Quantitative risk assessment of listeriosis from traditional brazilian minas artisanal semi-hard and fresh soft cheeses

Traditional Minas cheeses are very popular in Brazil. These cheeses are of two types: artisanal ripened semi-hard cheeses (HC) produced with raw milk and refrigerated fresh soft cheeses (SC) generally produced with pasteurized milk. Purpose: This study estimates the risk of listeriosis due to HC and SC consumption using quantitative microbial risk assessment (QMRA). Methods: QMRAs were developed for both cheese types. The HC model contained a module for predicting Listeria monocytogenes decline during ripening. The SC model contained a refrigerated storage module for predicting L. monocytogenes growth during storage. HC modeling scenarios varied L. monocytogenes starting concentration over -2.4 to 6 log CFU/ml in raw milk and three ripening times (4, 22, and 60 days). SC modeling scenarios varied L. monocytogenes starting concentration (-2.4 to 4 log CFU/ml in milk). Inclusion of antilisterial lactic acid bacteria (LAB) was also examined. Simulations (100,000 iterations per scenario) were carried out using the @Risk add-in for Excel. Results: Aging HC reduced risk, and risk was influenced by L. monocytogenes starting concentration. Aging 22 days with inhibitory LAB reduced risk more than 4 million-fold when L. monocytogenes was assumed to be 6 log CFU/ml in raw milk, and was lower risk than HC made without LAB and with a starting concentration of 1 log CFU/ml in raw milk. Inclusion of inhibitory LAB reduced risk of listeriosis from SC, but not as dramatically as for HC. Relative risk to vulnerable populations reduced 4.4 and 3.4-fold when L. monocytogenes starting concentrations were 1 log and 4 log CFU/ml in milk, respectively. Significance: The results of the QMRA predict that consumption of contaminated Minas cheeses can present a high risk of listeriosis, especially for vulnerable populations. Scenario analyses indicate that aging of HC and inclusion of LAB in HC and SC are effective risk mitigation measures.info:eu-repo/semantics/publishedVersio

External Load Affects Ground Reaction Force Parameters Non-uniformly during Running in Weightlessness

Long-term exposure to microgravity induces detrimefits to the musculcskdetal system (Schneider et al., 1995; LeBlanc et al., 2000). Treadmill exercise is used onboard the International Space Station as an exercise countermeasure to musculoskeletal deconditioning due to spaceflight. During locomotive exercise in weightlessness (0G), crewmembers wear a harness attached to an external loading mechanism (EL). The EL pulls the crewmember toward the treadmill, and provides resistive load during the impact and propulsive phases of gait. The resulting forces may be important in stimulating bone maintenance (Turner, 1998). The EL can be applied via a bungee and carabineer clip configuration attached to the harness and can be manipulated to create varying amounts of load levels during exercise. Ground-based research performed using a vertically mounted treadmill found that peak ground reaction forces (GRF) during running at an EL of less than one body weight (BW) are less than those that occur during running in normal gravity (1G) (Davis et al., 1996). However, it is not known how the GRF are affected by the EL in a true OG environment. Locomotion while suspended may result in biomechanics that differ from free running. The purpose of this investigation was to determine how EL affects peak impact force, peak propulsive force, loading rate, and impulse of the GRF during running in 0G. It was hypothesized that increasing EL would result in increases in each GRF parameter

Quantitative risk assessment of Listeria monocytogenes in traditional Minas cheeses: the cases of artisanal semi-hard and fresh soft cheeses

