PREVALENCE OF COLIFORMS AND STAPHYLOCOCCUS AUREUS IN HANDS OF FOOD HANDLERS OF AN OPEN-AIR MARKET IN THE MUNICIPALITY OF VITÓRIA-ES

Abstract

Introduction: Foodborne diseases (FDs) represent a major public health issue and negatively reflect on the health of populations and on the economic development of countries, leading to labor incapacity and costs with treatments and hospitalization. Objective:  Assess the importance of professional food handlers in open-air markets in the spread of microbial pathogens. Method: The samples consisted of biological material collected from both hands of 17 vegetable handlers working in an open-air market in the municipality of Vitoria. The material was analyzed for isolation and identification of total and heat-resistant coliforms, Escherichia coli and Staphylococcus aureus followed by the antimicrobial susceptibility testing. Results: It was possible to verify the presence of total and heat-resistant coliforms and strains of Staphylococcus aureus. No Escherichia coli strain was isolated. Conclusion: The results indicate that food handlers in open-air markets play an important role as potential disseminator of pathogenic bacteria. The study indicates the need to widen the practices of sanitary and hygiene control of these workers, as well as encourage them to maintain proper and regular hand washing, make available clean running water in their workplace and teach them to assign tasks so as to separate money from food.

Introduction

Foodborne diseases (FDs) represent a major public health issue and negatively reflect on the health of populations and on the economic development of countries, leading to labor incapacity and costs with treatments and hospitalization. In Brazil, between 1999 and 2004, 3,410,048 hospitalizations due to FDs were recorded, with an average of 568,341 cases and 8,427 thousand deaths per year.^1,2

According to the Atlanta Center for Disease Control and Prevention (CDC), most of the FDs are linked to the inadequate handling of food. Therefore, food handlers play an important role in the transmission of foodborne diseases, partly due to poor personal hygiene and domestic habits, besides a deficiency in hygiene and environmental control.^3,4

The collective food services have grown globally, and in Brazil this market serves about 2 million workers. Thus, a lack of hygiene control of the food sold by these handlers represents a major obstacle in the introduction of control measures to control FDs.⁵

The transmission of infectious diseases through hands was described 120 years ago by Semmelweis, but it was Price who actually studied the type of skin bacteria, classifying them into “resident and transitory”.⁶

The bacteria of the genus Staphylococcus are Gram-positive motionless cocci, part of the family Micrococcaceae and, due to dividing themselves in different layers, they are formed in grape-like clusters when analyzed in a microscope. They are facultative anaerobic bacteria, catalase producers under anaerobic conditions. They are also important pathogens for humans since they are responsible for a wide range of systemic diseases, including skin infections, soft tissues, bones, urinary tract besides food poisoning, the latter the scope of this study.

The species Staphylococcus aureus, besides being the most virulent, is often associated to staphylococcus diseases, whether they be food-based or not, which makes this group the target of several studies.^7,8 Its colonies are of a golden yellow coloration as a result of carotenoid pigmentation that are formed during the growing phase, hence the name of the species.⁸

Men and animals are the main recipients of S. aureus. In humans, the nasal cavity is the main habitat of these bacteria and, from there, they may reach the skin as well as wounds, air, water, soil, milk and any surface or object that has had contact with a human.⁷ Approximately 15% of healthy adults are persistent carriers of S. aureus in the nasopharynx.⁸ Thus, the nasal carriers that work handling food become important sources of food contamination, and potential causative agents of food poisoning.

Food poisoning through S. aureus is one of the most common FDs, being an intoxication rather than an infection, since the symptoms are caused by the ingestion of food containing the bacterial toxin, and not by the direct action of the microorganism.^7,8 Such toxins, known as enterotoxins, are heat-resistant, which makes them an asset to the food industry, considering that most foods undergo a thermal treatment during its processing, which would not inactivate the toxin if present, making the food more prone to causing some kind of poisoning. The enterotoxins can cause vomiting, diarrhea, inflammation with the development of enteritis, making up a wide spectrum of symptoms.⁷

Staphylococcus become a meaningful part of the resident microbiota in many individuals and, due to the pathogenicity of some strains and capacity to produce enterotoxins, it is important to eliminate them by washing the hands. However, individuals with a relatively low educational level are not often aware of the importance of such hygiene habits.⁶

