Molecular Characterization of <em>Mycobacterium</em> spp. Isolated from Cattle and Wildlife in Poland

Although Poland is officially tuberculosis (TB) free, meaning that less than 0.1% of her cattle herd is TB-positive, the problem of bovine TB in Poland may be re-emerging: its presence has recently been confirmed in domestic and companion animals, wildlife such as the European bison, and even humans. The aim of this chapter was to review all reports of bovine TB in Poland described to date, with particular emphasis on molecular studies, and determine further research directions. These studies include a range of molecular methods for diagnosis, including genotyping, spoligotyping and MIRU- VNTR; such methods successfully identifies a tuberculosis-positive European bison as the source of wild boar infection in the Bieszczady Mountains based on its spoligotype. This chapter argues that identified trains should be better archived, as such records would allow detailed epidemiological investigations and shed greater light on the activity of Mycobacterium spp. The current epidemiological situation in Poland highlights the need for further studies to determine epidemiological links and confirm possible routes of transmission based on whole genome sequencing; this need is accentuated by the zoonotic potential of such infections and the endangered species at risk

Biopsy and tracheobronchial aspirates as additional tools for the diagnosis of bovine tuberculosis in living European bison (bison bonasus)

CITATION: Didkowska, Anna et al. 2020. Biopsy and tracheobronchial aspirates as additional tools for the diagnosis of bovine tuberculosis in living European bison (bison bonasus). Animals, 10(11):2017, doi:10.3390/ani10112017.The original publication is available at: https://www.mdpi.comENGLISH ABSTRACT: The diagnosis of bovine tuberculosis (BTB) in living wildlife remains a complex problem, and one of particular importance in endangered species like European bison (Bison bonasus). To identify infection and avoid the unnecessary culling of such valuable individuals, current best practice requires the collection and culture of material from living animals, as mycobacteria isolation remains the gold standard in BTB diagnosis. However, such isolation is challenging due to the need for the immobilization and collection of appropriate clinical material, and because of the sporadic shedding of mycobacteria. In the present study, we evaluated the potential of sampling for the detection of BTB in a group of seven living European bison suspected of being infected with Mycobacterium caprae. The specimens were collected both as swabs from the nasal and pharyngeal cavities, tracheobronchial aspirates (TBA), ultrasound-guided biopsies from lateral retropharyngeal lymph nodes, and post mortem, from mandibular, retropharyngeal and mediastinal lymph nodes. Clinical samples were tested for mycobacterial species via mycobacteriological culture and PCR. M. caprae was isolated from collected material in two out of four living infected individuals (TBA, biopsy) and mycobacterialDNAwas detected in three out of four (TBA, pharyngeal swab) bison. This is the first report of isolation of M. caprae in living European bison. Our findings demonstrate the value of diagnostic tests based on both molecular testing and culture in European bison and confirm the respiratory shedding of viable M. caprae in this host species.Publisher's versio

Prevalence of <i>Mycobacterium tuberculosis</i> complex infections in animals. Part I. Small ruminants

Animal tuberculosis is a highly infectious disease caused by acid fast organisms of the Mycobacterium tuberculosis complex (MTBC). Tuberculosis in cattle and other animal species is mainly caused by Mycobacterium bovis and, to a lesser extent, Mycobacterium caprae. Tuberculosis is a chronic, debilitating disease characterized by the formation of tubercles, most commonly observed in the lymph nodes, lungs, pleura, peritoneum, and less often in the intestines, liver and spleen. Despite decades of eradication efforts and an overall downward trend, the incidence of cases in cattle is not uniform and clusters of infection are still common. Other species are presumed to play a key role in maintaining the disease in the environment. Therefore, tuberculosis control measures should not be cattle limited but include small ruminants, as evidenced by the increased incidence of tuberculosis in sheep and goats, especially in endemic multi-host systems

Prevalence of <i>Mycobacterium tuberculosis</i> complex infections in animals. Part II. Companion animals

Tuberculosis is a chronic, infectious disease, that can affect humans as well as farm, wild and companion animals. Due to close and frequent contacts with humans, companion animals pose the greatest threat to the public health if infected with mycobacteria. Dogs and cats are the most generally owned pet animals. Although not highly sensitive to infection, new cases of tuberculosis in this group of animals are reported every year in Europe. The most common source of infection in pet animals are free-living animals that are reservoirs of tuberculosis. The clinical symptoms most often relate to emaciation and respiratory disorders. However, less common symptoms can also develop, such as diarrhea, vomiting and ulcerative skin wounds. Dogs and cats, can be effective vectors of pathogen transmission to other animal species, as well as to humans. Therefore, if disease is suspected, the animal should be tested to confirm or rule out infection with Mycobacterium tuberculosis complex but intradermal tuberculin tests, effective in the diagnosis of many animal species, are useless for canines and felids. The current in vivo diagnostics is based mainly on the IFN-γ test, tissue biopsy and PCR

