than tissues (milk and urine of cows, feeding water, scrapings from the shed, fodder and others) resulted in M. bovis being detected in three samples (scrapings from shed walls). By the spoligotyping method M. bovis subsp. caprae was found in six selected isolates originating from two cows, two heifers and two bulls. It may therefore be assumed that there was one source of infection in the herd, which was the first infected old cow. In comparison with 3 176 spoligotypes in the existing database RIVM (National Institute of Public Health and the Environment, Bilthoven, e Netherlands) and literary data it was found that this spoligotype was also found in Sweden, Bel- gium, Great Britain, Spain, Poland, Germany and the Czech Republic. It was impossible to determine the source of M. bovis subsp. caprae of the first infected cow on the basis of results from database and from anamnestic data. Green fodder coming from the farmer's pastures near a forest could be considered as a possible source of M. bovis from wild ruminants like red deer (Cervus elaphus), which was found infected with bovine tuberculosis in another district of the Czech Republic in 1991.
Spoligotyping was used to examine IS6110-positive DNA of 26 Mycobacterium bovis, M. bovis BCG and M. bovis subsp. caprae non-viable isolates stored up to 10 years. All of these isolates were previously identified by biochemical tests and all 17/17 tested isolates were earlier found virulent for guinea pigs. In total seven spoli- gotypes, designated S1-S7, were detected and compared with the spoligotypes of 3 176 isolates in the database of the National Institute of Public Health and the Environment (RIVM) in Bilthoven, the Netherlands. A Neotype M. bovis strain, isolated in 1965 in the USA and thereafter stored in e Czechoslovak National Collection of Type Cultures (My 310/87) since 1987 was of an identical spoligotype S4 with the original reference M. bovis strain from the USA. e M. bovis isolates from capybara's (Hydrochoerus hydrochaeris) imported from Germany to the Czech Republic in 1989, as well as cattle isolates from 1966, 1991 and 1994, were of the most common type S1. Also a human isolate from 1981, a M. bovis BCG vaccine strain and clinical M. bovis BCG isolates from three children with post-vaccinal complications were of this most predominant spoligotype. e four unique spoligotypes S2, S3, S5 and S6 were identified in M. bovis isolates from cattle in the years 1965, 1996 and 1967 in the Czech Republic, respectively, but also in isolates from farmed red deer (Cervus elaphus) from 1991 and in cattle isolates from Slovakia from the year 1992. e scarcely occurring spoligotype S7, which is typical for M. b. caprae was detected in the Czech Republic from farmed red deer (1999), cattle isolates (1966, 1991, 1995) and in a strain isolated from an 80-year-old man (1999). Several strains isolated in each of three outbreaks in cattle herds were examined. Identical spoligotypes were detected in two outbreaks and different causal agents ( M. bovis of spoligotype S1 and M. b. caprae of spoligotype S7) were identified in two cows from the third outbreak. e results confirm an effective control of bovine tuberculosis in the Czech Republic and Slovakia during 1959-1968, because previously circulating spoligotypes were successfully eradicated. e data also suggest other reservoirs of bovine tuberculosis may exist among free-living wild animals.
45 Bovine tuberculosis, caused by Mycobacterium bovis (M. bovis) is considered as the serious epizootics of animals and humans (Thoen and Steele, 1995; Grange, 1996). Beyond food producing animals, mainly cattle, the bovine tuberculosis is known to have a wide range of hosts. Although the incidence of the disease in the industrially advanced countries of Europe has declined dramatically since the completion of the eradication programmes, the infection is considered as serious epi-zootic diseases which are subject to mandatory an-nouncement and obligatory eradication. Thus when dealing with bovine tuberculosis one should bear in ABSTRACT: The post-eradication incidence of bovine tuberculosis in seven Central European Countries (Bosnia and Herzegovina, Croatia, Czech Republic, Hungary, Poland, Slovakia and Slovenia) was studied between 1990–1999 years. The majority of cattle to the age of 24 months were screened by tuberculin skin test on annual basis. Tuberculous lesions observed during meat inspection at abattoirs were further laboratory examined by direct microscopy, cultivation and histology for the presence of mycobacteria. Data describing the incidence of the disease in animals for the whole period were obtained from all countries except Bosnia and Herzegovina, where data were obtained after the year 1996. Between 1990 and 1999 years, bovine tuberculosis was diagnosed in a total of 1 084 cattle herds. Nine hundred and seventy five (89.9%) outbreaks of the disease were reported in small herds (≤10 cows) and 109 (10.1%) outbreaks in large cattle herds (>10 cows). The last outbreak of bovine tuberculosis was diagnosed in Slovakia, Slovenia, the Czech Republic, Croatia and Bosnia and Herze-govina in 1993, 1993, 1995, 1999 and 1999, respectively. Bovine tuberculosis was diagnosed due to the proper quarantine in Slovenia in 37 fattening bulls imported from two foreign European countries before animals were introduced to the targeting farms.
