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TLATEMOANI
Academic Research Journal
Edited by Eumed.net
No. 36 - April 2021.
Spain
ISSN: 19899300
magazine.tlatemoani@uaslp.mx

Reception date: October 01, 2020

Acceptance date: April 01, 2021

ADVANCES IN FOOD MICROBIOLOGY

ADVANCES IN FOOD MICROBIOLOGY

Authors:

Maria Luisa Carrillo Inungaray

maluisa@uaslp.mx

Ricardo López Aportela

ricardoenglad2015@gmail.com

Ana Patricia Fernández Villedas

Paty_s97@hotmail.com

Guicela Ramírez Bernal

guicela.ramirez@uaslp.mx

Autonomous University of San Luis Potosí

ABSTRACT
The use of microorganisms in food processing has been practiced since the
oldest cultures, although their function was not yet known by manufacturers.
Microorganisms can be the best allies in the food industry, but they can also
contribute to food spoilage and cause illness. This work collects relevant
information on food microbiology published in the last five years. Only some of
the most recent advances in the areas of testing, microbial control,

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disinfection processes, functions of microorganisms in the food and food
industry. Although there is still much to discover, it can be said that, thanks to the
progress of food microbiology and new technologies, it is now possible to enjoy
safe food.

Keywords: Microbiological, antimicrobial, disinfection, functional foods,

probiotics, foodomics analysis.

ABSTRACT

The use of microorganisms in food processing has been practiced since the
oldest cultures, although their function was not yet known by manufacturers.
Microorganisms can be the best allies in the food industry, but they can also
contribute to food spoilage and cause illness. This work collects relevant
information on food microbiology published in the last five years. Only some of
the most recent advances in the areas of analysis, microbial control, disinfection
processes, functions of microorganisms in the food and food industry are
presented. Although much remains to be discovered, it can be said that, thanks
to the progress of food microbiology and new technologies, it is now possible to
enjoy safe food.

Keywords: Microbiological analysis, antimicrobial, disinfection, functional

foods, probiotics, foodomics.

INTRODUCTION

Long before microorganisms were known, foods such as leavened bread,

fermented milk, wine and beer were made. The manufacturers then did not know
or understand the process, and their work was by hit and miss.

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 The date on which Food Microbiology begins to be recognized as an independent
discipline cannot be precisely defined; But it could be said that there are two facts
responsible for this independence: the reports of diseases caused by food, and the
increase in international trade, since the food was produced in different continents,
sometimes from endemic areas of enteric diseases, which that endangered the health of
the population and also caused economic losses.

In the 1960s there was a growing recognition of foodborne illness and an increase
in microbiological tests to detect the causative agent. In this environment, the
International Commission on Microbiological Food Specifications was founded in order to
reduce the incidence of pathogenic microorganisms, prevent food spoilage and facilitate
its commercialization. Although its recommendations do not have an official status, they
are considered to develop standards.

As in other sciences, a lot of knowledge has been generated in food

microbiology, which fortunately is published and we can have access to it. When
reviewing articles from the last five years, published in Science Direct, Directory
of Open Access Journals, Scopus, Web of Science and Google Scholar, it is easy to
see that there is a lot of information on food microbiology, so only some of the
most recent advances in the areas of analysis, microbial control, disinfection
processes, functions of microorganisms in the food and food industry.

Microbiological analysis

In relation to the microbiological analysis, both of food and surfaces, in the

last decades the so-called rapid methods for the identification of pathogens have
been developed, which constitute an alternative for the industry to ensure the
safety of its products and take rapid corrective actions. .

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 Among these methods, the use of immunoassays guarantees faster results
in a more reproducible and cost-effective way, compared to culture-based
techniques. Among the rapid methods based on antigen-antibody reactions,
ELISA is undoubtedly the most widely adopted when assessing microbiological
risk due to Brucella abortus, Yersinia enterocolitica and Escherichia coli
O157: H7 from food (Tilocca et al., 2020).