This study estimated the risk of listeriosis from Brazilian cheese consumption using quantitative microbial risk assessment (QMRA). Risks associated to consumption of two cheese types were assessed: artisanal ripened semihard cheese (produced with raw milk) and refrigerated fresh soft cheese (produced with pasteurized milk). The semi-hard cheese model predicted Listeria monocytogenes growth or decline during ripening, while the soft cheese model predicted pathogen growth during refrigerated storage. Semi-hard cheese modeling scenarios considered L. monocytogenes starting concentration from −2.4 to 6 log CFU/mL in raw milk and three ripening times (4, 22 and 60 days). Soft cheese modeling scenarios considered L. monocytogenes starting concentration from −2.4 to 4 log CFU/mL in milk. The inclusion of anti-listerial lactic acid bacteria (LAB) in cheeses was also examined. Risk of listeriosis due to consumption of soft cheese was 6000 and 190 times greater than that of semi-hard cheese, for general and vulnerable populations, respectively. Aging semi-hard cheese reduced risk, and risk was influenced by L. monocytogenes starting concentration. Aging cheese with inhibitory LAB for 22 days reduced risk over 4 million-fold when L. monocytogenes was assumed to be 6 log CFU/mL in raw milk. The inclusion of inhibitory LAB also reduced risk of listeriosis due to soft cheese consumption, but not as much as for semi-hard cheese. QMRA results predicted that consumption of contaminated cheeses can carry a high risk of listeriosis, especially for vulnerable populations. Scenario analyses indicated that aging of semi-hard cheese and inclusion of antimicrobial LAB mix in semi-hard and soft cheeses are effective risk mitigation measures.The authors thank to Fundação de Amparo à Pesquisa do Estado de São Paulo for the financial support (Grants 2014/14891-7 and 2016/ 09346-5) and to Conselho Nacional de Desenvolvimento Científico e Tecnológico (Grant # 302763/2014-7 and #305804/2017-0), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Grant #33003017027P1). Dr. Gonzales-Barron wishes to acknowledge the financial support provided by the Portuguese Foundation for Science and Technology (FCT) through the award of a five-year Investigator Fellowship (IF) in the mode of Development Grants (IF/00570).info:eu-repo/semantics/publishedVersio

Loading Configurations and Ground Reaction Forces During Treadmill Running in Weightlessness

Studies have shown losses in bone mineral density of 1-2% per month in critical weight bearing areas such as the proximal femur during long-term space flight (Grigoriev, 1998). The astronauts currently onboard the International Space Station (ISS) use a treadmill as an exercise countermeasure to bone loss that occurs as a result of prolonged exposure to weightlessness. A crewmember exercising on the treadmill is attached by a harness and loading device. Ground reaction forces are obtained through the loading device that pulls the crewn1ember towards the treadmill surface during locomotion. McCrory et al. (2002) found that the magnitude of the peak ground reaction force (pGRF) during horizontal suspension running, or simulated weightlessness, was directly related to the load applied to the subject. It is thought that strain magnitude and strain rate affects osteogenesis, and is a function of the magnitude and rate of change of the ground reaction force. While it is not known if a minimum stimulus exists for osteogenesis, it has been hypothesized that in order to replicate the bone formation occurring in normal gravity (1 G), the exercise in weightlessness should mimic the forces that occur on earth. Specifically, the pGRF obtained in weightlessness should be comparable to that achieved in 1 G

Behavior of Listeria monocytogenes in the presence or not of intentionally-added lactic acid bacteria during ripening of artisanal Minas semi-hard cheese

The fate of Listeria monocytogenes during ripening of artisanal Minas semi-hard cheese, as influenced by cheese intrinsic properties and by autochthonous (naturally present) or intentionally-added anti-listerial lactic acid bacteria (LAB) was modeled. Selected LAB strains with anti-listerial capacity were added or not to raw or pasteurized milk to prepare 4 cheese treatments. Counts of LAB and L. monocytogenes, pH, temperature and water activity were determined throughout cheese ripening (22 days, 22±1ᵒC). Different approaches were adopted to model the effect of LAB on L. monocytogenes: an independent approach using the Huang primary model to describe LAB growth and the linear decay model to describe pathogen inactivation; the Huang-Cardinal [pH] model using the effect of pH variation in a dynamic tertiary approach; and the Jameson-effect with Nmax tot model which simultaneously describes L. monocytogenes and LAB fate. L. monocytogenes inactivation occurred in both treatments with added LAB and inactivation was faster in raw milk cheese (−0.0260 h−1) vs. pasteurized milk cheese (−0.0182 h−1), as estimated by the linear decay model. Better goodness-of-fit was achieved for the cheeses without added LAB when the Huang primary model was used. A faster and great pH decline was detected for cheeses with added LAB, and the Huang-Cardinal [pH] model predicted higher pathogen growth rate in cheese produced with raw milk, but greater L. monocytogenes final concentration in pasteurized milk cheese. The Jameson-effect model with Nmax tot predicted that LAB suppressed pathogen growth in all treatments, except in the treatment with pasteurized milk and no LAB addition. The Huang-Cardinal [pH] model was more accurate in modeling L. monocytogenes kinetics as a function of pH changes than was the Jameson-effect model with Nmax tot as a function of LAB inhibitory effect based on the goodness-of-fit measures. The Jameson-effect model may however be a better competition model since it can more easily represent L. monocytogenes growth and death. This study presents crucial kinetic data on L. monocytogenes behavior in the presence of competing microbiota in Minas semi-hard cheese under dynamic conditions.This work was supported by The São Paulo Research Foundation – FAPESP [grant numbers 2014/14891–7 and 2016/09346–5]; CNPq - The National Council for Scientific and Technological Development [grant numbers 302763/2014–7 and 305804/2017–0]; and CAPES –Coordination for the Improvement of Higher Education Personnel [Finance Code 001].info:eu-repo/semantics/publishedVersio