The coliforms have been initially described as potential microorganisms indicators of fecal contamination in fresh water.⁹ The use of Escherichia coli, one of its representatives, as an indicator of fecal contamination in water was proposed in 1892, provided that this microorganism exclusively lives in the intestine tract of humans and hot-blooded animals.⁷

Over the years, such concept has become broader, and nowadays the research on coliforms, besides used for water quality evaluation, has also been used for the evaluation of poor hygiene and sanitary conditions while handling, producing and preparing foods. Such microorganisms are classified as total coliforms and heat-resistant coliforms.^10,11

The group of total coliforms is composed of bacteria of the family Enterobacteriaceae and include all the bacteria that are Gram-negative, rod-like, non-sporogenic, aerobic or facultative anaerobic, able to ferment lactose with the production of gas when incubated at 35-37°C (95-98.6°F) for 24-48 hours. It includes about 20 species of bacteria belonging to the genuses Escherichia, Klebsiella, Enterobacter and Citrobacter. Out of the latter, only Escherichia coli has as a primary habitat the intestinal tract of humans and animals. The others, besides being found in feces, are also present in other environments, such as in plants and soil. Consequently, the presence of total coliforms in food does not necessarily indicate a recent fecal contamination or the occurrence of enteropathogens.^7,12,13

The heat-resistant coliforms, previously referred to as fecal coliforms, correspond to the total coliforms that are able to continuously ferment lactose with the production of gas, when incubated at 44.5%-45.5ºC (112.1-113.9 ºF) for 24 hours. Under these conditions, around 90% of the E. coli cultures are positive, while among the other genuses, only some strains of Enterobacter and Klebsiella maintain such characteristic. In this way, the research on heat-resistant coliforms or E. coli in food provides solid information on product hygiene and efficiently indicates an occasional presence of entheropathogens.^7,13

Among the bacteria of admittedly fecal habitat, E. coli is the most known and easily differentiated among the non-fecal members. Several ones from its lineage are proven to be pathogenic to humans, able to cause serious infections and even death. These pathogenic strains are classified according to their effects in the recipient. The categories that cause intestinal infection are collectively called E. coli diarrheagenic, and may be classified as: enteropathogenic E. coli (EPEC), enterotoxigenic (ETEC), enteroinvasive (EIEC), enterohemorrhagic (EHEC) e enteroaggregative (EAEC).¹² Data reported by the Center for Disease Control on food poisoning and infection surges in the US food service indicate food handlers accounting for 26% of the responsibility for such surges.¹⁴ Considering that in Brazil there are few lines of research assessing the occurrence of microorganisms in food handlers, despite being a relevant and current issue, the present study was then designed to investigate total and heat-resistant coliforms (Escherichia coli) and Staphylococcus aureus in the hands of stallholders handling in natura food and working at open-air markets.

Method

The samples of the present study are composed of biological material collected from 17 vegetable handlers working in an open-air market in the municipality of Vitória, state of Espírito Santo, Brazil. The professionals who agreed on being submitted to the research had to sign the Term of Informed Consent at the moment of the interview that preceded the material collection.

Nail bed sample

Each research participant had the material from both hands, including nail bed, collected with sterilized Swab, packaged and transported in a Stuart^TM medium as per described in the Manual of Clinical Microbiology for the Control of Infection Related to Assistance and Healthcare, identified with a number for the subject, and taken to the Microbiology Laboratory at the Escola Superior de Ciências da Santa Casa de Misericórdia in Vitória (EMESCAM) for analysis.¹⁵

The Stuart^TM medium presents a nutritive composition able to guarantee the survival of microorganisms, although it considerably hinders their multiplication due to a lack of nitrogen source in the medium.¹⁵

Sample analysis

The material was analyzed so as to isolate and identify Total Coliforms, Heat-resistant Coliforms, Escherichia coli and Staphylococcus aureus by using specific selection medium followed by the performance of confirmatory tests for identification and susceptibility to antimicrobials. The storage and analysis were carried out at the Microbiology Laboratory at the Escola Superior de Ciências da Santa Casa de Misericórdia in Vitória (EMESCAM), according to the current norms of Biosafety.

