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DOI 10.1007/s00217-005-0125-8O R I G I NA L PA P E R Mar´ıa Elvira L´opez-Caballero · ´ Oscar Mart´ınez- ´ Alvarez · Mar´ıa Carmen G´omez-Guill´en · Pilar Montero Quality of Norway lobst

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DOI 10.1007/s00217-005-0125-8

O R I G I NA L PA P E R

Mar´ıa Elvira L´opez-Caballero ·

´

Oscar Mart´ınez- ´ Alvarez ·

Mar´ıa Carmen G´omez-Guill´en · Pilar Montero

Quality of Norway lobster (Nephrops norwegicus) treated

with a 4-hexylresorcinol-based formulation

Received: 12 May 2005 / Revised: 12 July 2005 / Accepted: 16 July 2005 / Published online: 25 October 2005

C

 Springer-Verlag 2005

Abstract In the present work, the effect on

biochem-ical indexes and microbial growth was studied in

Nor-way lobster (Nephrops norwegicus), using a formulation

containing 4-hexylresorcinol (0.1 and 0.05%) in

combi-nation with organic acids (citric, ascorbic and acetic) and

chelating agents (ethylenediaminetetraacetic acid [EDTA]

and di-sodium di-hydrogen pyrophosphate [PPi]) Lobsters

treated with 4% of a commercial formula based on

sul-phites were used for control purposes The treatment with

4-hexylresorcinol-based formulations delayed the increase

in K-value and total volatile bases, while evolution of pH

and trimethylamine was similar regardless of the treatment

No relation was found between biochemical and

micro-biological indexes Regarding microflora, although

com-mercial sulphites slightly slowed the growth of seafood

spoiler organisms, as Shewanella putrefaciens and

lumi-nescent colonies, these organisms were not found in a very

high number (∼6 log cfu/g) at the end of storage

More-over, the formulation containing 4-hexylresorcinol 0.1%

appeared to stimulate the growth of lactic acid bacteria

The sensory quality of lobster, in terms of melanosis,

re-mained with a good appearance for 12 days Formulations

based on 4-hexylresorcinol preserved the quality and could

therefore replace the traditional sulphites during storage of

Norway lobster

Keywords 4-Hexylresorcinol-based formulation

Sulphites Spoilage Lobster

Introduction

Melanosis and microbial growth are the two main causes

of spoilage in fresh crustaceans Crustaceans are usually

M E L´opez-Caballero ( ) · ´O Mart´ınez- ´Alvarez ·

M C G´omez-Guill´en · P Montero

Instituto del Fr´ıo (CSIC), Jos´e Antonio Novais,

10, 28040 Madrid, Spain

e-mail: ifrel54@if.csic.es

Tel.: +34-91-5445607

Fax: +34-91-5493627

treated with melanosis inhibitors immediately after har-vesting to prevent black spot appearance, since it starts with oxygen contact Microbiological spoilage assumes impor-tance during the storage of crustaceans In this connection,

it was of interest to search for compounds presenting a dou-ble action, as a melanosis inhibitor and as an antimicrobial Sulphites are the most widely and effective additive used

to prevent melanosis in crustaceans, although several health problems (most frequently asthma) related to the use of sulphites have been reported in the last two decades [1,2] The antimicrobial effect of sulphites has been described [3

6] and this effect depends on the type of microorganisms [7] Moreover, bisulphites were shown to degrade several toxins from moulds [8] However, the antibacterial effect during the iced storage of cultured prawns was little [9,10]

It is well known that the effect of antimelanosics is spe-cific for each species, requiring adequate doses and formu-lations [11, 12] In the search for alternative compounds

to sulphites, 4-hexylresorcinol (4-HR) has been reported

as a substitute both inhibiting melanosis [2,13] and as an-timicrobial [10] The effective doses of 4-hexylresorcinol differs depending on physiological state, season, method

of application, etc The species, however, is one of the most important of these factors Montero et al [14] found that, regardless of the season, a concentration of 0.25% 4-hexylresorcinol was effective at extending shelf-life of pink shrimp Further studies have revealed that combina-tions of 4-hexylresorcinol with acids and chelants allowed doses of 4-hexylresorcinol to be diminished in order to

pre-vent melanosis in shrimp (Parapenaeus longirostris) [14] However, there are no studies dealing with shelf-life using these formulas in other crustaceans

