University of Management and Technology, Lahore

Seafood processing 3(2-1)

Seafood Processing
FT-405

Dr. Momna Rubab

School of Food and Agricultural Sciences

1
 Lipid Oxidation in Seafood
• Lipid oxidation is associated with early postmortem changes in
the fish tissue
• Lipid Oxidation is initiated by removal of a proton from the
central carbon of the unsaturated fatty acid, usually a pentadiene
moiety of the fatty acid, and formation of a lipid radical
• The latter reacts quickly with atmospheric oxygen making a
peroxy-radical (LOO’)
• The chain reaction involving the peroxy-radical results in the
formation of hydroperoxides that are readily broken down and
catalyzed by heavy metal ions to secondary products

Lipid
Oxidation
Lipid Oxidation

• The reactions are favored by activation of haemoproteins

and increase in free iron, while antioxidants negatively
influence the oxidation
• Lipid oxidation is comparatively more during frozen storage
than during chilling (0 to 2 ◦C) storage, and can be both
nonenzymatic as well as enzymatic
• Enzymes such as lipoxygenase, peroxidase, and microsomal
enzymes from animal tissues can also potentially initiate lipid
peroxidation producing hydroperoxides
• The breakdown of hydroperoxides into aldehydes, ketones,
and alcohols causes development of off-flavors
Oxidation of PUFA’s
Lipid Oxidation

• Fish lipids, rich in n-3 polyunsaturated fatty acids, are very

susceptible to oxidation, giving rise to n-3 aldehydes that
cause distinctive oxidative off-flavors
• Oxidative compounds include cis-4-heptenal, trans-2-
heptenal, trans-2-cis-4-heptadienal, etc. and also 1,5-octadien-
3-ol, 1-octen-3-ol, and hexanal
• Lipoxygenases are concentrated in the skin tissue and
remain active for upto 48 h of chilled storage
• The pro-oxidative activity due to haem proteins continued
longer than that due to lipoxygenase
Lipid Oxidation

• The most abundant degradation products of the

hydroperoxides formed from arachidonic and
docosahexaenoic acids are 12- and 16-hydroxy acids
• The extent of lipid oxidation can be reduced by glutathione
peroxidase, which reduces unstable lipid hydroperoxides to
nonradical, stable products, which are inactive in the
oxidative chain propagating mechanism
• Other enzymes useful in this respect are superoxide
dismutase and catalase, which remove superoxides from the
peroxidation mechanism
Lipid Oxidation

▪ Fish lipids are also prone to hydrolysis by lipases with the

formation of free fatty acids

▪ Lipid hydrolysis is more in ungutted than in gutted fish,

probably due to the involvement of lipases present in the
digestive enzymes

▪ Cellular phospholipases are known to hydrolyze the lipids,

particularly, phospholipids that leads to increased oxidation
of the hydrolyzed lipids
Fish as Substrate for Microbial
Growth

• Important factors contributing to the microbiological

complexity of seafood are:

▪ Specific as well as non-specific contamination of the live

animal from the environment and of products during
processing

▪ Growth conditions for microorganisms due to specific

intrinsic and extrinsic factor (température, pH,
microbial interactions etc.)
Factor Influence the Microbial
Spoilage
• There are several important specific intrinsic factors in fish
which greatly influence the microbiology and spoilage:

▪ The poikilotherm nature of the fish and its aquatic environment

▪ Post mortem pH in the flesh (usually > 6.0)

▪ Presence of large amounts of Non-protein-nitrogen (NPN)

▪ Presence of trimethylamine oxide (TMAO) as part of the NPN

fraction
Microbial Spoilage
▪ The fish muscle is sterile at the time of catch
▪ Contaminated by surface as well as intestinal bacteria, and
bacteria from water, equipment, and humans during handling
and processing
▪ Microorganisms are found on the outer surfaces (skin and
gills) and in the intestines of live and newly caught fish
▪ The microflora of fish from temperate waters is dominated
by psychotropic, aerobic or facultative anaerobic Gram
negative, rod-shaped bacteria and in particular Psuedomonas,
Moraxella, Acinetobacter, Shewanella, Flavobacterium, Vibrio,
Photobacterium, and Aeromonas spp
▪ The Gram positive organisms that are isolated from fish
include Bacillus, Micrococcus, Clostridium, Lactobacillus,
Coryneforms, and Brochothrix
Microbial Spoilage

▪ In polluted waters, high numbers of Enterobacteriaceae may

be found
▪ During chilled storage, there is a shift in bacterial types, with
psychotrophic Pseudomonas and Shewanella spp. dominating
after 1 to 2 weeks of storage
▪ At higher storage temperatures, for example, 20◦C, the
microflora, which will ultimately grow on the products, are
mesophilic ones including Bacillus and Micrococcus spp
▪ Bacterial spoilage of fish was essentially caused by organisms
belonging to Pseudomonas and Alteromonas spp.,
Aeromonas, and Proteus spp. were found to cause proteolysis
in shrimp
Microbial Spoilage

