DRY FERMENTED SEAFOOD: PROPERTIES, NUTRITIONAL VALUES, MICROBIAL

SAFETY OF SELECTED STARTER CULTURE.

Name

[DATE]
 1
DRY FERMENTED SEAFOOD

Abstract
The need to enhance food preservation methods has seen the embracing of sun-drying and
fermentation methods. These approaches have ensured an improved shelf life besides improving
the palatability of the different dishes. In this paper, we will look at fermenting seafood products
by using microbial as starter cultures.

The study mostly employed the mixed-methods approach to gather the data on the different
aspects and progressions involved in producing fermented seafood products. Participants were
asked about their preferences between fresh and fermented fish, and they gave reasons for their
choices. Also, quantitative methods were used mostly in the laboratory settings to measure the
proliferation of the starter cultures, timelines for successful fermentation, and various
microorganisms' resultant production.

The study has shown the microbiological, physicochemical, and sensory alterations during
seafood products' fermentation. There is a reduction in the cumulative PH of the sample as
organic acid produced. There were noted changes in the flavor and aroma in the fermented
product, the biochemical modifications in the fermented foods to influence food quality. The
paper focused on the changes in texture, the formation of flavor, and how the biogenic amines
accumulate in fermented seafood products.

This paper has also addressed the problems encountered and the future research orientation
regarding seafood fermentation. To ensure a robust food security and humanity's health, future
studies into enhancing seafood fermentation methods will prove valuable. Comprehensive in
vivo studies are needed before there are bio-active fermented seafood products for man's health
benefits.
 2
DRY FERMENTED SEAFOOD

Table of Contents
Abstract............................................................................................................................................1
1.0 Overview of the dried seafood product and usage.....................................................................3
2.0 Type of microorganism use as starter culture in dried fermented seafood product and their
effect on the final product................................................................................................................4
2.1 Fermented fish and microorganisms......................................................................................7
2.1.1 Use of microorganisms as starter cultures.......................................................................7
2.2 Effects of using microorganisms as starter cultures in dried fermented seafood...................9
2.2.1 Effects on flavor............................................................................................................10
2.2.2 Effects on taste..............................................................................................................11
2.2.3 Effects on color.............................................................................................................12
3.0 Factor influencing the choice of a starter culture....................................................................13
3.1 Physiochemical Changes......................................................................................................15
3.2 Microbiological Changes.....................................................................................................16
3.3 Sensory changes of dried fermented seafood using different yeasts or bacteria in
comparison to dried seafood products.......................................................................................18
4.0 Starter culture inoculation mode in the fermented dry seafood production process...............18
4.1 Starter Microbial Culture in Fermented seafood production (Lactic acid bacteria (LAB). .19
4.1.1 Materials and Methods..................................................................................................19
4.1.2 Results and Discussion..................................................................................................20
4.1.3 Summary.......................................................................................................................21
5.0 Effect of the interaction between selected starter culture and flavor precursors in   fermented,
dried seafood..................................................................................................................................23
6.0 Influence of starter culture on the Biogenic Amines in dry fermented seafood......................26
7.0 Conclusion...............................................................................................................................29
8.0 References................................................................................................................................30
 3
DRY FERMENTED SEAFOOD

1.0 Overview of the dried seafood product and usage.

Drying fish in the sun is vital in fish preservation and other seafood products in different parts of
the globe. Drying has been a go-to method widely practiced with the most common mode of
drying being sun drying. According to Mansur et al. (2013), sun drying is the oldest known
method of preserving foodstuffs generally. Dried fish has emerged as a crucial source of
nutrients, especially to people living in coastal areas due to its unique taste and flavor and also
due to its availability. According to Amit et al. (2017), drying the fish solely in the sun or by
integration with salting has resulted in palatable seafood for people who prefer food possessing a
characteristic taste, texture, and flavor. The dried seafood products are considered an excellent
source of vitamins, minerals, and proteins for different people in different geographical settings.
The composition of amino acid in dried fish is higher than in eggs (Chalamaiah et al. 2012). This
study has proven the importance of dried fish products over fresh fish.

Dried seafood does not require refrigeration due to its characteristic longer shelf life, according
to a study done by Immaculate et al. (2013). The increased shelf-life has seen an increased
allocation of resources to process dried fish and other seafood products (Patterson et al. 2018).
About 10% of the total seafood harvested is preserved through drying. Millions of people
involved in fishing and dried fish and retailing have reaped immense benefits from this
preservation method. There is a sequential increase in the demand for dried seafood products
worldwide, which has translated to enhanced profits. The business and practice of drying fish
have evolved from subsistence level operation to a flourishing business. The figure below
represents the process of preparing a dried seafood product "Sheetal Shutki" in Bangladesh:
 4
DRY FERMENTED SEAFOOD

Fig1. Process of preparing dried fermented seafood (fish) in Bangladesh.

2.0 Type of microorganism use as starter culture in dried fermented seafood product and
their effect on the final product.
Fermentation of seafood is a traditional method used in fish preservation and other
seafood products to increase the shelf life, organoleptic and nutritional attributes, and the safety
of the seafood. Fermented fish and seafood are popularly consumed in Southeast Asia and form
an integral part of most food cultures globally. The process of seafood fermentation has been in
practice since ancient technology by our ancestors to overcome the perishable nature of fish;
hence it forms an important industry in most countries. It is a source of exciting microbes. This
review focuses on the types of microorganisms use as starter cultures and how they affect
seafood products. This work mainly emphasizes the microorganisms' contribution to the
 5
DRY FERMENTED SEAFOOD
fermentation process and their effects on flavor, color, and taste. The review has utilized various
approaches in evaluating the composition and functionality of microbial in fermented seafood.

The fermentation process of the seafood is prepared and assisted by the preparation of
starter cultures. Starter cultures represent a microbiological culture that consists of a cultivated
medium that is well colonized by microorganisms responsible for fermentation, as defined by
Fiorda et al. (2017). A cultivation medium might include a grain, seed, or nutrient liquids.
Microbial starter cultures use commercially considered as food ingredients. They can sell as
preparations and formulations of a concentration of either a single microbial species or strains
such as media components that are carried from pieces of fermentation in which they are stored
and standardized for their survival and suitability for food processing applications (Peppler, &
Perlman, (2014).

Products Microorganisms involved in seafood fermentation

 6
DRY FERMENTED SEAFOOD
Fermented,  Lc. Lactis
dried, and  S. carnosus
smoked fish  Strep. faecalis
products.  Lb. plantarum
 Sarcina sp.
 Lb. pobuzihii
 Corynebacterium sp
 Lb. fermentum
 Tetragenococus halophilus subsp. flandriensis
 Lb. amyamylophilus
 Pseudomonas sp
 Lb. coryniformis
 Halococcus sp
 Ent. Faecium
 Halobacterium salinarium
 Ent. Faecalis
 H. cutirubrum
 Bacillus subtilis
 Clostridium irregular
 B indicus
 Azorhizobium caulinodans
 Micrococcus sp.
 Candida sp
 Staphylococcus cohnii subsp. Cohnii
 Saccharomycopsis sp.

Table1. Microorganisms used in the fermentation of seafood products.

 7
DRY FERMENTED SEAFOOD
2.1 Fermented fish and microorganisms
In maintaining the character of the fermented products, renovation through the utilization
of microbiota in improving the nutritional qualities of a product with the microbiota of traditional
products has played an enormous role in ensuring a degree of quality and safety of the Fermented
products is met (Beena Divya et al., 2012). Fermented products are of high value biologically,
sometimes exceeding those of unfermented ones described by Selhub, Logan, & Bested (2014).
This section focuses on various areas on fermented products microorganisms as they are used as
starter cultures.

Table 2. Nutritional composition of fermented seafood (fish).

2.1.1 Use of microorganisms as starter cultures.

Fermentation of seafood products with microorganisms as starter cultures can subdivide
into two groups: (1) fermented seafood products with the addition of carbohydrates and (2)
fermented seafood products without carbohydrates. Microorganisms play a significant role in the
 8
DRY FERMENTED SEAFOOD
fermentation of dry seafood products when used as starter cultures. Zaman et al. (2011)
described the starter cultures' part in inhibiting biogenic amines production. Šušković et al.
(2010) stated that they are essential in stimulating antimicrobial activity against target pathogenic
microorganisms.