The aim of this work was to evaluate the effectiveness on Norway lobster of 4-hexylresorcinol-based formulations to prevent the spoilage during chilled storage

Material and methods

The species used was Norway lobster (Nephrops norvegi-cus) caught by trawl off the southern coast of Spain

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(Ayamonte, Huelva) harvested in November 2004

Av-erage and standard deviation sizes, and weights were

approximately 19.6± 1.7 cm (including the tail, carapace

and clawed legs) and 46± 6 g, respectively Lobsters were

placed alive on board the ship in polystyrene boxes and

covered with ice The boxes were taken by refrigerated

truck to the Instituto del Fr´ıo (Madrid), where most of the

lobsters arrived still alive within 12–16 h after capture The

Norway lobsters were separated in lots and treated with

several formulations (w/w) Based on previous studies

[14], two formulations with 0.1 or 0.05% 4-hexylresocinol

(H6250, Sigma Chemical Co., St Louis, Mo, USA)

combined with citric acid (0.5%), ascorbic acid (0.5%),

acetic acid (0.3%), EDTA (500 mg/kg) and disodium

dihydrogen pyrophosphate (1.5%) were selected (R-0.1

lot from 0.1% 4-hexylresorcinol and R-0.05 lot from

formulation containing 0.05% 4-hexylresorcinol)

The additives were dissolved in slightly salted water

(3.5% NaCl) and sprayed on the surface of the crustaceans

A third lot was treated by dust with a commercial

sulphite-based product (Melaplus M, Turco SA, Barcelona, Spain)

with sulphites (approximately 13%) and acids (citric and

ascorbic) (CS, control lot), at the concentration normally

used by fishermen (around 4%) After treatment, lobsters

were placed in perforated polystyrene boxes, covered with

ice and stored at 2◦C A first control at day 0 of chilled

storage was performed on Norway lobsters with no

addi-tives Further analyses on the crustaceans with additives

were done during chilled storage

pH

Approximately 5–10 g of muscle was homogenized with a

double quantity (g/ml) of distilled water After 5 min at

am-bient temperature, pH was determined with a pHm93

pH-meter and a combined pH electrode (RadiopH-meter,

Copen-hagen, Denmark) The experiments were repeated at least

in triplicate

K-value

Adenosine 5-triphosphate (ATP) and its breakdown

prod-ucts were determined by ionic exchange chromatography

according to Saito et al [15] Results were expressed as

µmol/g wet weight flesh K-value was calculated as

per-centage of the ratio between inosine (Ino)+ hypoxantine

(Hx) to all ATP-related products

Total volatile basic nitrogen (TVB-N)

Total volatile basic nitrogen determinations were carried

out in triplicate over the storage period using the method

of Antonacopoulos and Vyncke [16] Ground sample (10

± 0.1 g) were weighed in a suitable container and

homog-enized for 2 min with 100 ml 6% perchloric acid

solu-tion After filteration, the extract was alkalinized with 20%

sodium hydroxide solution and submitted to steam distil-lation The volatile base components were absorbed by an acid receiver and determined by titration of the absorbed bases All analyses were performed at least in triplicate

Trimethylamine (TMA-N) TMA-N was determined using the Dyer method modified

by Tozawa, as described in the AOAC [17] Results were expressed in mg of TMA-N per 100 mg of sample

Biogenic amines Agmatine (Agm), cadaverine (Cad), histamine (His), pu-trescine (Put) and tyramine (Tyr) were determined in 10% aqueous trichloroacetic acid extract by the post-column method described by Ritchie [18] Detection was carried out by fluorescence measurement Results of three repli-cates were expressed as mg of amine per kilogram (mg/kg)

of wet sample

Microbiological assays

At least 12 prawns per batch were used for microbiological analysis A total amount of 10 g of muscle was collected and placed in a sterile plastic bag (Sterilin, Stone, Staffordshire, UK) with 90 ml of buffered 0.1% peptone water (Oxoid, Basingstoke, UK) in a vertical laminar-flow cabinet (mod AV 30/70 Telstar, Madrid, Spain) After

1 min in a Stomacher blender (model Colworth 400, Seward, London, UK), appropriate dilutions were prepared for the following microorganism determinations: (i) total bacterial counts (TBC) on spread plates of Iron Agar 1% NaCl incubated at 15 ◦C for 3 days; (ii) H2S-producers organisms, as black colonies, on spread plates of Iron Agar 1% NaCl incubated at 15◦C for 3 days; (iii) luminescent bacteria on spread plates of Iron Agar 1% NaCl incubated

at 15 ◦C for 5 days; (iv) Pseudomonas on spread plates

of Pseudomonas Agar Base (Oxoid) with added CFC (cetrimide, fucidine, cephalosporine) supplement for

