FOODBORNE PATHOGENS AND DISEASE Volume 11, Number 6, 2014 ª Mary Ann Liebert, Inc. DOI: 10.1089/fpd.2013.1712
Risky Consumption Habits and Safety of Fluid Milk Available in Retail Sales Outlets in Vic¸osa, Minas Gerais State, Brazil Fabio Alessandro Pieri,1,2 Monique Colombo,1 Carolina Milner Merhi,1 Vinı´cius Augusto Juliati,1 Marcello Sebe Ferreira,1 Marcelo Antoˆnio Nero,3 and Luis Augusto Nero1
Abstract
This study aimed to assess raw milk consumption habits in the urban population of Vic¸osa, Minas Gerais, Brazil, and the microbiological safety and quality of the fluid milk available in retail sales outlets in the same region. A simplified questionnaire regarding raw milk consumption was applied to the persons responsible for food acquisition in 411 residences. The regular consumption of raw milk was observed by 18.5% of the interviewers, and lack of knowledge of possible risks related to this food product. Microbiological safety and quality were assessed for raw (n = 69), pasteurized (n = 80), and ultra-high-temperature (UHT)-treated milk (n = 80) by analyzing the counts of mesophilic aerobes, coliforms, and Escherichia coli, and detection of Listeria monocytogenes and Salmonella spp.; raw milk samples were also subjected to enumeration of coagulasepositive Staphylococcus. Concerning raw milk, 59.4% of the samples were considered as produced in inadequate hygienic conditions, 5.8% of the samples presented counts of coagulase-positive Staphylococcus lower than 100 colony-forming units (CFU)/mL, and no samples presented with positive results for L. monocytogenes or Salmonella spp. All pasteurized and UHT milk samples presented with low counts of mesophilic aerobes and coliforms, while L. monocytogenes and Salmonella spp. were absent. The data demonstrated that raw milk was consumed by the population studied. Despite the absence of potential hazards, raw milk was of poor hygienic quality, in contrast with the processed fluid milk available in retail sales outlets that was safe and of good hygienic quality, highlighting the suitability of pasteurized and UHT milk for human consumption.
Introduction
T
he control and monitoring of foodborne diseases are a challenge for public health agencies in several countries (Angelillo et al., 2000; Kennedy et al., 2005). Particular concern is related to the consumption of animal-origin foods, mainly raw, as they are susceptible to contamination by zoonotic agents and foodborne pathogens in distinct steps of production and processing (Angelillo et al., 2000; De Buyser et al., 2001). In this context, raw milk and related products are considered relevant sources of such pathogenic agents, mainly due to the usual belief that these foods are healthier and more natural than processed foods, commonly observed in several countries including Brazil (Katafiasz and Bartlett, 2012; Nero et al., 2004). Currently in Brazil, retail sales of raw milk are not allowed, and restrictive rules and guidelines were published in order to provide proper monitoring of its quality and control of spe1
Departamento Departamento Valadares, Minas 3 Departamento nambuco, Brazil. 2
cific animal diseases (Brasil, 2002, 2011). However, it can be observed that the consumption of raw milk and its products is common in distinct Brazilian regions (Cruz and Menasche, 2014; Nero et al., 2003; Olival et al., 2002; personal observation). Despite having observed this scenario in Brazil, there are no official data that associate the consumption of contaminated foods with foodborne diseases. It is difficult to estimate the actual number of outbreaks of food poisoning caused by the consumption of raw milk in Brazil, because only a small number of foodborne disease cases and outbreaks are confirmed, due to both the major deficiency of epidemiological surveillance services and the lack of awareness of the population regarding this risk (Franco et al., 2003). Studies show that even in developed countries where the consumption of raw milk is restricted to a small portion of the population (CDC, 2007), this product or cheese made from raw milk is responsible for about 70% of foodborne disease outbreaks
de Veterina´ria, Universidade Federal de Vic¸osa, Vic¸osa, Minas Gerais, Brazil. de Cieˆncias Ba´sicas da Sau´de, Caˆmpus Governador Valadares, Universidade Federal de Juiz de Fora, Governador Gerais, Brazil. de Engenharia Cartogra´fica, Universidade Federal de Pernambuco, Centro de Tecnologia e Geocieˆncias, Recife, Per-
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related to dairy products (Robinson et al., 2011). Thus, these numbers may represent a very worrying percentage in countries like Brazil, where raw milk is consumed by a great part of the population. In this context, studies that describe the consumer habits of a specific population are valuable for determining a profile of risky habits, which can be used as guidance for proper control and strategic policies concerning consumption of raw foods. In addition, evaluation of the safety and quality of raw foods is relevant and provides scientific data to assess the hazards associated with these products. In this context, the present study aimed to determine the risky consumption habits regarding raw milk by the population in Vic¸osa, Minas Gerais (MG) state, Brazil, and to assess the quality and safety of the fluid milk available in retail sales. Materials and Methods Consumption habits of the population
The present study was conducted in the urban area of Vic¸osa city, located in MG state in the southeastern region of Brazil (Fig. 1). In the 2008 census (Secretaria de Planejamento of Vic¸osa municipality, personal communication), 14,586 residences were registered in the urban area of Vic¸osa; this value was used as a reference to determine the sampling size of the epidemiological study (EpiInfo 3.3.2., Centers for Disease Control and Prevention, Atlanta, GA). In this study, the following parameters were considered: 30% as the expected frequency of raw milk consumption; 25–35% as the acceptable frequency; and 95% as the confidence level. Then, the urban area of Vic¸osa was divided into 10 areas based on established municipal districts, and the total sampling number was divided proportionally to the number of
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residences in each region. In each region, the sampled residences were chosen based on the established sampling size to obtain a homogeneous distribution. To provide a characterization of the consumption of raw milk, a simplified questionnaire was applied in the selected residences. The questionnaire that was applied to the person responsible for food acquisition in each residence included basic demographic data and information about raw milk consumption, knowledge of risk-related foods, and basic knowledge of food inspection practices (Table 1). During the interviews, the study residences were georeferenced using a Global Positioning System (eTrex Vista Cx, Garmin International Inc., Olate, KS) that provided coordinates accurate to within 10–15 m. The frequencies of the recorded responses were calculated using EpiInfo 3.3.2. software (CDC). The resulting data were plotted in a cartographic representation of Vic¸osa city, and thematic maps were created using MapInfo Professional 10.0 software (Pitney Bowes Inc., Troy, NY) to provide a clear visualization of the recorded parameters. The epidemiological investigation included in this part of the study was submitted to the Ethics Committee for Research in Humans of the Universidade Federal de Vic¸osa, and was approved according the protocol 060/2006. Microbiological safety and quality of fluid milk
Samples of raw milk (n = 69), pasteurized milk (n = 80), and UHT-treated milk (n = 80) were obtained from retail sales in Vic¸osa, MG, Brazil. Raw milk samples were obtained from distinct suppliers that usually delivered this product directly to consumer homes, without refrigeration. Processed fluid milk samples were obtained from retail stores in their current storage conditions (pasteurized: refrigerated, UHT: room
FIG. 1. Location of Minas Gerais state in Southeastern region of Brazil (A); Location of Vic¸osa municipality in Minas Gerais state (B); The urban area of Vic¸osa city, divided into 10 regions and the number of residences included in the study for each region (C).
