Ole Mejlholm

Antimicrobial effect of essential oils on the seafood spoilage micro-organism Photobacterium phosphoreum in liquid media and fish products.

Aims: To evaluate the antimicrobial effect of nine essential oils (EO) on P. phosphoreum and determine the effect of oregano oil on the shelf‐life of modified atmosphere‐packed (MAP) cod fillets.

Importance of Photobacterium phosphoreum in relation to spoilage of modified atmosphere‐packed fish products

Occurrence and growth of Photobacterium phosphoreum were studied in 20 experiments with fresh fish from Denmark, Iceland and Greece. The organism was detected in all marine fish species but not in fish from fresh water. Growth of P. phosphoreum to high levels (>107 cfu g−1) was observed in most products and the organism is likely to be of importance for spoilage of several modified atmosphere‐packed (MAP) marine fish species when stored at chill temperatures. Some microbiological methods recommended for control of fish products by national and international authorities are inappropriate for detection of psychrotolerant and heat‐labile micro‐organisms like P. phosphoreum. These methods have been used in many previous studies of MAP fish and this could explain why, contrary to the findings in the present study, P. phosphoreum in general was not detected previously in spoiled MAP fish.

Shelf life and safety aspects of chilled cooked and peeled shrimps (Pandalus borealis) in modified atmosphere packaging

Aims:  To evaluate the growth of Listeria monocytogenes and shelf life of cooked and peeled shrimps in modified atmosphere packaging (MAP).

Application of an iterative approach for development of a microbial model predicting the shelf-life of packed fish

An iterative approach was used to develop a microbial model for shelf-life prediction of cod fillets packed in modified atmospheres. The effect of temperature (0-15 degrees C) and CO2 (0-100%) on growth of the specific spoilage organism, Photobacterium phosphoreum, was studied in packed cod and in liquid media. P. phosphoreum was a dominant part of the spoilage microflora of packed cod stored at the extremes of the range of conditions studied. The organism is therefore likely to be important for spoilage and the development of a microbial model within this domain seems relevant. A liquid medium was developed to provide growth kinetics of P. phosphoreum similar to those observed in packed cod. Using this medium, the effect of temperature and CO2 on the maximum specific growth rate of Photobacterium phosphoreum was determined by absorbance measurements and modelled by a square root equation and by a polynomial equation. Product validation studies were carried out during summer and winter using naturally contaminated packed cod fillets which were stored at constant and at changing temperatures. The shelf-life of the packed fillets was predicted on the basis of the initial numbers of P. phosphoreum, product temperature profiles and the level of CO2 in the modified atmosphere. The average deviations between shelf-life determined by sensory evaluation and shelf-life predicted by the square root equation and by the polynomial equation were 17% and 9%, respectively.

Predicting growth rates and growth boundary of Listeria monocytogenes - An international validation study with focus on processed and ready-to-eat meat and seafood.

The performance of six predictive models for Listeria monocytogenes was evaluated using 1014 growth responses of the pathogen in meat, seafood, poultry and dairy products. The performance of the growth models was closely related to their complexity i.e. the number of environmental parameters they take into account. The most complex model included the effect of nine environmental parameters and it performed better than the other less complex models both for prediction of maximum specific growth rates (micro(max) values) and for the growth boundary of L. monocytogenes. For this model bias and accuracy factors for growth rate predictions were 1.0 and 1.5, respectively, and 89% of the growth/no-growth responses were correctly predicted. The performance of three other models, including the effect of five to seven environmental parameters, was considered acceptable with bias factors of 1.2 to 1.3. These models all included the effect of acetic acid/diacetate and lactic acid, one of the models also included the effect of CO(2) and nitrite but none of these models included the effect of smoke components. Less complex models that did not include the effect of acetic acid/diacetate and lactic acid were unable to accurately predict growth responses of L. monocytogenes in the wide range of food evaluated in the present study. When complexity of L. monocytogenes growth models matches the complexity of foods of interest, i.e. the number of hurdles to microbial growth, then predicted growth responses of the pathogen can be accurate. The successfully validated models are useful for assessment and management of L. monocytogenes in processed and ready-to-eat (RTE) foods.

Modeling and predicting the growth of lactic acid bacteria in lightly preserved seafood and their inhibiting effect on Listeria monocytogenes.