Pseudomonas spp (Oxoid) incubated at 25◦C for 48 h; (v)

Enterobacteriaceae on double-layered plates of Violet Red

Bile Glucose agar (VRBG, Oxoid) incubated at 30◦C for

48 h [after first adding 5 ml of Tryptone Soy Agar (Merck, Darmstadt, Germany) and incubating at room temperature for 1 h]; (vi) lactic acid bacteria on double-layered plates

of MRS Agar (Oxoid) incubated at 30 ◦C for 72 h All microbiological counts are expressed as the log of the colony-forming units per gram (log cfu/g) of sample All analyses were performed in triplicate

Overall appearance Over the 12-day storage period, a group of trained panel-lists routinely evaluated lobster overall appearance every

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2 days (ten individuals per treatment per evaluation),

scor-ing melanosis accordscor-ing to a scale from 0 to 10, where 0

corresponded to total absence of melanosis and the

maxi-mum 10 corresponded to the point of rejection established

at approximately 25% presence of blackspot on the surface

of the crustacean

Statistical analyses

The significance of differences between mean values was

evaluated using two-way ANOVA Statistical processing

was by the SPSS 12.0 computer program (Chicago, Illinois,

USA) The level of significance setting was p≤0.05

Results and discussion

Before treatments, the pH of lobster was 7.2 (Fig.1) This

value is similar to that found in Norway lobster caught in

Grand Sole [19] During storage, an increase in pH was

ob-served in all lots due to the production of basic compounds

related to spoilage [20] Despite the R-0.05 registering of

the highest values from day 7, all lots obtained pH around

8.3 at the end of the period (p >0.05, Table1)

The K-value is a freshness index showing the extent of

enzymatic activity-related nucleotides spoilage in fish and

shellfish Ogawa et al [21] established the K-value as the

only reliable index of freshness in lobster tails Initially,

K-value for lobster registered 18% (Fig.2) Mendes et al

[22] reported a K-value <10% at 0 and 72 h after catch in

the same species (N norvegicus) At the end of 12 days,

control lobster (CS lot) obtained a final value of ∼=60%

(p >0.05) The formulations based on 4-hexylresorcinol

delayed degradation of nucleotides during 5–7 days

Table 1 Analyses of variance of pH, K-value, TVB-N and TMA-N

Indexes Lots Days of storage

R-0.05 a/x a/x a/y b/y a/z

R-0.05 a/x a/x a/x a/x a/x

R-0.05 a/x a/y a/xy a/xy b/z

R-0.05 a/x a/x b/y a/yz b/z

R-0.1: 0.1% 4-hexylresorcinol; R-0.05: 0.05% 4-hexylresorcinol;

CS: 4% commercial sulphites Description of the formulas in M&M.

Different letters (a, b, c .) in the same column indicate significant

differences (p≤0.05) as a function of treatment; different letters (x,

y, z .) in the same row indicate significant differences (p≤0.05) as

a function of storage time

Fig 1 pH (average± SD) in lobster stored at (2 ± 1) ◦C for 12 days.

R-0.1: 0.1% 4-hexylresorcinol; R-0.05: 0.05% 4-hexylresorcinol; CS: commercial sulphites Description of the formulas in M&M

Fig 2 K-value (average± SD) in lobster stored at (2 ± 1) ◦C

for 12 days R-0.1: 0.1% hexylresorcinol; R-0.05: 0.05% 4-hexylresorcinol; CS: commercial sulphites

(Fig 2) K-value increased in both 4-HR formulas, first

moderately until day 5, then rapidly, reaching on day 12 around 50 and 60% for R-0.05 and R-0.1, respectively (Table1) Yamanaka and Shimada [23] found a K-value

of approximately 20–25% in spiny lobster at the stage of

initial decomposition The K-value mentioned by those

authors in spiny lobster is near that found initially in the present work As described below for some other biochem-ical indexes (TVB-N, Fig.3), noteworthy differences were found between our results and those described in the liter-ature; variability could be then related to species, season, geographic area, etc In this connection, Shimada et al [24] reported ATP changes in the muscle of three species of lobster (spiny, rock and American lobster), each of which exhibited different patterns and was temperature related Changes in total volatile bases are shown in Fig 3 Initially, lobsters registered 27.5 mg/100 g This amount is considerably higher than<10 mg/100 g reported by Fatima

and Qadri [25] in Pakistani lobster (Panulirus polyphagus)

and higher than≈15 mg/100 g found in Taiwanese shrimp

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Fig 3 Total volatile basic nitrogen, TVB-N (average± SD) in

lob-ster stored at (2 ± 1) ◦C for 12 days R-0.1: 0.1% 4-hexylresorcinol;

