Journal of Food and Nutrition Research, 2017, Vol. 5, No.

10, 729-735
Available online at http://pubs.sciepub.com/jfnr/5/10/2 Science and
Education Publishing DOI:10.12691/jfnr-5-10-2

Microbiological and Physicochemical Qualities of

Sushi and Sashimi from Japanese Restaurants in
Brazil
Camila Moura Batista1, Maria Luiza Rezende Ribeiro1, Mauricio Junio Franco de Souza1, Liana
Jayme Borges1, Tnia Aparecida Pinto de Castro Ferreira 1, Maria Cludia Porfirio Andr2,*
1 2
Faculdade de Nutrio, Universidade Federal de Gois, Goinia, GO, Brasil Instituto de Patologia
Tropical e Sade Pblica, Universidade Federal de Gois, Goinia, GO, Brasil *Corresponding
author: [email protected]
Abstract Japanese cuisine is known for its nutritional and sensory quality. Sushi and sashimi are highly perishable
and widely consumed foods in Brazil. The production and storage of these foods under appropriate conditions are
critical factors in ensuring consumer safety. This study was carried out to evaluate the microbiological and
physicochemical conditions of sushi and sashimi sold in Japanese restaurants as well as their handling conditions.
Samples of sushi (36) and sashimi (35) and 112 swabs of food handlers were collected from 18 Japanese restaurants of
the central region, Brazil. Temperature, pH and Total Volatile Nitrogen Bases (in sashimi) of food samples were
determined. Thermotolerant coliforms, coagulase positive Staphylococcus, Vibrio parahaemolyticus, Bacillus cereus (in
sushi) counts and the detection of Salmonella sp. were also performed. Staphylococcus aureus and Escherichia coli
were investigated in the swabs samples. The average temperature of the food samples at all the restaurants was higher
than the recommended value as well as 63.9% of the sushi and 54.3% of the sashimi samples had above recommended
pH. All sashimi samples had content within legal limits of Total Volatile Nitrogen Bases. Four samples (5.6%) were
positive for thermotolerant coliforms, and in three of them (4.2%), E. coli was isolated. High counts of Bacillus cereus
were found in 22.2% of the sushi samples. Coagulase positive Staphylococcus, Vibrio parahaemolyticus, and
Salmonella sp. were not found in the samples analysed. In 9.0% of the clinical samples, thermotolerant coliforms were
detected and E. coli was found in 5.4% (n = 6) of them. Staphylococcus aureus was not detected on any food handler.
The high storage temperature of food, the lack of heat treatment and the microbial contamination found may lead the
present microorganisms to reach high counts until consumption, and therefore represent risk to consumers. Thus,
effective prevention by supervisory bodies is essential.
Keywords: fish, microbiological quality, pH, Bacillus cereus
Cite This Article: Camila Moura Batista, Maria Luiza Rezende Ribeiro, Mauricio Junio Franco de Souza, Liana
Jayme Borges, Tnia Aparecida Pinto de Castro Ferreira, and Maria Cludia Porfirio Andr, Microbiological and
Physicochemical Qualities of Sushi and Sashimi from Japanese Restaurants in Brazil. Journal of Food and Nutrition
Research, vol. 5, no. 10 (2017): 729-735. doi: 10.12691/jfnr-5-10-2.

Japanese cuisine consists mainly of fresh, natural foods,

especially fish and vegetables. The main dishes are the
1. Introduction sushi, seasoned and rolled in a filling with cooked rice,
and the sashimi, fish fillets in natura cut into thin slices
The fish consumption in Brazil has increased in recent [2].
years and exceeds 12 kg/inhabitant/year, which is the Fish meat and fish products are highly perishable.
minimum quantity recommended by the World Health These foods are subject to contamination by pathogenic
Organization [1]. This is due mainly to its nutritional and spoilage bacteria from capture to consumption.
quality. Fish is an excellent source of animal protein, has Intrinsic characteristics are very important, as the high
anti-inflammatory action, and significant amounts of water activity, pH close to neutral and an environment rich
magnesium, phosphorus, and vitamins A and D. In in nutrients that is optimal for bacterial growth. Extrinsic
addition, being a food rich in polyunsaturated fatty acids factors also facilitate the deterioration of fish, such as the
and lower cholesterol levels, it reduces the risk of type of capture and transport, the extensive manipulation
developing cardiovascular diseases [2]. that these foods are subjected to, and hygienic conditions
The globalization of eating habits has generated rapid and temperature for storage and handling. Therefore may
spread of the raw fish consumption, a traditional Japanese pose risk to consumers health [4].
food. Thus, several restaurants specialized in Japanese In Brazil, the Ministry of Agriculture, Livestock, and
cuisine have sprung up in western cities. However, the Supply recommends physicochemical analysis to evaluate
restaurants do not always meet the minimum requirements the quality and freshness of the fish. Determination of pH
of location, time, and temperature of food exposition [3]. and Total Volatile Nitrogen Bases (TVNB) are very useful
730 Journal of Food and Nutrition Research