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Table 1. Simplified Description of the Epidemiological Questionnaire About Consumption of Noninspected Foods of Animal Origin Applied to the Responsibility of Foods Acquisition in the Residences from the Urban Area of Vic¸osa, Minas Gerais, Brazil Questions 1. Basic identification Area, geographic coordinators 2. Milk consumption Raw milk consumption Knowledge about related hazards 3. Additional information Identification of inspection stamp
Possible answers Detailed description Yes/no Yes/no Yes/no
Results Consumption habits of the population
The initial sample size defined by the study parameters was 316 residences. However, to guarantee homogeneous distribution of the interviews, a total of 411 residences were visited (Fig. 1). The responses to each question are shown in Table 2. These results document the risks related to raw milk consumption in the urban area of Vic¸osa. Identification of inspection stamps on food was reported by only 153 individuals responsible for food acquisition; this result may be a matter of concern for public health agencies. The thematic maps created from the study data on raw milk consumption, knowledge of the health risks of these habits, and checks for inspection stamps are presented in Figure 2. Microbiological safety and quality of fluid milk
temperature); all processed fluid milk samples were produced by dairy industries regularly inspected by federal inspection. All samples were kept under refrigerated conditions until laboratory analysis, when they were subjected to microbiological analysis to count mesophilic aerobes, coliforms, and E. coli, and to detect L. monocytogenes and Salmonella spp. In addition, raw milk samples were also subjected to enumeration of coagulase-positive Staphylococcus. All samples were subjected to a 10-fold dilution series in NaCl 0.85% (wt/vol) and selected dilutions were plated on Petrifilm AC and Petrifilm EC (3M Microbiology, St. Paul, MN), followed by incubation at 35C for 24 and 48 h, when the colonies were counted. Final results were expressed as CFU/mL. L. monocytogenes was detected according to the protocol described in the Bacteriological Analytical Manual (BAM) (FDA, 2010), by transferring 25 mL of each sample to Listeria Enrichment Broth (Oxoid Ltd., Basingstoke, England) and incubating at 30C for 24–48 h. Aliquots of the obtained cultures were streaked on Palcam agar (Oxoid) and Oxford agar (Oxoid) and incubated at 35C for 24–48 h. When present, suspected colonies of Listeria spp. were subjected to biochemical identification according to the FDA (2010). Salmonella spp. were investigated as described in the BAM (FDA, 2010), by transferring 25 mL of each sample to lactose broth (Oxoid) and incubating at 35C for 16 h. An aliquot of 1.0 mL was transferred to tetrathionate broth (Oxoid) and incubated at 35C for 24 h, and 0.1 mL was transferred to Rappaport-Vassiliadis broth (Oxoid) and incubated at 42.5C for 24 h. Aliquots of the obtained cultures were streaked on xylose lysine deoxycholate agar (Oxoid), Brilliant Green Agar (Oxoid), and Hektoen enteric agar (Oxoid), followed by incubation at 35C for 24 h. Presumptive Salmonella spp. suspected colonies were plated on triple sugar iron agar (Oxoid) and lysine iron agar (Oxoid) and incubated at 35C for 24 h. Cultures that showed typical reactions were subjected to serological identification with flagellar and somatic polyvalent antisera (Probac Brazil, Sa˜o Paulo, SP, Brazil). Coagulase-positive Staphylococcus were enumerated according to ISO 6888-2 (ISO, 1999). Pour plates were produced in duplicate for selected dilutions using rabbit plasma fibrinogen agar (BioMe´rieux S.A., Marcy l’Etoile, France), followed by incubation at 35C for 48 h, when typical colonies were counted. The results were expressed as CFU/mL.
The results of the microbiological analyses of samples of raw, pasteurized, and UHT milk are shown in Table 3. None of the samples showed a positive result for L. monocytogenes or Salmonella spp. The obtained data demonstrate the overall quality of the fluid milk available in retail sales outlets in Vic¸osa, MG, Brazil. Discussion
The results of the epidemiological questionnaire applied in the urban area of Vic¸osa identified the primary risky feeding habits related to raw milk consumption in the population (Table 2). Identifying associations between feeding habits and risk factors is the most efficient strategy for controlling foodborne disease outbreaks and cases (Tauxe et al., 2010). This strategy has been adopted by various countries to prevent these diseases (Lin et al., 2005; McMeekin et al., 2006; Broner et al., 2010; Wang et al., 2010). Studies of associations between feeding habits and risk factors have been conducted in other Brazilian regions (Olival et al., 2002; Nero et al., 2003), with similar results concerning raw milk. Using the thematic maps as a reference, it is easy to locate the specific city areas where the consumption of raw milk was more prevalent. For example, consumption of raw milk at frequencies higher than 25% was clearly identified in regions I, VII, VIII, and IX (Fig. 2). Relatively high consumption levels of raw milk have also been observed in similar studies
Table 2. Frequencies of Positive Responses to the Epidemiological Questionnaire Administered in 411 Residences in the Urban Area of Vic¸osa City; The Respondents Were the Persons Responsible for Acquisition of Food
Questions 1. Basic identification Area, geographic coordinators 2. Milk and dairy consumption Raw milk consumption Knowledge about related hazards 3. Additional information Identification of inspection stamp
Frequencies of positive answers/interviews Not applicable 76/411 183/411 153/411
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FIG. 2. Thematic maps of Vic¸osa city plotted showing (A) raw milk consumption in each region of the city; (B) knowledge about raw-milk–related hazards; (C) population habits in checking for inspection stamps in milks before acquisition; and (D) relation between consumption of raw milk and knowledge about raw-milk–related hazards. in other Brazilian regions (Olival et al., 2002; Nero et al., 2003). In areas I, VII, VIII, and IX, the data also revealed higher frequencies of a lack of knowledge about raw milk– related risks (Fig. 2). Finally, it was observed that, in general, fewer than 50% of those responsible for food acquisition checked for inspection stamps on the obtained milk (Fig. 2); this is a matter of concern due to the hazards that noninspected food can carry (Franco et al., 2013). As a result of analysis of the thematic maps, ‘‘raw milk consumption’’ and ‘‘knowledge of raw milk-related risks’’ answers were plotted together on a dot map (Fig. 2). Again, the data demonstrated that raw milk consumption can be associated with a lack of knowledge of the related risks (Fig. 2). The consumption of raw milk is a challenge to public health agencies because it is strongly associated with personal beliefs about health and, specifically, about the beneficial effects of raw milk on consumers (Katafiasz and Bartlett, 2012). Thus, visual representation of data can add
relevant support when statistical analysis is not able to detect significant relationships between risk-related habits and the occurrence of foodborne or other diseases (Broner et al., 2010; Wang et al., 2010). The knowledge of consumer feeding habits is a key point in controlling the occurrence of foodborne diseases (Grace et al., 2008), and information about the quality and hazards of related foods must also be considered for a holistic assessment of the risk for a specific population. Based on the obtained data (Table 3), it can be observed that 43 (62.3%) samples of raw milk presented mesophilic aerobe counts higher than 100,000 CFU/mL, the initial reference parameter for adequacy established by the Brazilian Ministry of Agriculture (Brasil, 2002), and a usual reference value of good manufacturing practices proposed by Robinson (2002). However, considering the current quality parameter for mesophilic aerobes in raw milk of 600,000 CFU/mL, from an update by the Brazilian Ministry of Agriculture (Brasil,
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Table 3. Numbers of Samples of Raw Milk Marketed Informally, Pasteurized and Ultrahigh Temperature (UHT) Milks in Vic¸osa, with Different Contamination Levels of Hygiene Indicators Milk Raw (n = 69)
Pasteurized (n = 80)
UHT (n = 80)
Count range (CFU/mL)
Mesophilic
Coliforms
Escherichia coli
Coagulase- positive Staphylococcus
< 100 100–1000 1000–10,000 10,000–100,000 100,000–1,000,000 > 1,000,000 < 100 100–1000 1,000–10,000 10,000–100,000 100,000–1,000,000 > 1,000,000 < 100 100–1000 1000–10,000 10,000–100,000 100,000–1,000,000 > 1,000,000
0 0 8 18 26 17 53 26 1 0 0 0 80 0 0 0 0 0
4 4 12 9 8 32 76 4 0 0 0 0 80 0 0 0 0 0
38 12 13 4 2 0 80a 0 0 0 0 0 80b 0 0 0 0 0
4 6 19 26 13 1 NT NT NT NT NT NT NT NT NT NT NT NT
a
E. coli count lower than 10 colony-forming units (CFU)/mL (n = 2), and lower than 1 CFU/mL (n = 78). E. coli count lower than 1 CFU/mL (n = 80). NT, not tested.