Microorganisms significantly impact the Fermented seafood products, which have

diversified uses both traditionally and commercially—many organisms belonging to the Lactic
Acid Bacteria (LAB). The bacteria have a far-reaching metabolic activity and can fully inhabit
some habitats (Giraffa et al., 2010). It includes the genus Lactobacillus, the predominant flora in
som-fak is LAB, including lactobacillus pentosus and Lactobacillus Plantarum. Som-fak is a
seafood product prepared from the amalgamation of fish, rice, garlic, and sucrose. Yang et al.
(2020) identified Lactococcus lactis up. Lactis as the best stater in the fermentation of air-dried
Megalobrama amblycephala, a name for a fermented fish in china. Lactobacillus sakei was the
predominant out of 61 starter cultures of LAB in the fermented fish product (chouguiyu) in
china. Other identified microorganisms included species of Vagococcus, Lactococcus,
micrococcus, Enterococcus, and staphylococcus as determined by (Martínez-Álvarez et al.,
2017). Pediococcus pebtosaceus and L. Plantarum was present as starter cultures in grass carp
sausages (Nie et al., 2014). Koyanagi et al. (2011) found that Pediococcus and Lactobacillus
were present in the fermentation of Sushi in japan (Narezushi).

Vital halotolerant microorganisms or halophilic bacteria are isolated and characterized to

help in the standardization of seafood products quality since the production of fish-type products
using the genus Pediococcus intensifies the rates of processing and production consistency,
improve sensing characteristics and microbial quality hence shortening fermentation time
according to a study done by Speranza et al. (2017).

Apart from Lactobacillus, which dominates the market for fermentation of seafood, other
species of bacteria were identified in dried fermented seafood products. Staphylococcus spp is
the primary starter culture identified in cincalok/or Malaysian fermented shrimp, as stated by
Hajar and Hamid (2013). Enterococcus faecalis and Enterococcus faecium bacteriocin-producing
strains that lack the decarboxylase activity of amino-acid were found in fermented seafood
products (shrimp, mussel, and fish) in Thai (Valenzuela et al., 2010). Following the presence of
Lactobacillus bacteria, Pseudomonas, Bacillus, Staphylococcus, Candida, Klebsiella,
 9
DRY FERMENTED SEAFOOD
Debaromyces, and Pediococccus were present in enam nesetaake (fermented fish-carbohydrates
meal containing yams, ginger, onions, salt, and fish.

Molds are used in the fermentation of seafood products as starter cultures. For instance,
Pehtze surimi's fermentation is done using Actinomucor Elegans XH- 22 (Zhou et al., 2014). On
the other hand, douchie starter culture and koji molds (containing live Aspergillus oryzae) are
used in the fermentation of silver carp fish as studied by Kasankala et al. (2012).

Researches show that microbial are used both as starters and as probiotics in the dried
fermented seafood products. A study by Kanno et al. (2012) found that Leuconostoc
mesenteroides 1RM3 Lactibaliccus Plantarum 7FM10, which were among five isolates types of
LAB (that can ferment and scavenge DPPH- and O2), showed synergistic effects on saba-
narezushi (fermented chub mackerel with rice). The two genera of LAB can be used as probiotics
starters. Kuda et al. (2014) identified pediococcus pentosaceus, L. Plantarum, and Lactococcus
lactis microorganisms from samples of fermented fish and intestines of the fish. These
microorganisms showed their potential to be used as starters and functionality with antioxidant
and anti-inflammatory properties. Singh et al. demonstrated a microbiota diversity present in
samples of hentak ngari and tungtap, products of fermented seafood in northeast India. (, 2018).
In the examples, they identified LAB (L. lactis subsp. cremoris L. Plantarum, Lactobacillus
coryniformis subsp. Torquens, Lactobacillus amylophilus, Enterococcus faecium, and
Lactobacillus fructosus), which showed a robust probiotic character (hydrophobicity).
Endospore-forming rods (both Bacillus pumilus and Bacillus subtilis), yeasts (Candida and
Saccharomycopsis), and aerobic coccal strains (Micrococcus) were also identified. These strains
could not produce biogenic amines (cadaverine, putrescine, tyramine, and histidine) because they
were isolated.

2.2 Effects of using microorganisms as starter cultures in dried fermented seafood.

Proteinaceous diets such as dried fermented seafood products such as fish are associated
with a significant hazard from pathogenic bacteria such as the vibrio spp., production of
physiologically active amines, and the presence of parasitic worms (Shikongo-Nambabi, &
Schneider (2012). For unheated seafood in anaerobic conditions, a concern occurs in the growth
of Clostridium botulinum and its toxin production. However, through lactic fermentation, the
pathogenic bacteria's development will be inhibited in high-salt, and low-salt lactic fermentation
 10
DRY FERMENTED SEAFOOD
of seafood products once prepared in the appropriate concentration of salt and proper pH. The
table below shows the pH of samples of seafood samples fermented with LAB as starter cultures.

Table 2. pH of samples fermented with LAB

Histamine is a type of physiologically active amines, formed through the bacterial

decarboxylation of histidine. Histamine can be included in large quantities and might cause
poisoning in different seafood products. Jet-gal is a high-salt fermented seafood product used as
additives in making Kimchi and a side dish. It is obtained from viscera and salts' muscles and
contains precursor amino acids of biogenic mines in large. Some studies suggest that the addition
of garlic and glycine can inhibit decarboxylase activity of amino acid in Myoelchi-joet (made of
anchovies). In the research done by Zhou et al. (2016), as noted, a starter culture treated with
garlic extract saw a reduction of the contents of cadaverine and tyramine by 18% and 30.9%,
respectively. Compared to the control, Contents of putrescine, histamine, spermicide, tyramine,
and cadaverine saw a reduction of 32.6%, 93.2%, 100%, 100%, and 78.4%, respectively. Glycine
records the most vigorous inhibitory activities on the production of biogenic amine.

In the use of microorganisms as starter cultures in dried fermented seafood products,

there are various identifiable effects on flavor, taste, and color. Such results can be
discussed below as follows:

2.2.1 Effects on flavor

In Asia, fermented fish products include salt-fermented products such as fish plates,
cured fish, and fish sauce. Fermentation by microorganisms as starter cultures is done when the
salt concentration is below 20%. When the concentration of salt is higher than 20% of the total
 11
DRY FERMENTED SEAFOOD
weight, the growth and multiplication of microorganisms, i.e., pathogenic and putrefactive, are
prevented. In a low salt concentration below 20% of the total value, the old lactic fermentation
method is used to preserve fish by adding carbohydrates (Riaz, 2019). In the lactic fermentation
method, ingredients such as sugar, rice, millet, and flour are used as the source for carbohydrates,
as described in the study by Zhang et al. (2020). Microorganisms responsible for the
fermentation process of dried fish products also contribute to aroma and flavor formation.

Alternatively, in low-salt concentration, the dried fish can be fermented with vinegar at
low temperatures. This method is widely used in Scandinavian countries. Vinegar is a food
product made from acetic acid bacteria commonly found in fruits and is used in fermentation.
Vinegar adds flavor to fermented fish products and is aromatized with spices and herbs.

I mixed starter cultures of Lactobacillus casei subsp. Casei- 10001, p. pentosaceus-

ATCC33316, Staphylococcus xylosus – 12, and L. Plantarum- 15 were fouQ7 3OOU nd to

enhance minced silver cap muscles in terms of qualities; functionality, flavor, nutritional value,

and digestibility according to Zheng et al., 2013.

Activities of the gut bacteria in the belly cavity of an eviscerated fish cause off-flavors'
production, as stated by Safaeian & Khanzadi. On the other hand, Prabhakar. (2020) explained
that the metabolic activities of a fish's existing microorganisms react to cause spoilage upon the
death of a fish aided by the oxidation process. As a result, the fish loses its freshness, which can
be determined by the change in smell and visual analysis—the smell changes from no smell to an
intense off-odor.

2.2.2 Effects on taste.

Upon the death of a fish, the microorganism present goes through a metabolic activity
that affects the fish's taste in four phases of spoilage. The fish develops a sweet, sea-weedy taste,
and a slightly metallic taste in the first grade. A neutral taste follows in the second phase, while a
sour taste dominates the proceeding phases in quality three and enhances the flavor in the final
grade.
 12
DRY FERMENTED SEAFOOD

Fig 1. Effect of microorganisms' starter cultures on the taste of dried fermented seafood
products.

The above process analyzed by the quality scoring methods and represented the complex
mixture of chemical, physical, biochemical, and microbiological actions strongly influenced by
storage.