[5]. Other methods commonly used to access the quality content present in food (mg N/100 g of fish), and consists
of fresh fish, include sensory and microbiological analysis in the precipitation of protein nitrogen with trichloroacetic
[6]. acid (Synth) . The liquid portion is separated after
The pH of fish and other marine products is neutral, centrifugation, alkalized, and subsequently titrated with
which is a favorable condition for microorganisms to standard acid solution in the presence of mixed indicator.
develop [7]. TVNB are volatile compounds present in fish The standards used were those established by the
meat and are indicative of the free nitrogen content in the Brazilian Agricultural Protection Department of the
food, which is a chemical index used for the quality Ministry of Agriculture, Livestock and Supply, which
control of fish [5]. establishes the value of 30 mg N/100 g of sample as
Food handlers also affect the quality of the final maximum TVNB for fresh fish [5]. To classify samples as
product, especially raw or unprocessed products. More having excellent freshness, the content of 5 to 10 mg
than 200 diseases can be transmitted to people through the N/100 g is required; as reasonable freshness state, TVNB
ingestion of food contaminated with microorganisms and between 15 to 25 mg N/100 g; as in the process of
contamination can occur because of poor food-handling putrefaction, TVNB between 30 to 40 mg N/100 g; and as
practices [8]. deteriorated, TVNB 50 mg N/100 g of sample,
In view of the increased consumption of Japanese food according Ogawa and Maia [13].
and the lack of the information about the risks that it can
represent, this study aimed to evaluate the microbiological 2.3. Microbiological Analyses
and physicochemical aspects of sushi and sashimi sold in
Japanese restaurants, as well as their preparation conditions. The sushi and sashimi microbiological analyses
followed the procedures described by the Food and Drug
Administration [14] and all analyses were performed in
2. Materials and Methods duplicate. For these samples, 25 g were diluted in 225 mL
of 0.1% buffered peptone water and homogenized for 60s
2.1. Sampling in a stomacher. Then, 10-fold dilutions of the homogenates
were made in the same diluent [14].
The study included all 18 Japanese restaurants (10 Total and thermotolerant coliforms were enumerated on
served la carte and 8 were self-service buffets), located Violet Red Bile Agar (VRBA) incubated at 37C for 24
in the city of Goiania, capital of Goias State, Brazil, with -48h and subsequent confirmative tests were performed in
an operating license from the Sanitary Surveillance of the brilliant green bile 2% lactose broth (Oxoid) at 37C for
city at the time of the survey. 24h. Thermotolerant coliforms were determined in EC
From each restaurant, four types of food were selected broth (Oxoid) incubated at 45.5C for 24 h and E. coli
such as Philadelphia sushi (with salmon and cream confirmation was performed by Gram staining and the
cheese); California sushi (with kani, cucumber, and IMViC tests [14].
mango); salmon sashimi and sea bass sashimi. Two Coagulase positive Staphylococcus were enumerated on
hundred and fifty grams (250 g) of each sample were Baird-Parker agar (Oxoid) supplemented with egg yolk
aseptically collected, when available, and transported in tellurite emulsion, incubated at 37C for 48 h. After
ice to the laboratory in sterile plastic bags. The growth, presumptive S. aureus colonies were counted and
temperatures of the samples were measured using digital tested for catalase, coagulase and thermonuclease (TNase)
thermometer ranging from -32C to 380C. production and anaerobic fermentation of glucose and
In addition, swab samples of nostrils and hands of food mannitol [14].
handlers directly involved in the preparation of sushi and For determination of V. parahaemolyticus the MPN
sashimi were obtained after written informed consent technique was performed in alkaline peptone water and
from the participants. Then, the swabs were inoculated subsequent inoculation in TCBS (Thiosulfate Citrate Bile
into tubes containing BHI broth (Brain Heart Infusion) Sacarose) agar. After incubation at 35-37C for 24h,
and transported to the laboratory. typical colonies were purified and the confirmatory tests
were performed [14].
2.2. Physicochemical Analyses All samples were analysed for the presence of
Salmonella spp. after pre-enrichment in buffered peptone
The pH of sushi and sashimi samples was measured -water and enrichment in tetrathionate broth (Oxoid) and
with a digital potentiometer (pH Meter TEc-2, Tecnal, Rapapport-Vassiliads broth (Oxoid). Enrichment cultures
Piracicaba, Brazil) in 5 g of crushed and homogenized were streaked on xylose lysine deoxycholate (XLD) agar,
sample with 50 mL of distilled water [9]. bismuth sulphite agar and Hektoen enteric agar (Oxoid)
The values considered acceptable for fish, which were and the typical colonies were identified by biochemical
established by the Regulation for Industrial and Sanitary tests [14].
Inspection of Animal Products (RIISPOA) [10], are pH < For sushi samples the Bacillus cereus counts were
6.5 in the inner flesh. For evaluating the pH of sushi, pH < enumerated on mannitol-egg yolk-polymyxin (MYP, Oxoid)
4.6 was considered acceptable because of the acidification agar incubated at 37C for 48h and biochemical tests [14].
of rice used in the preparation [11]. To evaluate the microbiological quality of sushi and
The determination of TVNB content in sashimi was sashimi, Brazilian legislation [15] establishes limits of 102
performed according to the methodology proposed by CFU/g for thermotolerant coliforms; 5 x 10 3 CFU/g for
Saway da Silva et al. [12] and Brazilian legislation [5]. coagulase-positive Staphylococci, 103 CFU/g for Vibrio
This method allows quantification of the free nitrogen parahaemolyticus, and absence in 25g for Salmonella sp.
 Journal of Food and Nutrition Research 731