122 National programmes for the elimination of Myco-bacterium bovis infection in cattle were successfully completed in seven Central European countries (Bosnia and Herzegovina, Croatia, Czech Republic, Hungary, Poland, Slovakia, and Slovenia) in the second half of the 20th century (Table 1). In the subsequent post-elimination period, the incidence of bovine tu-berculosis in cattle decreased and reached zero level in some countries, or sporadic outbreaks of bovine tu-berculosis in cattle were recorded in Slovakia and Slovenia in 1993, in the Czech Republic in 1995, in Croatia and Bosnia and Herzegovina in 1998 (Pavlik et al., 2002; Table 1).
Paratuberculosis or Johne's disease is a chronic intestinal disease caused by Mycobacterium avium subsp. paratuberculosis, which continues to spread in agricultural species. Control of paratuberculosis is challenging and should not be underestimated. Due to the long incubation period of the infection, disease is largely subclinical in domesticated livestock. Hence, direct effects on animal productivity and welfare are often masked and may appear insufficient to justify large investments in control programmes by individual farmers, livestock industries or governments. Furthermore, in some countries the main effects of the disease are indirect, resulting from the impact of market discrimination against herds and flocks known to be infected, or from the control measures enforced to reduce transmission. In such circumstances, producers may be unwilling to co-operate with surveillance that may detect infection in herds or flocks. As control programmes are rarely successful in eliminating the infection from a herd or flock in the short term without an aggressive and costly programme, financial and community support assists producers to deal with the challenge. Successful prevention and control depends on animal health authorities and livestock industries acquiring a good understanding of the nature and epidemiology of infection, and of the application of tools for diagnosis and control. Building support for control programmes under the leadership of the affected livestock industries is critical, as programmes are unlikely to be successful without ongoing political will, supported by funding for research, surveillance and control.
Six isolates of Mycobacterium avium of genotype dnaJ+ IS901− IS1311+ IS1245+ and serotypes 6 (n = 1), 6/9, (n = 2), and 9 (n = 3) were obtained within a 5-month period from a human immunodeficiency virus-negative patient treated for tuberculosis.
The isolates were identified with PvuII restriction fragment length polymorphism (RFLP) analysis as a single IS1311 RFLP type and six different IS1245 RFLP types. Six separate colonies/clones obtained by subculture from each of the six isolates were tested for MICs of a set
of 10 drugs. This report documents the appearance of isolates that are resistant to antimycobacterial drugs as the duration
of therapy increases. Because isolates recovered from the patient following longer duration of treatment were more likely
to be resistant to more antimycobacterial drugs, we would conclude that there was selection for antimycobacterial drug-resistant
isolates. Analyses of all 36 clones identified three IS1311 and 22 IS1245 types forming three clusters. Tests of 105 environmental samples collected in the home and the work place of the patient
yielded 16 mycobacterial isolates, of which one M. avium from soil was of genotype dnaJ+ IS901+ IS1311+ IS1245+ and serotype 2, and the second M. avium from a vacuum cleaner was of genotype dnaJ+ IS901− IS1311+ IS1245+ and serotype 9. Overall analyses of the results did not reveal any relation between serotype, RFLP type, and drug susceptibility.
Based on the course of the disease in the patient and different serotypes, IS1311 and IS1245 RFLP types of isolates of M. avium we suppose represent polyclonal infection.
To study the dissemination of Mycobacterium bovis subsp. caprae, 79 European isolates from cattle, humans, and other hosts were examined by spoligotyping and IS6110 restriction 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.
