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May | June 2013
They are what they eat - Enhancing
the nutritional value of live feeds with
microalgae
The International magazine for the aquaculture feed industry
International Aquafeed is published six times a year by Perendale Publishers Ltd of the United Kingdom.
All data is published in good faith, based on information received, and while every care is taken to prevent inaccuracies,
the publishers accept no liability for any errors or omissions or for the consequences of action taken on the basis of
information published.
©Copyright 2013 Perendale Publishers Ltd.All rights reserved.No part of this publication may be reproduced in any form
or by any means without prior permission of the copyright owner. Printed by Perendale Publishers Ltd. ISSN: 1464-0058
INCORPORATING
f ish farming technolog y
www.oj-hojtryk.dk
Die and roll re-working machines
O&J Højtryk A/S
Ørnevej 1, DK-6705
Esbjerg Ø
CVR.: 73 66 86 11
Phone: +45 75 14 22 55
Fax: +45 82 28 91 41
mail: info@oj-hojtryk.dk
O&J HØJTRYK A/S endeavours to be a powerful, vigorous
and energetic company. With our customers needs and the
immediate environment as our point of departure, we want
to be a trendsetter in our core area of business - Mechanical
Engineering - Re-working of Dies and Rollers for the
manufacture of Feedstuffs and Biopellets, as well as the sale
and delivery of wearing parts in connection
with Roller Re-working - as well as to
remain open to new initiatives and
business opportunities.
O&J HØJTRYK A/S has drawn up a
strategy plan setting out guidelines for
how we as an organisation must conduct
and develop ourselves in order to make
our mark, not just in the present, but also in the
future.
O&J HØJTRYK A/S must achieve success by adapting
to the conditions of the market more quickly
and efficiently than our competitors. As a service
organisation we must, in relation to our customers,
attach great importance to our own vitality and efficiency
by constantly living up to the ever-growing demands made on
the services we provide.
“Today the most costly and
perhaps least understood live
food are the unicellular algae”
- Dhert & Sorgeloos 1995
L
ive	 feeds	 are	 often	 essential	 for	
larval	 fish.	 Live	 feeds	 are	 proven	
to	 be	 essential	 first-feed	 for	 many	
larval	 fish,	 essentially	 all	 those	 that	
hatch	 from	 small	 eggs	 with	 limited	 yolk	
reserves	 and	 often	 immature	 feeding	 and	
digestive	functions.	Live	feeds	provide	larval	
fish	with	essential	nutrients	that	are	naturally	
‘microencapsulated’	 in	 bite-sized	 packages.	
They	 include	 a	 high	 proportion	 of	 easily-
assimilated	 free	 amino	 acids	 and	 free	 fatty	
acids,	 as	 well	 as	 digestive	 enzymes	 and	
beneficial	 bacterial	 microfloras	 in	 the	 gut	
contents	of	the	prey.	The	swimming	activity	
of	live	prey	also	stimulates	feeding	responses	
in	 larval	 fish,	 a	 vital	 concern	 because	 small	
larvae	with	very	limited	metabolic	reserves	
can	 quickly	 starve	 if	 they	 do	 not	 promptly	
begin	feeding	actively.	
The	natural	live	foods	of	such	larvae	are	of	
course	microplankton,	both	zooplankton	and	
(although	often	not	appreciated)	phytoplank-
ton.	 Natural	 zooplankton	 assemblages	 are	
often	highly	diverse	and	may	include	protozoa,	
rotifers,	arroworms,	microcrustaceans	such	as	
copepods,	and	eggs	and	larvae	of	nearly	every	
group	 of	 marine	 animals	 including	 sponges,	
coelenterates,	 polychaetes,	 various	 crusta-
ceans,	 molluscs,	 echinoderms,	 and	 even	 fish.	
This	diverse	array	of	prey	organisms	supplies	
multiple	sources	of	essential	nutrients.	But	it	
can	be	very	difficult	to	obtain	sufficient	natural	
plankton	 to	supply	the	needs	 of	a	hatchery,	
and	natural	plankton	can	introduce	predators,	
parasites	 and	 pathogens.	 Hatchery-cultured	
live	feeds	are	therefore	the	only	practical	and	
safe	feed	for	many	larval	fish.