The presence of Escherichia coli and Staphylococcus temperature of the sashimi samples from the 18
aureus was also determined in food handlers by restaurants ranged from -1.2C to 19.2C, but no
inoculating samples of hand and nasal swabs previously restaurant presented the average temperature of the
incubated at 37C for 24 h in BHI broth, on EMB agar samples below the limit established by law (5C).
surface and Salt Mannitol agar, respectively. Confirmation Of the 18 restaurants evaluated, in 13 (72.2%) (Figure 2)
was performed by standard tests [16]. and 10 (55.6%) (Figure 3), the average pH of sushi and
sashimi, respectively, was above the recommended by
2.4. Ethics legislation. Of the 36 samples of sushi evaluated, the average
pH was 5.04 and ranged from 4.35 to 6.94 (Figure 2).
This study was approved by the Ethics Committee of Twenty-three samples (63.9%) showed pH > 4.6, which
the Federal University of Goias (CEP/UFG) under the is above the limit set for classification of this food as
number 360/11. suitable for consumption [11]. In 35 sashimi samples, pH
values ranged from 5.06 to 8.73 with average of 6.59
(Figure 3), and 54.3% of these samples showed pH > 6.5;
3. Results therefore, above the maximum limit established by the law
[10].
The 71 collected samples included 36 sushi According to the average values of TVNB (mg
(Philadelphia and California types) and 35 sashimi TVNB/100 g) observed, all the sashimi samples analysed
(salmon and sea bass). were within the recommended legal limits, with free
The average temperature of the sushi and sashimi nitrogen level < 30 mg N/100 g [5]. There was no
obtained from 18 restaurants was 14.1C ( Figure 1). The significant difference between the levels of free nitrogen
temperatures of sushi ranged from 11.6C to 27.5C. No found in the two types of evaluated sashimi, ranging from
sample was at the temperature recommended by the 5.09 to 10.49 mg N/100 g in sea bass sashimi and from
Brazilian legislation (5.0C) [17]. 5.49 to 8.69 mg N/100 g in salmon sashimi. The samples
From 35 samples of sashimi tested, 30 of them (85.7%) also presented excellent freshness (between 5 to 10 mg
were at temperature above the permissible limit. The N/100 g) [13].

Figure 1. Average temperatures (C) of sushi and sashimi obtained from Japanese restaurants

Figure 2. Dispersion of the averages and standard deviation of pH of the sushi collected in Japanese restaurants
732 Journal of Food and Nutrition Research

Figure 3. Dispersion of the averages and standard deviation of pH of sashimi collected in Japanese restaurants

Table 1. Frequency of Indicator Microorganisms in Sushi and Sashimi of Japanese Restaurants

Total coliforms Thermotolerant coliforms E. coli B. cereus

Type of sushi/sashimi Samples collected
n* % n % n % n %
Philadelphia sushi 18 9 50.0 2 11.1 2 11.1 5 27.8
California-type sushi 18 5 27.8 0 0.0 0 0.0 3 16.7
Salmon sashimi 18 8 44.4 1 5.6 1 5.6 - -
Sea bass sashimi 17 7 41.2 1 5.9 0 0.0 - -
Total 71 29 40.8 4 5.6 3 4.2 8 22.2

*n = number of positive samples.

