The present study highlights the transboundary nature of tuberculosis (TB) in alpacas and the failure of current antemortem testing protocols to identify TB-free alpaca herds and individuals for exportation. The tuberculin skin test (TST) failed to identify Mycobacterium bovis-infected animals prior to movement from the United Kingdom (UK) to Poland. This study describes the use of four serological assays [Enferplex Camelid TB, dual-path platform (DPP) VetTB and BovidTB assays, and multi-antigen print immunoassays (MAPIAs)] to detect TB in an alpaca herd with negative TST results. The breeding in Poland purchased alpacas for several years from the UK with the last group arriving in May 2018. In July 2018, two sick alpacas from the centre were hospitalized in a veterinary clinic and both died of TB a few weeks later. In November 2018, 20 alpacas remaining in this M. bovis-affected herd were euthanized and samples were collected. The study population included 20 M. bovis-infected and 20 uninfected alpacas, but only 15 infected animals were tested by all serology tests. The DPP VetTB and DPP BovidTB assays detected antibodies in 14 of the 20 infected alpacas, with results confirmed by MAPIA, and in none (MAPIA and DPP BovidTB) or one (DPP VetTB) of the 20 uninfected animals. None of the infected alpacas tested positive using the Enferplex assay. In addition, the group included three orphans and two cria-dam pairs, which provided an opportunity to analyse immune aspects of cria-mother relationships in this herd. The results suggest high susceptibility of this host species to M. bovis infection and rapid progression to disease. The serological tests used in this study offer useful tools for the detection of M. bovis infection in TST and Enferplex test non-reactive alpacas. These tests should be further evaluated for implementation into TB management and control strategies for camelid species. K E Y W O R D S alpaca, dual-path platform assay, Enferplex Camelid TB test, MAPIA, Mycobacterium bovis, tuberculin skin test | 1307 KRAJEWSKA-WĘDZINA Et Al.
Introduction. Bovine tuberculosis is a chronic contagious disease caused by Mycobacterium bovis or Mycobacterium caprae. Before widespread action conducted in Poland between 1959-1975 to combat bovine tuberculosis (BTB), about 40% of all tuberculosis cases in pigs was caused by the bovine bacillus. At the present time, correctly carried out, long-term control of cattle has resulted in cases of bovine tuberculosis in pigs and humans being extremely rare and sporadic. In pigs, tuberculosis is most often caught in a slaughterhouse during slaughter. Materials and method. Samples came from pigs kept on the farm. Traditional bacteriological methods on solid media (Stonebrink, LJ with pyruvate) supported by the semi-automatic, liquid indicative culture method (MGIT) and PCR test were applied in targeted studies. The GenoType Mycobacterium MTBC and CM tests (Hain Lifescience, Germany) were used to additionally confirm that isolated strains classification was used. Results. Strains of mycobacteria were isolated from all examined pigs. Mycobacterium bovis was determined by real time PCR and Hain Genotype methods. Conclusions. In order to effectively fight against BTB, all animals on farms should be tested, regardless of species, while the milk of suspected cows should be utilized without being used for feed. It is important to adapt the current legal regulations to the current epidemiological situation.
The most numerous group of bacteria in the genus Mycobacterium is the nontuberculous mycobacteria. Currently, over 200 species of bacteria have been classified as belonging to this group, of which approximately 30 are pathogenic to humans and animals. Mycobacterium kansasii complex numbers among these pathogenic species. The submandibular lymph nodes of a wild boar shot by a hunter were examined in order to confirm or exclude infection with bacteria of the genus Mycobacterium. In culture, a bacterial isolate was obtained after 12 days of incubation on Petragnani and Stonebrink media. A multiplex PCR clearly indicated that the isolate was a nontuberculous mycobacterium. The results of species identification attempts via both molecular biology methods and mass spectrometry confirmed that the isolated strain belonged to MKC. The described case of a wild boar infection with MKC is the first documented case in Poland and only the second in Europe, and in confirming the presence of this pathogen among free-living animals, this report implies that MKC is of great concern. Our research elucidates some specifics of wild boar mycobacteriosis and may be used to instill awareness in the public of the dangers of dressing hunt prey or consuming its meat in ignorance of safe procedures, which can contribute to the transmission of the pathogen to humans.
Introduction The study highlights the transboundary nature of tuberculosis (TB) in alpacas and the failure of current ante-mortem testing protocols (the tuberculin skin and Enferplex Camelid TB tests) to identify TB-free alpaca herds and individuals for export. Our research and the available literature indicate that the alpaca (Vicugna pacos) is extremely susceptible to Mycobacterium bovis infection, and that testing periodicity fails to take into account that animals do not manifest disease symptoms for a long time. The skin test failed to identify Mycobacterium bovis infection in two alpacas prior to their movement from the UK to Poland. The animals were purchased by a breeding centre in Poland, and were then shown at an international animal exhibition. The last owner of the alpacas before their deaths from TB bought the infected animals unwittingly in order to run rehabilitation activities with disabled children on his farm. Material and Methods Thoracic lymph node, lung and liver tissue samples obtained at necropsy were examined histopathologically after Ziehl–Neelsen staining. Tissue samples were homogenised and mycobacteria present there were cultured on Stonebrink’s medium during a 6-week incubation. A commercial test using polymorphism of the chromosomal direct repeat region provided species identification and additional identification was by spacer oligonucleotide typing and mycobacteria interspersed repetitive unit–variable number tandem repeat analysis with a gel electrophoresis protocol. Results The microbiological examination confirmed multiorgan TB caused by the SB0666 spoligotype of Mycobacterium bovis. Conclusion Due to the suboptimal performance of current diagnostic tests for TB in alpacas, there is a risk that infected animals may be moved unwittingly. A risk of TB spread associated with the international movement of alpacas is implied by this study.
Since 2009, Poland has been recognized as a country officially free of bovine tuberculosis (BTB). However, new outbreaks are each year quoted. In many countries it has been shown that badgers (Meles meles) are a vector of Mycobacterium bovis/caprae (M.bovis/caprae) and a source of bovine tuberculosis for many domestical species, mainly for cattle. The aim of the presented study was to determine, for the first time in Poland, the occurrence of tuberculosis in badgers in areas where the disease occurs in cattle. Tissue samples were examined by classical microbiology methods, mycobacteria growth indicator tube (MGIT), and real time PCR. A total of 155 samples from 31 badgers were examined. In any case Mycobacterium bovis/caprae infection has not been diagnosed. This indicates that badgers probably are not a vector of bovine tuberculosis in Poland.
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