b
2011), 41 (59.4%) samples would be considered as having been produced under inadequate hygienic conditions. The overall poor quality of the raw milk can be confirmed by the observed high counts of coliforms (Table 3): only 4 (5.8%) raw milk samples presented counts lower than 100 CFU/mL, a reference parameter proposed by Robinson (2002) to indicate good hygienic practices. Despite the observed poor hygienic quality of raw milk samples (Robinson, 2002; Jay et al., 2005), 38 (55.1%) presented E. coli counts lower than 100 CFU/mL, and none of the samples presented positive results for L. monocytogenes or Salmonella spp. In contrast, only 4 (5.8%) raw milk samples presented counts of coagulase-positive Staphylococcus lower than 100 CFU/mL. The presence of coagulase-positive Staphylococcus at high concentrations may suggest the presence of enterotoxigenic strains capable of producing different types of enterotoxins with a potentially pathogenic effect on consumers (Carmo et al., 2002; Boerema et al., 2006; Srinivasan et al., 2006), but can also be considered as a good indicator of improper handling. Regarding the quality and safety of raw milk, similar results were also observed by studies conducted in distinct Brazilian regions (Moraes et al., 2005; Nero et al., 2005; Rosa and Queiros, 2007; Mattos et al., 2010), including studies that considered raw milk samples available for retail sales without official inspection (Beloti et al., 1999; Mendes et al., 2010). The obtained data confirmed the poor microbiological quality of the raw milk produced and available for retail sales in Vic¸osa, MG, Brazil, and demonstrated that, in general, this product does not fit the minimum required microbiological standards for reception and processing in processing industries (Robinson, 2002; Brasil, 2011). It is noteworthy that no relevant microbiological hazards were identified in the raw milk samples. Coagulase-positive Staphylococcus was detected at high levels in various samples (Table 3), and even then, it was not possible to associate
them directly with the presence of enterotoxins or the ability to produce them (Veras et al., 2008). However, the absence of these pathogens does not indicate that that raw milk is safe for consumers. Staphylococcal intoxication is caused by eating foods containing enterotoxins produced by enterotoxigenic Staphylococcus, of which S. aureus is the most prevalent etiological agent. Such poisoning is one of the most common types of foodborne illness (Le Loir et al., 2003). Staphylococcal food poisoning is attributed to ingestion of enterotoxins that are produced and released by bacteria in the food during their multiplication. These enterotoxins are heat stable and may remain in the food after cooking, favoring intoxication (Ostyn et al., 2010). Despite the observed overall poor quality of raw milk, the obtained counts for pasteurized and UHT milks indicated good production practices and proper processing in dairy industries (Table 3). As reference, the Brazilian legislation for processed fluid milk available for consumption establishes the control of thermotolerant coliforms (maximum of four most probable number per milliliter) and Salmonella spp. (absence), and for UHT milk, the absence of pathogenic and spoilage bacteria (without reference values) (Brasil, 2001). Considering the obtained data (Table 3), all pasteurized and UHT milk samples complied with the established reference parameters. In addition, all samples presented low counts of aerobic mesophiles, coliforms, and the absence of L. monocytogenes, confirming the adequacy of the processing in dairy industries. A study conducted by Bersot et al. (2010) identified 36 of 150 UHT milk samples with mesophilic aerobe counts higher than 100 CFU/mL, indicating inadequacy of processing according to Brazilian legislation. The absence of Salmonella spp. and E. coli can be considered a good indicator of the safety of the processed fluid milk available for retail sales in Vic¸osa, MG, Brazil, demonstrating the relevance of heat treatment of milk (Agarwal et al., 2012).
RAW MILK CONSUMPTION, QUALITY, AND SAFETY Conclusions
The data obtained allowed accurate identification of the risk-related habits in the population of the urban area of Vic¸osa, MG, Brazil regarding the consumption of raw milk. They demonstrated a lack of knowledge of the risks of raw milk, as well as a lack of knowledge regarding the need to verify food inspection stamps before consumption. As complementary information, the poor hygienic quality of the raw milk available for retail sales was demonstrated, but there was an absence of potential hazards with this product. In contrast, the processed fluid milk available for retail sales presented high hygienic quality and proper safety, highlighting the suitability of pasteurized and UHT milk for human consumption. Acknowledgments
The authors thank Conselho Nacional de Desenvolvimento Cientı´fico e Tecnolo´gico (CNPq), Coordenac¸a˜o de Aperfeic¸oamento Pessoal de Nı´vel Superior (CAPES), and Pesquisa do Estado de Minas Gerais (FAPEMIG). Disclosure Statement
No competing financial interests exist. References
Agarwal A, Awasthi V, Dua A, Ganguly S, Garg V, Marwaha SS. Microbiological profile of milk: Impact of household practices. Indian J Public Health 2012;56:88–94. Angelillo IF, Viggiani NMA, Rizzo L, Bianco A. Food handlers and foodborne diseases: Knowledge, attitudes, and reported behavior in Italy. J Food Prot 2000;63:381–385. Beloti V, Barros MAF, Souza JA, Nero LA, Santana EHW, Balarin O, Curiaki Y. Avaliac¸a˜o da qualidade do leite cru comercializado em Corne´lio Proco´pio, Parana´: Controle do consumo e da comercializac¸a˜o. Semina 1999;20:12–15. Bersot LS, Galva˜o JA, Raymundo NKL, Barcellos VC, Pinto JPAN, Maziero MT. Avaliac¸a˜o microbiolo´fica e fı´sico-quı´mica dos leite UHT produzidos no Estado do Parana´, Brasil. Semina 2010;31:645–652. Boerema JA, Clemens R, Brightwell G. Evaluation of molecular methods to determine enterotoxigenic status and molecular genotype of bovine, ovine, human and food isolates of Staphylococcus aureus. Int J Food Microbiol 2006;107:192–201. Brasil, ANVISA. Resoluc¸a˜o-RDC no 12, Aprova o regulamento te´cnico sobre padro˜es microbiolo´gicos para alimentos. January 2, 2001. In: Dia´rio Oficial da Unia˜o 2001, January 10, 1:45. (in Portuguese). Brasil, MAPA. Instruc¸a˜o normativa n 51, Aprova o Regulamento Te´cnico de Produc¸a˜o, Identidade e Qualidade do Leite tipo A, o Regulamento Te´cnico de Identidade e Qualidade de Leite Cru Refrigerado, o Regulamento Te´cnico de Identidade e Qualidade de Leite Pasteurizado e o Regulamento Te´cnico da Coleta de Leite Cru Refrigerado e seu Transporte a Granel. September 18, 2002. In: Dia´rio Oficial da Unia˜o 2002, September 20, 1:64. (in Portuguese). Brasil, MAPA. Instruc¸a˜o normativa n 62, Aprova os Regulamentos Te´cnicos de Produc¸a˜o, Identidade e Qualidade do Leite tipo A, do Leite Pasteurizado e do Leite Cru Refrigerado e o Regulamento Te´cnico da Coleta de Leite Cru Refrigerado e seu Transporte a Granel. December 29, 2011. In: Dia´rio Oficial da Unia˜o 2011, December 30, 1:24. (in Portuguese).