The frequency of indicators and pathogenic Nasal and hands swabs were collected from 56 food
microorganisms isolated from sushi and sashimi are shown in handlers, totalizing 112 samples. In 10 (9.0%) samples
Table 1. Of the 18 restaurants evaluated, 88.9 % (n=16) obtained from eight (14.3%) food handlers, thermotolerant
presented at least one sample with high counts of total coliforms were detected, with 50% in nasal samples and
coliforms. Of the 71 sushi and sashimi samples collected, 50% in hands samples. In six (5.4%) samples, E. coli was
40.8% (n=29) were positive for this indicator microorganism, identified (Table 2). Staphylococcus aureus was not
with counts ranging from 103 to 8.8 x 105 CFU/g. detected in any sample obtained from handlers.
Of samples with high scores for coliforms, 5.6% (n=4)
were positive for thermotolerant coliforms, which included
two Philadelphia sushi, one salmon sashimi, and one sea bass 4. Discussion
sashimi. The scores ranged from 4.8 x 10 2 to 8.8 x 105
CFU/g. Of these, E. coli was isolated in three (4.2%). This study found an average temperature of the
Thirty-six sushi samples were analysed, of these, eight evaluated preparations (14.1C) much higher than the
(22.2%) showed positive results and high score of temperature recommended by Brazilian legislation (5C)
Bacillus cereus ranging from 103 to 5.0 x 104 CFU/g. [17]. Similar results were observed in other Brazilian
Among the contaminated preparations, five (27.8%) were cities such as Fortaleza/CE, where 100% of sushi and 90%
Philadelphia sushi and three (16.7%) were California-type of the sashimi evaluated were above the recommended
sushi. Coagulase-positive Staphylococcus, Vibrio temperatures [18]. In Porto Alegre/RS, the average of
parahaemolyticus and Salmonella sp. were not identified exposure temperatures reached 16.6C [19], and in Rio de
in any sample analysed. Janeiro/RJ an average temperature was observed of
27.15C and 15.8C in samples of sushi and sashimi,
Table 2. Frequency of indicator microorganisms in nasal and hand
respectively [20]. High storage temperatures of sushi
samples of food handlers from Japanese restaurants
(18.0C) were also observed in studies conducted in
Thermotolerant
E. coli Taiwan [21] and in Australia (15C and 9.6C) [22].
Samples origin Samples coliforms
collected Maintaining food highly manipulated such as sushi and
n* % n %
Nasal swabs 56 5 8.9 2 3.6
sashimi at recommended temperature by Brazilian
legislation proved to be a challenge in this study. The
Hand swabs 56 5 8.9 4 7.1
exposure of these foods at improper temperatures favors
Total 112 10 8.9 6 5.4
the microbial multiplication and increases the risk of
n* = number of positive samples. foodborne diseases (FBD) outbreaks.
 Journal of Food and Nutrition Research 733