R-0.05: 0.05% 4-hexylresorcinol; CS: commercial sulphites

(Metapenaeopsis barbata) [26], in shrimp from India (P

in-dicus and M monoceros) [27] and in cultured tiger prawns

[9] On the contrary, some authors [19, 28, 29] reported

initial TVB-N concentrations higher than 25 mg/100 g in

good quality crustaceans As mentioned above,

discrep-ancy between those values was to be assumed since our

experiment was started with most of the lobsters still alive

In the present work, all lots slightly increased TVB-N

con-centration until day 7; from this moment onwards a sharp

enlargement was observed in the control lot, registering

around 64 mg/100 g at the end of the storage (p≤0.05,

Table 1) Muscle (40 mg/100 g) is considered the

upper limit unfit for human consumption (Directive

91/493/CEE) The R-0.1 lot reached this figure at the end

of the storage period

Originally, the TMA-N concentration was at the trace

level (Fig.4) A very slight increase was noted after 5 days

in all the lots, R-0.05 and CS lots registering similar values

(around 0.6 mg/100 g), while 1.4 mg/100 g was reached

by R-0.1 (p≤0.05, Table1) Despite some differences, the

three batches ranged between 1 and 1.5 mg/100 g on day

12 In this regard, TMA-N value of 1.5 mg/100 g could

be indicative of spoilage of lobster tails [25] The TMA-N

levels were lower than those found in Norway lobster [19]

and in lobster P polyphagus [25] Taking into account the

low quantity of TMA-N found in our investigation (Fig.4),

according to Vanderzant et al [30], ammonia is supposed

to be the primary component included in the total volatile

fraction (Fig.3)

Biogenic amines are produced by decarboxylase

activ-ity of both endogenous and microbial enzymes on free

amino acids, and they have been proposed as a chemical

index of seafood quality [31] Table2shows the biogenic

amines in Norway lobster Histamine and tyramine were

under threshold limit throughout the entire storage period

Agmatine was the principal amine found from day 7

on-wards, registering around 25.9± 5.7 mg/kg for R-0.1 at

the end of the period Agmatine was found to be the

pre-dominant biogenic amine during iced shrimp storage [32,

33] Its presence is attributed to the decarboxylation of free

Fig 4 Trimethylamine (TMA-N) (average± SD) in lobster stored

at (2 ± 1) ◦C for 12 days R-0.1: 0.1% 4-hexylresorcinol; R-0.05:

0.05% 4-hexylresorcinol; CS: commercial sulphites

arginine, amino acid presented in high concentration in crustaceans [34] High levels of putrescine and cadaverine were found during spoilage of lobster and shrimp [35] In the present work, cadaverine and putrescine were detected

in R-0.1 lot on day 12 but in a low concentration (Table2) Biogenic amines were not detected in control lobster for at least during 7 days

Initially, the total bacteria count was 4 log cfu/g (Fig.5) This figure is in agreement with that reported by some authors in lobster tails [25,36] The three batches evolved

similarly and a lag phase was observed for 5 days (p >0.05,

Table3) The batches then resumed growth reaching about

7 log cfu/g for 4-hexylresorcinol lots, and 7.7 log cfu/g in control lot at the end of storage

During early storage, counts for H2S-producing microor-ganisms were similar to the TBC, and a lag phase was