As of 10 October, 1968, bovine tuberculosis in cattle was eliminated in the Czech Republic within the framework of national elimination programme (1959 to 1968). The postelimination period (1969 to 1996) was typical by Vanishing of infection source reservoirs during several following years. Currently only sporadic cases are recorded. in the period of the years 1969 to 1996 bovine tuberculosis was newly detected in 369 farms of cattle (42 small farms with up to 9 dairy cows and 327 larger farms with more than 10 dairy cows). No occurrence of bovine tuberculosis was found in the years 1981, 1987 to 1990, 1993 and 1996. In the remaining years of the period between 1980 and 1996, there were always maximum 3 outbreaks of bovine tuberculosis in cattle detected per year. The rate of infected animals out of the total size of herds was very low and did not exceed 5 to 10% of animals. In the years 1970 to 1996 the infection with Mycobacterium bovis was also diagnosed in total of 119 animals (zoological gardens, wild animals, small farms) and in 10 samples of milk. In the sense of OIE definition (International Animal Health Code) the territory of the Czech Republic. is free from bovine tuberculosis (prevalence up to 0.2% of infected herds of cattle). In human population in the years 1969 to 1996 the spread of M. tuberculosis was recorded in totally 77 739 newly infected persons and the infection with M. bovis in 476 patients. In 1981 to 1996 the prevalence of bovine tuberculosis ranged in absolute figures between 4 and 20 patients of higher age groups - above 50 years (0.04 to 0.20 per 100 000 inhabitants). The incidence of this disease in absolute figures was 3 to 19 patients (0.05 to 0.18 per 100 000 inhabitants).
The prevalence of Rhodococcus equi and atypical mycobacteria in the lymph nodes of pigs (n = 1 382) and cattle (n = 231) without clinical signs was studied in the Czech Republic over the period 1996-1998. R. equi alone was isolated from 7.4% of pigs, and in a mixed infection with atypical mycobacteria in another 2.3% of pigs and 1.7% of cattle. The frequency of R. equi was higher (p = 0.01) in pigs than in cattle. Atypical mycobacteria alone were more frequently isolated from pigs - 37.2% positive findings than from cattle - 28.2% positive findings (p = 0.01). Of the total of 546 mycobacterial strains isolated from pigs, 96.2% belonged to the Mycobacterium avium complex, and 3.8% (21 strains) belonged to other species (5 strains M. chelonae, 6 strains M. terrae, 2 strains M. phlei, 6 strains M. fortuitum and 2 strains not identified). A total of 65 mycobacterial strains belonging only to M. avium complex were identified in cattle. Of 492 pigs, R. equi alone was isolated from submaxillary lymph nodes at frequency of 19.5%, atypical mycobacteria were isolated at 30.1% and mixed infection at 4.9%. On the other hand, of the total of 639 pigs, R. equi alone was isolated from mesenteric lymph nodes in 0.5% of animals; atypical mycobacteria in 42.6% and mixed infection in 0.8% of pigs. The isolation of R. equi from submaxillary lymph nodes was more frequent (p = 0.01) than isolation from mesenteric lymph nodes. Examinations of lymph nodes from 218 pigs without tuberculoid nodules (group S1) resulted in isolation of R. equi in 2.8% of animals compared with significantly higher (p = 0.01) isolation from 703 pigs with caseated tuberculoid nodules (group S2) which was 13.7%. No R. equi was isolated in the 461 animals with calcified tuberculoid nodules (group S3). The detection rate of atypical mycobacteria increased in these groups S1, S2 and S3, the values being 19.3, 37.3 and 45.6%, respectively, and those of R. equi and/or atypical bacteria from pigs were 23.4, 45.8 and 54.9%, respectively. The difference between the groups was highly significant (p = 0.01). The differences between the groups were highly significant (p = 0.01). Of a total of 765 pig and cattle farms R. equi was isolated from 93 farms. Contact of swine on these farms with horses or their faeces was confirmed only in 19 (20.4%) cases.