A cardinal parameter model was developed to predict the effect of diacetate, lactate, CO2, smoke components (phenol), pH, NaCl, temperature, and the interactions between all parameters on the growth of lactic acid bacteria (LAB) in lightly preserved seafood. A product-oriented approach based on careful chemical characterization and growth of bacteria in ready-to-eat seafoods was used to develop this new LAB growth model. Initially, cardinal parameter values for the inhibiting effect of diacetate, lactate, CO2, pH, and NaCl-water activity were determined experimentally for a mixture of LAB isolates or were obtained from the literature. Next, these values and a cardinal parameter model were used to model the effect of temperature (T(min)) and smoke components (P(max)). The cardinal parameter model was fitted to data for growth of LAB (mu(max) values) in lightly preserved seafood including cold-smoked and marinated products with different concentrations of naturally occurring and added organic acids. Separate product validation studies of the LAB model resulted in average bias and accuracy factor values of 1.2 and 1.5, respectively, for growth of LAB (mu(max) values) in lightly preserved seafood. Interaction between LAB and Listeria monocytogenes was predicted by combining the developed LAB model and an existing growth and growth boundary model for the pathogen (O. Mejlholm and P. Dalgaard, J. Food Prot. 70:70-84). The performance of the existing L. monocytogenes model was improved by taking into account the effect of microbial interaction with LAB. The observed and predicted maximum population densities of L. monocytogenes in inoculated lightly preserved seafoods were 4.7 and 4.1 log CFU g(-1), respectively, whereas for naturally contaminated vacuum-packed cold-smoked salmon the corresponding values were 0.7 and 0.6 log CFU g(-1) when a relative lag time of 4.5 was used for the pathogen.

Modeling and predicting the growth boundary of Listeria monocytogenes in lightly preserved seafood.

The antimicrobial effect of diacetate and lactate against Listeria monocytogenes was evaluated in challenge tests with vacuum-packaged or modified atmosphere packaged (MAP) cold-smoked salmon, marinated salmon, cold-smoked Greenland halibut, marinated Greenland halibut, and gravad salmon. MAP cold-smoked salmon with the addition of 0.15% (wt/wt) diacetate prevented the growth of L. monocytogenes for more than 40 days at 8 degrees C, whereas the addition of 0.15% (wt/wt) diacetate reduced the growth rate of the pathogen in MAP cold-smoked Greenland halibut. This difference between the two types of products was explained by a higher content of naturally occurring lactate in cold-smoked salmon (0.77 to 0.98%, wt/wt) than in cold-smoked Greenland halibut (0.10 to 0.15%, wt/wt). In fact, the addition of 0.15% (wt/wt) diacetate and 0.75% (wt/wt) lactate to MAP cold-smoked Greenland halibut prevented the growth of L. monocytogenes for more than 45 days at 8 degrees C. A mathematical model that included the effect of diacetate, lactate, CO2, smoke components, nitrite, pH, NaCl, temperature, and interactions between all these parameters was developed to predict the growth boundary of L. monocytogenes in lightly preserved seafood. The developed growth boundary model accurately predicted growth and no-growth responses in 68 of 71 examined experiments from the present study as well as from literature data. Growth was predicted for three batches of naturally contaminated cold-smoked salmon when a no-growth response was actually observed, indicating that the model is fail-safe. The developed model predicts both the growth boundary and growth rate of L. monocytogenes and seems useful for the risk management of lightly preserved seafood. Particularly, the model facilitates the identification of product characteristics required to prevent the growth of L. monocytogenes, thereby making it possible to identify critical control points, and is useful for compliance with the new European Union regulation on ready-to-eat foods (EC 2073/2005).

Development and validation of an extensive growth and growth boundary model for Listeria monocytogenes in lightly preserved and ready-to-eat shrimp.