2.2.3 Effects on color

The use of microorganisms as starter cultures in fermented, dried seafood has several
effects on seafood color. (Ikapet, 2017) stated; flesh on the belly cavity affected by activity of the
gut bacteria to cause discoloration of fish.

Upon the death of a fish, for instance, a metabolic activity through the bacteria inside the
fish's system reacts to cause spoilage, hence the need for fermentation. Microorganisms affect
the color of the fish, which can be determined after a short time upon the fish's death through
visual analysis. Identifiable changes in the shade of fish include a shift from pink and shiny color
to brown and slimy on the gills, a shift from iridescent to dull and bleached with a slime of
bacteria for the skin, and a change bright and elastic to flat and soft on the flesh (Prager, 2011).

Property of the fish/seafood Effect of the starter cultures

Flavor The sweet, sea-weedy aroma in phase 1, the
 13
DRY FERMENTED SEAFOOD
little flavor in grade 2, bitter-off flavors,
cabbage-like, ammoniac, sulfurous smells in
level 3 intensification of the bitter-off flavors
in step 4.
Taste Delicate sweet in phase 1, neutral taste in
grade 2, sour taste in step 3, and a concentrated
sour taste in grade 4.
Color Discolored the fish; change from a shiny pink
color to a slimy brown color on the gills. A
shift from an iridescent skin to a dull and
messy bleached skin, and a change from a
bright elastic flesh to soft and pale flesh.

3.0 Factor influencing the choice of a starter culture

During the pursuit of ideal starter cultures to be used in dry fermented seafood, Sornplang et al.
(2016) recommend the studying of the proneness of microorganisms derived from fermented
seafood products and the antibiotics. However, this does not mean the study of microorganisms
is restricted to searching for an ideal starter culture. According to Barbieri et al. (2019), the US
Food and Drug Administration (FDA) has recommended the use of starters considered as GRAS
(Generally Regarded As Safe) because they can curtail the proliferation of detrimental
microbiota such as spoilage microorganisms and pathogenic.

The criterion used in the selection of starter cultures should take into consideration the raw
material to use, the requirements of food safety, the inherent properties of the strain, and the
quality attributes. There is a lot of attention given to the application of starter culture in the
fermentation of seafood products. The use of starter cultures is a vital and sustainable approach
in the efficient preservation of seafood products with a lot of advantages technology-wise.
According to Wang et al. (2013), arrays of microorganisms are utilized as starter cultures in
curtailing the development of pathogenic microbes and subduing the growth of microorganisms
that aid in spoilage. Different starter cultures can be used in various products based on consumer
and technological specifications.

Starter cultures work by rapid matrix acidification or by producing bacteriocins. Bacteriocins are
antimicrobial peptidic toxins produced by the bacteria to inhibit the growth of similar bacterial
strains and are an ideal replacement of chemical preservatives in the fermentation of seafood
products. The safety of the seafood products may be jeopardized by foodborne pathogens such as
Listeria spp, which may cause serious health complications to humans. The antimicrobial activity
 14
DRY FERMENTED SEAFOOD
of the numerous microorganisms used as culture starters should be extensively studied to ensure
the safety of the finished fermented fish products.

In the selection of starter cultures for fermentation, e.g., LAB, their efficacy must be considered.
The selection criteria will depend on the following parameters:

a) Ability to release fast acidification and production of antimicrobial compounds

Fermentation using a starter, e.g., LAB, relies on eliminating fermentable carbohydrates,

oxygen consumption, organic acid production, and the subsequent decrease in the Ph. A
vital factor in selecting the starter culture is that it should be capable of swift and robust
production of organic acid that will alter the pH of below 4.0 inside the 24 hours
(Saithong et al. 2010). Starter cultures have been found to produce organic acids like
diacetyl and ethanol. According to a study by Zeng et al. (2014), the swift production of
the organic acid aids in the pathogenic and flora inhibition and leads to the enhancement
of the shelf life and microbial integrity of the fermented seafood products.

b) Capacity to control the original microbiota

Another attribute of a starter culture is that during the seafood fermentation, it can dominate the
local pathogenic populations. This domination is achieved by the relatively high exponential
growth of the starter culture during the fermentation and the release of bacteriocins with
incompatible properties (Sonsa-Ard et al. 2015).

c) Capacity to impart probiotic effects.

Although the study into the benefits accrued from starter culture, i.e., LAB, have presented the
researchers with doubts, their probable probiotic attributes, which include antimicrobial
characteristics and their survival in bile and acids, must be considered in adherence with the
selection criteria for starter cultures (Beganović et al. 2014). For curtailing the proliferation of
pathogenic bacteria, probiotic bacteria have proven valuable, and its ability to survive in the
upper gastrointestinal tracts, its tolerance to acids, and bile is the parameter that is considered.

d) Capability to hydrolyze the starch

The ideal starter cultures work by hydrolyzing the present starch in the seafood product sample
(Songre-Ouattara et al. 2009). The starter culture with the most significant completion of the
hydrolysis process will thus be preferred the fermentation process. It is important to note that the
hydrolysis enhances the taste of the seafood product as it usually leads to an increase in the
density of the nutrient.

e) Capability to have a solid stability

This characteristic refers to the feasibility and the metabolic undertakings of the dried starter
culture. According to Yao at al. (2014), it is of crucial importance that for the successful
 15
DRY FERMENTED SEAFOOD
production of starter cultures, the stability of the cultures during the drying should be stable and
so is the storage for long-term aims. According to Rezac et al. (2018), to accrue the health
benefits from fermented seafood samples, the probiotic microorganisms should be feasible in the
range of 10⁶ - 10⁸ CFU/g of the food sample.

3.1 Physiochemical Changes

According to a study by Mueda (2015), the physiochemical attributes of raw and fermented
seafood products at varying stages of the fermentation process are analyzed periodically to
determine or assess the pH, moisture content, and acidity. With the use of a digital pH meter
(NIG, Model No. 333), the PH of the product can be determined, and the moisture content can be
assessed and determined by the aid of the thermogravimetric method and then presented as a
percentage of moisture. In this method, there is the derivation of the moisture content by
calibrating the product weight change while still heating the product in a controlled environment
until no further weight is observed. The homogenized fish product is titrated against 0.1 NAOH
with phenolphthalein to determine its acidity.

A type of fish called silver barb, also scientifically known as Barbonymus gonionotus, is usually
preferred in the fermented state due to the enhanced nutritional attributes, the unique aroma, and
flavor. During the fermentation of fish, it is mixed with salt, garlic, and sticky rice and then
wrapped tightly in a plastic bag until a sour tasted is achieved by the aid of the indigenous
microorganisms. After the progression of the fermentation progress, LAB is identified as the
principal bacterial species; it produces lactic acid by utilizing carbohydrates. Thus the quality of
the resulting fermented fish is customarily related to the pH and acidity of the end product.

The table below shows the physicochemical changes during the successful fermentation of silver
barb during the 3rd day to the 60th day.

Sample Moisture Content (%) pH Titrable Acidity

Raw fish 69.1 7.63 0.14
3rd Day 64.0 6.7 0.22
10th Day 57.5 6.36 0.29
20th Day 48.9 5.8 0.43
30th Day 39.2 5.5 0.50
40th Day 34.8 5.8 0.43
50th Day 28.2 6.3 0.37
Final product 24.42 6.4 0.26
(60th Day)

In a study by Saithong et al. (2010), he posits that since the available carbohydrates are
fermented to produce organic acids, i.e., lactic acid, this leads to a considerable reduction in the
pH. The decrease in pH is the main preservation factor in fermented seafood and other general
 16
DRY FERMENTED SEAFOOD
fermented food products. In the table illustrated, a gradual reduction in the pH indicates excellent
acidification of the raw material (fish) via lactic acid bacteria fermentation.

There is a sequential reduction in the moisture content of the raw fish due to the osmotic process.
The salt wrapped tightly with the fish product helps in the osmotic loss of water from the fish as
the cell membranes are selectively permeable, allowing the penetration of salt in the cell. The
water in the cells is pushed out automatically until an osmotic pressure is reached.

As it's evident in the table, there are changes in the total acidity; this affects the stability of the
sauce due to the ability of lactic acid content to make the sauce to last for long. The amount of
lactic acid produced will increase exponentially as the fermentation goes on. The availability of
nutrients/food during the process of fermentation supports the growth of lactic acid. The primary
role of lactic acid bacteria is to produce organic acids, i.e., acetic acid, lactic acid, and other
metabolites. The produced lactic acid also creates a distinctive flavor of organic acid.