Staphylococcus aureus isolation and identification

Specific medium seeding

For the isolation of S. aureus, the samples were seeded in a Hypertonic Mannitol Agar (HMA) medium and incubated in a bacteriological incubator at 35ºC±1ºC (95ºF) for 48 hours. After this period, there was a growth of microorganisms resistant to the presence of high saline concentrations (7.5%), such as bacteria of the genus Staphylococcus. As there was growth of Staphylococcus aureus, it has been observed the color alteration of the culture medium, from pinkish to yellow, due to the turning of the pH indicator (phenolsulfonphthalein/PSP), resulting from the fermentation of mannitol. In case of no mannitol fermentation (S. non-aureus), the medium color remained unaffected (mannitol negative).^16,17

The suggestive colonies of Staphylococcus aureus have been seeded in a new plate containing a HMA medium for the pure and subsequent performance of confirmatory tests of identification.

Identification confirmatory tests

GRAM coloration

Initially, the GRAM coloration of the suggestive colonies of Staphylococcus aureus present in the material was carried out according to the following process: fixate the material sample on the slide. Cover the slide with the Crystal Violet dye for 1 minute. Wash the slide in running water. Cover the slide with Lugol for 1 minute. Wash the slide in running water. Remove dye excess with alcohol/acetone for about 20 seconds. Wash the slide in running water. Cover the slide with safranin for 30 seconds. Wash the slide in running water. Dry the slide. With an optical microscope, in the presence of S. aureus, cocci have been identified as grape-like clusters of a purplish color, due to Gram coloration.¹⁷

Catalase Test

This experiment has allowed the differentiation of the catalase-producer microorganisms (Staphylococcus spp.) from the non-producers (Streptococcus spp.).¹⁷

The suggestive colonies of Staphylococcus spp. were placed on a glass slide and, afterwards, a drop of 3% hydrogen peroxide was added. When positive, the immediate presence of bubbles was observed due to the conversion of the hydrogen peroxide into water and gaseous oxygen, which confirms the presence of Staphylococcus spp. The non-emergence of bubbles rules out the possibility of staphylococcal colonies.^16,17

DNase test

This test verifies the production of the deoxyribonuclease enzyme, which degrades acid nucleic (DNA). The test is useful to differentiate Serratia, from the genus Enterobacter, Staphylococcus aureus from the coagulase-negative Staphylococcus and Moraxella catarrhalis from the Neisseria species.¹⁷

The suspicious colonies were seeded with a platinum loop in circular movements on a DNase Agar plate. The plates were placed in an inverted position and incubated at 35±2°C (95±35.6°F) for 48 hours. After incubation, the cultivation was covered with a 1N hydrochloric acid solution (HCl). The acid was allowed to penetrate the entire surface of the medium for 2 minutes and a reaction was observed. Upon showing a clear zone encompassing the growth areas on the DNase Agar, after the addition of 1N HCl, a positive reaction was confirmed. By contrast, the negative reaction was indicated by any clouding or precipitation around the colonies.¹⁷

Disc diffusion test for antimicrobial susceptibility assessment

In order to determine the susceptibility pattern to oxacillin, isolates of S. aureus were used, taken from the pure sample from the plates with a positive result for them. The antimicrobial susceptibility test (AST), based on the agar diffusion principle with paper filter discs through the Bauer-Kirby method¹⁷

The method consists in obtaining the bacterial inoculum containing 10⁸ CFU/mL, corresponding to the 0.5 MacFarland Scale standard. Therefore, each tested lineage was inoculated in tubes containing 5mL of a 0.85% saline solution, with the use of a previously heated bacterial loop, in a sufficient amount to reach the 0.5 standard clouding of the mentioned scale. With the aid of a sterile swab, the culture was seeded in Petri plates containing Mueller-Hinton agar until obtaining a uniform smear. After smear drying, discs with selected antibiotics were applied to the surface of the culture medium, with the aid of a sterile pickup, in accordance with the recommendation of the Clinical Laboratory Standards Institute. After that, the plates were incubated at 37ºC±1ºC  (98.6ºF) for 24 hours.¹⁸

Oxacillin susceptibility assessment through disc diffusion test

For the assessment of presence of the methicillin/oxacillin resistant S. aureus (MRSA/ORSA), with the use of cefoxitin, the disc fusion test was performed, based on the Kirby-Bauer method, according to the methodology described above. However, instead of using the oxacillin discs, the cefoxitin discs were used (30 µg). For the interpretation of the inhibition halos, the data described in Table 1 were used.¹⁷

Table 1 – Staphylococcus aureus screening: ORSA/MRSA with cefoxitin disc

Microorganism	Inhibition halo (mm)
S. aureus	≤ 21*	≥ 22*
SCN	≤ 24*	≥ 25**

SCN: Staphylococcus coagulase-negative; *report as resistant oxacillin; **report as sensitive oxacillin. The cefoxitin result is not reported in the result.