It was observed that 63.9% of the sushi samples were temperatures that allow spores to germinate and multiply.
inadequate as regards the pH (4.6 maximum) with an Contamination was observed in 11.3% of the sushi, more
average of 5.04. The average pH was also high on analysis than found in Seattle (4.7%) and similar to that of Taiwan,
carried out in Taiwan, which obtained pH from 4.51 to 18.2% (n=4) [23].
6.11 [23], and in the state of Rio de Janeiro, which The Brazilian health legislation does not establish
obtained pH of 5.75 in sushi and sashimi from the city parameters for B. cereus in sushi, so the studies developed
Duque de Caxias and 5.92 in the city of Rio de Janeiro in Brazil did not investigate the presence of this
[20]. microorganism. However, we observed in this study the
The average pH of the assessed sashimi was 6.59, and importance in considering the determination of this
the pH of 54.3% of the samples were above the value microorganism in the routine analysis of this food. In all
recommended (6.5) [10]. In Portugal, 40% of fresh fish restaurants evaluated, the sushi was not stored at the
analysed showed pH above the allowed values [24]. recommended temperature and 63.9% of the samples did
Damasceno [25] detected values closer to the not have proper pH. This demonstrates the risk that
recommended in Belo Horizonte/MG, Brazil, where consumers of this food are exposed daily in the various
18.0% of fish samples showed pH > 6.5. establishments that sell this kind of food.
The determination of pH is an important factor in The absence of coagulase positive Staphylococcus,
assessing the quality and conservation status of fish and Vibrio parahaemolyticus, and Salmonella sp. is a positive
its derivatives. If the pH is near to neutrality (pH=7.0), the factor, because these microorganisms are pathogens and
risk of microbial multiplication increases, whereas their intake can cause FBD in consumers. The absence of
microorganisms multiply preferably at pH 6.5 to 7.5 [9]. these microorganisms has also been observed by other
As for the physicochemical deterioration measured by authors in Brazil and in other countries like Australia
free nitrogen concentration (TVNB) of sashimi samples, [22,28,31].
these samples were found to have good freshness, with The acidification of the rice by vinegar and the
values lower than 30 mg N/100 g [5]. Most of the reduction of water activity by the addition of salt and
nitrogenous component of fish is part of proteins and sugar inhibit the multiplication of microorganisms.
reflects the characteristics of the skeletal muscle of the However, as shown by other authors, even at acidic pH,
fish. Other studies evaluating this parameter also found the sushi showed higher contamination than sashimi. This
satisfactory results including Damasceno [25] who fact can be explained by excessive handling of rice, most
analysed salmon, and Cartonilho and Jesus [26] who of the time without using gloves and without proper hand
analysed tambaqui. hygiene, as well as the inadequate storage temperature of
Coliforms bacteria in high amounts do not necessarily rice [18].
indicate that this is a pathogenic microorganism and that There are no data on the Brazilian legislation on
there is a risk of FBD. However, its determination in high maximum tolerance for thermotolerant coliforms and E.
amounts indicates improper hygienic and sanitary coli in clinical samples of handlers, but the presence in
conditions as well as failures in food processing. Coliform hands and nostrils reveals poor hygiene, fecal
contamination was identified in 40.8% of the samples, a contamination, and transferability of this and other
result that corroborates with the study conducted in enteropathogens to highly manipulated foods [32].
Taiwan. [21] In a study conducted in Recife/PE, 100.0% It should be noted that during the visit and sample
of the evaluated sashimi samples were contaminated by collection in the presence of inspectors of the Health
these bacteria [27]. Surveillance, the food handlers washed their hands and
The presence of thermotolerant coliforms is indicative disinfected extensively with 70% alcohol before sample
of recent fecal contamination and the possible presence of collection. Despite this, coliforms and E. coli were found,
pathogens. This group consists mostly of E. coli but some which indicates that the contamination could be much
other enterics such as Klebsiella can also ferment lactose larger in routine working conditions.
at these temperatures and therefore, be considered as fecal The correct washing and hands hygiene are the most
coliforms [14]. This study detected the presence of these effective measures to eliminate pathogenic microorganisms.
bacteria in 5.6% of the samples, values much lower than Microbiological analyses of handlers gloves in Teixeira de
those found in other studies in Brazil, such as 25.0% in Freitas, BA found thermotolerant coliforms on 73.3 % of
Brasilia/DF [28], and 66.7% in Porto Alegre/RS [19], as gloves and E. coli on 6.7% of the samples [32].
well as in other countries like Taiwan (68.2%) [23]. This highlights the need to improve the Good
E. coli was isolated in 4.2% (n=3) of samples, similar Manufacturing Practices to ensure consumer safety. It was
to other studies [23,27,29]. However, it was lower than observed that there is an urgent need for compliance with
that observed by Basti et al. [30], who isolated this the basic conditions of food preservation, which in the
microorganism in 30.8% of the evaluated food. case of sushi and sashimi refers to the control of
This study found no statistically significant relationship temperature and pH.
between the high temperatures of the samples and
contamination by thermotolerant coliforms as observed in
other studies [3,19]. 5. Conclusions
Bacillus cereus, due to the resistance of its spores, is
widely distributed in nature. For this reason, easily The most significant microbiological contamination
contaminated foods such as cereals can trigger episodes of found in the samples was by coliforms, indicating, once
food poisoning. High levels of these bacteria indicate more, conditions of inadequate handling; and B. cereus,
survival during rice cooking and storage at incorrect whose detection is not required by current legislation.
This
734 Journal of Food and Nutrition Research

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 Kualitas Mikrobiologis dan Fisikokimia Sushi
dan Sashimi dari Restoran Jepang di Brazil
Camila Moura Batista 1, Maria Luiza Ribeiro Rezende 1, Mauricio Junio Franco de Souza 1,
Liana Jayme Borges 1, Tania Aparecida Pinto de Castro Ferreira 1, Maria Cludia Porfirio
Andr 2, *