495
Broner S, Torner N, Dominguez A, Martinez A, Godoy P, Working Group Study Outbreaks A. Sociodemographic inequalities and outbreaks of foodborne diseases: An ecologic study. Food Control 2010;21:947–951. Carmo LS, Dias RS, Linardi VR, Sena MJ, Santos DA, Faria ME, Pena EC, Jett M, Heneine LG. Food poisoning due to enterotoxigenic strains of Staphylococcus present in Minas cheese and raw milk in Brazil. Food Microbiol 2002;19:9–14. [CDC] Centers for Disease Control and Prevention. Foodborne Diseases Active Surveillance Network (FoodNet): Population Survey Atlas of Exposures, 2006–2007. Atlanta: Centers for Disease Control and Prevention, 2007, 28 pp. Cruz FT, Menasche R. Tradition and diversity jeopardised by food safety regulations? The Serrano Cheese case, Campos de Cima da Serra region, Brazil. Food Pol;2014:45:116–124. De Buyser ML, Dufour B, Maire M, Lafarge V. Implication of milk and milk products in food-borne diseases in France and in different industrialised countries. Int J Food Microbiol 2001;67:1–17. [FDA] Food and Drug Administration. BAM—Bacteriological Analytical Manual. Silver Spring, MD: U.S. Food and Drug Administration, 2010. Franco BDGM, Landgraf M, Destro MT, Gelli DS. Foodborne Diseases in Southern South America. In: International Handbook of Foodborne Pathogens. Milliotis M, Bier J (eds.). New York: Marcel Dekker, 2003, pp. 733–743. Franco MM, Paes AC, Ribeiro MG, de Figueiredo Pantoja JC, Santos AC, Miyata M, Leite CQ, Motta RG, Listoni FJ. Occurrence of mycobacteria in bovine milk samples from both individual and collective bulk tanks at farms and informal markets in the southeast region of Sao Paulo, Brazil. BMC Vet Res 2013;9:85. Grace D, Randolph T, Olawoye J, Dipelou M, Kang’ethe E. Participatory risk assessment: A new approach for safer food in vulnerable African communities. Dev Pr 2008;18:4611–4618. [ISO] International Organization for Standardization. ISO 6888-2. Microbiology of Food and Animal Feeding Stuffs—Horizontal Method for the Enumeration of Coagulase Positive staphylococci (S. aureus and Other Species): Part 1. Technique Using Rabbit Plasma Fibrinogen Agar Medium. Geneva: International Organisation of Standardisation, 1999. Jay JM, Loessner MJ, Golden DA. Modern Food Microbiology, 7th ed. New York: Food Science Text Series, 2005. Katafiasz AR, Bartlett P. Motivation for Unpasteurized Milk Consumption in Michigan, 2011. Food Prot Trends 2012;32: 124–128. Kennedy J, Jackson V, Blair IS, McDowell DA, Cowan C, Bolton DJ. Food safety knowledge of consumers and the microbiological and temperature status of their refrigerators. J Food Prot 2005;68:1421–1430. Le Loir Y, Baron F, Gautier M. Staphylococcus aureus and food poisoning. Genet Mol Res 2003;2:63–76. Lin CTJ, Jensen KL, Yen ST. Awareness of foodborne pathogens among US consumers. Food Qual Pref 2005;16:401–412. Mattos MR, Beloti V, Tamanini R, Magnani DF, Nero LA, Barros MAF, Pires EMF, Paquereau BPD. Qualidade do leite cru produzido na regia˜o do agreste de Pernambuco, Brasil. Semina 2010;31:173–182. McMeekin TA, Baranyi J, Bowman J, Dalgaard P, Kirk M, Ross T, Schmid S, Zwietering MH. Information systems in food safety management. Int J Food Microbiol 2006;112: 181–194. Mendes CG, Sakamoto SM, Silva JBA, Ja´come CGM, Leite AI. Ana´lises fı´sico-quı´micas e pesquisa de fraude no leite infor-
496
mal comercializado no municı´pio de Mossoro´, RN. Rev Cienc Anim Bras 2010;11:349–356. Moraes CR, Fuertefria AM, Zaffari CB, Conte M, Rocha JPAV, Spanamberg A, Valente P, Corc¸a˜o G, Costa M. Qualidade microbiolo´gica de leite cru produzido em cinco municı´pios do Estado do Rio Grande do Sul, Brasil. Acta Sci Vet 2005;33:259–265. Nero LA, De Mattos MR, Beloti V, Barros MAF, Netto DP, Pinto JPAN, De Andrade NJ, Silva WP, Franco BDGM. Hazards in non-pasteurized milk on retail sale in Brazil: Prevalence of Salmonella spp, Listeria monocytogenes and chemical residues. Braz J Microbiol 2004;35:211–215. Nero LA, Mattos MR, Beloti V, Barros MAF, Pinto JPAN, Andrade NJ, Silva WP, Franco BDGM. Leite cru de quatro regio˜es leiteiras brasileiras: Perspectivas de atendimento dos requisitos microbiolo´gicos estabelecidos pela Instruc¸a˜o Normativa 51. Cienc Tecnol Aliment 2005;25:191–195. Nero LA, Maziero D, Bezerra MMS. Ha´bitos alimentares do consumidor de leite cru de Campo Moura˜o—PR. Semina 2003;24:21–26. Olival AA, Spexoto AA, Campos DFS, Ferreira F, Fonseca LFL, Santos MV, Dias RA. Ha´bitos de consumo do leite informal, associado ao risco de transmissa˜o de doenc¸as no municı´pio de Pirassununga, SP. Hig Alim 2002;16:35–40. Ostyn A, De Buyser ML, Guillier F, Groult J, Fe´lix B, Salah S, Delmas G, Hennekinne JA. First evidence of a food poisoning outbreak due to staphylococcal enterotoxin type E, France, 2009. Euro Surveil 2010;15:ID19528. Robinson RK. Dairy Microbiology Handbook: The Microbiology of Milk and Milk Products, 3rd ed. New York: WileyInterscience, 2002. Robinson T, Scheftel J, Smith K. Sporadic reportable enteric pathogen cases reporting domestic raw milk consumption, Minnesota, 2001–2010. In: Proceedings of the 100th Annual
PIERI ET AL.
Meeting of the International Association for Food Protection; 2011; Milwaukee, WI. Abstract T4-10. Rosa LD, Queiros MI. Avaliac¸a˜o da qualidade do leite cru e resfriado mediante a aplicac¸a˜o de princı´pios do APPCC. Cienc Tecnol Aliment 2007;27:422–430. Srinivasan V, Sawant AA, Gillespie BE, Headrick SJ, Ceasaris L, Oliver SP. Prevalence of enterotoxin and toxic shock syndrome toxin genes in Staphylococcus aureus isolated from milk of cows with mastitis. Foodborne Pathog Dis 2006; 3:274–283. Tauxe RV, Doyle MP, Kuchenmuller T, Schlundt J, Stein CE. Evolving public health approaches to the global challenge of foodborne infections. Int J Food Microbiol 2010;139:S16– S28. Veras JF, Carmo LS, Tong LC, Shuppe JW, Cummings C, Santos DA, Cerqueira MMCOP, Cantini A, Nicoli JR, Jett M. A study of the enterotoxigenicity of coagulase negative and coagulase-positive staphylococcal isolates from food poisoning outbreaks in Minas Gerais, Brazil. Int J Infect Dis 2008;12:410–415. Wang LG, Xu YY, Wang Y, Dong SC, Cao ZD, Zhou W, Sun HL, Huo DH, Zhang H, Sun YS, Huang LY, Yuan ZQ, Zeng DJ, Song HB. The epidemiological investigation and intelligent analytical system for foodborne disease. Food Control 2010;21:1466–1471.