Source: Oplustil (2010)

Isolation and identification of total coliforms and heat-resistant coliforms

Presumptive test for coliforms

The swab used for collection was introduced and agitated in a tube containing 10mL of Lauryl Sulfate Tryptose (LST) with an inverted Durham tube. The LST contains lactose and the observation of growth with gas production stemming from lactose, after 24-48 hours at 35ºC±1ºC (95ºF), is considered suspicious of the presence of coliforms.¹⁹

The LST tubes were incubated at 35±0.5ºC (95±32.9ºF) for 24/±2h and growth with gas production was observed. Tubes that showed growth and production of gas were considered positive. In case negative, it was reincubated until completing 48±2h, and the reading was repeated.¹⁹

Presence confirmation of Total Coliforms

For total coliform confirmation, a loopful of each suspect tube was transferred to a 2% Brilliant Green Bile Broth (BG) and incubated at 35±0.5ºC (95±32.9ºF) for 24/±2h. Growth with gas production was observed. Tubes that showed growth and production of gas were considered positive. In case negative, it was reincubated until completing 48±2h, and the reading was repeated. If once again negative, the sample was considered negative for total coliforms.¹⁹

Presence confirmation of Heat-resistant Coliforms

For the confirmation of heat-resistant coliforms, a loopful of each suspect tube was transferred to a E. coli (EC) Broth tube and incubated at 45.5±0,5ºC (113.9±32.9ºF) for 24/±2h. The tubes remained submerged in water up to a height higher than the surface of the culture. The procedure did not exceed 30 minutes between inoculation and transference to the inoculation bath.¹⁹

After this period, growth with gas production was observed. Tubes that showed growth and production of gas were considered positive. In case positive, the sample was considered positive for heat-resistant coliforms, being necessary to perform a confirmatory test for E. coli. In case negative, the sample was considered negative for heat-resistant coliforms.¹⁹

1. coli presence confirmation – Culture in selective/specific medium

The positive EC tubes for heat-resistant coliforms were suspicious of the presence of E. coli. In this case, they were submitted to a confirmatory test.

A handful of each tube was streaked (streaking technique) in Levine-Eosin Methylene Blue Agar (L-EMB), in a differential selective group to distinguish E. coli from the other heat-resistant coliforms. The plates were incubated at 35±1ºC (95±33.8ºF) for 24±2h and, after this period, the growth of typical colonies of E. coli, nucleoid colonies with a black center were observed, with or without a metallic glow.¹⁹

The suggestive E. coli colonies were seeded on a new plate containing a L-EMB medium for pure isolation and subsequent performance of confirmatory identification tests.

1. coli presence confirmation – biochemical proofs

In case of growth of typical colonies, two well isolated colonies from each plate were transferred to Standard Agar Tubes (PCA) for counting, then inclined and incubated at 35±1ºC (95±33.8ºF) for 24±2h. Given that, pure colonies were obtained and submitted to the GRAM coloration and Citrate Test.

Citrate Test

A loopful with a light culture inoculum was inoculated in a Koser citrate broth, incubated at 35ºC±1ºC (95ºF) for 96 hours while growth was observed (positive test) or not (negative test). The E. coli strains are citrate-negative.¹⁹

Statistical Analysis

Type of study: descriptive out of a series of cases, with the carrying out of descriptive statistics.

Inclusion and exclusion criteria

Subjects that participated of the Project: (1) older than 18 years old; (2) handlers of in natura food at an open-air market in the municipality of Vitória, Espírito Santo; and (3) those that agreed on joining the study by signing the Term of Informed Consent.