1
Faculdade de Nutrio, Universidade Federal de Gois, Goinia, GO, Brasil 2 Instituto de
Patologia Tropical e Sade Pblica, Universidade Federal de Gois, Goinia, GO, Brasil *
Sesuai penulis: [email protected]

Masakan Jepang abstrak dikenal karena kualitas gizi dan sensorik. Sushi dan sashimi sangat makanan yang
mudah rusak dan banyak dikonsumsi di Brasil. Produksi dan penyimpanan makanan ini dalam kondisi yang tepat
merupakan faktor penting dalam memastikan keamanan konsumen. Penelitian ini dilakukan untuk mengevaluasi
kondisi mikrobiologis dan fisikokimia sushi dan sashimi yang dijual di restoran Jepang serta kondisi
penanganannya. Sampel sushi (36) dan sashimi (35) dan 112 tangkai penangan makanan dikumpulkan dari 18
restoran Jepang di kawasan tengah, Brasil. Suhu, pH dan Total Nitrogen Basa Volatile (dalam sashimi) sampel
makanan ditentukan. Coliform tahan panas, koagulase positif Staphylococcus, Vibrio parahaemolyticus, Bacillus
cereus (di sushi) jumlah dan deteksi Salmonella sp. juga dilakukan Staphylococcus aureus dan Escherichia coli
diselidiki dalam sampel swab. Suhu rata-rata sampel makanan di semua restoran lebih tinggi dari nilai yang
disarankan dan juga 63,9% sushi dan 54,3% sampel sashimi di atas pH yang direkomendasikan. Semua sampel
sashimi memiliki kandungan dalam batas legal Total Volatile Nitrogen Bases. Empat sampel (5,6%) positif untuk
coliform tahan panas, dan dalam tiga dari mereka (4,2%), E. coli diisolasi. Jumlah tinggi dari Bacillus cereus
ditemukan di 22,2% dari sampel sushi. Koagulase Staphylococcus positif, Vibrio parahaemolyticus, dan
Salmonella sp. tidak ditemukan dalam sampel yang dianalisis. Di 9,0% klinis sampel, koliform tahan panas yang
terdeteksi dan E. coli ditemukan pada 5,4% (n = 6) dari mereka. Staphylococcus aureus tidak terdeteksi pada
setiap penjamah makanan. Suhu penyimpanan makanan yang tinggi, tidak adanya perlakuan panas dan
Kontaminasi mikroba yang ditemukan dapat menyebabkan mikroorganisme sekarang mencapai jumlah yang
tinggi sampai konsumsi, dan oleh karena itu merupakan risiko bagi konsumen. Dengan demikian, pencegahan
yang efektif oleh badan pengawas sangat penting.

Kata kunci: ikan, kualitas mikrobiologis, pH, Bacillus cereus

Cite Pasal ini: Camila Moura Batista, Maria Luiza Ribeiro Rezende, Mauricio Junio Franco de Souza, Liana
Jayme Borges, Tania Aparecida Pinto de Castro Ferreira, dan Maria Cludia Porfirio Andr, mikrobiologi dan
fisiko Kualitas Sushi dan Sashimi dari Restoran Jepang di Brasil. Journal of Nutrition Research, vol Makanan
dan. 5, tidak 10 (2017): 729-735. doi: 10.12691 / jfnr-5-10-2.

1. Perkenalan
Konsumsi ikan di Brazil telah meningkat dalam beberapa tahun terakhir dan melebihi 12 kg / penduduk / tahun,
yang merupakan jumlah minimum yang direkomendasikan oleh Organisasi Kesehatan Dunia [1]. Hal ini terutama
disebabkan oleh kualitas nutrisinya. Ikan adalah sumber protein hewani yang sangat baik, memiliki tindakan anti-
inflamasi, dan jumlah magnesium, fosfor, dan vitamin A dan D. yang signifikan. Sebagai tambahan, menjadi
makanan yang kaya akan asam lemak tak jenuh ganda dan menurunkan kadar kolesterol, ini mengurangi risiko
mengembangkan penyakit kardiovaskuler [2].
 Globalisasi kebiasaan makan telah menghasilkan penyebaran cepat konsumsi ikan mentah, makanan tradisional
Jepang. Dengan demikian, beberapa restoran khusus masakan Jepang bermunculan di kota-kota barat. Namun,
restoran tidak selalu memenuhi persyaratan minimum lokasi, waktu, dan suhu eksposisi makanan [3].
 Masakan Jepang terutama terdiri dari makanan alami dan segar, terutama ikan dan sayuran. Hidangan utama
adalah sushi, berpengalaman dan digulung dalam mengisi dengan nasi, dan sashimi, ikan fillet di natura potong
menjadi irisan tipis [2].