Address correspondence to: Luis Augusto Nero, DSc Departamento de Veterina´ria Universidade Federal de Vic¸osa Campus UFV 36570-000, Vic¸osa, MG, Brazil E-mail: [email protected]
Risky Consumption Habits and Safety of Fluid Milk Available in Retail Sales Outlets in Vic¸osa, Minas Gerais State, Brazil Fabio Alessandro Pieri,1,2 Monique Colombo,1 Carolina Milner Merhi,1 Vinı´cius Augusto Juliati,1 Marcello Sebe Ferreira,1 Marcelo Antoˆnio Nero,3 and Luis Augusto Nero1
Abstract
This study aimed to assess raw milk consumption habits in the urban population of Vic¸osa, Minas Gerais, Brazil, and the microbiological safety and quality of the fluid milk available in retail sales outlets in the same region. A simplified questionnaire regarding raw milk consumption was applied to the persons responsible for food acquisition in 411 residences. The regular consumption of raw milk was observed by 18.5% of the interviewers, and lack of knowledge of possible risks related to this food product. Microbiological safety and quality were assessed for raw (n = 69), pasteurized (n = 80), and ultra-high-temperature (UHT)-treated milk (n = 80) by analyzing the counts of mesophilic aerobes, coliforms, and Escherichia coli, and detection of Listeria monocytogenes and Salmonella spp.; raw milk samples were also subjected to enumeration of coagulasepositive Staphylococcus. Concerning raw milk, 59.4% of the samples were considered as produced in inadequate hygienic conditions, 5.8% of the samples presented counts of coagulase-positive Staphylococcus lower than 100 colony-forming units (CFU)/mL, and no samples presented with positive results for L. monocytogenes or Salmonella spp. All pasteurized and UHT milk samples presented with low counts of mesophilic aerobes and coliforms, while L. monocytogenes and Salmonella spp. were absent. The data demonstrated that raw milk was consumed by the population studied. Despite the absence of potential hazards, raw milk was of poor hygienic quality, in contrast with the processed fluid milk available in retail sales outlets that was safe and of good hygienic quality, highlighting the suitability of pasteurized and UHT milk for human consumption.
Introduction
T
he control and monitoring of foodborne diseases are a challenge for public health agencies in several countries (Angelillo et al., 2000; Kennedy et al., 2005). Particular concern is related to the consumption of animal-origin foods, mainly raw, as they are susceptible to contamination by zoonotic agents and foodborne pathogens in distinct steps of production and processing (Angelillo et al., 2000; De Buyser et al., 2001). In this context, raw milk and related products are considered relevant sources of such pathogenic agents, mainly due to the usual belief that these foods are healthier and more natural than processed foods, commonly observed in several countries including Brazil (Katafiasz and Bartlett, 2012; Nero et al., 2004). Currently in Brazil, retail sales of raw milk are not allowed, and restrictive rules and guidelines were published in order to provide proper monitoring of its quality and control of spe1
Departamento Departamento Valadares, Minas 3 Departamento nambuco, Brazil. 2
cific animal diseases (Brasil, 2002, 2011). However, it can be observed that the consumption of raw milk and its products is common in distinct Brazilian regions (Cruz and Menasche, 2014; Nero et al., 2003; Olival et al., 2002; personal observation). Despite having observed this scenario in Brazil, there are no official data that associate the consumption of contaminated foods with foodborne diseases. It is difficult to estimate the actual number of outbreaks of food poisoning caused by the consumption of raw milk in Brazil, because only a small number of foodborne disease cases and outbreaks are confirmed, due to both the major deficiency of epidemiological surveillance services and the lack of awareness of the population regarding this risk (Franco et al., 2003). Studies show that even in developed countries where the consumption of raw milk is restricted to a small portion of the population (CDC, 2007), this product or cheese made from raw milk is responsible for about 70% of foodborne disease outbreaks
de Veterina´ria, Universidade Federal de Vic¸osa, Vic¸osa, Minas Gerais, Brazil. de Cieˆncias Ba´sicas da Sau´de, Caˆmpus Governador Valadares, Universidade Federal de Juiz de Fora, Governador Gerais, Brazil. de Engenharia Cartogra´fica, Universidade Federal de Pernambuco, Centro de Tecnologia e Geocieˆncias, Recife, Per-
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related to dairy products (Robinson et al., 2011). Thus, these numbers may represent a very worrying percentage in countries like Brazil, where raw milk is consumed by a great part of the population. In this context, studies that describe the consumer habits of a specific population are valuable for determining a profile of risky habits, which can be used as guidance for proper control and strategic policies concerning consumption of raw foods. In addition, evaluation of the safety and quality of raw foods is relevant and provides scientific data to assess the hazards associated with these products. In this context, the present study aimed to determine the risky consumption habits regarding raw milk by the population in Vic¸osa, Minas Gerais (MG) state, Brazil, and to assess the quality and safety of the fluid milk available in retail sales. Materials and Methods Consumption habits of the population
The present study was conducted in the urban area of Vic¸osa city, located in MG state in the southeastern region of Brazil (Fig. 1). In the 2008 census (Secretaria de Planejamento of Vic¸osa municipality, personal communication), 14,586 residences were registered in the urban area of Vic¸osa; this value was used as a reference to determine the sampling size of the epidemiological study (EpiInfo 3.3.2., Centers for Disease Control and Prevention, Atlanta, GA). In this study, the following parameters were considered: 30% as the expected frequency of raw milk consumption; 25–35% as the acceptable frequency; and 95% as the confidence level. Then, the urban area of Vic¸osa was divided into 10 areas based on established municipal districts, and the total sampling number was divided proportionally to the number of
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residences in each region. In each region, the sampled residences were chosen based on the established sampling size to obtain a homogeneous distribution. To provide a characterization of the consumption of raw milk, a simplified questionnaire was applied in the selected residences. The questionnaire that was applied to the person responsible for food acquisition in each residence included basic demographic data and information about raw milk consumption, knowledge of risk-related foods, and basic knowledge of food inspection practices (Table 1). During the interviews, the study residences were georeferenced using a Global Positioning System (eTrex Vista Cx, Garmin International Inc., Olate, KS) that provided coordinates accurate to within 10–15 m. The frequencies of the recorded responses were calculated using EpiInfo 3.3.2. software (CDC). The resulting data were plotted in a cartographic representation of Vic¸osa city, and thematic maps were created using MapInfo Professional 10.0 software (Pitney Bowes Inc., Troy, NY) to provide a clear visualization of the recorded parameters. The epidemiological investigation included in this part of the study was submitted to the Ethics Committee for Research in Humans of the Universidade Federal de Vic¸osa, and was approved according the protocol 060/2006. Microbiological safety and quality of fluid milk
Samples of raw milk (n = 69), pasteurized milk (n = 80), and UHT-treated milk (n = 80) were obtained from retail sales in Vic¸osa, MG, Brazil. Raw milk samples were obtained from distinct suppliers that usually delivered this product directly to consumer homes, without refrigeration. Processed fluid milk samples were obtained from retail stores in their current storage conditions (pasteurized: refrigerated, UHT: room
FIG. 1. Location of Minas Gerais state in Southeastern region of Brazil (A); Location of Vic¸osa municipality in Minas Gerais state (B); The urban area of Vic¸osa city, divided into 10 regions and the number of residences included in the study for each region (C).
