Detection of Mycobacterium tuberculosis complex in milk using polymerase chain reaction (PCR)

Pinto, Angela Di; Ciccarese, Giuseppina; Tony, Forte Vito; Bijo, Bizena; Fatmira, Shehu; Giuseppina, Tantillo

doi:10.1016/j.foodcont.2005.04.019

Cited by 11 publications

(13 citation statements)

References 16 publications

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“…This could be explained by the fact that cattle with BTB may not show any clinical signs even during the advanced stages of the disease [6]. Furthermore, the signs may not be noticed by the veterinarians or cattle owners because of negligence.…”

Section: Resultsmentioning

confidence: 99%

“…BTB remains a worldwide problem, with more than 50 million cattle estimated to be infected [ 5 ]. The disease is an important zoonosis which poses a significant threat to humans as it can be transmitted through consumption of contaminated milk and close contact with infected cattle [ 6 ]. M. bovis accounts for approximately 0.5–1.5% of the entire human TB cases in some developed countries [ 7 ].…”

Section: Introductionmentioning

confidence: 99%

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Molecular Detection and Drug Resistance of Mycobacterium tuberculosis Complex from Cattle at a Dairy Farm in the Nkonkobe Region of South Africa: A Pilot Study

Silaigwana

Green

Ndip

2012

IJERPH

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Mycobacterium tuberculosis complex (MTBC) causes tuberculosis (TB) in humans and animals. We investigated the presence of MTBC in cattle milk and its drug resistance using polymerase chain reaction (PCR). Two hundred samples (100 mL each) were obtained from a dairy farm in the Nkonkobe region of South Africa. The samples were processed using the modified Petroff method. DNA was isolated using a Zymo Bacterial DNA kit and amplified using Seeplex® MTB Nested ACE assay. The Genotype® Mycobacterium tuberculosis-multidrug resistantplus (MTBDRplus) assay was used to perform drug susceptibility and detection of mutations conferring resistance to isoniazid (INH) and rifampicin (RIF). Eleven samples tested positive for MTBC DNA using the Seeplex® MTB Nested ACE assay. The Genotype® MTBDRplus assay showed that 10/11 samples were resistant to both INH and RIF i.e., multi-drug resistant (MDR). The most and least frequent rpoB mutations detected in RIF resistant samples were H526Y (9/10) and D516V (2/10) respectively. None of the INH resistant samples harbored mutations in the katG gene. However, all of them harbored the T8A mutation in the inhA gene. These results have clinical and epidemiological significance and calls for further studies and necessary actions to delineate the situation.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Molecular Detection and Drug Resistance of Mycobacterium tuberculosis Complex from Cattle at a Dairy Farm in the Nkonkobe Region of South Africa: A Pilot Study

Silaigwana

Green

Ndip

2012

IJERPH

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“…However, these nutrients can also serve as substrates for pathogenic microorganisms. The traditional consumption of homemade dairy products, and especially cheeses, that are composed of non-heat-treated milk poses a serious risk to public health [1]. It is estimated that more than 90 percent of all of the cases of illness related to the consumption of dairy products are of bacterial etiology, and at least 21 different diseases of this type have been identified [2].…”

Section: Introductionmentioning

confidence: 99%

Occurrence of mycobacteria in bovine milk samples from both individual and collective bulk tanks at farms and informal markets in the southeast region of Sao Paulo, Brazil