Table 2 Biogenic amines (mg/kg) in lobster stored at (2 ± 1) ◦C

for 12 days Amines Lots Days of storage

Agmatine R-0.1 <1.46 <1.46 11.55 25.99 ± 5.77

R-0.05 <1.46 <1.46 <1.46 –

Cadaverine R-0.1 <0.96 <0.96 <0.96 3.30

R-0.05 <0.96 4.54 ± 1.51 <0.96 –

Histamine R-0.1 <8.96 <8.96 <8.96 <8.96

R-0.05 <8.96 <8.96 <8.96 –

Putrescine R-0.1 <5.31 <5.31 <5.31 8.66 ± 2.06

R-0.05 <5.31 <5.31 <5.31 –

Tyramine R-0.1 <11.41 <11.41 <11.41 <11.41

R-0.05 <11.41 <11.41 <11.41 –

CS <11.41 <11.41 <11.41 –

R-0.1: 0.1% 4-hexylresorcinol; R-0.05: 0.05% 4-hexylresorcinol; CS: commercial sulphites Description of the formulas in M&M (–) Not determined

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Table 3 Analyses of variance of total bacteria count, H2S-producing

microorganisms, luminous colonies, Pseudomonas spp., lactic acid

bacteria and Enterobacteriaceae

Microorganisms Lots Days of storage

Total viable count R-0.1 a/x a/x a/y a/z a/v

R-0.05 a/x b/y a/x b/z a/v

CS a/x a/xy a/y b/z b/v H2S-producers R-0.1 a/x a/xy a/y a/z a/v

R-0.05 a/x b/x a/y b/y a/z

CS a/x c/xy a/x b/y a/z Luminous colonies R-0.1 a/x a/y a/z a/v a/z

R-0.05 a/x b/y a/z b/v b/v

R-0.05 a/x b/x b/y b/y ab/z

Lactic acid bacteria R-0.1 a/x a/y a/z a/v a/w

R-0.05 a/x b/y b/z b/v b/w

R-0.05 a/x a/x a/y b/x a/z

CS a/x a/y a/z b/xz b/v R-0.1: 0.1% 4-hexylresorcinol; R-0.05: 0.05% 4-hexylresorcinol;

CS: 4% commercial sulphites Description of the formulas in M&M.

Different letters (a, b, c .) in the same column indicate significant

differences (p≤0.05) as a function of treatment; different letters (x,

y, z .) in the same row indicate significant differences (p≤0.05) as

a function of storage time

Fig 5 Total bacteria count (TBC), (average± SD) in lobster stored

at (2 ± 1) ◦C for 12 days R-0.1: 0.1% 4-hexylresorcinol; R-0.05:

0.05% 4-hexylresorcinol; CS: commercial sulphites

prolonged for 5 days (Fig 6) Lobsters treated with

4-hexylresorcinol, especially R-0.1 lot, registered higher

val-ues than the control on day 7 (p≤0.05) These

microor-ganisms reached about 7 log cfu/g at day 12 in the three

lots

The genera Pseudomonas together with S putrefaciens

composed the spoilage association under aerobic iced

stor-age of fish [37] Initially, it was present in a concentration of

3.5 log cfu/g and showed a moderate growth during storage

(Fig.7, Table3) On day 12, all lots obtained slightly over

5 log cfu/g (p≤0.05); therefore this group is not considered

to dominate at the last stages of spoilage Although counts

Fig 6 H2S-producer organisms (average ± SD) in lobster stored

at (2 ± 1) ◦C for 12 days R-0.1: 0.1% 4-hexylresorcinol; R-0.05:

0.05% 4-hexylresorcinol; CS: commercial sulphites

(2 ± 1) ◦C for 12 days R-0.1: 0.1% 4-hexylresorcinol; R-0.05:

0.05% 4-hexylresorcinol; CS: commercial sulphites

for pseudomonads were related to total count at the first stage of storage, H2S-producing microorganisms consti-tute the major part of the total bacteria especially from day

6 In this connection, Chinivasagam et al [34] reported that

Pseudomonas fragi spoiled sulphite-treated prawns, and is

the major spoilage organism in tropical prawns [38] The luminescent colonies were not found (detection limit

<2 log cfu/g) until day 3 onwards (Fig.8) During storage, even 2 log cycles of difference were found between control

lot and the others containing 4-hexylresorcinol (p≤0.05)

On day 12, R-0.1 registered the lowest count for this type of

bacteria, presumably considered as Photobacterium phos-phoreum [39] Mart´ınez- ´Alvarez et al [10] found that sul-phites inhibited this type of bacteria during the storage of pink shrimp Despite the counts obtained on day 7, this fact could not be clearly shown throughout the studied period (Fig.8)