The organs of 30 insectivorous mammals and 62 rodents from areas inhabited by people or livestock where cattle paratuberculosis or mycobacterial infections of swine had been found to occur were examined by cultivation during the monitoring of occurrence and spread of mycobacterioses in cattle and swine. Mycobacteria were found in the organs of 3 insectivores (10%) and 6 rodents (9.7%).Mycobacterium chelonae was isolated from the organs of the lesser white-toothed shrew (Crocidura suaveolens) and the common vole (Microtus arvalis), andM. vaccae andM. avium subsp.avium (IS901
+, serotype 1) from the organs of the common shrew (Sorex araneus). M. avium subsp.avium (IS901
+, serotype 1) was also isolated from the organs of the yellow-necked mouse (Apodemus flavicollis). Slow-growing mycobacteria of group III (according to Runyon) were isolated from the organs of the mouse (Mus musculus sensu lato) and the yellow-necked mouse (A. flavicollis). These findings had no connection with the epizootological situation in the nearby livestock.M. fortuitum was isolated from the organs of the common vole (M. arvalis) caught in a field within easy reach of a swine breeding heard.M. fortuitum was also identified in the lymph nodes and droppings of this swine herd, as well as in the straw, scrapings from the floor of stalls, troughs and banisters, as well as from larvae and imagoes of dipterous insects. These results demonstrate the possibility that insectivores and small rodents can spread the causative agents of mycobacteria in wild and domestic animals.
Mycobacteria were isolated from 14 (4.5%) of 314 samples, containing 7791 adult Diptera, which were collected in the Czech Republic and Slovakia in 1997-2000. These flies were collected from three cattle herds with paratuberculosis, two pig herds with mycobacterial infections and one farm that kept both cattle and pigs and that did not have problems of mycobacterial infections. Mycobacterium intracellulare was isolated from Eristalis tenax Linnaeus (Diptera: Syrphidae) captured from a pig herd. Mycobacterium avium ssp. avium (serotype 8) was isolated from flies of the genera Drosophila Fallen (Diptera: Drosophilidae) and Musca Linnaeus (Diptera: Muscidae) originating from a pig herd. Mycobacterium spp. were isolated from Musca spp. and Mycobacterium fortuitum was isolated from dung flies of the genus Scatophaga Meigen (Diptera: Scatophagidae), Musca spp. and Stomoxys calcitrans Linnaeus (Diptera: Muscidae) captured in the same herd. Mycobacterium scrofulaceum was isolated from S. calcitrans from the farm with both cattle and pigs. Mycobacterium avium ssp. paratuberculosis was isolated from Scatophaga spp. collected from pastures grazed by one of the cattle herds and from Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae) and Lucilia caesar Linnaeus (Diptera: Calliphoridae) captured in a slaughterhouse, where cattle infected with paratuberculosis were slaughtered. Mycobacterium phlei was isolated from flies of the genus Lucilia captured at a waste bin. These data indicate that mycobacteria may be spread by adult flies that have been in contact with material contaminated with these pathogens.
This study surveys 2,593,348 cattle slaughtered between 1996 and 2000, and further investigates 571 (0.02%) animals found to have tuberculous lesions. Culture of 346 randomly selected tissue samples from animals younger (n = 215) and older (n = 131) than 2 years, isolated mycobacteria from 91 animals (26.3%). These included 74 Mycobacterium avium subsp. avium isolates of IS901+ and IS1245+ genotype and serotype 2, 13M. avium subsp. hominissuis isolates of IS901- and IS1245+ genotype and serotypes 8 (n = 7) and 4 (n = 6), two M. chelonae, one M. avium subsp. paratuberculosis (RFLP type B-C1), and one M. terrae. Culture of mesenteric lymph node samples obtained 66 isolates of M. avium complex (MAC) and four isolates of other mycobacterial species. M. bovis was significantly absent from all samples. Mycobacteria were more frequently (P = 0.01) isolated from tissues of animals under 2 years (34.4%) than animals over 2 years (13.0%). IS901 and IS1245 RFLP methods were used to type 17 randomly selected MAC isolates, virulent after intramuscular inoculation of pullets, from 17 different cattle herds. These revealed 11 distinct IS901 RFLP types and three IS1245 RFLP profiles. Polyclonal infection of individual animals was detected by IS901/IS1245 typing in 2 of the 17 selected isolates.
Mycobacterial catalases peroxidases and superoxide dismutase and their effects on virulence and isoniazid-susceptibility in mycobacteria
- M J O Bartos
- Falkinhamiii
- Pavlik