Another analysis technique is fluorescence microscopy, which was used to

demonstrate that the use of a fluorescent dye provides adequate staining of both
the cell wall and the nucleus of Lactobacillus acidophylus
(Page et al., 2020).

Although there is no evidence that food can transmit the SARS-CoV-2 virus that
causes the COVID disease, many companies in the food sector want to prevent the
spread of the virus in their facilities and among their employees.

Researchers from the Higher Council for Scientific Research (CSIC) are currently
working on the European project called COVID-19 BEAMitup, which consists of designing
a molecular detection method for the SARS-CoV-2 on surfaces food processing. East
automated device I know
It will be placed directly in the food processing facility and will provide results in less than
an hour without the need for complex instrumentation or specialized personnel. This
system offers the advantage that it can be used in situ and that the results are obtained
very quickly (Ruiz, 2020).

Microbial control

Another area in which progress has been made is in the control of

microorganisms. And what is the purpose of controlling them? For safety
reasons; to avoid disease and increase the shelf life of food.

We know that the growth of microorganisms can be controlled by

traditional and non-traditional methods. The former can be grouped into
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physical, chemical or biological methods and their various combinations and the
latter contemplate: electrical pulses, ohmic heating, obstacle technology and high
hydrostatic pressures, among others.

In recent years, food preservation has been evaluated using biological

methods, which has generated new paradigms for food safety. Such is the case of
bacteriophages, viruses that infect and multiply in bacteria. Its applications have
been studied in the decontamination of fresh products, in the disinfection of
equipment and surfaces in contact with food, as biocontrol, and as a natural
preservative to extend the shelf life of perishable products. There are products on
the market that have FDA approval for the use of bacteriophages against
pathogens, such as Listeria monocytogenes, Salmonella and E. coli ( Jorquera

et al., 2015).

Contamination of food by pathogens can occur, from farms to factories

and retail, food services and warehousing, originating from different sources,
such as raw food, materials, operators and environmental conditions of the plant.
manufacturing.

Therefore, the growth of pathogenic microorganisms cannot always be

controlled. As Dziedzinska proved et al. ( 2018) who examined field-grown
strawberries, the strawberry farm environment, and fresh strawberries from
markets for bacterial, viral, and protozoan pathogens. Through culture and PCR
they looked for Listeria monocytogenes, Cronobacter sp. and Escherichia coli in
fresh strawberries. All the farms studied applied preventive measures such as
drip irrigation, avoided organic fertilizers and the use of gloves so that workers
reduce the contamination of strawberries. Despite this, certain pathogens were
found that, even in low concentrations, can be a source of infection for
consumers. Therefore, its presence in strawberries is particularly

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importance, since they are mostly consumed fresh and without any thermal
processing.

There are many studies on unconventional antimicrobials. One of them is

the one made by Sanguiñedo et al. ( 2019) who evaluated the antimicrobial
potential of silver nanoparticles prepared using the fungus
Punctularia atropurpurascens, front Staphyloccocus aureus, Escherichia coli, and
Candida albicans. The results demonstrated the antimicrobial potential of silver
nanoparticles for bacteria and of agri-food interest.

Another advance for the control of microorganisms is in foods ready for

consumption or those that are prepared for services of cattering, since they
always have the risk of containing pathogens, so it is necessary to avoid the
temperature range called the danger zone (5 to 57 ° C), conditions that exist
during the cooling of food. Riccia et al. ( 2020) proposed an alternative to cold
storage before serving. They kept the prepared dishes at more than 70 ° C, for
several hours, and not only for less than 2 h before consumption, as is usually
done. With this practice, the development of L. monocytogenes and E. coli,
including O157: H7.

In pursuit of knowledge to prevent the survival of pathogens in food, Zhuosheng

et al. ( 2020) monitored the survival of Escherichia coli Shiga toxin producer (STEC),
Salmonella spp. and Listeria monocytogenes in nuts. Sulfur dioxide treated and
untreated sun dried apricots were used. The results demonstrated that pathogens can
survive in nuts and that factors affecting their survival include the type of pathogen and
the use of sulfur dioxide.