Daging ikan dan produk ikan sangat mudah rusak. Makanan ini terkena kontaminasi oleh bakteri patogen dan
pembusukan dari penangkapan ke konsumsi. Karakteristik intrinsik sangat penting, karena aktivitas airnya tinggi,
pH mendekati netral dan lingkungannya kaya akan nutrisi yang optimal untuk pertumbuhan bakteri. Faktor
ekstrinsik juga memfasilitasi kemerosotan ikan, seperti jenis penangkapan dan pengangkutan, manipulasi ekstensif
yang menyebabkan makanan ini, dan kondisi dan suhu higienis untuk penyimpanan dan penanganan. Oleh karena
itu dapat menimbulkan risiko bagi kesehatan konsumen ' [4].

Di Brasil, Kementerian Pertanian, Peternakan, dan Pasokan merekomendasikan analisis fisikokimia untuk
mengevaluasi kualitas dan kesegaran ikan. Penentuan pH dan Total Volatile Nitrogen Bases (TVNB) sangat
bermanfaat
730 Jurnal Penelitian Makanan dan Gizi

[5]. Metode lain yang umum digunakan untuk mengakses kualitas ikan segar, termasuk analisis sensorik dan
mikrobiologi [6].

PH ikan dan produk laut lainnya adalah netral, yang merupakan kondisi yang menguntungkan bagi
mikroorganisme untuk mengembangkan [7]. TVNB adalah senyawa volatil hadir dalam daging ikan dan adalah
indikasi dari kandungan nitrogen bebas dalam makanan, yang merupakan indeks kimia yang digunakan untuk
kontrol kualitas ikan [5].

Penangan makanan juga mempengaruhi kualitas produk akhir, terutama produk mentah atau yang belum diproses.
Lebih dari 200 penyakit dapat ditularkan ke orang melalui konsumsi makanan yang terkontaminasi dengan
mikroorganisme dan kontaminasi dapat terjadi karena praktik makanan-penanganan miskin [8].

Mengingat meningkatnya konsumsi makanan Jepang dan kurangnya informasi tentang risiko yang dapat diwakili,
penelitian ini bertujuan untuk mengevaluasi aspek mikrobiologis dan fisikokimia sushi dan sashimi yang dijual di restoran
Jepang, serta kondisi persiapannya.

2. Bahan dan Metode

2.1. Contoh
Penelitian ini melibatkan semua 18 restoran Jepang (10 dilayani la carte dan 8 buffet swalayan), yang terletak
di kota Goiania, ibukota Goias Negara, Brasil, dengan izin operasi dari Sanitary Surveillance kota pada saat itu dari
survei
Dari masing-masing restoran, empat jenis makanan dipilih seperti sushi Philadelphia (dengan salmon dan krim
keju); California sushi (dengan kani, mentimun, dan mangga); salmon sashimi dan ikan bass sashimi. Dua ratus
lima puluh gram (250 g) setiap sampel dikumpulkan secara aseptik, bila tersedia, dan diangkut dalam es ke
laboratorium dengan kantong plastik steril. Suhu sampel diukur dengan menggunakan termometer digital mulai dari
-32 C sampai 380 C.

Selain itu, sampel lubang hidrat dari lubang hidung dan tangan penangan makanan yang terlibat langsung dalam
persiapan sushi dan sashimi diperoleh setelah mendapat informed consent dari para peserta. Kemudian, penyeka
diinokulasi ke dalam tabung berisi kaldu BHI (Brain Heart Infusion) dan diangkut ke laboratorium.

2.2. Analisis Fisikokimia

PH sushi dan sashimi sampel diukur dengan potensiometer digital (pH meter TEC-2, Tecnal, Piracicaba, Brazil)
di 5 g sampel hancur dan dihomogenisasi dengan 50 mL air suling [9].

Nilai dianggap dapat diterima untuk ikan, yang didirikan dengan Peraturan untuk Industri dan Inspeksi Sanitasi
Produk Hewan (RIISPOA) [10], adalah pH <6,5 pada daging bagian dalam. Untuk mengevaluasi pH sushi, pH <

4,6 dianggap dapat diterima karena pengasaman beras yang digunakan dalam penyusunan [11].

Penentuan kadar TVNB dalam sashimi dilakukan sesuai dengan metodologi yang diusulkan oleh Saway da Silva
dkk. [12] dan undang-undang Brasil [5]. Metode ini memungkinkan kuantifikasi nitrogen bebas
konten hadir dalam makanan (mg N / 100 g ikan), dan terdiri dalam pengendapan nitrogen protein dengan asam
trikloroasetat (Synth ). Bagian cairan dipisahkan setelah sentrifugasi, alkalized, dan selanjutnya dititrasi dengan
larutan asam standar dengan adanya indikator campuran.

