**1. Introduction**

Most of the analysis techniques described in this chapter were recommended by the Agência Nacional de Vigilância Sanitária (the National Health Surveillance Agency) (ANVISA) [1], including the Official Analytical Methods for the Microbiological Analysis and Control of Products from Animal and Water Sources [2], those of the American Public Health Association; described in the fourth edition of the Compendium of Methods for the Microbiological Examination of Foods [3–5], the International Commission on Microbiological Specifications for Foods [6, 7]; the Food and Drug Administration (FDA), recommended by the Ministry of Agriculture for the analysis of foods of animal origin, in accordance with Normative Instruction number 62, dated August 26th, 2003 [2]; the Food Safety and Inspection Service of the US Department of Agriculture [8], the Association of the Official Methods of Analysis of AOAC International [9], the Bacteriological Analytical Manual (FDA) [10], the Microbiology Laboratory Guidebook [11] and the latest editions of the International Organization for Standardization [12, 13].

Among the various parameters that indicate the quality and safety of honey, the most important are those that define its microbiological characteristics. Honey, as with any other raw material of vegetable or animal origin, naturally presents microbial contaminants of commercial importance formed by microorganisms adapted to the characteristics of the honey, such as high-sugar content, low acidity and the presence of natural antimicrobial substances. Because of these characteristics, the microbial load in honey is generally low, below 10<sup>2</sup> CFU/g, and can even reach 10<sup>3</sup> –10<sup>4</sup> CFU/g. Consequently, it can cause undesirable changes by reducing the shelf-life of the product. It presents floral indicators of the possible presence of pathogenic microorganisms, and so can be harmful to the health of the consumer. Protecting food products from any kind of contamination or adulteration which can cause harm to public health or economic disorder is a global concern [8] and specific methods of analysis are required to evaluate this type of raw material. Moreover, the risks represented by the poor handling conditions used by workers responsible for the harvest, extraction and preparation of this product require effective interventions and procedures to minimize these risks [14].

Aiming to control the quality of honey, the World Trade Organization recommends the adoption of standards, guidelines and norms developed by Codex Alimentarius—the revised codex standard for honey 2001 [15]. This is an international public agency created by the Food and Agricultural Organization (FAO) and the World Health Organization (WHO) [16], both of which form part of the United Nations Organization (UNO). MERCOSUL GMC resolution n° 15 1994 approved the Technical Regulations for the Identity and Quality of Honey, based on resolutions n° 18 (1992) and n° 91 (1993) of the Common Market Group [17], in which honey can contain a maximum of 100 colony forming units of fungus per gram (CFU/g). Normative instruction no 11 approved, on 20 October 2000, the Technical Regulations for the Identity and Quality of Honey [18] and normative instruction no 3 dated January 19th 2001 approved the Technical Regulations for the Identity and Quality of bee apitoxin, beeswax, royal jelly, lyophilized royal jelly, bee pollen, propolis and propolis extract [19], as previously microbiological standards had not been established for these apiculture products. To ensure the credibility of the results however, some steps must be observed. The methods in this chapter are described as simply as possible in order to be accessible to fully qualified professionals, lab technicians and students with varying levels of education and training. This chapter provides comprehensive material presented in a didactic manner, with texts and diagrams that facilitate understanding. The basic techniques of microbiology described are accompanied by a brief overview of the microorganism researched in order to provide a solid theoretical basis, which will be of great value for understanding the method and interpretation of results. This chapter, therefore, aims to present the most commonly used techniques for assessing the microbiological characteristics of honey to identify its contaminant flora, its significance and its control in this type of food.