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Table 1. Simplified Description of the Epidemiological Questionnaire About Consumption of Noninspected Foods of Animal Origin Applied to the Responsibility of Foods Acquisition in the Residences from the Urban Area of Vic¸osa, Minas Gerais, Brazil Questions 1. Basic identification Area, geographic coordinators 2. Milk consumption Raw milk consumption Knowledge about related hazards 3. Additional information Identification of inspection stamp
Possible answers Detailed description Yes/no Yes/no Yes/no
Results Consumption habits of the population
The initial sample size defined by the study parameters was 316 residences. However, to guarantee homogeneous distribution of the interviews, a total of 411 residences were visited (Fig. 1). The responses to each question are shown in Table 2. These results document the risks related to raw milk consumption in the urban area of Vic¸osa. Identification of inspection stamps on food was reported by only 153 individuals responsible for food acquisition; this result may be a matter of concern for public health agencies. The thematic maps created from the study data on raw milk consumption, knowledge of the health risks of these habits, and checks for inspection stamps are presented in Figure 2. Microbiological safety and quality of fluid milk
temperature); all processed fluid milk samples were produced by dairy industries regularly inspected by federal inspection. All samples were kept under refrigerated conditions until laboratory analysis, when they were subjected to microbiological analysis to count mesophilic aerobes, coliforms, and E. coli, and to detect L. monocytogenes and Salmonella spp. In addition, raw milk samples were also subjected to enumeration of coagulase-positive Staphylococcus. All samples were subjected to a 10-fold dilution series in NaCl 0.85% (wt/vol) and selected dilutions were plated on Petrifilm AC and Petrifilm EC (3M Microbiology, St. Paul, MN), followed by incubation at 35C for 24 and 48 h, when the colonies were counted. Final results were expressed as CFU/mL. L. monocytogenes was detected according to the protocol described in the Bacteriological Analytical Manual (BAM) (FDA, 2010), by transferring 25 mL of each sample to Listeria Enrichment Broth (Oxoid Ltd., Basingstoke, England) and incubating at 30C for 24–48 h. Aliquots of the obtained cultures were streaked on Palcam agar (Oxoid) and Oxford agar (Oxoid) and incubated at 35C for 24–48 h. When present, suspected colonies of Listeria spp. were subjected to biochemical identification according to the FDA (2010). Salmonella spp. were investigated as described in the BAM (FDA, 2010), by transferring 25 mL of each sample to lactose broth (Oxoid) and incubating at 35C for 16 h. An aliquot of 1.0 mL was transferred to tetrathionate broth (Oxoid) and incubated at 35C for 24 h, and 0.1 mL was transferred to Rappaport-Vassiliadis broth (Oxoid) and incubated at 42.5C for 24 h. Aliquots of the obtained cultures were streaked on xylose lysine deoxycholate agar (Oxoid), Brilliant Green Agar (Oxoid), and Hektoen enteric agar (Oxoid), followed by incubation at 35C for 24 h. Presumptive Salmonella spp. suspected colonies were plated on triple sugar iron agar (Oxoid) and lysine iron agar (Oxoid) and incubated at 35C for 24 h. Cultures that showed typical reactions were subjected to serological identification with flagellar and somatic polyvalent antisera (Probac Brazil, Sa˜o Paulo, SP, Brazil). Coagulase-positive Staphylococcus were enumerated according to ISO 6888-2 (ISO, 1999). Pour plates were produced in duplicate for selected dilutions using rabbit plasma fibrinogen agar (BioMe´rieux S.A., Marcy l’Etoile, France), followed by incubation at 35C for 48 h, when typical colonies were counted. The results were expressed as CFU/mL.
The results of the microbiological analyses of samples of raw, pasteurized, and UHT milk are shown in Table 3. None of the samples showed a positive result for L. monocytogenes or Salmonella spp. The obtained data demonstrate the overall quality of the fluid milk available in retail sales outlets in Vic¸osa, MG, Brazil. Discussion
The results of the epidemiological questionnaire applied in the urban area of Vic¸osa identified the primary risky feeding habits related to raw milk consumption in the population (Table 2). Identifying associations between feeding habits and risk factors is the most efficient strategy for controlling foodborne disease outbreaks and cases (Tauxe et al., 2010). This strategy has been adopted by various countries to prevent these diseases (Lin et al., 2005; McMeekin et al., 2006; Broner et al., 2010; Wang et al., 2010). Studies of associations between feeding habits and risk factors have been conducted in other Brazilian regions (Olival et al., 2002; Nero et al., 2003), with similar results concerning raw milk. Using the thematic maps as a reference, it is easy to locate the specific city areas where the consumption of raw milk was more prevalent. For example, consumption of raw milk at frequencies higher than 25% was clearly identified in regions I, VII, VIII, and IX (Fig. 2). Relatively high consumption levels of raw milk have also been observed in similar studies
Table 2. Frequencies of Positive Responses to the Epidemiological Questionnaire Administered in 411 Residences in the Urban Area of Vic¸osa City; The Respondents Were the Persons Responsible for Acquisition of Food
Questions 1. Basic identification Area, geographic coordinators 2. Milk and dairy consumption Raw milk consumption Knowledge about related hazards 3. Additional information Identification of inspection stamp
Frequencies of positive answers/interviews Not applicable 76/411 183/411 153/411
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FIG. 2. Thematic maps of Vic¸osa city plotted showing (A) raw milk consumption in each region of the city; (B) knowledge about raw-milk–related hazards; (C) population habits in checking for inspection stamps in milks before acquisition; and (D) relation between consumption of raw milk and knowledge about raw-milk–related hazards. in other Brazilian regions (Olival et al., 2002; Nero et al., 2003). In areas I, VII, VIII, and IX, the data also revealed higher frequencies of a lack of knowledge about raw milk– related risks (Fig. 2). Finally, it was observed that, in general, fewer than 50% of those responsible for food acquisition checked for inspection stamps on the obtained milk (Fig. 2); this is a matter of concern due to the hazards that noninspected food can carry (Franco et al., 2013). As a result of analysis of the thematic maps, ‘‘raw milk consumption’’ and ‘‘knowledge of raw milk-related risks’’ answers were plotted together on a dot map (Fig. 2). Again, the data demonstrated that raw milk consumption can be associated with a lack of knowledge of the related risks (Fig. 2). The consumption of raw milk is a challenge to public health agencies because it is strongly associated with personal beliefs about health and, specifically, about the beneficial effects of raw milk on consumers (Katafiasz and Bartlett, 2012). Thus, visual representation of data can add
relevant support when statistical analysis is not able to detect significant relationships between risk-related habits and the occurrence of foodborne or other diseases (Broner et al., 2010; Wang et al., 2010). The knowledge of consumer feeding habits is a key point in controlling the occurrence of foodborne diseases (Grace et al., 2008), and information about the quality and hazards of related foods must also be considered for a holistic assessment of the risk for a specific population. Based on the obtained data (Table 3), it can be observed that 43 (62.3%) samples of raw milk presented mesophilic aerobe counts higher than 100,000 CFU/mL, the initial reference parameter for adequacy established by the Brazilian Ministry of Agriculture (Brasil, 2002), and a usual reference value of good manufacturing practices proposed by Robinson (2002). However, considering the current quality parameter for mesophilic aerobes in raw milk of 600,000 CFU/mL, from an update by the Brazilian Ministry of Agriculture (Brasil,
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Table 3. Numbers of Samples of Raw Milk Marketed Informally, Pasteurized and Ultrahigh Temperature (UHT) Milks in Vic¸osa, with Different Contamination Levels of Hygiene Indicators Milk Raw (n = 69)
Pasteurized (n = 80)
UHT (n = 80)
Count range (CFU/mL)
Mesophilic
Coliforms
Escherichia coli
Coagulase- positive Staphylococcus
< 100 100–1000 1000–10,000 10,000–100,000 100,000–1,000,000 > 1,000,000 < 100 100–1000 1,000–10,000 10,000–100,000 100,000–1,000,000 > 1,000,000 < 100 100–1000 1000–10,000 10,000–100,000 100,000–1,000,000 > 1,000,000
0 0 8 18 26 17 53 26 1 0 0 0 80 0 0 0 0 0
4 4 12 9 8 32 76 4 0 0 0 0 80 0 0 0 0 0
38 12 13 4 2 0 80a 0 0 0 0 0 80b 0 0 0 0 0
4 6 19 26 13 1 NT NT NT NT NT NT NT NT NT NT NT NT
a
E. coli count lower than 10 colony-forming units (CFU)/mL (n = 2), and lower than 1 CFU/mL (n = 78). E. coli count lower than 1 CFU/mL (n = 80). NT, not tested.