Another work carried out to find new ways to control microbial growth in
food was carried out by Sarjit et al. ( 2020) who studied the survival of Salmonella
after heat exposure in a model meat juice system. Exposure to heat consisted of a
gradual heat treatment at 70 ° C, followed by a heat shock, to

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investigate whether fat, fatty acids, iron, and pH contribute to the effect of heat
against Salmonella. They concluded that heat shock reduces Salmonella more
effectively than gradual heating of red meat, that fat content can protect
Salmonella against the effect of heat, and that the influence of genotypic factors
can correlate with Salmonella survival. .

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 Pathogens in food, in addition to being controlled by physical methods,
can also be achieved by adding salts to food. Heads-Pizarro et al. ( 2018)
evaluated the antimicrobial activity of different

sodium and potassium salts of aliphatic and aromatic carboxylic acids on

Leuconostoc mesenteroides, Lactobacillus plantarum, Enterococcus faecalis,
Candida albicans, Pseudomonas aeruginosa, Salmonella enteritidis and Listeria
monocytogenes. The antimicrobial effect of aliphatic acid salts was greater for
butanoic> hexanoic> octanoic> decanoic acid, while a greater effect of benzoic>
gallic acid was observed in aromatic acids.
> caffeic (Figure 1). And that in general, sodium salts were more inhibitory than
potassium salts (p <0.05).

Figure 1. Effect of aliphatic and aromatic carboxylic acids on bacteria (Information taken from
Heads-Pizarro et al., 2018)

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 To control the development of microorganisms in food, the food industry uses
predictive microbiology as a tool. For which models are used that are generated from the
study of microbial ecology.

The food industry has programs that predict the growth of bacteria of interest.
One of them is the Pathogen Modeling Program, that uses mathematical models to
predict the growth of a certain microorganism and according to the different factors,
predict the number of CFUs that will be reached in a certain time. This allows to know
when the microbiological limits that mark the sanitary specifications for a particular food
will be exceeded.

Disinfection methods in the food industry

The food industry is one of the most productive sectors in many countries,
and meat, bakery and dairy companies, among many others, have very well
defined cleaning processes. The use of incorrect procedures can lead to cross
contamination of food and consequently to spoilage or the transmission of
foodborne pathogens.

For this reason, several strategies have been developed in order to obtain
a good disinfection of surfaces and products; however, microbial resistance has
emerged against commonly used products.

In the food industry, the presence of biofilms generates a serious hygienic-

sanitary problem, causing points of contamination that are difficult to control
during the production process. These are biological structures where
microorganisms grow attached to surfaces and embedded in extracellular
matrices that they themselves synthesize.

In many cases biofilms initiated by microorganisms with high colonization

capacity favor the inclusion of pathogenic microorganisms on surfaces. The
following stages are carried out in its formation:

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 1) Presence of planktonic cells

2) Later, these are grouped into microcolonies that initiate adhesion

3) They can initiate the synthesis of extracellular polymeric substances, which

form the biofilm, in this stage the cell proliferation continues, thus initiating
the adhesion capacity.

4) The biofilm maturation process is carried out favoring intercellular

communication.

5) The release of planktonic bacteria for the formation of other bacterial

communities

The implications of biofilms in the food industry are enormous and represent
a potential threat to all surfaces, since there are phenotypes resistant to cleaning
and sanitizing operations (Martínez et al.,
2020).

Conventional methods of cleaning and disinfection of equipment and surfaces

are effective for the control of bacteria in the free state (or planktonic state).
However, on many occasions they are ineffective for the elimination of biofilms,
since the cells that compose it are more resistant to stress conditions and
antimicrobial agents than cells in the free state.

Traditional antibiofilm methods are limited, alternative methods are currently

being developed, such as coatings with antimicrobial agents or coatings to
modify the surface composition and methods based on plasma-polymerization
technology using atmospheric plasma equipment ( Múgica et al., 2018).