3.2 Microbiological Changes

For all seafood fermentations aided by the injection of the starter cultures, there is a steady, sharp
increase in the cumulative number of lactic acid bacteria just after 24 hours of fermentation.
When the Silver barb is introduced to the single starter culture, there are different patterns.
 17
DRY FERMENTED SEAFOOD
Figure: Shows the population of LAB and TPC during Silver barb fermentation. The data
representation is the average of the two independent trials. The standard deviation is represented
by the bars.

The number of lactic acid bacteria in the silver barber inoculated with a single starter culture
decreased slightly from the 48th hour until the termination of fermentation. The number of LAB
at the end was 3.0 x 10⁹ CFU/g. There was a similar pattern of growth in the total plate counts
(TPC) during the same fermentation process (Above fig B). The aerobic plate counts initially in
the two fermentation batches were the same in both sets; after 24 hours, the bacteria started
proliferating. In the control fermentation batch, the aerobic plate count was much lower than in
the standardized silver barb inoculated with a starter culture. From these observations, it's
concluded that lactic acid bacteria dominated the fermentation process of vaccinated silver barb
fish but not in the control sample.

The study assumed that yeast and bacteria were responsible for the process of fermentation. In
the infancy of the fermentation, the total yeast count grew from 4 Log CFU g¯¹ on the first day,
on the second day it was 6Log CFU g¯¹ and finally decreased to 3 Log CFU g¯¹ on the last day
of fermentation. The growth of the total count was explained as resulting from fierce competition
by the lactic acid bacteria, which were growing fast. The development of lactic acid bacteria led
to the production of organic acids that work as preservatives. The simple sugars availed were the
result of the fermented rice; it provided energy for the microorganisms aiding in the
fermentation.

3.3 Sensory changes of dried fermented seafood using different yeasts or bacteria in
comparison to dried seafood products.
In most East-Asia parts, the main ingredient and source of condiments have been the fish sauce;
it imparts on the umami taste, enhancing the salinity in the food. This attribute has found fish
sauce used as a temporary alternative to sodium chloride to prepare food. This attribute has
presented vital options for consumers with health complications, such as in salt-prohibited
consumers. There are no studies that have been done to assess the degree in which efforts to
reduce the NaCl content can compromise the uniqueness of the food, i.e., taste and influence the
consumer preferences. The study has hypothesized that for successful reduction of salt content,
fish sauce can be used as an alternative to salt without affecting the consumer's palatability and
acceptance.

The study also sought to rate the three sensory attributes of the fish sauce. The features are the
saltiness, deliciousness, and intensity of the taste. The involved fermenters provided the
descriptions of the fermented silver barb fish; there was a description of the alterations in the
color, texture, and aroma and sought to know if the process of fermentation had gone to
completion. As a part of the periodical observations, there was a presentation of a triplet of
questions:
 18
DRY FERMENTED SEAFOOD
 What was the observed color of the fermented seafood product after the completion of
the fermentation process?
 Had the expected texture of the seafood product been possible?
 Had the desired aroma of the seafood product achieved?

Before starting the fermentation, the fish samples had a dominant smell of fresh and seawater
fish. Beginning the 7th day and peaking by the 24th, fermented fish's sour smell began to appear.
Also, the spoiled, intense aroma of the fermented fish persisted until the fermentation's end day.
The putrid smell also masked the fish's salinity starting on the 12 th day, where the sourness had
eclipsed the saltiness until the last day of observations. The application of a colorimeter assessed
the ensuing colors of the dry fermented silver barb fish. The assessed data is presented in Lab
color space where the observable colors alternated from red to yellow.

4.0 Starter culture inoculation mode in the fermented dry seafood production process
Fermentation is a method that is used to preserve foods for organoleptic and nutritional values.
Seafood products are essential sources of integral food worldwide (Jinhong Zang et al., 2020). In
addition to that, these fermented seafood products provide sources of essential microbes used in
food processing industries. This work clarifies microbes and microorganisms' contribution to
fermented seafood and emphasizes updating the manufacturing processes for fermented seafood
globally. A substantial number of methods and approaches have been incorporated to understand
and determine the composition of microbes in the seafood and their relevance in fermentation
processes. In this paper, we try to understand the starter inoculation mode in the fermented dry
seafood production process using bacteria and yeast in a series of tests.

According to T. Ohshima (2014), fermented seafood is any fishery product that has goes through
a series of enzymatic or microbiological activities. The starter culture used in the processes is
prepared from living microorganisms deliberately to aid the beginning of fermentation to infer
changes in the specimens' properties, chemically and sensory properties, so that a homogenous
product is produced. Starter culture inoculation mode is the use of prepared living
microorganisms to assist the fermentation of dry seafood.

During the production of fermented seafood, large populations of required microbial organisms,
referred to as starter cultures, are used (Hosseini and Pilevar, 2017). Honzel, W. H. (2002) stated
that these starter cultures could be derived from microbiotas pre-existing in the specimen; in this
case, seafood, or else subjecting selected starter cultures under a controlled environment to
ensure monitored, procedural methods. In comparison, impulsive fermentation has a higher trait.
Impulsive fermentation displays a better quality due to specific yeast and bacterium (Leroy.F. et
al., 2006). According to Xu et al. (2010) publication, these starter cultures comprise Lactic acid
that produces lactic acid bacteria (LAB), and micrococci bacteria are used as healing agents,
mold, and yeast.
 19
DRY FERMENTED SEAFOOD
4.1 Starter Microbial Culture in Fermented seafood production (Lactic acid bacteria
(LAB)

4.1.1 Materials and Methods

This review sought to evaluate the effects of starter cultures in microbiological, physicochemical,
and sensory characteristics of seafood products. The research used single starter culture, one
bacteria (Lactococcus lactis M10), mixed starter culture comprised of one bacteria and yeast
(Lactococcus lactis M10 and W. cibaria M3), and another sample without any starter culture,
which acts as a control. The study aimed at shortening the fermentation period by starter culture
inoculation.

In an experiment done by C.H Lee (2010), nine LAB bacteria were isolated from a
seafood fish and used to inject the fish. Then the sensory evaluations were performed. For 18hrs
and at 37⁰C, the two bacteria; Lactococcus lactic M10 and W. Cibaria M3, Lactococcus lactis,
and W. cibaria M3, were grown in MRS broth. The individual cell excrements were harvested by
separating particles from the suspension and later washed twice in alkaline-based water (saline
water) and then re-suspended in sterile water. The fish specimen under scrutiny were prepared
according to ancient methods and then divided into four batches. The batches of saltwater and
fish inoculated with various starter cultures, including L (Lactococcus lactis M10), W (W.
cibaria M3), LW (lactococcus lactis M10 and W. cibaria M3,1:1), and a batch of control
specimens. The curing liquid and each of the starter cultures mixed thoroughly, and the
containers sealed and kept below the temperature for a week.

4.1.2 Results and Discussion

Changes in Lactic Acid Bacteria during the fermentation

The incentive pH varied from 6.81 to 6.97 in the control specimen and injected samples
from this data in the table. It was discovered that during the procedure, the alkalinity values
gradually decreased and ranged between 6.47 and 6.58 in all specimen samples
 20
DRY FERMENTED SEAFOOD
The acceptability of the product was evaluated to determine sensory changes with
inoculation or without. These changes included; outward form, smell, flavor, and tactile
appearance across all samples. In the four instances, no changes in the exterior look discovered
(p<0.05), leading to the conclusion that injection did not affect texture and appearance. (Cruzen.
Et.al, 2014) stated that the activity of endogenous enzymes in the specimen was the reason for
the observations made in this study and attributed to seafood products' tenderness. Hence, the
study safely concluded that the use of starter cultures did not affect both outward look or flavor

According to (Zeng et al. (2018), the growth of volatile compounds essential for the
distinct smell in fermented seafood products is mostly from microbiotas in preference to the
specimens' enzymes. This result led to the conclusion that the injection of Lactococcus lactis
M10 and W. cibaria M3 were crucial players in improving quality and acceptability while
delayed seafood products to expire quickly at the same time.

The table above shows the sensory changes by injecting various starter cultures and control.