Modelling growth and decline in a two-species model system: Pathogenic Escherichia coli O157:H7 and psychrotrophic spoilage bacteria in milk

Producción CientíficaShiga toxin-producing Escherichia coli O157:H7 is a food-borne pathogen and the major cause of hemorrhagic colitis. Pseudomonas is the genus most frequent psychrotrophic spoilage microorganisms present in milk. Two-species bacterial systems with E. coli O157:H7, non-pathogenic E. coli, and P. fluorescens in skimmed milk at 7, 13, 19, or 25 °C were studied. Bacterial interactions were modelled after applying a Bayesian approach. No direct correlation between P. fluorescens’s growth rate and its effect on the maximum population densities of E. coli species was found. The results show the complexity of the interactions between two species in a food model. The use of natural microbiota members to control foodborne pathogens could be useful to improve food safety during the processing and storage of refrigerated foods

Quantitative Microbial Risk Assessment of Antibacterial Hand Hygiene Products on Risk of Shigellosis

ABSTRACT There are conflicting reports on whether antibacterial hand hygiene products are more effective than nonantibacterial products in reducing bacteria on hands and preventing disease. This research used new laboratory data, together with simulation techniques, to compare the ability of nonantibacterial and antibacterial products to reduce shigellosis risk. One hundred sixtythree subjects were used to compare five different hand treatments: two nonantibacterial products and three antibacterial products, i.e., 0.46% triclosan, 4% chlorhexidine gluconate, or 62% ethyl alcohol. Hands were inoculated with 5.5 to 6 log CFU Shigella; the simulated food handlers then washed their hands with one of the five products before handling melon balls

Selection of indigenous lactic acid bacteria presenting anti-listerial activity, and their role in reducing the maturation period and assuring the safety of traditional Brazilian cheeses

Artisanal raw milk cheeses are highly appreciated dairy products in Brazil and ensuring their microbiological safety has been a great need. This study reports the isolation and characterization of lactic acid bacteria (LAB) strains with anti-listerial activity, and their effects on Listeria monocytogenes during refrigerated shelf-life of soft Minas cheese and ripening of semi-hard Minas cheese. LAB strains (n ¼ 891) isolated from Minas artisanal cheeses (n ¼ 244) were assessed for anti-listerial activity by deferred antagonism assay at 37 C and 7 C. The treatments comprised the production of soft or semi-hard Minas cheeses using raw or pasteurized milk, and including the addition of selected LAB only [Lactobacillus brevis 2-392, Lactobacillus plantarum 1-399 and 4 Enterococcus faecalis (1-37, 2-49, 2-388 and 1-400)], L. monocytogenes only, selected LAB co-inoculated with L. monocytogenes, or without any added cultures. At 37 C, 48.1% of LAB isolates showed anti-listerial capacity and 77.5% maintained activity at 7 C. Selected LAB strains presented a bacteriostatic effect on L. monocytogenes in soft cheese. L. monocytogenes was inactivated during the ripening of semi-hard cheeses by the mix of LAB added. Times to attain a 4 log-reduction of L. monocytogenes were 15 and 21 days for semi-hard cheeses produced with raw and pasteurized milk, respectively. LAB with anti-listerial activity isolated from artisanal Minas cheeses can comprise an additional barrier to L. monocytogenes growth during the refrigerated storage of soft cheese and help shorten the ripening period of semi-hard cheeses aged at ambient temperature.The authors thank the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) for financial support (Grants 13/ 20456-9, 14/14891-7, 15/25641-4, 16/21041-5, 16/12406-0 and 16/ 09346-5). Dr. Gonzales-Barron also acknowledges the financial support provided by the Portuguese Foundation for Science and Technology (FCT) through the award of an Investigator Fellowship (IF) in the mode of Development Grants (IF/00570). A.S. Sant'Ana acknowledges the financial support of “Conselho Nacional de Desenvolvimento Científico e Tecnológico” (CNPq) (Grant #302763/ 2014-7) and CAPES (Grant #33003017027P1).info:eu-repo/semantics/publishedVersio