Use of live feeds in aquaculture
By	far	the	most	commonly-used	live	feeds	
in	 hatcheries	 are	 rotifers	 (Brachionus	 spp.)	
and	brine	shrimp	(Artemia)	(Conceição	et	
al.	2010),	with	some	use	of	copepods	such	
as	 species	 of	 Acartia,	 Calanus,	 Tisbe,	 and	
Parvocalanus.	 Although	 copepods	 gener-
ally	 provide	 better	 nutritional	 value,	 their	
culture	 presents	 so	 many	 difficulties	 that	
they	are	not	commonly	used	in	hatcheries	
(Drillet	et	al.	2006,	2011).	Rotifers	can	read-
ily	 be	 mass-cultured	 at	 high	 densities	 and	
can	double	their	numbers	in	a	day.	Rotifers	
are	 smaller	 than	 newly-hatched	 Artemia,	
which	 can	 be	 too	 large	 for	 some	 larvae.	
Artemia are	most	convenient	because	their	
resting	 eggs	 (cysts)	 can	 be	 purchased	 and	
hatched	 when	 needed,	 but	 newly-hatched	
Artemia nauplii	 do	 not	 begin	 to	 feed	 until	
after	the	first	molt,	so	their	nutritional	value	
depends	entirely	on	the	nutritional	environ-
ment	 of	 the	 previous	 wild	 generation	 that	
produced	 the	 eggs.	 One	 study	 found	 that	
the	 content	 of	 the	 important	 omega-3	
Poly-Unsaturated	 Fatty	 Acid	 (PUFA)	 EPA	
in	Artemia cysts	from	the	same	source	can	
vary	as	much	as	44-fold	(Dhert	&	Sorgeloos	
1995).	Such	variations	mean	that	the	nutri-
tional	 content	 of	 newly	 hatched	 Artemia
may	 be	 largely	 unknown,	 and	 only	 after	
the	first	molt	can	their	nutritional	value	be	
improved	by	feeding.
It	 is	 important	 to	 understand	 that	 nei-
ther	 Brachionus	 rotifers	 nor	 Artemia are	
truly	 marine	 organisms.	 Rather	 they	 are	
found	 in	 ‘saline’	 habitats,	 which	 are	 mostly	
inland	 environments	 with	 often	 extreme	
seasonal	 variations	 in	 temperature,	 salinity,	
and	 even	 availability	 of	 water.	 Adaptation	
to	 such	 extreme	 conditions	 has	 endowed	
these	 species	 with	 characteristics	 that	 are	
very	useful	in	aquaculture,	such	as	tolerance	
of	a	wide	range	of	culture	conditions,	rapid	
asexual	 reproduction	 by	 parthenogenesis	
(Brachionus),	and	formation	of	resistant	rest-
ing	cysts	(Brachionus	and	Artemia).	They	are	
also	relatively	omnivorous	and	do	not	have	
stringent	 nutritional	 requirements,	 and	 so	
can	be	fed	on	low-cost	feeds	such	as	yeast,	
starch,	rice	bran,	and	dried	Spirulina	(cyano-
bacteria).
It	may	be	no	surprise	that	feeding	larvae	
only	 one	 or	 two	 species	 of	 hatchery-pro-
duced	live	feeds	might	not	provide	adequate	
nutrition.	 But	 the	 underlying	 cause	 of	 such	
nutritional	inadequacy	is	often	the	low	quality	
of	 the	 low-cost	 food	 sources	 used	 to	 pro-
duce	the	live	feeds.	It	is	therefore	necessary	
to	 choose	 carefully	 the	 food	 sources	 used	
for	 hatchery-produced	 live	 feeds	 if	 they	 are	
to	 provide	 adequate	 nutritional	 support	 for	
larval	fish.
They are what they eat
Enhancing the nutritional value of
live feeds with microalgae
by Eric C Henry PhD, research scientist, Reed Mariculture Inc., USA
12 | InternatIonal AquAFeed | May-June 2013
FEATURE
Innovations for a better world.
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fu.buz@buhlergroup.com, www.buhlergroup.com
Fatten up your bottom line. Bühler high-performance animal and aqua feed production
systems are used by leading companies around the world. These producers know they
can rely not just on the technology itself, but also on the support that accompanies it. A
service combining local presence with global expertise both lowers feed mill operating
costs and increases capacity utilization. To find out more, visit www.buhlergroup.com
Limitations of formulated feeds
for live feed production
Formulated	feeds	offer	low	cost	and	con-
venience,	 but	 they	 have	 fundamental	 short-
comings.	 Zooplankton,	 including	 rotifers	 and	
Artemia,	can	feed	only	on	micro	particles	of	
appropriate	size	(from	bacteria	to	10	µm	for	
Brachionus	 [Baer	 et	 al.	 2008,	 Vadstein	 et	 al.	