Considerations

The existing risks for the professionals in the procedures of biological material collection were: (1) nail bed bleeding, which can be spontaneously solved, except in specific occasions; and (2) occasional possibility of contamination for professionals. However, all the material used in the procedures was sterilized according to the norms of the Ministry of Health.

After analysis, the results were handed in individually and under absolute confidentiality to each subject in a previously arranged and informed date.

The biological material has been disposed of in accordance with the protocol of the Health Service Residue Management Program (PGRSS), following the Board of Trustee’s Resolution (RDC) no. 306/2004 by the Brazilian Health Surveillance Agency (ANVISA), implemented in the laboratories of the Microbiology and Parasitology disciplines at EMESCAM. The set of information and work data will be stored for five years in the archives of the Microbiology and Parasitology disciplines of EMESCAM.

Ethical and Biosafety commitment

This project has been approved by the Ethics and Research Committee (ERC) of the Escola Superior de Ciências da Santa Casa de Misericórdia in Vitória (EMESCAM), complying with the requirements of Resolution no. 446 as of December 2012 by the Brazilian National Health Council. All food handlers from the open-air market, subjects of research, were made aware of the project’s goals, that is, to outline the prevalence of coliforms and Staphylococcus aureus present in their nail beds. They were also informed that the results are classified, their privacy guaranteed and that they would be granted access to the research results if they wished so.

The biological material was only collected by the research subject after signing of the Term of Informed Consent. The biological samples obtained have been stored. As previously described in the material and methods section, the samples will not be used for unplanned means as per the Term of Informed Consent without a new authorization from the volunteering handler and from the ERC.

Results

Assessment of the presence of Staphylococcus aureus and its susceptibility to oxacillin

Samples from both hands were collected from 17 stallholders, adding up to 34 samples. Among these samples, 20 Staphylococcus aureus lineages (58.8%) were isolated. 8 samples from the left hand and 12 from the right hand were isolated. In 7 (41.2%) stallholders, S. aureus was isolated in both hands. The lineages assessed in vitro as opposed to oxacillin showed susceptibility in 18 (90%) samples and resistance in 2 (20%) samples (Table 2).

 

 

Table 2 – Isolate numbers of S. aureus in the hands of open-air market food handlers. Assessment of sample susceptibility to oxacillin

Microorganism	Right hand	Left hand	Both hands	Susceptibility to oxacillin
S. aureus	8	12	7	18

Source: The author

 

 

Assessment of the presence of contamination indicators. Presence of coliforms.

Among the studied stallholders, it was possible to verify the presence of total coliforms in 7 (20.5%) samples, and heat-resistant in 3 (8.8%) samples in their hands (Table 3). However, the presence of E. coli, an important indicator of fecal contamination, was not verified in the hands of food handlers.

 

Table 3 – Number of contamination indicators isolates in the hands of food handlers in open-air market

Indicator	Right hand	Left hand	Both hands
Total coliforms	4	3	2
Heat-resistant coliforms	1	2	1
Escherichia coli	0	0	0

Source: The author

Discussion

The presence of S. aureus and coliforms is considered an important indicator of improper conduct in the handling of foods.⁹ Their presence in the hands of food handlers makes evident a worrisome situation in the environment studied, since the presence of these microorganisms, associated to the food handlers’ poor hygiene and sanitary conditions, may cause food poisoning.²⁰ Such fact confirms the hypothesis of the present research and showcases the importance of food handlers in the transmission of FDs.

Food handlers, whether they operate in the industry or food businesses, are important driving sources of Staphylococcus aureus due to the fact they are mostly asymptomatic carriers. Whether it be in preparation, transportation, distribution or commercialization of foods, the presence of this microorganism is an indicative of poor hygiene and sanitary conditions and interpreted as an indicative of contamination from the food handlers’ skin, mouth and/or respiratory system.^9,20

For growing in temperature ranging from 7-47.8ºC (44.6-118.04ºF) and for producing toxins between 10-46ºC (50-114.8ºF), these enterotoxins are thermostable at 100ºC (212ºF) for up to 30 minutes, besides being resistant to hydrolysis from the gastric enzymes and the jejunum, turning out to be perfect for causing foodborne diseases. Thus, once the food had been contaminated by the staphylococcus producer of enterotoxins, and the toxins had been produced in a sufficient amount, not even a food mild reheating and exposition to gastric acid will suffice to avoid poisoning.⁸