4.1.3 Summary
This experiment's findings showed the injection of starter cultures could speed up the process
and are aggressive interactions between Lactococcus lactis M10 and W. cibaria M3. Firstly,
flavor formation was depended on lactis M10 and later W. Cibaria M3. Hence, observed that the
use of these two LABs fosters flavor enrichment, quality improvement, and generally favorable
reception and shortening fermentation time in large scale production of dry fermented seafood
products. Therefore, the co-inoculation of Lactoc. lactis M10 and W. cibaria M3 in seafood
products could enhance the flavor formation, improve quality properties, overall acceptance, and
shorten fermentation time in large scale production of dry fermented seafood products.

In the book, LWT-Food Science, 2008 pg569-580), research on the effect of injection of
various LABs, the fermentation and quality of Som-fug from bigeye evaluated. In this method,
 21
DRY FERMENTED SEAFOOD
Pediococcus acidilactici at 10^4 CFU/g injected in the som-fug, and the control set up. They
showed greater acceptability than those inoculated with Lactobacillus Plantarum and
Pedococcuc pentosaceus at either 10^4 or 10^6 CFU/g and the power (without injection). During
the procedure, Pediococcus acidilactici fermented earlier than the control set up. This
fermentation was attributed to a more significant alkalinity decrease and the production of lactic
acid. Based on the alkalinity desired (pH 4.5), the process was done within two days for the
bacterium-induced specimen and one and a half days for the sample without the cultures.

Som-fug injected with LABs was generally more rigid and showed higher adherence
properties than the control. From the result, injection with P. acidilactici resulted in a faster
fermentation process and improvements in quality. Therefore, the bacterium is used as a
potential culture starter in producing fermented dry seafood. Besides, Hosseini and Pilevar
(2017) reported that the level acid and bacitracin go up, and this attributes to more significant
populations of L. curvatus and L. sakei, that subdues maturations disadvantageous pathogens.

According to (Maria Joao Fraqueza et al. 2017), another critical seafood starter culture
was composed of coagulase-negative staphylococci (CNS). These re the Gram-positive cocci
group, and the genus belonging to the genera Staphylococcus and Kocuria is one of most
essential cultures. They also stated that at the early stages of the procedure, the microbiota's
outward look is primarily due to yeasts. This, however, showed that as the yeast composition
decreases, the molds outgrow the yeasts and entirely colonize the finished product. In some
cases, it is considered an early indication of spoilage. Yeasts are the endogenic properties of the
microbiota existing on fermented seafood products. The environment in which they thrive and
the seafood used as a natural resource triggers yeast growth since they naturally exist on fresh
seafood. The recurrent genera are Candida, Rhodotorula, Debaryomyces, and Trichosporon
(Mario Joao Fraqueza,2017). In their publication, (Mi-Ju Kim et al., 2020), the white colony-
forming yeast in seafood products were critical elements in the fermentation conditions and
metabolite changes in seafood products. These indicate a reduction in seafood product quality.

Their publication investigated the result of disparate fermentation temperatures all below
room temperatures and. The four species of C. sake, K. servazii, K. barnetti, and Debaryomyces
hancenii primly detected at ten ⁰C.

From research by (Mi-Ju Kim et al., 2020), under the open packaging, the white colony-
forming yeast (WCFY) population's diversity was more significant than that under the closed
packaging condition. However, the nonvolatile production by the varying WCFY community did
not differ majorly in both states at all temperatures. In contrast, glycerol levels in the samples
hosting WCFY increased relative to the control specimen (0 days). From these observations, an
indication that the temperature and air exposure can change the diversity of WCFY on the
surface of the seafood surface.
 22
DRY FERMENTED SEAFOOD
According to (Di Zhang et al., 2010), a study on one of the most preferred seafood, yellowfin
Tuna, has high economic value because of its eccentric flavor and augmentation. The impact of
volatile and nonvolatile compounds on the taste of seafood products was investigated (Yang et
al., 2016. The research concluded that the flavor components in seafood products highly
depended on the method of production used.

From the research done on this paper on starter culture inoculation in the fermentation of dry
seafood production processes, the impact of bacteria-based inoculation mode surpasses the yeast-
based way. The culture injection method of fermentation is a popular approach used by seafood
products' production to augment fermented seafood products (de Almeida et al., 2018; Xu et al.,
2018). Compared to impulsive fermentation, injection of neophyte starter culture into fermented
seafood products has more, such as delayed degradation time, reduced spoilage, improvement of
taste, and enriching the work's quality. Lactic acid bacteria (LAB) in various fermentation
processes, for instance, fish, have been widely applied in manufacturing industries (Zeng et al.,
2013; Gao et al., 2018).

Dry fermented seafood products are low-calorie protein sources and are considered t have
high vitamin content. Due to their sensory properties, the seafood protein is more comfortable to
digest because seafood has less connective tissue (Navarrete-de-Toro et al.,2015). In addition to
that, closed canned seafood products preserve their native tastes and flavor depending on the
processing method used (Di Zhang et al., 2010). However, in the same study (Hosseini and
Pilevar, 2017), it was discovered that Enterococcus species are a threat to consumers who have
large intakes of canned degraded dry seafood products. The species are conceptually responsible
for five types of side effects: systemic infections, harmful metabolic activities, and risks of
adjuvant side effects, excessive retroactivity in susceptible individuals, and peril of gene transfer.
Therefore, this paper recommends investigating the corollary effects and toxic elements before
using starter cultures in seafood and dry seafood products.

5.0 Effect of the interaction between selected starter culture and flavor precursors in  
fermented, dried seafood
Starter cultures are microbial relevant for promoting and guiding the fermentation of meat
products. This section focuses more on lactic acid bacteria and yeast, although other starter
cultures such as molds and coagulase-negative staphylococci (CNS) exist. The production of
antimicrobial substances such as bacteriocins and rapid matrix acidification is applicable to
achieve the increased safety of meat products' fermentation. Alternatively, starters help to
shorten the ripening period and standardize product properties. However, the following
microbiological may jeopardize the security of fermented meat products foodborne pathogens.
Consequently, various chemical hazards can jeopardize fermented seafood products' wellness;
they include nitrosamines, biogenic amines, mycotoxins, and polycyclic aromatic hydrocarbons
(PAH). Nitrogenous compounds such as biogenic amines emanate from the decarboxylation of
specific amino acids. Furthermore, their formation may be due to some microorganisms. Starters
 23
DRY FERMENTED SEAFOOD
can inhibit the accumulation of biogenic amines (BA) by decreasing PH, preventing amino acid
decarboxylative ability microorganisms from growing. To prevent pathogenic and spoilage
bacteria, a PH of five and below is required. Starters achieve BA production reduction by
competing with the non-starter microbiota and autochthonous through the ripening and storage
process. The formation/accumulation of BA reduced by the strains of Lactobacillus sakei and
Lactobacillus Plantarum. Subsequently, the degradation of BA in types of seafood can be due to
debaryomyces hansenii and staphylococcus xylosus strains. PAH products of incomplete
combustion of organic contain multiple aromatic rings, mostly in wood used for smoking meat.
However, the elimination of PAH by the starter culture is a procedure that causes common
misunderstandings. Recently, a mixed-strain culture applies to seafood fermentation. This
method is because seafood production has widened, making it an essential branch of the world's
fermented food industry. Therefore, there is a need to develop an influential starter culture in
seafood production to improve the fermentation process.

Flavor precursors are products which do not necessarily have flavoring properties in them.
Therefore, after being added intentionally, the flavor they produce is due to other components'
reactions or breaking down during food processing. They include amino acids, carbohydrates,
and oligopeptides. They contain functions such as giving a specific flavor to a product and
replacing the original flavor, which could have lost during food processing, especially in
fermented seafood.

According to Bassi et al. (2015), in sugar fermentation, lactic acid production is enhanced by
starter cultures. Therefore, growth rates of undesirable microorganisms are eliminated through
the development of unfavorable conditions. Reis et al. (2012) provide that the sugar fermentation
process produces hydrogen peroxide, ethanol, reuterin, acetic and propionic acids, and
antimicrobial peptides bacteriocins.