et al. 2013

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BackgroundMycobacterium spp. is one of the most important species of zoonotic pathogens that can be transmitted from cattle to humans. The presence of these opportunistic, pathogenic bacteria in bovine milk has emerged as a public-health concern, especially among individuals who consume raw milk and related dairy products. To address this concern, the Brazilian control and eradication program focusing on bovine tuberculosis, was established in 2001. However, bovine tuberculosis continues to afflict approximately 1,3 percent of the cattle in Brazil. In the present study, 300 samples of milk from bovine herds, obtained from both individual and collective bulk tanks and informal points of sale, were cultured on Löwenstein-Jensen and Stonebrink media. Polymerase chain reaction (PCR)-based tests and restriction-enzyme pattern analysis were then performed on the colonies exhibiting phenotypes suggestive of Mycobacterium spp., which were characterized as acid-fast bacilli.ResultsOf the 300 bovine milk samples that were processed, 24 were positively identified as Mycobacterium spp.Molecular identification detected 15 unique mycobacterial species: Mycobacterium bovis, M. gordonae, M. fortuitum, M. intracellulare, M. flavescens, M. duvalii, M. haemophilum, M. immunogenum, M. lentiflavum, M. mucogenicum, M. novocastrense, M. parafortuitum, M. smegmatis, M. terrae and M. vaccae. The isolation of bacteria from the various locations occurred in the following proportions: 9 percent of the individual bulk-tank samples, 7 percent of the collective bulk-tank samples and 8 percent of the informal-trade samples. No statistically significant difference was observed between the presence of Mycobacterium spp. in the three types of samples collected, the milk production profiles, the presence of veterinary assistance and the reported concerns about bovine tuberculosis prevention in the herds.ConclusionThe microbiological cultures associated with PCR-based identification tests are possible tools for the investigation of the presence of Mycobacterium spp. in milk samples. Using these methods, we found that the Brazilian population may be regularly exposed to mycobacteria by consuming raw bovine milk and related dairy products. These evidences reinforces the need to optimize quality programs of dairy products, to intensify the sanitary inspection of these products and the necessity of further studies on the presence of Mycobacterium spp. in milk and milk-based products.

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“…Taking unboiled/raw milk could be a potential source of infection to humans as described above since infected cows excrete M. bovis via milk [54]. Polymerase chain reaction conducted from Albanian milk samples in Tirana university revealed the presence of the MTC gene in 36/50 (72%) of the tested samples would support the possibility of ingesting the disease causing agent [55].…”

Section: Discussionmentioning

confidence: 99%

Tuberculosis Infection in Cattle and Cattle Owners in North Eastern Parts of Ethiopia

Mengistu¹,

Enquselassie²

2015

Biol Med (Aligarh)

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Introduction: Ethiopia is one of the highest TB burden countries in the world. Transmission of M. tuberculosis between humans is well known, however, little is known about the transmission of M. tuberculosis complex species between human and animals, in particular cattle. The purpose of this study was, therefore, to investigate the status of bovine tuberculosis infection in cattle owned by tuberculosis suspected households and isolate M. tuberculosis complex species from sputum of chronic coughers in North Wollo Zone of the Amhara National Regional State, Ethiopia. Method:A cross-sectional study was conducted in selected Kebeles of North Wollo Zone. An intradermal tuberculin skin test was carried out on 381 cattle and sputum samples collected from 124 chronic coughers. Sputum samples were cultured and positive ones were identified by polymerase chain reaction using RD4 and RD9 as a marker. Result:Of the 124 sputum samples cultured, 4/ 124 (3.2%) were positive. Polymerase Chain Reaction using has confirmed that 2/4 (50%) were found to be M. tuberculosis while the rest was atypical Mycobacterial species. Of the 381 tested cattle 5/381 (1.31%) and 10/38 (2.63%) were found positive according to manufacturer recommendation (with a cutoff value > 4 mms) and a cutoff value > 2 mms, respectively. About 67% (6/9) of the individuals who owned positive cattle were having the habit of drinking raw milk. Among the ten positives, five of them resided in lowland (Kolla) areas (1300-1500 mabs). However, none of the owners of tuberculin positive cattle were found to be TB positive. Conclusion:This study has shown 3.2% of culture positivity and of these two were found to be M. tuberculosis. The sputum sample culturing from suspected chronically coughing individuals would help detection of the TB causative agents. According to the current findings BTB is considerable in Ethiopia and cattle should be seen as a potential source of zoonotic TB to humans. B io lo g y and M e d ic in e

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Detection of Mycobacterium tuberculosis complex in milk using polymerase chain reaction (PCR)

Cited by 11 publications

References 16 publications

Molecular Detection and Drug Resistance of Mycobacterium tuberculosis Complex from Cattle at a Dairy Farm in the Nkonkobe Region of South Africa: A Pilot Study

Molecular Detection and Drug Resistance of Mycobacterium tuberculosis Complex from Cattle at a Dairy Farm in the Nkonkobe Region of South Africa: A Pilot Study

Occurrence of mycobacteria in bovine milk samples from both individual and collective bulk tanks at farms and informal markets in the southeast region of Sao Paulo, Brazil

Tuberculosis Infection in Cattle and Cattle Owners in North Eastern Parts of Ethiopia

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