Milk and milk products have been utilized by humans for many thousands of years. With the advent of metagenomic studies, our knowledge on the microbiota of milk and milk products, especially as affected by the environment, production, and storage parameters, has increased. Milk quality depends on chemical parameters (fat and protein content and absence of inhibitory substances), as well as microbial and somatic cells counts, and affects the price of milk. The effects of hygiene and effective cooling on the spoilage microbiota have shown that proteolytic and lipolytic bacteria such as Pseudomonas or Acinetobacter spp. predominate the spoilage bacterial populations. These bacteria can produce heat-stable proteases and lipases, which remain active after pasteurization and thus can spoil the milk during prolonged storage. Additionally, milk can become contaminated after pasteurization and therefore there is still a high demand on developing better cleaning and sanitation regimes and equipment, as well as test systems to (quantitatively) detect relevant pathogenic or spoilage microorganisms. Raw milk and raw milk cheese consumption is also increasing worldwide with the growing demand of minimally processed, sustainable, healthy, and local foods. In this context, emerging and re-emerging pathogens once again represent a major food safety challenge. As a result of global warming, it is conceivable that not only microbiological risks but also chemical risks relating to presence of mycotoxins or plant toxins in milk will increase. Herein, we provide an overview of the major microbial hazards occurring in the 21st century.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/1541-4337.12568

Microbial quality and safety of milk and milk products in the 21st century.

Mycobacteria include obligate and opportunistic pathogens that cause significant human and animal disease. The burden of tuberculosis has been largely reduced in developed territories but remains a huge problem worldwide. The significance of nontuberculous mycobacteria is growing considerably, especially in developed regions with higher life expectancy and more therapy-related immunosuppressed individuals. Due to their robustness mycobacteria can contaminate animal products by direct transmission from infected individuals or by environmental contamination during processing. The situation at market level is poorly known. Most studies analyzing commercially available foods are limited to a small or local scale and mainly focused on a particular mycobacterial species. There is a need to investigate if animal products that have passed the established controls to be for sale at main supermarkets could represent a route of contact with any mycobacteria. Thus, our goal was to study the prevalence of mycobacteria in these foods to assess if this could represent a source of human exposure. Five stores from the main supermarket chains in Spain were selected. 138 dairy and 119 meat products were purchased. All were processed using culture and multiplex real-time PCR methods. Additional molecular methods were used to specifically identify any positive result. Mycobacterium avium subsp. hominissuis (2), M. avium subsp. avium (1) and M. fortuitum (1) were isolated from powdered infant formula, chicken sausage, ground beef and mortadella cold cut, respectively. Mycobacterial DNA (M. avium, M. tuberculosis complex and other nontuberculous mycobacteria) was detected in 15% of dairy products and 2% of meat products. These results show that the prevalence of viable mycobacteria in foods of animal origin obtained at the supermarket was not substantial although a considerable proportion of them contained mycobacterial DNA. Contact with mycobacteria through this route could be ensured over time. Further investigation is necessary to determine the real impact of foodborne mycobacterial exposure on human health and identify critical points in the food production system to enable setting up more stringent control measures.

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Detection of mycobacteria by culture and DNA-based methods in animal-derived food products purchased at Spanish supermarkets.

Microbial safety status of Serro artisanal cheese produced in Brazil.

BACKGROUND The World Health Organization (WHO) has classified human zoonotic tuberculosis (TB) due to Mycobacterium bovis as a neglected issue in the developing world. In a recent cross-sectional study in Brazil, three of 189 TB patients presented with a coinfection of M. bovis and M. tuberculosis and were selected as cases for this study. OBJECTIVE The aim was to evaluate risk factors (RF) for zoonotic TB in an urban area of Brazil in order to guide preventive programmes. METHODS A matched case-control study was carried out nested within a cross-sectional study. For each of the three cases, 14 age- and sex-matched controls (TB due to M. tuberculosis) were selected. FINDINGS Zoonotic potential exposures (ZE) and extrapulmonary TB (EPTB) were independently associated with zoonotic TB in multivariate analyses. CONCLUSIONS ZE by occupation and consumption of raw milk and derivative products that place individuals in direct and indirect contact with animals and their excretions/secretions increase the risk for zoonotic TB in Brazil, especially among those with EPTB. Therefore, measures such as efficient control of bovine TB, distribution of pasteurised milk and its derivative products, and the diagnosis and monitoring of zoonotic TB in humans are essential steps, especially in developing countries where bovine TB is enzootic, and further studies are necessary.