b
2011), 41 (59.4%) samples would be considered as having been produced under inadequate hygienic conditions. The overall poor quality of the raw milk can be confirmed by the observed high counts of coliforms (Table 3): only 4 (5.8%) raw milk samples presented counts lower than 100 CFU/mL, a reference parameter proposed by Robinson (2002) to indicate good hygienic practices. Despite the observed poor hygienic quality of raw milk samples (Robinson, 2002; Jay et al., 2005), 38 (55.1%) presented E. coli counts lower than 100 CFU/mL, and none of the samples presented positive results for L. monocytogenes or Salmonella spp. In contrast, only 4 (5.8%) raw milk samples presented counts of coagulase-positive Staphylococcus lower than 100 CFU/mL. The presence of coagulase-positive Staphylococcus at high concentrations may suggest the presence of enterotoxigenic strains capable of producing different types of enterotoxins with a potentially pathogenic effect on consumers (Carmo et al., 2002; Boerema et al., 2006; Srinivasan et al., 2006), but can also be considered as a good indicator of improper handling. Regarding the quality and safety of raw milk, similar results were also observed by studies conducted in distinct Brazilian regions (Moraes et al., 2005; Nero et al., 2005; Rosa and Queiros, 2007; Mattos et al., 2010), including studies that considered raw milk samples available for retail sales without official inspection (Beloti et al., 1999; Mendes et al., 2010). The obtained data confirmed the poor microbiological quality of the raw milk produced and available for retail sales in Vic¸osa, MG, Brazil, and demonstrated that, in general, this product does not fit the minimum required microbiological standards for reception and processing in processing industries (Robinson, 2002; Brasil, 2011). It is noteworthy that no relevant microbiological hazards were identified in the raw milk samples. Coagulase-positive Staphylococcus was detected at high levels in various samples (Table 3), and even then, it was not possible to associate
them directly with the presence of enterotoxins or the ability to produce them (Veras et al., 2008). However, the absence of these pathogens does not indicate that that raw milk is safe for consumers. Staphylococcal intoxication is caused by eating foods containing enterotoxins produced by enterotoxigenic Staphylococcus, of which S. aureus is the most prevalent etiological agent. Such poisoning is one of the most common types of foodborne illness (Le Loir et al., 2003). Staphylococcal food poisoning is attributed to ingestion of enterotoxins that are produced and released by bacteria in the food during their multiplication. These enterotoxins are heat stable and may remain in the food after cooking, favoring intoxication (Ostyn et al., 2010). Despite the observed overall poor quality of raw milk, the obtained counts for pasteurized and UHT milks indicated good production practices and proper processing in dairy industries (Table 3). As reference, the Brazilian legislation for processed fluid milk available for consumption establishes the control of thermotolerant coliforms (maximum of four most probable number per milliliter) and Salmonella spp. (absence), and for UHT milk, the absence of pathogenic and spoilage bacteria (without reference values) (Brasil, 2001). Considering the obtained data (Table 3), all pasteurized and UHT milk samples complied with the established reference parameters. In addition, all samples presented low counts of aerobic mesophiles, coliforms, and the absence of L. monocytogenes, confirming the adequacy of the processing in dairy industries. A study conducted by Bersot et al. (2010) identified 36 of 150 UHT milk samples with mesophilic aerobe counts higher than 100 CFU/mL, indicating inadequacy of processing according to Brazilian legislation. The absence of Salmonella spp. and E. coli can be considered a good indicator of the safety of the processed fluid milk available for retail sales in Vic¸osa, MG, Brazil, demonstrating the relevance of heat treatment of milk (Agarwal et al., 2012).
RAW MILK CONSUMPTION, QUALITY, AND SAFETY Conclusions
The data obtained allowed accurate identification of the risk-related habits in the population of the urban area of Vic¸osa, MG, Brazil regarding the consumption of raw milk. They demonstrated a lack of knowledge of the risks of raw milk, as well as a lack of knowledge regarding the need to verify food inspection stamps before consumption. As complementary information, the poor hygienic quality of the raw milk available for retail sales was demonstrated, but there was an absence of potential hazards with this product. In contrast, the processed fluid milk available for retail sales presented high hygienic quality and proper safety, highlighting the suitability of pasteurized and UHT milk for human consumption. Acknowledgments
The authors thank Conselho Nacional de Desenvolvimento Cientı´fico e Tecnolo´gico (CNPq), Coordenac¸a˜o de Aperfeic¸oamento Pessoal de Nı´vel Superior (CAPES), and Pesquisa do Estado de Minas Gerais (FAPEMIG). Disclosure Statement
No competing financial interests exist. References
Agarwal A, Awasthi V, Dua A, Ganguly S, Garg V, Marwaha SS. Microbiological profile of milk: Impact of household practices. Indian J Public Health 2012;56:88–94. Angelillo IF, Viggiani NMA, Rizzo L, Bianco A. Food handlers and foodborne diseases: Knowledge, attitudes, and reported behavior in Italy. J Food Prot 2000;63:381–385. Beloti V, Barros MAF, Souza JA, Nero LA, Santana EHW, Balarin O, Curiaki Y. Avaliac¸a˜o da qualidade do leite cru comercializado em Corne´lio Proco´pio, Parana´: Controle do consumo e da comercializac¸a˜o. Semina 1999;20:12–15. Bersot LS, Galva˜o JA, Raymundo NKL, Barcellos VC, Pinto JPAN, Maziero MT. Avaliac¸a˜o microbiolo´fica e fı´sico-quı´mica dos leite UHT produzidos no Estado do Parana´, Brasil. Semina 2010;31:645–652. Boerema JA, Clemens R, Brightwell G. Evaluation of molecular methods to determine enterotoxigenic status and molecular genotype of bovine, ovine, human and food isolates of Staphylococcus aureus. Int J Food Microbiol 2006;107:192–201. Brasil, ANVISA. Resoluc¸a˜o-RDC no 12, Aprova o regulamento te´cnico sobre padro˜es microbiolo´gicos para alimentos. January 2, 2001. In: Dia´rio Oficial da Unia˜o 2001, January 10, 1:45. (in Portuguese). Brasil, MAPA. Instruc¸a˜o normativa n 51, Aprova o Regulamento Te´cnico de Produc¸a˜o, Identidade e Qualidade do Leite tipo A, o Regulamento Te´cnico de Identidade e Qualidade de Leite Cru Refrigerado, o Regulamento Te´cnico de Identidade e Qualidade de Leite Pasteurizado e o Regulamento Te´cnico da Coleta de Leite Cru Refrigerado e seu Transporte a Granel. September 18, 2002. In: Dia´rio Oficial da Unia˜o 2002, September 20, 1:64. (in Portuguese). Brasil, MAPA. Instruc¸a˜o normativa n 62, Aprova os Regulamentos Te´cnicos de Produc¸a˜o, Identidade e Qualidade do Leite tipo A, do Leite Pasteurizado e do Leite Cru Refrigerado e o Regulamento Te´cnico da Coleta de Leite Cru Refrigerado e seu Transporte a Granel. December 29, 2011. In: Dia´rio Oficial da Unia˜o 2011, December 30, 1:24. (in Portuguese).