The precise mechanism of the toxic activity is still not fully understood, but it is believed to be necessary around 10⁵–10⁶ CFU of S. aureus per gram of food so the toxin can be produced in such a level to be able to cause poisoning. Normally the bacteria are found in a small amount, which makes an infection difficult, since the multiplication is necessary.^7,8

In this study it was possible to prove the presence of Staphylococcus aureus in the hands of 13 out of the 17 assessed stallholders, being this presence observed in one or in both hands, adding up to 20 positive samples. Surprisingly, two of these samples have shown resistance to methicillin, which makes these results increasingly important.

The first time an antimicrobial drug was clinically used was against a sample of S. aureus, following the discovery of the penicillin. The drug met the needs accordingly until the 1960s, when resistant isolates started to be resistant to this antimicrobial. In order to tackle this problem, the methicillin was created, a synthetic beta-lactam that was resistant to the action of the beta-lactamases produced by the S. aureus. However, due to its capacity to rapidly develop resistance, it was not long for reports to show on samples also resistant to this antimicrobial, which have been referred to as methicillin resistant Staphylococcus aureus (MRSA), characterized by the resistance to all beta-lactam antimicrobial.^21,22

Until 1980, MRSA reports consisted of isolated cases, but after 1982 epidemic strains were described as multiresistant, able to colonize and cause infection surges, becoming a widely known cause for morbidity and mortality around the world.²³ Initially, these infections were limited to hospitals, but in the last years infections associated or acquired in a community have increasingly occurred in healthy subjects with no identifiable risk factor. That is alarming since patients infected by MRSA are five times likely to die than patients infected by methicillin sensitive S. aureus (MSSA), which makes the MRSA one of today’s key health issues.^21,22

Some studies have reported the detection of genes that codify staphylococcus enterotoxins among lineages of MRSA.  This way, it can be concluded that this group can also be involved in food poisoning surges, provided the ingestion of the toxin ingestion takes place. However, it is important to consider that the resistance to the methicillin is not a relevant factor for the production of enterotoxins. The presence of MRSA in food may not represent a greater risk of food poisoning.²³

Considering that the S. aureus is widely present in nature, its elimination from the environment becomes virtually impossible. The handling of by humans, one of the recipients of this bacterium, already increases a risk of contamination, being necessary to take measures to avoid its transmission.^7,24 Therefore, stallholders are tentative sources of contamination and dissemination of S. aureus, especially of resistant strains, and its presence in their hands represents poor hygiene conditions at the workplace. Hence, it will only be possible to reduce the rates of infections and contaminations through prevention and control of the transmission of multiresistant organisms. Washing the hands, a simple attitude, can help tackle this public health issue.^22,24

Regarding the presence of coliforms, their presence in 9 out of 17 stallholders has been observed, which means that more than 50% of them had the bacterium in their hands. It represents a substantial number of research subjects that had contact with enterobacteria, leaving room for a possible contamination to the handled food.

Coliforms constitute a group of bacteria present in feces and in the environment. They are generally subdivided into two groups: the total ones, which are from the environment and used as indicators of food hygiene quality, and the heat-resistant ones, which originate from recent fecal contamination and are used as sanitary quality indicators of foods.²⁵

The presence confirmation of total coliforms occurred in 5 different stallholders, while the presence of the heat-resistant ones was confirmed in 2 stallholders. The rate of fecal coliforms indicates the amount of organisms from human feces, that is, fecal contamination. This definition, at first, mainly focused on microorganisms from the intestinal tract. The group of total coliforms includes at least three genuses, Escherichia, Enterobacter and Klebsiella, out of which Enterobacter and Klebsiella include non-fecal strains. Due to this reason, the presence of fecal coliforms in food is less representative, as an indication of fecal contamination as opposed to the direct numbering of E. coli, though much more meaningful than the presence of total coliforms given the high incidence of E. coli in the fecal group.²⁶

The presence confirmation of Escherichia coli is of great importance since it represents a major indicator of fecal presence. However, despite the presence of heat-resistant and total coliforms have been confirmed in this study, positive samples in a selective medium of Escherichia coli have not been found. In doing so, regarding this important indicator, the results have been considered satisfactory, thus allowing these subjects to safely commercialize their products. The food handlers, subjects of this research, have been considered tentative transmitters of gastrointestinal infectious diseases, given that Staphylococcus aureus or total and heat-resistant coliforms had been isolated in some of the food handlers. Such data show the necessity to improve the good practices of hygiene and sanitary control for these workers.