Figure9shows the lactic acid bacteria in MRS agar At first, the count for this group was below the detection limit (<1 log cfu/g) The growth of these genera is supposed

to be favoured by the slight acid conditions caused by the components included in the tested formulations (i.e ascor-bic acid, citric acids, etc.) However, the three batches did not exceeded 4–5 log cfu/g at the end of the period Simi-larly, counts for R-0.1 lot were higher even in 2 log cycles

from day 5 (p≤0.05, Table3) It is important to note that the

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Fig 8 Luminiscent bacteria (average ± SD) in lobster stored at

(2 ± 1) ◦C for 12 days R-0.1: 0.1% 4-hexylresorcinol; R-0.05:

0.05% 4-hexylresorcinol; CS: commercial sulphites

Fig 9 Lactic acid bacteria (average ± SD) in lobster stored at

(2 ± 1) ◦C for 12 days R-0.1: 0.1% 4-hexylresorcinol; R-0.05:

0.05% 4-hexylresorcinol; CS: commercial sulphites

formulation containing 4-hexylresorcinol (0.1%) appeared

not to inhibit, but to stimulate slightly the growth of

lac-tic acid bacteria The inhibition of lactobacilli activity by

sulphites during ripening of salami was reported [5]

Counts for Enterobacteriaceae at first registered

2.3 log cfu/g (Table 3), (Fig 10) This group, which

in-dicates the sanitary quality of the product, was below the

maximum recommended by the Microbiological Spanish

Standards [40] Because of the low temperature during iced

storage, enterobacteria remained at 4 log cfu/g in all lots at

the end of the study

During the storage of lobster in ice, there was no clear

connection between biochemical and microbiological

re-sults, probably because of the activity of endogenous

en-zymes [41] that took important part in spoilage On the

contrary, increments in total bacteria counts correlated well

with the rise in TVB-N and TMA-N during spoilage of rock

lobster [42] The resorcinol-based formulas delay the

in-crement of biochemical indexes, especially K-value and

TVB-N (Figs 2 and 3, respectively) Nevertheless,

mi-crobiological results showed that the effect of sulphites

and 4-hexylresorcinol formulas on the mentioned bacterial

groups was similar and all the lots registered related counts

at the end of the storage (Figs.5 10)

Fig 10 Enterobacteriaceae (average ± SD) in lobster stored at (2 ± 1) ◦C for 12 days R-0.1: 0.1% 4-hexylresorcinol; R-0.05:

0.05% 4-hexylresorcinol; CS: commercial sulphites Regarding overall appearance, no development of black spots was observed during 7 days of storage, regardless

of the treatment, following which a slight increase in melanosis appearance was noted CS lot scaled 42% while R-0.1 and R-0.05 obtained 8% of melanosis at the end of storage 4-HR formulas were thus more effective (about 33%) than commercial sulphites in delaying melanosis Applewhite et al [43] found that 4-hexylresorcinol (1 min dip in 100 mg/kg) inhibited melanosis on spiny lobster during iced storage for 7–12 days Accordingly, formulas based on 4-HR assayed in this work prevented melanosis and the reddening in the gut region, which is an indicator of the advanced spoilage in shrimp treated with sulphites [44] Some authors found a connection between melanosis and microbial growth in crustaceans Thus, colour formation

(melanin) due to strains of P fragi may occur if prawns

are not properly chilled [34] Regarding this, Nokodemusz

et al [45] reported that the presence of microorganisms

(e.g Proteus spp., Pseudomonas, etc.) and the H2S pro-duced reacted with metals of the lobster shell resulting in black discoloration In the present study, counts for pseu-domonads and enterobacteria were≤5 log cfu/g, therefore they did not predominate during spoilage In previous stud-ies, the relation between melanosis and microbial growth was not shown during the storage of prawns [13], but the authors do not discard this hypothesis and further studies

in this subject are needed

In summary, formulations based on 4-hexylresorcinol (at least 0.05%) in combination with organic acids and chelating agents can satisfactorily replace the use of the traditional sulphites, and even improve the quality, during iced storage of lobster The 4-hexylresorcinol formulas reduced some biochemical indexes and could have favoured the growth of lactic acid bacteria In addition, the resorcinol formulas maintained a good appearance of lobsters for 12 days

Acknowledgements The authors wish to thank the Consejer´ıa de

Agricultura y Pesca de la Junta de Andaluc´ıa (Project CSIC-2000-664) and the European Union (Project CRUSTAMEL, FAIR-Life-CRAFT/001/1312) for co-financing the research Author L´opez-Caballero is under contract Ram´on y Cajal, MEC-CSIC.

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