Li et al. ( 2020) studied Shikonin, a bioactive compound found

in the roots of Lithospermum erythrorhizon, and examined its antibiofilm activity against
Listeria monocytogenes and on the key virulence factors of this bacteria. The results
indicated that shikonin could be used as an agent

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alternative to combat biofilm formation and infection by L. monocytogenes.

Microorganism functionality

One way to study microorganisms is according to their function in the food industry.
Some microorganisms are of interest because their biological properties make them
competent for the preparation of functional foods or as starters for the preparation of
other products.

It should be remembered that functional foods are those that have been modified in
their composition, and that provide a health benefit, superior to that of the nutrients that
it traditionally contains. Among the components that give functional classifications to
foods are antioxidant vitamins, omega 3, fiber, prebiotics and probiotics (Days et al.,
2017).

In relation to these functional foods are probiotics, which are live microorganisms
that, when administered in adequate quantities, confer benefits to the health of the
consumer (FAO / WHO, 2002). The main genera used as probiotics are Lactobacillus and
Bifidobacterium. Currently, foods containing probiotics represent an important part of
the functional food market with an emphasis on the dairy industry. However, before
planning the development of foods with probiotics, the challenges involving the proper
selection of these microorganisms must be overcome, as probiotics are expected to show
antimicrobial activity against pathogenic bacteria, having antibiotic resistance and being
capable of stimulating the host's immune system.

Here are some works related to the functionality and biotechnological application of
microorganisms in the food industry.

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 Ruiz et al. ( 2017) evaluated the inhibitory capacity of Lactobacillus spp.,
against pathogens implicated in foodborne diseases (FAD), such as Escherichia
coli O157: H7, Salmonella spp. and Staphylococcus aureus.

Heredia et al. ( 2017) evaluated the mechanisms of action and activity

antimicrobial against pathogens in cheeses. They found that the bacteriocins
produced by lactic acid bacteria have the ability to preserve dairy products.
Bacteriocins are peptides of ribosomal origin that act by forming pores in the cell
membrane of bacteria, causing apoptosis. Therefore, bacteriocins could be used
as a sanitary control in the cheese industry.

Ruiz et al. ( 2019) isolated 21 species of bacteria belonging to the genus

Enterococcus, Fructobacillus, Lactobacillus, Lactococcus, Leuconostoc and Weissella
of wild flowers and fruits. They concluded that LABs of floral and fruit origin have
functional properties that validate their potential use in the manufacture of
fermented fruit-based products, laying the foundations for the design of new
functional foods with this study.

Escobar et al. ( 2020) isolated a lactic bacteria resistant to digestion in vitro, in

pineapple tepache, a traditional Mexican fermented drink. They found that L.
pentosus is resistant to digestion in vitro, which can serve as a starter culture for
the production of dairy foods. This is the first report on the isolation of a lactic
acid bacteria resistant to digestion in vitro from the pineapple tepache.

Vallejo et al. ( 2020) isolated two strains of lactic acid bacteria from the
intestinal content of the Patagonian mussel, which were characterized by
phenotypic and molecular tests. This is the first report on the isolation of
bacteriocinogenic strains of Enterococcus hirae of Patagonian mussel. The high
inhibitory activity and the absence of virulence indicate that these
microorganisms could be applied in biotechnological areas such as food
biopreservation or probiotic formulations.

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 Foodomics

Another area that has gained momentum in the area of microbiology is

foodomics, a new science with a holistic approach. In recent years, whole genome
sequencing (WGS) technologies have been improved, prompting the use of omics
tools including genomics, transcriptomics, proteomics, and metabolomics to
achieve food safety. These techniques have overcome some deficiencies in
current molecular subtyping methods, which is a way of classifying
microorganisms at more specific levels, allowing them to pass the classification
barrier by species and placing them in more intimate categories such as
subspecies, et al., 2016), which improves the detection of foodborne bacteria
outbreaks.

Although genomics, transcriptomics, and proteomics are powerful enough to

generate a large amount of data, more knowledge has yet to be generated in
metabolomics, due to the wide diversity of metabolites. On the other hand, each
omic technology has operated in a fragmented way, so that, in the future, it will
be necessary to integrate the data generated by these technologies (Quiroga,
2016).