In many fermented fish products, LAB is the most dominant microorganisms. The main essence
of lactic acid and low ph is to help in the preservation of seafood. In LAB application, specific
flavor enzymes are inherent even though they are free from other enzymatic activities of the
complete pathway. However, some strain, such as cheese starter, does not have features for the
application. Therefore, proteolytic activities, good acidifying, and good flavor can achieve by
combining industrial strains and selected strains to use such strains better and surpass their
problems. It was found by Franciosi et al. (2009) that different strains could influence the
formation of flavor components. Strains can complement each other in cases where enzymes are
limited in a particular pathway. According to Baka et al. (2012), there was a decrease of
enterobacteria In low acid fermented sausage from after the usage of Lactobacillus sakei,
Lactobacillus Plantarum, and Lactobacillus curvatus conducted separately. According to
Casquete et al. (2012), the application of strains of pediococcus acidilactici and staphylococcus
vitulinus inoculates in salchichon, a traditional Iberian dry-sausage found to be able to prevent
enterobacteria and coliforms. The growth of strains of staphylococcus aureus, Lactococcus lactis
 24
DRY FERMENTED SEAFOOD
spp. Lactis and Lactococcus casei spp. The inhibition of Casei is possible, where salami
inoculation was conducted (Cenci-Goga et al. 2012).

In seafood fermentation, the air-drying method is more suitable than freeze-drying and vacuum
drying methods. When sterilized rice grains used as fillers in the air-drying process, it qualifies it
as the best starter culture in seafood products. The addition of air-dried starters during the
fermentation period influences changes to microbial. The differences are that, in seafood
fermentation, LAB can yield desirable characteristics compared to other bacteria. The sufficient
decrease of ph. in seafood products is due to the growth of the LAB level. However, comparing
both mixed and single starter cultures, LAB growth in both does not show a significant
difference at the beginning of the fermentation process. The safety of seafood products depends
on the need for bacteria such as Escherichia coli, and LAB inhibits other pathogenic bacteria as a
starter culture. LAB inoculated fermentation was proven to eliminate pathogenic bacteria in
seafood such as fish sausage, salami, fermented fish, and Balinese dry fermented sausage.
However, Salmonella spp cannot suppress when a single starter culture used is L. Plantarum. At
the same time, Escherichia coli cannot reduce wholly in L. reuteri single starter culture.

In contrast, mixed starter culture application in seafood fermentation leads to complete

elimination and suppression of Escherichia coli and salmonella spp. Therefore, using a mixed
starter culture to inhibit the growth of undesirable bacteria in seafood (O'Callaghan et al. 2012).
In terms of acid contents, the natural fermentation process produces less compared to a starter
culture.

Another major group found in seafood products is yeast. According to Singracha et al. (2017),
yeast produces inhibitory substances such as alcohol, which reduces salt levels. However, it
poses a danger to the consumers due to the high salt levels even though yeast survival depends
on the salt concentration. The main yeasts in the fermentation of seafood are saccharomyces
cerevisiae and C. holmii in this process. Consequently, during injection, the metabolism of yeast
reduces glucose content. According to Chen and Liu (2016), glycolysis in the cytoplasmic matrix
helps glucose conversion into pyruvic acid.

Moreover, ethanol and carbon dioxide produced in the alcoholic fermentation of glucose.
Therefore, ethanol plays various roles in yeast fermentation, whereby it acts as an odorant but as
a precursor to an ethyl ester and as a flavor carrier (LU et al., 2017). Since yeast's growth is not
affected by the increased acidity levels during fermentation, prove that these yeasts are tolerant
of acid and salt. Furthermore, yeast also helps in the improvement of smell in miso paste and soy
sauce. In seafood products, flavor and sensory characteristics of various metabolites undergo
enhancements. However, it is not clear how the above yeasts play a role in developing a taste for
seafood products. Furthermore, the enrichment of the right flavors in seafood products is through
yeasts' fermentation; it reduces the sweaty smell of fishy, rotten, and fecal notes in seafood.
 25
DRY FERMENTED SEAFOOD
The ph of seafood products regulation is by lactic acid produced in the opening stages of
fermentation. The cooked rice added to the fermentation acts as a substrate in acid production
for LAB. Furthermore, when conducting the sensory evaluation of seafood products after using
the LAB method, it shows that the use of LAB impacted seafood products' quality more than that
which uses the natural fermentation process. This distinction encompasses the following factors
texture, color, odor, and appearance of the sea products.

In terms of color, LAB starter cultures influence changes in the color of seafood products. These
color changes vary with the stage in which the fermentation process has reached. The more
observed colors are green, red, white, and yellow. Lightness is another practical change that
occurs in starter culture inoculates. Lightness increases after 24 hours of fermentation, which
after that, decreases until the end of fermentation.

Consequently, in LAB inoculate fermentation, a green color is observed at the early stages of
fermentation. After 48 hours, the green color turns into the red that remains until the
fermentation process. Furthermore, the yellow color remains until the close of the fermentation
process in LAB starter cultures. In seafood products, inoculated with starter culture shows
whiteness after 48 hours of fermentation.

Microbial population changes in seafood, where the selected starter culture is LAB for
inoculation, increases after 24 hours of fermentation compared to other in control methods. After
that, the number of LAB decreases after 48 hours till the completion of the fermentation process.
However, when a single starter culture can produce seafood, it yields different growth patterns—
the growth after 48 hours till the completion of the fermentation process, the number of LAB
decreases. Additionally, mixed starter culture causes an increase in the number of labs at 24
hours, and they remain stable until the termination of the process.

6.0 Influence of starter culture on the Biogenic Amines in dry fermented seafood.
Primary amines are organic bases with less active volatile compounds synthesized by microbial
metabolisms in the raw materials' enzymes. These elements are nonvolatile, meaning they do not
evaporate readily at room temperature but do so at higher temperatures. Microbial synthesis can
also obtain biogenic amines by reducing the corresponding amino acid's carbon, except
physiological polyamines (Vidal-Carou et al., 2007).

The figure below shows the composition of amines

 26
DRY FERMENTED SEAFOOD

Figure two shows the biological breakdown of polyamines

The essential amino acids in seafood, such as histamine, are among the crucial biogenic amino
acids produced from corresponding free amines such as histidine (Fig 2). Putrescine, for
instance, has spermidine and spermine (Zarei et al., 2011). During the biological formation of
histamine, histidine is vital because it is the derived element for histamine's biosynthesis (Table
1). According to a study by (Ezzar et al., 2015), the histidine concentration was below three
percent of amino acid concentration in natural and fermented seafood products, Ikan pekasam.
The most common Biogenic amine found in seafood is octopamine and dopamine and the other
amines (Naila, 2012). However, in the same publication, it was reported that high intakes of
biogenic amines in fermented seafood products cause hazardous risks and health implications
that triggers hypertensive reactions in the body.
 27
DRY FERMENTED SEAFOOD
This study observed the influence of starter cultures on these vital amines (Biogenic amines,
BAs). Amines' formation was arguably higher in experimental degradation without starter
cultures than when inoculated with cultures. Som-fug is a fermented fish product from Thailand
injected with Lactic acid bacteria starters. The build-up of these bio amines in som-fug declines
significantly by the incorporation of the leading LABs.

A study on the hygienic quality in dry fermented seafood products revealed that it is vital to
reduce the formation of injurious microbes and thus comprises a critical point in curbing the
synthesize of microbial pollutants in fermented seafood products (Naila et al., 2010). Additional
technological advancement engineered to reduce the microbe activity since hygiene is not
sufficient to do this. Injection of these cultures is a possible advancement strategy used
quantitatively to influence the build-up of these essential amines during the seafood degradation
process. The mode used by starter cultures is on the bases of eradicating the unwanted growth of
harmful aminogenic intrinsic bacteria.

Genus's cultures (LAB and CNS) are uniquely chosen as inoculators to adhere to some specified
technological procedures. Research by (Naila et al., 2010) also suggested that the ability to
determine if the lack of amino acid decarboxylase and competition for raw materials were ideal
for curbing amine production.

Some species of lactic acid bacteria and cocci stated to be unworthy microbiotas in the
breakdown of carbon and nitrogen in the amino acids, according to Linares et al. (2011). They
also noted that not all species and genera could break down the carbons and nitrogen since some
only produced specifically the primary amines and are not dependent on their strain. Due to this
reason, it is profound to do a sequential evaluation of the monogenic process when choosing the
right starter culture to use.

Several studies have experimented on ways to use these starter cultures in commercial lines to
inflate the breakdown of amino acids during the fermentation of sampled dry seafood products.
Through summarized research by (Lu et al.,2010; Baka et al., 2011) demonstrated benefits in
using controlled cultures to control biogenic amine build up. However, (Parente et a., 2001), in
their research, did not illustrate the benefits of fermented seafood.

The studies carried out on the same publications show that varying biogenic amine populations
depended on the bacterial species injected. In addition to that, they also stated that not all starters
had the capability to decrease the synthesis of biogenic amino acid. In general observations, the
acid bacteria injected as a starter exhibited more significant effects than the starters containing
the CNS.