Most of the people dealing with food lacks hygiene and sanitary knowledge to handle it, not being aware that they might be asymptomatic carriers of pathogenic microorganisms. As a consequence, this fact makes room for poor hygiene practices employed by unskilled subjects, posing a greater risk of food contamination.²⁰

The health condition of the food handlers, as well as their hygiene habits, directly affect the final quality of product. They must be trained and prepared to perform their job. It is of utmost importance to highlight that hygiene in handling and the proper preparation techniques of food contribute to decrease the risks of the FDs.²⁰

Conclusion

All things considered, it is concluded that the best habits must be encouraged, such as regular hand washing, toilet availability with clean and running water at the market site, and also teach them to assign tasks among co-workers so as to separate money from food.

References

1. Centers for Disease Control and Prevention. Diagnosis and management of foodborne illnesses: a primer for physicians and other health care professionals. Morbidity and mortality weekly report, 2004 April 16;53(no. Rr-4):1-33.
2. Secretaria de vigilância em saúde. Vigilância epidemiológica das doenças transmitidas por alimentos no Brasil, 1999-2004. Ministério da saúde, Brasil. Disponível em URL: http://bvsms.saude.gov.br/bvs/periodicos/boletim_eletronico_epi_ano05_n06.pdf
3. Food and Drug Administration. Bad bug book, foodborne pathogenic microorganisms and natural toxins handbook.
4. Nolla AC, Cantos GA. Ocorrência de enteroparasitas em indivíduos que manipulam alimentos em Florianópolis SC, Brasil. Revista Ciências da Saúde. 2002; 21: 27–31.
5. Abercem Notícias. Boletim informativo de Associação Brasileira de Empresas de refeições coletivas. São Paulo: 1990.
6. Almeida RCC, Kuaye AY, Serrano AM, Almeida PF. Avaliação e controle da qualidade microbiológica de mãos de manipuladores de alimentos. Rev Saúde Pública. 1995; 29(4):290-4.
7. Franco BDGM, Landgraf M. Microbiologia dos Alimentos. São Paulo: Atheneu, 2008. p.182.
8. Murray PR, Rosenthal KS, Pfaller MA. Microbiologia Médica. 7th ed. Rio de Janeiro: Elsevier, 2014. p. 33.
9. Pedrosa AC, Sylvestre SHZ, Fernandes GFR. Avaliação microbiológica das mãos de manipuladores de alimentos de uma cozinha piloto do município de Pirangi-SP. Int J Med Sci Clin Invent. 2015;2(07):1126–34. http://valleyinternational.net/index.php/our-jou/theijsshi/archive/133-theijsshi-volume-2-issue-7-july-2015