Yang et al., ( 2020) developed a detection method based on the

amplification of recombinase polymerase (APR) to Vibrio vulnificus
combining it with a lateral flow strip (LFS) for the visualized signal. The method
addressed the empV A conservative that encodes extracellular metalloproteinase
and completed detection in 35 minutes at a conveniently low temperature of 37 °
C. It showed good specificity and excellent sensitivity of 2 copies of the genome
or 10ˉ¹̄ colony forming unit (CFU) per reaction, or 1 CFU / 10 g in fortified food
samples. This combined APR-LFS method is well suited for detection in situ from
V. vulnificus.

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 Day et al., ( 2015) demonstrated that the PCR detection method for the
detection of Listeria monocytogenes It is highly sensitive and specific in infant
formula formulations and lettuce, which establishes a rapid identification time of
20 and 48 h for presumptive and confirmatory identification, respectively. The
method is a promising alternative to many currently used q-PCR detection
methods that employ traditional selective media for enrichment of contaminated
food samples. Macrophage enrichment of L. monocytogenes

removes PCR inhibitory food elements and contaminating food microflora

producing cleaner samples that increase the speed and sensitivity of detection.

Inns et al., ( 2015) studied an outbreak of Salmonella enteritidis phage type

14b (PT14b) in the UK between May and September 2014, where Public Health
England initiated an investigation to identify the source of infection and
implement control measures. During the same period, outbreaks caused by a
strain of Salmonella enteritidis with a specific profile of multilocus variable
number tandem repeat analysis (MLVA) in other Member States of the European
Union. Food tracing investigations in the UK and other affected European
countries linked the outbreaks to chicken eggs from a German company. They
carried out whole genome sequencing of isolates from UK and European cases,
implicated UK locals, and German eggs - the isolates were very similar. Combined
with the food trace information, this confirmed that the UK outbreak was also
linked to a German producer.

Thanks to foodomics studies, it has been possible to study Norovirus in the

human intestine as a model to evaluate its inactivation (Costantini et al.,
2018), some food toxins have been investigated (Rešetar et al., 2015), food and
microbiological risks of food have been evaluated (Tilocca et

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to the., 2020) and the genetic functions and metabolic properties of microbial
communities have been revealed (Xiao et al., 2020).

In recent years, the application of omics technologies, Metabolomics, has

opened up new perspectives, allowing to change and increase the objectives in
the study of food samples, for example, in the identification of microbial
metabolites. Although, like any emerging analytical methodology, it requires the
search and improvement of a series of experimental aspects, both from the point
of view of Metabolomics and the possible systems to study (Castrejón-Ferrer,
2017).

Prospective

Once we've seen these advances in food microbiology, we might ask ourselves
what's next? What are the prospects? There are new challenges. Table 1 shows a
list of some of the prospects for research in the food microbiology area.

Table 1. Prospects in food microbiology *

• Work on the homologation of rapid and molecular tests to achieve their insertion in the health
regulations of the different countries.
• Work on a microbiological alert system in packaged foods.
• Develop home pathogen identification kits so that consumers have a rapid alert and take
necessary action.
• Conduct further antimicrobial studies to study their safety, solubility, sensory properties (including
color and taste), as well as their stability in different food matrices.

• Generate vaccines by ingesting compounds that activate the immune system in a way directed
towards Gram negative bacteria; all this through the use of edible coatings.

• Continue with studies on pathogens in ready-to-eat foods, especially those pathogens with a low
infectious dose, such as viruses and parasites.
• In the area of foodomics, it will be sought that high-performance bioinformatics tools facilitate
the adoption of omic approaches in food safety so that they soon go beyond bacterial genomics.

* (Own elaboration)

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 CONCLUSIONS

After seeing all these advances and prospects in microbiology, it can be

considered a privilege to live in this historical point, where thanks to the progress
of science you can enjoy safe food. Continuing to cultivate microbiology, in the
interest of its advances, will allow us to have safer food and enjoy good health.

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