According to research by Latorre-Moratalla, during the injection of the mixed starter culture to a
biogenic amino acid, a descriptive rate was observed. After the addition of a single genus, the
composition of cadaverine was. In contrast, a single culture different from the one used before
injected in a similar result was way economic by a larger percentage. However, both strains,
 28
DRY FERMENTED SEAFOOD
when used simultaneously, reduced cadaverine by the most considerable portion. A mix of
cultures performs better than one culture, enabling the curb to form unwanted and wanted
bacterial groups (Naila et al., 2010).

With this regard to the commercial production, mixed varieties of starters are combined with
various LABs species (Gucukoglu and Kuplulu, 2010), and resulted in deprived amine
populations than those injected with a diverse traditional starter culture. Autochthonous cultures
consist of specific strains derived from fermented seafood products (Casquete et al., 2011). There
has been an implication that vaccination with the amine breakdown process improves the
deprivation of biogenic amine build-up produced during seafood degradation.

The extent of deprivation of biogenic build up attained by injection depends on variabilities that
influence the availability of intrinsic microbiotas and the engagement and attachment of the
added starter culture. The opposing amine starter's protective effect diminishes by unhygienic
raw materials and low quality of the same. A finding out that different environments and the type
of method mainly used affected reducing the discharge of opposing amine starter.

This review made some conclusions about methods used in genus selection purposed in
incorporation as cultures in the production of dry seafood products without biogenic amines. A
Mix of degradation cultures constitutes harmful amine strains of Lactic acids and cocci species,
which are very well related to seafood product degradation and were the best alternative to dry
seafood product processing. Good quality raw materials and most pleasing technological
environments are vital factors to ensure the actual performance of cultures and deprivation of
amino acids build-up in degraded dry seafood products.

7.0 Conclusion
Microorganisms play a pivotal role in the preservation of seafood products. They help maintain
the character of the fish and biologically enhance the nutritional value of the seafood products.
When used as starter cultures in the fermentation of fish or seafood products, microorganisms
play a vital role in inhibiting the production of biogenic amines that are responsible for spoilage.
They stimulate antimicrobial activity against target pathogenic organisms. LAB is the foremost
among microorganisms accountable for fermentation, and it helps in the standardization of
seafood’s quality. Other microorganisms used in dried fermented seafood include yeast, which
produces inhibitory substances, e.g., alcohol, for reducing salt concentration. Yeast helps in the
formation of aroma. Microorganisms utilize carbohydrates during the fermentation process to
produce organic acids, e.g., lactic acid from LAB, which reduces the pH. The quality of the
fermented seafood/ fish, therefore, is customarily related to the pH and acidity of the resulting
end product. The main preservation factor of the seafood and other fermented food products is
the reduction in pH.
 29
DRY FERMENTED SEAFOOD
Microorganisms have identifiable effects on the color, taste, and flavor of the dried fermented
seafood products. Noticeable changes in the color of the dried fermented seafood products
include red, green, white, yellow, and lightness of the fish. When selecting the appropriate
microorganisms to use as starter cultures, the raw material to use, safety requirements of the
food, properties of the strain that are inherent, and the attributes regarding the quality of the food
need to be considered. The type of starter culture to use for a particular seafood product depends
on the specifications of the consumer and the technology. In selecting the starter culture, a mixed
culture produces better results compared to a single starter culture.

A study on the proneness of the microorganisms obtained from fermented seafood products and
antibiotics is recommended. Also, Starter cultures that are generally regarded as safe (GRAS) are
recommended by the US Food and Drug Administration (FDA) due to their ability to curtail the
proliferation of pathogenic and other spoilage microorganisms.

Significant hazards associated with dried fermented seafood products include the growth of
pathogenic bacteria, biogenic amines, and the presence of parasitic worms. The development of
pathogenic bacteria, however, can be inhibited through high-and-low-salt fermentation by Lactic
Acid Bacteria (LAB) in the appropriate concentration and proper pH. The most vigorous
inhibitory activities of biogenic amine are from samples prepared with Glycine. Glycine reduces
the contents of cadaverine and tyramine by 18% and 30.9%, respectively. Further studies are
needed to assess the safety of the finished fermented seafood products on the antimicrobial
activity of the numerous microorganisms used as starter cultures. Seafood safety may be
jeopardized by foodborne pathogens like Listeria spp, thus causing severe complications to the
consumer. Additionally, following the extreme consumption of dried seafood products,
especially in East-Asia, presents diverse health complications to the consumers, e.g., in salt-
prohibited consumers.

Additionally, further study is needed to determine how the reduction of NaCl content can impact
the quality of the food in terms of taste and consumer preferences. This study has outlined the
substitution of salt consumption with the recommendation of fish sauce as the alternative. This
hypothesis targets to enhance the reduction of salt consumption and without affecting the
palatability and acceptance by the consumers. Other microorganisms on consumers include side
effects such as the risk of adjuvant, possible gene transfer, harmful metabolic activities, systemic
infections, and excessive retroactivity in susceptible individuals. As a result, an investigation on
the corollary effects and toxic elements is recommended before the use of starter cultures in dry
seafood products.
 30
DRY FERMENTED SEAFOOD

8.0 References.
Azam, M., Mohsin, M., Ijaz, H., Tulain, U. R., Ashraf, M. A., Fayyaz, A., & Kamran, Q. (2017).
Lactic acid bacteria in traditional fermented Asian foods. Pakistan Journal of
pharmaceutical sciences, 30(5).

Bassi, D., Puglisi, E., & Cocconcelli, P. S. (2015). Comparing natural and selected starter
cultures in meat and cheese fermentations. Current Opinion in Food Science, 2, 118-
122.

Beena Divya, J., Kulangara Varsha, K., Madhavan Nampoothiri, K., Ismail, B., & Pandey, A.
(2012). Probiotic fermented foods for health benefits. Engineering in Life Sciences,
12(4), 377-390.

Buckenhüskes, H. J. (1993). Selection criteria for lactic acid bacteria to use as starter cultures for
various food commodities. FEMS microbiology reviews, 12(1-3), 253-271.

Casquete, R., Benito, M. J., Martín, A., Ruiz-Moyano, S., Aranda, E., & Córdoba, M. G. (2012).
Microbiological quality of salchichón and chorizo, traditional Iberian dry-fermented
sausages from two different industries, inoculated with autochthonous starter cultures.
Food Control, 24(1-2), 191-198.

Cenci-Goga, B. T., Rossitto, P. V., Sechi, P., Parmegiani, S., Cambiotti, V., & Cullor, J. S.
(2012). Effect of selected dairy starter cultures on microbiological, chemical and sensory
characteristics of swine and venison (Dama dama) nitrite-free dry-cured sausages.
Meat science, 90(3), 599-606.

Chen, C. S., Pignatelli, B., Malaveille, C., Bouvier, G., Shuker, D., Hautefeuille, A., ... &
Bartsch, H. (1992). Levels of direct-acting mutagens, total N-nitroso compounds in
nitrosated fermented fish products, consumed in a high-risk area for gastric cancer in
southern China. Mutation Research/Fundamental and Molecular Mechanisms of
Mutagenesis, 265(2), 211-221.

Colombo, B., Sciarria, T. P., Reis, M., Scaglia, B., & Adani, F. (2016). Polyhydroxyalkanoates
(PHAs) production from fermented cheese whey by using a mixed microbial
culture. Bioresource technology, 218, 692-699.

Costa, N. E., O'Callaghan, D. J., Mateo, M. J., Chagrin, V., Castillo, M., Hannon, J. A., ... &
Beresford, T. P. (2012). Influence of an exopolysaccharide produced by a starter on milk
coagulation and curd syneresis. International dairy journal, 22(1), 48-57.
 31
DRY FERMENTED SEAFOOD
Florida, F. A., de Melo Pereira, G. V., Thomaz-Soccol, V., Rakshit, S. K., Pagnoncelli, M. G. B.,
de Souza Vandenberghe, L. P., & Soccol, C. R. (2017). Microbiological,
biochemical, and functional aspects of sugary kefir fermentation-A review. Food
Microbiology, 66, 86-95.

Franciosi, E., Settanni, L., Cavazza, A., & Poznanski, E. (2009). Biodiversity and technological
potential of wild lactic acid bacteria from raw cows' milk. International dairy journal,
19(1), 3-11.