Standar yang digunakan adalah yang didirikan oleh Brasil Perlindungan Pertanian Departemen Kementerian
Pertanian, Peternakan dan Pasokan, yang menetapkan nilai 30 mg N / 100 g sampel sebagai TVNB maksimum
untuk ikan segar [5]. Untuk mengklasifikasikan sampel memiliki kesegaran yang baik, kandungan 5 sampai 10 mg
N / 100 g diperlukan; sebagai keadaan kesegaran yang wajar, TVNB antara 15 sampai 25 mg N / 100 g; seperti
dalam proses pembusukan, TVNB antara 30 sampai 40 mg N / 100 g; dan sebagai memburuk, TVNB 50 mg N /
100 g sampel, menurut Ogawa dan Maia [13].

2.3. Analisis Mikrobiologi

Analisis sushi dan sashimi mikrobiologi mengikuti prosedur yang dijelaskan oleh Food and Drug Administration
[14] dan semua analisis dilakukan dalam rangkap dua. Untuk sampel ini, 25 g diencerkan dengan 225 mL air dogton
buffer 0,1% dan dihomogenisasi selama 60an dalam stomacher. Kemudian, 10 kali lipat pengenceran dari
homogenat dibuat dalam pengencer yang sama [14].

Koliform total dan termotoleran dicacah pada Violet Red Bile Agar (VRBA) yang diinkubasi pada suhu 37 C
selama 24-48 jam dan pengujian konfirmatori berikutnya dilakukan pada kaldu laktosa hijau terang (Oksida) 2%
hijau terang pada suhu 37 C selama 24 jam. Coliform tahan panas ditentukan di EC broth (Oxoid) diinkubasi pada
45,5 C selama 24 jam dan E. coli konfirmasi dilakukan oleh pewarnaan Gram dan tes IMViC [14].

Koagulase positif Staphylococcus dicacah di Baird-Parker agar (Oxoid) dilengkapi dengan kuning telur tellurite
emulsion, diinkubasi pada 37 C selama 48 jam. Setelah pertumbuhan, dugaan S. aureus koloni dihitung dan diuji
untuk katalase, koagulase dan thermonuclease produksi (TNase) dan fermentasi anaerob glukosa dan manitol [14].

Untuk penentuan V. parahaemolyticus teknik MPN dilakukan di dalam air pepton alkali dan inokulasi berikutnya
di TCBS (Tiosulfat Sitrat Bile Sacarose) agar. Setelah inkubasi pada 35-37 C selama 24 jam, koloni yang khas
adalah dimurnikan dan tes konfirmasi dilakukan [14].

Semua sampel dianalisis untuk kehadiran Salmonella spp. setelah pre-enrichment di buffered pepton-air dan
pengayaan kaldu tetrathionate (Oxoid) dan Rapapport-Vassiliads broth (Oxoid). Budaya pengayaan melesat di
xylose lysine deoxycholate (XLD) agar, bismuth sulfit agar dan Hektoen enteric agar (Oxoid) dan koloni yang khas
diidentifikasi dengan tes biokimia [14].

Untuk sampel sushi jumlah cereus Bacillus dicacah pada (MYP, Oxoid) agar manitol-kuning telur-polimiksin
diinkubasi pada 37 C selama 48 jam dan tes biokimia [14].

Untuk mengevaluasi kualitas mikrobiologis sushi dan sashimi, undang-undang Brasil [15]
menetapkan batas 10 2 CFU / g untuk coliform tahan panas; 5 x 10 3 CFU / g untuk
koagulase-positif Staphylococcus, 10 3 CFU / g untuk Vibrio parahaemolyticus, dan tidak
adanya di 25g untuk Salmonella sp.
Jurnal Penelitian Makanan dan Gizi 731
 Kehadiran Escherichia coli dan Staphylococcus aureus
juga ditentukan di penjamah makanan dengan
menginokulasikan sampel penyeka tangan dan hidung
yang sebelumnya diinkubasi pada suhu 37 C selama 24
jam dalam kaldu BHI, pada permukaan agar-agar EMB
dan agar Garam Mannitol. Konfirmasi dilakukan dengan
tes standar [16].

2.4. Etika
Studi ini disetujui oleh Komite Etika Universitas
Federal Goias (CEP / UFG) dengan nomor 360/11.