Short communication: detection of Mycobacterium avium subspecies paratuberculosis by polymerase chain reaction in bovine milk in Brazil (original) (raw)
Related papers
Foodborne Pathogens and Disease, 2010
Paratuberculosis, or Johne's disease, is caused by Mycobacterium avium subsp. paratuberculosis (Map), and it generates great economic losses for the dairy industry worldwide. In humans, Map has been associated with Crohn's disease. Mexico has unknown paratuberculosis prevalence, and yet, control programs have not been applied. This study aimed to determine the presence of Map in milk samples from seropositive goats and cows and bulk tank milk samples from herds previously designated Map-infected using indirect enzyme-linked immunosorbent assay. Map DNA was detected in 100% of the bulk tank milk samples of 14 bovine herds and 3 caprine flocks using a modified insertion sequence 900 polymerase chain reaction (PCR). Additionally, Map DNA was detected in 100% of the individual milk samples from 10 cows and 8 goats. Further, based on the findings of the experimental insertion sequence 900 PCR assessment, evaluation of bulk tank and individual milk samples through a type-specific PCR was performed, which confirmed our previous findings and revealed that 56.25% cow and 63.63% goat milk had concurrent infections of the C, I, and S types. Out of 14 bulk tank milk samples, 10 had viable mycobacteria. Paratuberculosis was detected at a high frequency in cow and goat milk, which suggests that raw milk ingestion represents a potential risk of Map infection.
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire, 2009
Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne's disease in cattle and other farm ruminants, and is also a suspected pathogen of Crohn's disease in humans. Development of diagnostic methods for MAP infection has been a challenge over the last few decades. The objective of this study was to investigate the relationship between different methods for detection of MAP in milk and fecal samples. A total of 134 milk samples and 110 feces samples were collected from 146 individual cows in 14 MAP-infected herds in southwestern Ontario. Culture, IS900 polymerase chain reaction (PCR) and nested PCR methods were used for detecting MAP in milk; results were compared with those of fecal culture. A significant relationship was found between milk culture, direct PCR, and nested PCR (P < 0.05). The fecal culture results were not related to any of the 3 assay methods used for the milk samples (P > 0.10). Although fecal culture showed a higher sensitiv...
2012
Johne's disease is a chronic intestinal infection in ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Infected livestock diffuses bacterium to the environment in different ways, of which milk is the most common way. Since a relationship is proposed between the causes of the Johne's disease in livestock and Crohn's disease in human being, a study on the presence of the bacterium in the milk can provide an important index to the rate of development of this infection in the livestock and, probably in human societies. The current study aimed to identify MAP infection in dairy cattle bred in the northern Iran. A total of 90 dairy cattle were tested by comparative tuberculin test and results of the positive, negative and suspicious cases were obtained in the forms of 3, 64 and 23, respectively. A nested PCR was also conducted using two pairs of primers to amplify a 194 bp fragment located on IS900 element to confirm the results obtained by the former meth...
[Mycobacterium avium subsp. paratuberculosis in food and its relationship with Crohn's disease]
PubMed, 2007
Paratuberculosis or Johne's disease is a chronic enteritis of the cattle and other small ruminant animals caused by Mycobacterium avium subsp. paratuberculosis. In Argentina, the strains were characterized in beef and dairy cattle and deer in different genetic patterns by molecular tools. M. avium subsp. paratuberculosis has been linked in men to a chronic inflammation of the intestine, named Crohn's disease. There is clinical and experimental evidence to link M. avium subsp. paratuberculosis with Crohn's disease by PCR, positive bacteriological culture from mother milk, blood and affected tissues by in situ hybridization. The milk and sub-products might be one of the possible infection sources and it has been suggested that M. avium subsp. paratuberculosis could resist pasteurization. Several works showed that this mycobacteria could be present in retail milk of countries such as United Kingdom, USA, Czech Republic, and recently in Argentina. M. avium subsp. paratuberculosis was associated with different dairy products and water for human consumption. Therefore, it is possible that these food sources may have a role for transmission. New investigations should emphasize the role of contaminated food and water in human infection around the world and determine the possible zoonotic role of M. avium subsp. paratuberculosis.
Mycobacterium Avium Subspeciae Paratuberculosis Identification in Milk by Means of IS900 PCR
Macedonian Journal of Animal Science, 2012
Between the numerous emerging pathogens, Mycobacterium avium subsp. paratuberculosis (MAP) reports a primary interest in its detection for its possible correlation with Crohn’s Disease in man (5). Milk and its subproducts may be very important in transmitting Mycobacterium avium subsp. paratuberculosis. The potential sanitary risk is related to the germ capacity in surviving to routine pasteurization treatments (1). These study objectives were the determination and the optimization of DNA extraction and purification assays to the direct Mycobacterium avium subsp. paratuberculosis (MAP) detection in milk, by means of PCR. The obtained results with the applied method are based in the milk and its subproducts chain inspective and control actions, relating to the increasing paratuberculosis etiological agent presence and the potential zoonotic transmitting.
Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne's disease (JD) in cattle, sheep, goats and other ruminants, and Crohn's disease in humans. MAPs are shed to external environment through feces and milk. The present study was aimed to evaluate the utility of milk as a non-invasive sample in stage II MAP infections in goats using IS900 polymerase chain reaction (PCR) and sequencing analysis. A total of 32 milk samples from lactating does were collected. Within these 32 milk samples, 15 were collected from pre-confirmed JD positive goats. By IS900 PCR, all the 15 (100%) known JD positive goat milk samples revealed the presence of MAP. However, no unknown goat was identified as MAP positive. The results of this study established the usefulness of milk as a non-invasive sample in screening and confirmation of stage II MAP infection in goats.
Polish Journal of Veterinary Sciences, 2014
Mycobacterium avium ssp. paratuberculosis (MAP) is the cause of chronic gastroenteritis in cattle called the Johne’s disease (JD). The disease causes significant economic losses in cattle production. MAP is also supposed to be involved in the Crohn’s disease and inflammatory bowel disease (IBD) in people. The detection of the cattle infection based on investigations of milk samples and evaluation of the capacity of the methods used to detect the disease was the objective of the present study. Following methods were applied for milk samples testing: detection of MAP in bacterial culture, detection of the specific IS-900 fragment of MAP in the genetic material isolated directly and detection of MAP antibodies. The results obtained were compared with the “golden standard” results, i.e. the isolation of MAP from the faeces. PQStat-the program for diagnostic reliability estimation, was used for evaluation of the sensitivity, specificity and predictive value. The method based on detection...
Acta veterinaria Scandinavica, 2014
Bovine paratuberculosis is an incurable chronic granulomatous enteritis caused by Mycobacterium avium subspecies paratuberculosis (MAP). The prevalence of MAP in the Swiss cattle population is hard to estimate, since only a few cases of clinical paratuberculosis are reported to the Swiss Federal Food Safety and Veterinary Office each year.Fecal samples from 1,339 cattle (855 animals from 12 dairy herds, 484 animals from 11 suckling cow herds, all herds with a history of sporadic paratuberculosis) were investigated by culture and real-time polymerase chain reaction (PCR) for shedding of MAP. By culture, MAP was detected in 62 of 445 fecal pools (13.9%), whereas PCR detected MAP in 9 of 445 pools (2.0%). All 186 samples of the 62 culture-positive pools were reanalyzed individually. By culture, MAP was grown from 59 individual samples (31.7%), whereas PCR detected MAP in 12 individual samples (6.5%), all of which came from animals showing symptoms of paratuberculosis during the study. ...
Journal of microbiology, immunology, and infection = Wei mian yu gan ran za zhi
Mycobacterium avium subsp paratuberculosis (MAP), excreted in the feces and milk, is reported to be not easily inactivated by pasteurization and thermal treatments as other bacteria infecting humans and animals do. The D values of all MAP strains tested were considerably higher than those published for other pathogens. Culturing techniques for this organism are labor intensive. Although an increasing amount of scientific evidence suggests that this organism can be responsible for at least some cases of Crohn's disease (CD), there is controversy about MAP being a cause of CD in humans. In general, although some studies have described an association between the presence of MAP and CD, the role of Mycobacterium species and MAP in the etiology of this human disease remains unestablished. Although published reports indicate that it may not be completely inactivated by pasteurization of milk, the effectiveness of increasing the time or temperature in the pasteurization process has not...
… clinical microbiology, 2007
bottles and cartons of commercially pasteurized cow's milk were obtained at random from retail outlets throughout the Czech Republic. During the same period, samples of raw milk and of milk that was subsequently subjected to a minimum of 71.7°C for 15 s in a local pasteurization unit were also obtained from two dairy herds, designated herds A and B, with low and high levels, respectively, of subclinical Mycobacterium avium subsp. paratuberculosis infection, and from one herd, herd C, without infection. Infection in individual cows in each herd was tested by fecal culturing. Milk samples were brought to the Veterinary Research Institute in Brno, Czech Republic, processed, inoculated onto Herrold's egg yolk slants, and incubated for 32 weeks. Colonies were characterized by morphology, Ziehl-Neelsen staining, mycobactin J dependency, and IS900 PCR results. M. avium subsp. paratuberculosis was cultured from 4 of 244 units (1.6%) of commercially pasteurized retail milk. M. avium subsp. paratuberculosis was also cultured from 2 of 100 (2%) cartons of locally pasteurized milk derived from infected herds A and B and from 0 of 100 cartons of milk from uninfected herd C. Raw milk from 1 of 10 (10%) fecal culture-positive cows in herd A and from 13 of 66 (19.7%) fecal culture-positive cows in herd B was culture positive for M. avium subsp. paratuberculosis. These findings confirm that M. avium subsp. paratuberculosis is present in raw milk from subclinically infected dairy cows. The culture of M. avium subsp. paratuberculosis in the Czech Republic from retail milk that had been pasteurized locally or commercially to the required national and European Union standards is in agreement with similar research on milk destined for consumers in the United Kingdom and the United States and shows that humans are being exposed to this chronic enteric pathogen by this route.