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into the host genome, with the successful infection always resulting in the death of their host (Hagens and Loessner 2007). Since the first report of the use of phage for detection by Ulitzur and Kuhn (1987), different strategies have been described for the detection of *Salmonella*. Generally, the majority of methods described involve measuring the activity of a reporter gene (generally, the luciferase *lux* genes from *Vibrio fisherii*), cloned into a vector carried by a phage, and expressed only after infection (Kuhn et al. 2002; Thouand et al. 2008). Luciferase genes have the enormous advantage in that background noise or photon emission is absent from food samples and the luminescence, when detected, reflects the presence of viable target bacteria. Other approaches include use of an ice nucleation reporter phage (Wolber and Green 1990); concentration by IMS followed by phage mediated release of adenylate kinase (AK) (Blasco et al. 1998; Wu et al. 2001); fluorescently labelled phage (Jiang et al 2009); and an IMS-bacteriophage plaque formation assay requiring the addition of a virucide to inactivate free phage particles (Fravrin et al. 2001). The usefulness of phagebased cell wall recognition proteins for magnetic capture has also been recently described utilizing cell-wall-binding domains (CBDs) highly specific for recognition and binding to target cells surfaces (Kretzer et al. 2007; Korndoerfer et al. 2006; Loessner et al. 2002). Paramagnetic beads coated with CBD molecules were shown to outperform commercially available antibody-based magnetic beads with respect to sensitivity and percent recovery (Kretzer et al. 2007). An extension to this approach has been the use of phage-tail-associated recognition proteins for the immobilization of gram-negative cells (Galikowska et al. 2011). For example, BioMerieux. has recently introduced *Salmonella* Up, an automated ELISA based VIDAS assay using a phage recombinant protein derived from specific bacteriophage tail fibers for the detection of *Salmonella* in food and food ingredients within 18-24 hours

Although at present commercial phage based detection systems are limited, the technology may circumvent the problem of viability presented by PCR, while promising to be more

**6. Conclusions and future perspectives on** *Salmonella* **detection methods** 

A wide range of methods for the detection of *Salmonella* has been developed in the last decade and significant progress has been made in sample preparation techniques for improved isolation and detection of *Salmonella* in foods and food ingredients. The use of immunomagenetic separation technique which separates target organisms from background flora, is now routinely applied in various diagnostic labs for a variety of foodborne pathogens including *Salmonella*. This technique has increased the sensitivity of the detection of *Salmonella* in various types of food and food ingredients as well as environmental samples with high levels of background. Similarly, the application of molecular methods, immunological methods, and bacteriophage detection systems for *Salmonella* is now routine in many diagnostic food microbiology labs. Novel technologies such as the application of biosensors, microarrays, and nanotechnology are currently in the research stage and these are likely to become available for routine testing of food and

The application of rapid methods for the detection, identification, and characterization of *Salmonella* provides a useful tool for assessment of the safety of food products when used in conjunction with foodsafety programs such as the Hazard Analysis Critical Control Point (HACCP) program for the assessment of raw materials and food ingredients used in food

after enrichment in a non-selective broth.

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**18** 

*Poland* 

**Detection of** *Salmonella* **spp. Presence in Food** 

The analysis of food products for presence of pathogenic microorganisms is one of the basic steps to control safety and quality of food. Development of new, fast, and reliable identification methods for biological threats are necessary to meet the safety standards of food products and risk management. *Salmonella* spp., a marker of food products safety, is

The standard culture methods to detect the presence of microorganisms in food products are well developed; although these methods require 4 to 5 days to obtain presumptive positive or negative results. These tests are time-consuming and can take up to 7 days depending on the realization of biochemical and serological confirmations. In addition, sensitivity of cultures can be affected by antibiotic treatment, inadequate sampling, and a small number of

Standardized classical culture methods are still in use by many labs, especially by regulatory agencies, because they are harmonized methods, looked at as the "gold standards" in food diagnostics and thus overall well accepted. These are important aspects in international trade and compliance testing. A serious drawback is that, although they demand no expensive infrastructure and are rather cheap in consumables, they are laborious to perform, demand large volumes usage of liquid and solid media and reagents, and encompass time-

As an alternative to time-consuming culture methods, several approaches have been developed to accelerate detection of pathogenic microorganisms in food products. In the present work, besides the standard method of *Salmonella* spp. detection in food products

The first stage of microbiological analysis of food consists in taking and preparing a sample for analyses. Incorrect sampling can lead to obtaining false negative or false positive results. When talking about taking samples, the term "representative sample" is often used. The sample should reflect the image of the product from which it originates as precisely as possible. It is quite easy to take a representative sample from liquid products, e.g. milk, if the milk has been sufficiently mixed before taking the sample. On the other hand, when the subject of examination is a product of high viscosity, with slow flow or of a heterogeneous structure, then it is very difficult to assess the microbiological quality of the entire batch (e.g. a barrel or a

**1. Introduction** 

widely distributed foodborne pathogen.

viable microorganisms in samples.

**2. Taking samples for tests** 

consuming procedures both in operation and data collection.

(ISO 6579:2003) some alternative detection methods have been presented.

*University of Warmia and Mazury in Olsztyn, Faculty of Food Sciences* 

Anna Zadernowska and Wioleta Chajęcka

*Chair of Industrial and Food Microbiology* 