Alpacas - a new breeding species in Poland. Part II. Breeding and nutrition

The article presents the second part of the series.The authors have focused on husbandry, breeding and feeding of alpacas. Alpacas must be well protected from rain, cold and high air temperature. When choosing this type of animals, you should consider how much profit you want to achieve from breeding. Will animals be treated as a source of income or a hobby? Alpacas live more than 20 years and their reproductive life is also long. Alpacas are not farm animals within the meaning of Polish regulations, and thus no additional permits or registration are required. However, possession of the herd should be reported to the appropriate County Veterinary Inspectorate. Alpacas are herd animals even the males, so it is strongly discouraged to isolate a single male from other alpacas. The herd gives them a sense of security that is fundamental to their welfare. Only within a herd can they live relaxed, graze peacefully, rest, sleep and build harmonious relationships. Lonely alpacas live under constant stress, which can lead to a serious illness or even death. They are pasture animals. 80-90% of alpacas food is roughage, i.e. hay, grass. It is quite difficult to create one perfect recipe for a good feeding program because a lot depends on the conditions on a given farm, on the quality of the soil, pasture and the hay quality.Here, we have also presented the breeding program and some aspects of veterinary surveillance for alpacas

Alpacas, new breeding species in Poland. Part IV. Delivery and neonatal care

The aim of this article was to present the specificity of alpacas reproductive physiology, different from other farm animals. Camelids are induced ovulators and seasonal breeders. Breeding females in good condition should give cria every year or nearly every year. In alpacas, early embryonic death and early fetal death is a significant and quite common problem, therefore early pregnancy diagnosis should be confirmed by ultrasound examination around 40–50 days and repeated at 80-90 days of pregnancy. Most parturitions take place between spring and autumn. Few reliable indications of pending delivery exist. Due to the anatomy of the fetus (exceptionally long neck and limbs), obstacles to labor are frequent so it is advisable to monitor the perinatal period and delivery with cameras. The breeder should carefully watch the cria in the first hours of life. It is standing usually 30 to 90 minutes after birth, then begins to suckle and releases meconium. Newborn alpacas rely on passive transfer of immunity through early colostrum intake. The quality of colostrum depends on several factors, including the immune status of the dam and udder health. The mammary gland secretes colostrum 24 hours after delivery and the concentration of IgG drops quickly in the secretion, so its early supply is crucial. The survival of cria depends largely on conditions in the first few days after birth

Infection of a Free-Living Wild Boar (<i>Sus scrofa</i>) with a Bacterium from the <i>Mycobacterium kansasii</i> Complex

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

Infection of a Free-Living Wild Boar (Sus scrofa) with a Bacterium from the Mycobacterium kansasii Complex

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

Ocena epizootyczna zakażeń Mycobacterium avium spp. paratuberculosis u bydła w Polsce w latach 2011-2020 jako potencjalny czynnik etiologiczny choroby Leśniowskiego-Crohna u ludzi

Background. Mycobacterium avium spp. paratuberculosis (MAP) is a bacterial germ and the etiologic agent in paratuberculosis also known as Johne's disease, a debilitating disease seen in animals. MAP is one of the triggers in the development of Crohn's disease in humans. Therefore the presence of these microorganisms in the environment and their prevalence in farm animals in relation to Crohn's disease is a new area of research. To date, it has not been evaluated in Poland. Material and methods. The materials for this study consisted of 14,468 samples of bovine serum across Poland. Determination of anti-MAP antibody levels was performed using an immunoenzymatic Paratuberculosis Screening Ab Test p/n P07130-5® (IDEXX, Westbrook, Maine 04092, United States). Results. The study consisted of 14,468 samples of bovine serum. Anti-MAP antibodies were identified in 349 cattle sera. Conclusions. Successful paratuberculosis screening programs can lead to a reduction of MAP in animal products. As the role of MAP in Crohn’s disease is not fully understood, future research should be conducted to address this knowledge gap and avoid a potential zoonotic public health problem.Wprowadzenie. Mycobacterium avium spp. paratuberculosis (MAP) zwany także prątkiem Johnego jest czynnikiem etiologicznym paratuberkulozy, wyniszczającej choroby występującej u zwierząt. Jednym z czynników niezbędnych do rozwoju choroby Crohna u ludzi jest obecność MAP, stąd też obecność w środowisku tych drobnoustrojów, a przede wszystkim ocena ich występowania u zwierząt gospodarskich w odniesieniu do choroby Crohna, stanowi nowy obszar badań, nieopisany dotąd w Polsce. Materiał i metody. Materiał do badań stanowiło 14468 bydlęcych surowic pochodzących z terenu całej Polski. Oznaczenie poziomu przeciwciał anty-MAP wykonano przy użyciu immunoenzymatycznego testu Paratuberculosis Screening Ab Test p/n P07130-5® (IDEXX, Westbrook, Maine 04092, United States). Wyniki. W badanej grupie 14468 surowic, przeciwciała anty-MAP potwierdzono u 349 sztuk bydła. Wnioski. Udane programy kontroli paratuberkulozy mogą prowadzić do zmniejszenia występowania MAP w produktach zwierzęcego pochodzenia. W związku z tym, że rola MAP nie jest do końca poznana w chorobie Crohna, przyszłe badania powinny zająć się tymi lukami, aby uniknąć potencjalnego odzwierzęcego problemu zdrowia publicznego