An existing cardinal parameter growth and growth boundary model for Listeria monocytogenes (O. Mejlholm and P. Dalgaard, J. Food Prot. 70:70-84 and 2485-2497, 2007) was expanded with terms for the effects of acetic, benzoic, citric, and sorbic acids to include a total of 12 environmental parameters and their interactive effects. The new model predicted growth rates (micro(max) values) of L. monocytogenes accurately with bias and accuracy factors of 1.0 and 1.5, respectively, for 16 batches of brined shrimp with benzoic, citric, and sorbic acids. Corresponding values of 0.9 and 1.2, respectively, were obtained for five batches of brined shrimp with acetic and lactic acids. Growth and no-growth responses of L. monocytogenes were also appropriately predicted with 88% correct prediction for 26 experiments with brined shrimp. The new model performed better than existing L. monocytogenes models with a comparable degree of complexity. The high number of environmental parameters, including six organic acids (acetic acid, benzoic acid, citric acid, diacetate, lactic acid, and sorbic acid), allows the new model to predict the effect of substituting one set of preserving parameters for another. The new model also allowed the distance between the growth boundary and specific product characteristics to be quantified by a psi value. This can be of practical importance in the development or reformulation of seafood with preserving parameters that prevent growth of L. monocytogenes and take variability in product characteristics into account.

Atmospheric pressure plasma produced inside a closed package by a dielectric barrier discharge in Ar/CO2 for bacterial inactivation of biological samples

The generation and evaluation of a dielectric barrier discharge produced inside a closed package made of a commercially available packaging film and filled with gas mixtures of Ar/CO2 at atmospheric pressure is reported. The discharge parameters were analysed by electrical measurements and optical emission spectroscopy in two modes of operation: trapped gas atmosphere and flowing gas atmosphere. Gas temperature was estimated using the OH(A–X) emission spectrum and the rotational temperature reached a saturation level after a few minutes of plasma running. The rotational temperature was almost three times higher in the Ar/CO2 plasma compared with an Ar plasma. The efficiency of the produced plasma for the inactivation of bacteria on food inside the closed package was investigated.

Microbial changes and growth of Listeria monocytogenes during chilled storage of brined shrimp (Pandalus borealis)

Thirteen storage trials and ten challenge tests were carried out to examine microbial changes, spoilage and the potential growth of Listeria monocytogenes in brined shrimp (Pandalus borealis). Shrimp in brine as well as brined and drained shrimp in modified atmosphere packaging (MAP) were produced and studied. Different recipes were used to study the effect of preserving parameters (organic acids, pH and NaCl) on growth of microorganisms and shelf life at 7-8 degrees C or 12 degrees C. Particularly, brines with different concentrations of (i) benzoic, citric and sorbic acids or (ii) acetic, citric and lactic acids were studied. Furthermore, the effect of adding diacetate to brined shrimp was evaluated. A single batch of cooked and peeled shrimp was used to study both industrially and manually processed brined shrimp with respect to the effect of process hygiene on microbial changes and the shelf life of products. Concentrations of microorganisms on newly produced brined shrimp from an industrial scale processing line were 1.0-2.3 log (CFU g(-1)) higher than comparable concentrations in manually processed samples. This resulted in a substantially shorter shelf life and a more diverse spoilage microflora of the industrially processed brined shrimp. In addition, shelf life of brined shrimp was affected by the types and concentrations of organic acids and by the storage temperature as expected. The effect of MAP was less pronounced. Eighty-two isolates from the spoilage microflora of brined shrimp were identified and they included 53 lactic acid bacteria, 6 coagulase negative Staphylococcus spp., 18 Pseudomonas fluorescens and 5 yeast isolates. After storage at 7 degrees C, P. fluorescens, Enterococcus-like isolates, E. malodoratus, Carnobacterium maltaromaticum, coagulase negative Staphylococcus spp. and Lactobacillus sakei constituted the dominating microflora of shrimp in brines that contained benzoic, citric and sorbic acids as preservatives. L. sakei dominated the spoilage microflora of brined and drained MAP shrimp, and of brined shrimp preserved using acetic, citric and lactic acids, irrespective of packaging conditions. Shrimp in brine with benzoic, citric and sorbic acids prevented growth of L. monocytogenes during more than 40 days at 7 degrees C when the preserving parameters resembled those of commercial products. However, small changes in the preserving parameters and, particularly, reduced concentrations of benzoic acid led to growth of L. monocytogenes in brined shrimp. The present study provides significant new information on microbial changes, shelf life and growth of L. monocytogenes in brined shrimp. This information can facilitate development of new and safe brined shrimp products.