10. Brito CS, Rossi DA. Bolores e leveduras, coliformes totais e fecais em sucos de laranja in natura e industrializados não pasteurizados comercializados na cidade de Uberlândia – MG. Biosci J. 2005; 21(1):133-40.
11. Abreu CO, Merlini LS, Begotti IL. Pesquisa de Salmonella spp, Staphilococcus aureus, coliformes totais e coliformes termotolerantes em carne moída comercializada no município de Umuarama – PR. Arq Ciênc Vet Zool. 2011;14(1):19-23.
12. Leite AMO, Franco RM. Coliformes totais e Escherichia coli em coxas de frango comercializados no Rio de Janeiro. R Bras Ci Vet. 2006;13(2):80-3.
13. Cardoso ALSP, Tessari ENC, Castro AGM, Kanashiro AMI, Gama NMSQ. Pesquisa de coliformes totais e coliformes fecais analisados em ovos comerciais no laboratório de patologia avícola de Descalvado. Arq Inst Biol São Paulo. 2001;68(1):19–22.
14. Silva Jr EA, Iaria ST, Andrade CR, Martins EA. Fundamentos para diagnóstico e prevenção das toxinfecções alimentares na cozinha industrial. São Paulo, 1990.
15. Anvisa. Manual de microbiologia clínica para o controle de infecção relacionada à assistência à saúde. Módulo 5: tecnologias em serviços de saúde: descrição dos meios de cultura empregados nos exames microbiológicos. Brasília: 2013. 95p.
16. Anvisa. Manual de microbiologia clínica para o controle de infecção relacionada à assistência à saúde. Módulo 6: detecção e identificação de bactérias de importância médica. Brasília: 2013. 150p.
17. Oplustil CP, Zoccoli CM, Tobouti NR, Sinto SI. Procedimentos básicos de Microbiologia Clínica. 3ª ed. São Paulo: Sarvier, 2010. 530p
18. Guimarães AG, Cardoso RCV, Azevedo PF, Menezes RB. Perfil de susceptibilidade antimicrobiana de bactérias isoladas de queijos coalho. Rev Inst Adolfo Lutz. São Paulo: 2012; 71(2):259-65.
19. Silva N, Junqueira VCA, Silveira NFA, Taniwaki MH, Santos RFS, Gomes RAR, et al. Manual de métodos de análise microbiológica de alimentos. 3ª Ed. São Paulo: Varela, 2007. 536p.
20. Machado JR, Marson JM, Oliveira ACS, Silva PR, Terra APS. Avaliação microbiológica das mãos e fossas nasais de manipuladores de alimentos da unidade de alimentação e nutrição de um Hospital Universitário. Medicina (Ribeirão Preto). 2009;42(4):461-5.

21. Gelatti LC, Bonamigo RR, Becker AP, D’azevedo PA. Staphylococcus aureus resistentes à meticilina: disseminação emergente na comunidade. An Bras Dermatol. 2009;84(5):501-6.
22. Ribeiro IF, Silva SFR, Leite da Silva S, Ribeiro TR, Rocha MMNP, Stolp AMV. Identificação de Staphylococcus aureus e Staphylococcus aureus resistente à meticilina em estudantes. Rev Ciênc Farm Básica Apl. 2014;35(2):301-4.
23. Cerqueira ES, Almeida RCC. Staphylococcus aureus resistente à Meticilina (MRSA) em alimentos de origem animal: Uma revisão sistemática. Rev Inst Adolfo Lutz. 2013;72(4):268–81.
24. Esteves ASMF. Perigos microbiológicos em Alheira: Principais vias de contaminação por Staphylococcus aureus, Clostridium perfringens e Salmonella spp. Dissertação (Doutorado em Ciências Veterinárias) – Universidade de Trás-Os-Montes e Alto Douro, 2005.
25. Franco BDGM, Landgraf M. Microbiologia dos alimentos. Rio de Janeiro: Atheneu, 2006.
26. Silva N, Neto RC, Junqueira VCA, Silveira NFA. Manual de métodos de análise microbiológica da água. São Paulo: Varela, 2005.

Authors

Vitor Furtado Macedo¹; Jonatas Goulart Zanardo²; Raisa Pedrini Coelho Lopes³; Haydêe Fagundes Moreira Silva de Mendonça⁴; Norma Lúcia Santos Raymundo⁵; Rodrigo Moraes⁶

 

^1,2 Undergraduate students of the Medical School at the Escola Superior de Ciências da Santa Casa de Misericórdia de Vitória – EMESCAM.

³ Medical Doctor graduated from the Escola Superior de Ciências da Santa Casa de Misericórdia de Vitória – EMESCAM.

⁴ Master of Natural Sciences from the Biological Sciences Post-graduation Program of the Federal University of Espírito Santo – UFES, Pharmacist, Professor of the Escola Superior de Ciências da Santa Casa de Misericórdia de Vitória – EMESCAM.

⁵ Master of Microbiology from the Federal University of Minas Gerais – UFMG, Medical Doctor, Associate Professor and Vice Coordinator of the Medical School of the Escola Superior de Ciências da Santa Casa de Misericórdia de Vitória – EMESCAM.

⁶ Master of Biological Sciences (Microbiology) from the Federal University of Minas Gerais – UFMG, Pharmacist, Coordinator of the Biomedicine Course at Faculdade Pio XII and Professor of the Escola Superior de Ciências da Santa Casa de Misericórdia de Vitória – EMESCAM.