495
Broner S, Torner N, Dominguez A, Martinez A, Godoy P, Working Group Study Outbreaks A. Sociodemographic inequalities and outbreaks of foodborne diseases: An ecologic study. Food Control 2010;21:947–951. Carmo LS, Dias RS, Linardi VR, Sena MJ, Santos DA, Faria ME, Pena EC, Jett M, Heneine LG. Food poisoning due to enterotoxigenic strains of Staphylococcus present in Minas cheese and raw milk in Brazil. Food Microbiol 2002;19:9–14. [CDC] Centers for Disease Control and Prevention. Foodborne Diseases Active Surveillance Network (FoodNet): Population Survey Atlas of Exposures, 2006–2007. Atlanta: Centers for Disease Control and Prevention, 2007, 28 pp. Cruz FT, Menasche R. Tradition and diversity jeopardised by food safety regulations? The Serrano Cheese case, Campos de Cima da Serra region, Brazil. Food Pol;2014:45:116–124. De Buyser ML, Dufour B, Maire M, Lafarge V. Implication of milk and milk products in food-borne diseases in France and in different industrialised countries. Int J Food Microbiol 2001;67:1–17. [FDA] Food and Drug Administration. BAM—Bacteriological Analytical Manual. Silver Spring, MD: U.S. Food and Drug Administration, 2010. Franco BDGM, Landgraf M, Destro MT, Gelli DS. Foodborne Diseases in Southern South America. In: International Handbook of Foodborne Pathogens. Milliotis M, Bier J (eds.). New York: Marcel Dekker, 2003, pp. 733–743. Franco MM, Paes AC, Ribeiro MG, de Figueiredo Pantoja JC, Santos AC, Miyata M, Leite CQ, Motta RG, Listoni FJ. Occurrence of mycobacteria in bovine milk samples from both individual and collective bulk tanks at farms and informal markets in the southeast region of Sao Paulo, Brazil. BMC Vet Res 2013;9:85. Grace D, Randolph T, Olawoye J, Dipelou M, Kang’ethe E. Participatory risk assessment: A new approach for safer food in vulnerable African communities. Dev Pr 2008;18:4611–4618. [ISO] International Organization for Standardization. ISO 6888-2. Microbiology of Food and Animal Feeding Stuffs—Horizontal Method for the Enumeration of Coagulase Positive staphylococci (S. aureus and Other Species): Part 1. Technique Using Rabbit Plasma Fibrinogen Agar Medium. Geneva: International Organisation of Standardisation, 1999. Jay JM, Loessner MJ, Golden DA. Modern Food Microbiology, 7th ed. New York: Food Science Text Series, 2005. Katafiasz AR, Bartlett P. Motivation for Unpasteurized Milk Consumption in Michigan, 2011. Food Prot Trends 2012;32: 124–128. Kennedy J, Jackson V, Blair IS, McDowell DA, Cowan C, Bolton DJ. Food safety knowledge of consumers and the microbiological and temperature status of their refrigerators. J Food Prot 2005;68:1421–1430. Le Loir Y, Baron F, Gautier M. Staphylococcus aureus and food poisoning. Genet Mol Res 2003;2:63–76. Lin CTJ, Jensen KL, Yen ST. Awareness of foodborne pathogens among US consumers. Food Qual Pref 2005;16:401–412. Mattos MR, Beloti V, Tamanini R, Magnani DF, Nero LA, Barros MAF, Pires EMF, Paquereau BPD. Qualidade do leite cru produzido na regia˜o do agreste de Pernambuco, Brasil. Semina 2010;31:173–182. McMeekin TA, Baranyi J, Bowman J, Dalgaard P, Kirk M, Ross T, Schmid S, Zwietering MH. Information systems in food safety management. Int J Food Microbiol 2006;112: 181–194. Mendes CG, Sakamoto SM, Silva JBA, Ja´come CGM, Leite AI. Ana´lises fı´sico-quı´micas e pesquisa de fraude no leite infor-
496
mal comercializado no municı´pio de Mossoro´, RN. Rev Cienc Anim Bras 2010;11:349–356. Moraes CR, Fuertefria AM, Zaffari CB, Conte M, Rocha JPAV, Spanamberg A, Valente P, Corc¸a˜o G, Costa M. Qualidade microbiolo´gica de leite cru produzido em cinco municı´pios do Estado do Rio Grande do Sul, Brasil. Acta Sci Vet 2005;33:259–265. Nero LA, De Mattos MR, Beloti V, Barros MAF, Netto DP, Pinto JPAN, De Andrade NJ, Silva WP, Franco BDGM. Hazards in non-pasteurized milk on retail sale in Brazil: Prevalence of Salmonella spp, Listeria monocytogenes and chemical residues. Braz J Microbiol 2004;35:211–215. Nero LA, Mattos MR, Beloti V, Barros MAF, Pinto JPAN, Andrade NJ, Silva WP, Franco BDGM. Leite cru de quatro regio˜es leiteiras brasileiras: Perspectivas de atendimento dos requisitos microbiolo´gicos estabelecidos pela Instruc¸a˜o Normativa 51. Cienc Tecnol Aliment 2005;25:191–195. Nero LA, Maziero D, Bezerra MMS. Ha´bitos alimentares do consumidor de leite cru de Campo Moura˜o—PR. Semina 2003;24:21–26. Olival AA, Spexoto AA, Campos DFS, Ferreira F, Fonseca LFL, Santos MV, Dias RA. Ha´bitos de consumo do leite informal, associado ao risco de transmissa˜o de doenc¸as no municı´pio de Pirassununga, SP. Hig Alim 2002;16:35–40. Ostyn A, De Buyser ML, Guillier F, Groult J, Fe´lix B, Salah S, Delmas G, Hennekinne JA. First evidence of a food poisoning outbreak due to staphylococcal enterotoxin type E, France, 2009. Euro Surveil 2010;15:ID19528. Robinson RK. Dairy Microbiology Handbook: The Microbiology of Milk and Milk Products, 3rd ed. New York: WileyInterscience, 2002. Robinson T, Scheftel J, Smith K. Sporadic reportable enteric pathogen cases reporting domestic raw milk consumption, Minnesota, 2001–2010. In: Proceedings of the 100th Annual
PIERI ET AL.
Meeting of the International Association for Food Protection; 2011; Milwaukee, WI. Abstract T4-10. Rosa LD, Queiros MI. Avaliac¸a˜o da qualidade do leite cru e resfriado mediante a aplicac¸a˜o de princı´pios do APPCC. Cienc Tecnol Aliment 2007;27:422–430. Srinivasan V, Sawant AA, Gillespie BE, Headrick SJ, Ceasaris L, Oliver SP. Prevalence of enterotoxin and toxic shock syndrome toxin genes in Staphylococcus aureus isolated from milk of cows with mastitis. Foodborne Pathog Dis 2006; 3:274–283. Tauxe RV, Doyle MP, Kuchenmuller T, Schlundt J, Stein CE. Evolving public health approaches to the global challenge of foodborne infections. Int J Food Microbiol 2010;139:S16– S28. Veras JF, Carmo LS, Tong LC, Shuppe JW, Cummings C, Santos DA, Cerqueira MMCOP, Cantini A, Nicoli JR, Jett M. A study of the enterotoxigenicity of coagulase negative and coagulase-positive staphylococcal isolates from food poisoning outbreaks in Minas Gerais, Brazil. Int J Infect Dis 2008;12:410–415. Wang LG, Xu YY, Wang Y, Dong SC, Cao ZD, Zhou W, Sun HL, Huo DH, Zhang H, Sun YS, Huang LY, Yuan ZQ, Zeng DJ, Song HB. The epidemiological investigation and intelligent analytical system for foodborne disease. Food Control 2010;21:1466–1471.
Address correspondence to: Luis Augusto Nero, DSc Departamento de Veterina´ria Universidade Federal de Vic¸osa Campus UFV 36570-000, Vic¸osa, MG, Brazil E-mail: [email protected]