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Risk factors for human Mycobacterium bovis infections in an urban area of Brazil

Zoonotic tuberculosis is a reemerging infectious disease in high-income countries and a neglected one in low- and middle-income countries. Despite major advances in its control as a result of milk pasteurization, its global burden is unknown, especially due the lack of surveillance data. Additionally, very little is known about control strategies. The purpose of this review was to contextualize the current knowledge about the epidemiology of zoonotic tuberculosis and to describe the available evidence regarding surveillance and control strategies in high-, middle-, and low-income countries. We conducted this review enriched by a One Health perspective, encompassing its inherent multifaceted characteristics. We found that the burden of zoonotic tuberculosis is likely to be underreported worldwide, with higher incidence in low-income countries, where the surveillance systems are even more fragile. Together with the lack of specific political commitment, surveillance data is affected by lack of a case definition and limitations of diagnostic methods. Control measures were dependent on risk factors and varied greatly between countries. This review supports the claim that a One Health approach is the most valuable concept to build capable surveillance systems, resulting in effective control measures. The disease characteristics and suggestions to implement surveillance and control programs are discussed.

/pdf/zoonotic-tuberculosis-in-humans-control-surveillance-and-the-1cugrh31ib.pdf

Zoonotic Tuberculosis in Humans: Control, Surveillance, and the One Health Approach.

Objective/background The present study was aimed at detecting Mycobacterium bovis in artisanal cheese using real-time quantitative polymerase chain reaction. Methods One hundred seven cheese samples (250 g) were purchased in 107 commercial establishments including neighborhood grocery stores, bakeries, and open-air markets from 19 municipalities of the state of Pernambuco, Brazil. Ten grams of each cheese sample were macerated with sterile saline solution in a sterile bag and DNA was extracted from 20 mg of the macerated material using the Wizard SV Genomic DNA Purification System. The quantitative polymerase chain reaction amplified a fragment corresponding to the region of difference 4 of M. bovis . Results Of the 107 samples analyzed, three (2.8%) were positive for M. bovis and their identities were confirmed by sequencing. This is perhaps the first report of the presence of M. bovis in artisanal cheese in the state of Pernambuco, Brazil. Conclusion The results of the present study highlight the need for improving sanitary measures during the production of artisanal cheese to prevent zoonotic tuberculosis in humans, resulting from the consumption of food contaminated with M. bovis .

Detection of Mycobacterium bovis in artisanal cheese in the state of Pernambuco, Brazil

The present study aimed to direct detect Mycobacterium bovis in milk (n = 401) and blood (n = 401) samples collected from 401 dairy cows of 20 properties located in the state of Pernambuco, Brazil, by real-time quantitative PCR (qPCR) targeting the region of difference 4 (RD4). Risk factors possibly associated with bovine tuberculosis (BTB) were also evaluated. Of the 802 samples analyzed, one milk (0.25 %) and eight blood (2 %) samples were positive for M. bovis in the qPCR and their identities were confirmed by sequencing. Animals positive for M. bovis were found in six (30 %) of the 20 properties visited. None of the risk factors evaluated were statistically associated with BTB. M. bovis DNA was detected in one milk sample what may pose a risk to public health because raw milk is commonly consumed in Brazil.

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Molecular detection of Mycobacterium bovis in cattle herds of the state of Pernambuco, Brazil

Paratuberculosis is a chronic and incurable disease that affects ruminants and other domestic animals. It is caused by Mycobacterium avium subsp. paratuberculosis (MAP) that may also be involved in some human diseases such as Crohn's disease, type 1 diabetes, sarcoidosis, multiple sclerosis, and Hashimoto's thyroiditis. The objective of this study was to investigate the occurrence of MAP DNA in samples of artisanal coalho cheese purchased in the State of Pernambuco. Forty samples of coalho cheese submitted to the Real Time Polymerase Chain Reaction (qPCR) technique were analyzed for the detection of the MAP region IS900. 11 (27.5%) were positive with a mean of 195.9 MAP colony forming unit (CFU) per gram of each sample, with a minimum of 30.3 CFU/g and a maximum of 324.2 CFU/g. Thus, this type of cheese that is one of the most consumed in this region of Brazil constitutes a source of human exposure to MAP. Further research in this area should be performed to evaluate the viability of the bacteria in this cheese type.

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Renata Duarte da Silva Cezar

Papers

Detection of Mycobacterium bovis in artisanal cheese in the state of Pernambuco, Brazil

Molecular detection of Mycobacterium bovis in cattle herds of the state of Pernambuco, Brazil

Occurrence of Mycobacterium avium subsp. paratuberculosis in coalho cheese in the State of Pernambuco, Brazil