▪ Some of the other major microbial spoilers include

Pseudomonas fluorescens, P. perolens, and Alteromonas
putrefactiueos
▪ Bacteria initially grow at the expense of soluble, low
molecular weight compounds present in the fish muscle and
slime that serve as nutrients
▪ Organisms belonging to Pseudomonas spp. grow more rapidly
in the protein-free fraction of fish press juice containing
soluble components than in the protein fraction devoid of
soluble compounds
 Microbial Spoilage

• Initially bacterial growth is on the surface, and when slime builds

up, conditions become more favorable for anaerobes
• With the exhaustion of nutrients, protein catabolism becomes
essential for the survival of microflora
• At terminal spoilage, only those organisms that secrete proteases
can survive in the fish muscle, which leads to a predominance of
proteolytic bacteria in spoiling protein foods including fish
Microbial Spoilage

▪ The contaminant organisms initially utilize lactic acid and

nonprotein-nitrogen compounds, particularly
trimethylamine-oxide (TMAO) with the liberation of
trimethylamine (TMA), dimethylamine (DMA), ammonia, and
other volatile compounds
▪ During 4 to 7 days of storage of fish in ice, a sharp reduction
in the content of free amino acids can be noticed without
any significant extent of proteolysis
Microbial Spoilage

• The bacterial counts increase to above 108 colony forming

units (CFU) per cm2 skin surface or per g tissue, causing
breakdown of proteins accompanied by increase in amino
acids and volatile sulfur compounds, such as mercaptans and
H2 S
• This stage is also marked by adverse changes in the
organoleptic quality of the fish
• The number of H2S producing bacteria could, thus, be taken
as a general index of the extent of proteolysis and hence,
spoilage
Microbial Spoilage

• Trimethylamine-oxide is a natural nontoxic compound,

generally associated with the osmoregulatory function of
marine fish
• After the death of the fish some of the bacterial species
such as Alteromonas, Proteus, Photobacterium, Vibrio, and S.
putrefaciens, and also intestinal bacteria of the
Enterobacteriaceae present are able to carry out anaerobic
respiration by using TMAO as an electron acceptor
• The bacterial enzyme TMAO-reductase reduces TMAO to
TMA (Trimethylamine)
• Formation of TMA depends primarily on the content of
TMAO in the fish
Microbial Spoilage
• Most marine animals contain TMAO in appreciable quantities,
freshwater fish do not contain TMAO but some species like the
Nile perch and tilapia contain TMAO
• Bacterial reduction of TMAO results in accumulation of about 10
to 25 mg TMA per 100 g muscle of spoiling fish, contributing to
the characteristic spoiled fish odor
• TMA, in addition to ammonia and other volatile amines, is a major
component of the total volatile basic nitrogen (TVBN)
compounds in the spoiled fish
• TMA production in many fish species is also paralleled by
bacterial production of Hx
• Many spoilage causing bacteria such as Shewanella putrefaciens and
Vibrio spp. also produce off-smelling volatile sulfur compounds
such H2S as methyl mercaptan, and dimethylsulfide, from sulfur
containing amino acids
Microbial Spoilage

• Shewanella putrefaciens and Psuedomonas spp. have been

recognized as the specific spoilage organisms (SSOs) in iced
fresh fish such as cod
• During the storage of seafood at particular conditions of
temperature, atmospheres, salt, aw, preservatives, etc.
• SSOs grow faster than the remaining microflora, and
eventually produce the metabolites responsible for off-flavors
and sensory product rejection
• Modified atmosphere-stored marine fish from temperate
waters are spoiled by the CO2 resistant Photobacterium
phosphoreum
Microbial Spoilage

▪ Fish products with high salt contents may spoil due to the
growth of halophilic bacteria, anaerobic bacteria, and yeast,
whereas in lightly salted fish, spoilage could be due to lactic
acid bacteria and certain Enterobacteriacea
▪ The number of four different SSOs, that is, S. putrefaciens, P.
phosphoreum, Brochothrix thermosphacta and lactic acid
bacteria have been shown to correlate significantly with the
remaining shelf life of the product
▪ Bacteria contribute most of the sour and putrid odors
characteristic of spoiled fish
Microbial Spoilage

• Humans are able to detect very small quantities of

certain obnoxious compounds and are less sensitive to
others
• Putrefactive bacteria produce amines, pungent acids,
and other highly undesirable by-products while others,
notably lactic bacteria, produce less obnoxious
compounds
• Conditions that favor the latter group will often extend
shelf life by suppressing the growth of less desirable
organism
• Equipment used to hold and process fish is readily
contaminated with the very bacteria most detrimental
to shelf life