Ghanbari, M., Jami, M., Domig, K. J., & Kneifel, W. (2013). Seafood biopreservation by lactic
acid bacteria–a review. LWT-Food Science and Technology, 54(2), 315-324.

Giraffa, G., Chanishvili, N., & Widyastuti, Y. (2010). Importance of lactobacilli in food and feed
biotechnology. Research in microbiology, 161(6), 480-487.

Hajar, S., & Hamid, T. H. T. A. (2013). Isolation of lactic acid bacteria strains Staphylococcus
piscifermentans from Malaysian traditional fermented shrimp cincaluk. International
Food Research Journal, 20(1), 125.

Ikape, S. I. (2017). Fish spoilage in the tropics: A review. Octa Journal of Biosciences, 5(2).

Kanno, T., Kuda, T., An, C., Takahashi, H., & Kimura, B. (2012). Radical scavenging capacities
of saba-narezushi, Japanese fermented chub mackerel, and lactic acid bacteria. LWT-
Food Science and Technology, 47(1), 25-30.

Kasankala, L. M., Xiong, Y. L., & Chen, J. (2012). Enzymatic activity and flavor compound
production in fermented silver carp fish paste inoculated with a douchi starter culture.
Journal of agricultural and food chemistry, 60(1), 226-233.

Koyanagi, T., Kiyohara, M., Matsui, H., Yamamoto, K., Kondo, T., Katayama, T., & Kumagai,
H. (2011). Pyrosequencing survey of the microbial diversity of 'narezushi,' an
archetype of modern Japanese Sushi. Letters in applied microbiology, 53(6), 635- 640.

Kuda, T., Kawahara, M., Nemoto, M., Takahashi, H., & Kimura, B. (2014). In vitro antioxidant
and anti-inflammation properties of lactic acid bacteria isolated from fish intestines and
fermented fish from the Sanriku Satoumi region in Japan. Food research international, 64,
248-255.

Li, C., Wu, J., Li, Y., & Dai, Z. (2013). Identification of the aroma compounds in stinky
mandarin fish (Siniperca cheats) and comparison of volatiles during fermentation and
storage. International journal of food science & technology, 48(11), 2429-2437.

Lu, L., Guo, X., & Zhao, J. (2017). A unified nonlocal strain gradient model for nanobeams and
the importance of higher order terms. International Journal of Engineering Science, 119,
265-277.
 32
DRY FERMENTED SEAFOOD
Martínez-Álvarez, O., López-Caballero, M. E., Gómez-Guillén, M. C., & Montero, P. (2017).
Fermented seafood products and health. In Fermented foods in health and disease
prevention (pp. 177-202). Academic Press.

Molnár, H., Bata-Vidács, I., Baka, E., Cserhalmi, Z., Ferenczi, S., Tömösközi-Farkas, R., ... &
Székács, A. (2018). The effect of different decontamination methods on the microbial
load, bioactive components, aroma and colour of spice paprika. Food Control, 83, 131-
140.

Navarrete-del-Toro, M. A., García-Carreño, F. L., Hernández-Cortés, P., Molnár, T., & Gráf, L.
(2015). Biochemical characterisation of chymotrypsin from the midgut gland of
yellowleg shrimp, Penaeus californiensis. Food Chemistry, 173, 147-155.

Nie, X., Zhang, Q., & Lin, S. (2014). Biogenic amine accumulation in silver carp sausage
inoculated with Lactobacillus Plantarum plus Saccharomyces cerevisiae. Food
Chemistry, 153, 432-436.

Parente, E., Cogan, T. M., & Powell, I. B. (2017). Starter cultures: general aspects. In Cheese
(pp. 201-226). Academic Press.

Peppler, H. J., & Perlman, D. (Eds.). (, 2014). Microbial technology: Fermentation technology.
Academic Press.

Prabhakar, P. K., Vatsa, S., Srivastav, P. P., & Pathak, S. S. (2020). A comprehensive review of
the freshness of fish and assessment: Analytical methods and recent innovations. Food
Research International, 109157.

Riaz, S. (2019). Antimicrobial Activity of Propolis Extracts on Microorganisms (Doctoral

dissertation, CAPITAL UNIVERSITY).

Safaeian, S., & Khanzadi, S. Microbiology of Fish and Seafood.

Selhub, E. M., Logan, A. C., & Bested, A. C. (2014). Fermented foods, microbiota, and mental
health: ancient practice meets nutritional psychiatry. Journal of physiological anthropology,
33(1), 2.

Shikongo-Nambabi, M. N. N. N., & Schneider, M. B. (2012). The role, isolation and

identification of Vibrio species on the quality and safety of seafood. Biotechnology and
Molecular Biology Reviews, 7(2), 16-30.

Silvestri, L. G., & Hill, L. R. (1965). Agreement between deoxyribonucleic acid base
composition and taxometric classification of gram-positive cocci. Journal of
Bacteriology, 90(1), 136-140.
 33
DRY FERMENTED SEAFOOD
Singh, S. S., De Mandal, S., Lalnunmawii, E., & Kumar, N. S. (2018). Antimicrobial,
antioxidant, and probiotics characterization of dominant bacterial isolates from traditional
fermented fish of Manipur, North-East India. Journal of food science and technology,
55(5), 1870-1879.

Singracha, P., Niamsiri, N., Visessanguan, W., Lertsiri, S., & Assavanig, A. (2017). Application
of lactic acid bacteria and yeasts as starter cultures for reduced-salt soy sauce (moromi)
fermentation. LWT, 78, 181-188.

Speranza, B., Racioppo, A., Beneduce, L., Bevilacqua, A., Sinigaglia, M., & Corbo, M. R.
(2017). Autochthonous lactic acid bacteria with probiotic aptitudes as starter cultures for
fish-based products. Food Microbiology, 65, 244-253.

Šušković, J., Kos, B., Beganović, J., Leboš Pavunc, A., Habjanič, K., & Matošić, S. (2010).
Antimicrobial activity–the essential property of probiotic and starter lactic acid bacteria.
Food Technology and Biotechnology, 48(3), 296-307.

Valenzuela, A. S., Benomar, N., Abriouel, H., Cañamero, M. M., & Gálvez, A. (2010). Isolation
and identification of Enterococcus faecium from kinds of seafood: antimicrobial
resistance and production of bacteriocin-like substances. Food Microbiology, 27(7), 955-
961.

Yang, J., Guo, J., & Yuan, J. (2008). In vitro antioxidant properties of rutin. LWT-Food Science
and Technology, 41(6), 1060-1066.

Yang, Z., Liu, S., Lv, J., Sun, Z., Xu, W., Ji, C., ... & Lin, X. (2020). Microbial succession and
the changes of flavor and aroma in Chouguiyu, a traditional Chinese fermented fish. Food
Bioscience, 100725.

Zaman, M. Z., Bakar, F. A., Jinap, S., & Bakar, J. (2011). Novel starter cultures to inhibit
biogenic amines accumulation during fish sauce fermentation. International journal of
food microbiology, 145(1), 84- 91.

Zang, J., Xu, Y., Xia, W., Yu, D., Gao, P., Jiang, Q., & Yang, F. (2018). Dynamics and diversity
of microbial community succession during fermentation of Suan yu, a Chinese traditional
fermented fish, determined by high throughput sequencing. Food Research International,
111, 565-573

Zang, J., Xu, Y., Xia, W., & Regenstein, J. M. (2020). Quality, functionality, and microbiology
of fermented fish: a review. Critical reviews in food science and nutrition, 60(7), 1228-
1242.
 34
DRY FERMENTED SEAFOOD
Zeng, X., Xia, W., Jiang, Q., & Yang, F. (2013). Effect of autochthonous starter cultures on
microbiological and Physico-chemical characteristics of Suan yu, a traditional Chinese
low salt fermented fish. Food Control, 33(2), 344-351.

Zhou, M., Chen, Y., Huang, H., Zhou, X., Liu, J., & Huang, G. (2016). Prognostic value of total
lesion glycolysis of baseline 18F-fluorodeoxyglucose positron emission
tomography/computed tomography in diffuse large B-cell lymphoma. Oncotarget, 7(50),
83544.

Zhou, X. X., Zhao, D. D., Liu, J. H., Lu, F., & Ding, Y. T. (2014). LWT-Food Science and
Technology, 59(1), 335-341.

Zhou, X., Qiu, M., Zhao, D., Lu, F., & Ding, Y. (2016). Inhibitory effects of spices on biogenic
amine accumulation during fish sauce fermentation. Journal of food science, 81(4),
M913-M920.