1993],	and	from	bacteria	to	28	µm,	with	the	
optimum	about	8-16	µm	for	Artemia [Makridis	
and	 Vadstein	 1999,	 Fernández	 2001]).	 It	 is	
difficult	 to	 produce	 dry	 feeds	 that	 provide	
uniform	particle	sizes,	and	even	when	uniform	
dry	 particles	 can	 be	 produced	 they	 can	 be	
subject	 to	 clumping	 when	 dispersed	 into	
water	for	feeding.	But	probably	the	most	criti-
cal	shortcoming	of	dry	feeds	is	rapid	leaching	
of	water-soluble	nutrients;	the	smaller	the	par-
ticle,	the	faster	nutrients	are	leached	out.	Not	
only	are	leached	nutrients	unavailable	to	the	
live	feeds,	they	can	cause	fouling	of	the	water.
Rotifer Brachionus
plicatilis
Algae concentrate (Reed
Mariculture Tetraselmis 3600)
May-June 2013 | InternatIonal AquAFeed | 13
FEATURE
Lipid	emulsions	of	high-PUFA	oils	may	be	
used	to	improve	the	fatty	acid	profile	of	live	
feeds.	Although	their	contents	are	not	subject	
to	leaching,	lipid	droplets	are	prone	to	stick	
to	surfaces,	including	the	walls	of	the	culture	
tank	and	the	live	feed	organisms	themselves.	
Lipid	 enrichment	 protocols	 therefore	 often	
must	 include	 a	 rinsing	 step	 to	 clean	 the	
rotifers	or	Artemia of	adhering	lipid	droplets,	
which	 would	 otherwise	 foul	 the	 larval	 tank.	
Short-term	 feeding	 of	 oil	 emulsions	 results	
in	 lipid-enriched	 rotifers	 with	 high	 EPA	 and	
DHA	 levels,	 but,	 they	 are	 prone	 to	 rapid	
loss	 of	 their	 gut	 contents	 and	 acquire	 an	
extreme	lipid:protein	ratio	(Dhert	et	al.	2001).	
Moreover,	it	has	been	shown	that	when	the	
rotifers	are	collected	on	screens,	as	they	are	
for	 rinsing,	 this	 mechanical	 stress	 can	 cause	
ejection	 of	 the	 nutritious	 gut	 contents	 that	
were	 ingested	 during	 enrichment	 feeding	
(Romero-Romero	&	Yúfera	2012),	defeating	
the	purpose	of	the	enrichment.
Advantages of microalgae
Microalgae	 are	 the	 base	 of	 the	 plankton	
food	web,	and	their	great	biochemical	diver-
sity	is	the	source	of	the	high	nutritional	value	
of	 natural	 zooplankton.	 As	 the	 natural	 food	
of	zooplankton,	microalgae	offer	a	number	of	
advantages	 over	 formulated	 feeds.	 They	 are	
natural	‘microencapsulation’	particles	bounded	
by	 a	 cell	 membrane	 that	 retains	 the	 nutri-
tious	contents.	They	naturally	contain	a	wide	
spectrum	of	nutritional	components,	such	as	
essential	amino	acids,	PUFAs,	sterols,	vitamins,	
and	phytopigments.	Different	species	provide	
a	 wide	 range	 of	 cell	 sizes	 and	 nutritional	
factors,	as	well	as	components	that	enhance	
digestion	 and	 immune	 functions	 (Guedes	
&	 Malcata	 2012).	 Some	 strains	 have	 been	
found	to	have	antibacterial	effects	(Austin	&	
Day	1990,	Kokou	et	al.	2012,	Regunathan	&	
Wesley	2004).
Selecting the right microalgae
Although	 hundreds	 of	 microalgae	 strains	
have	 been	 tested	 as	 feeds	 for	 aquaculture,	
fewer	than	20	are	in	widespread	use	(Guedes	
&	 Malcata	 2012).	 Because	 these	 strains	
vary	 so	 greatly	 in	 their	 nutritional	 profiles,	
careful	 consideration	 is	 necessary	 in	 order	
to	 select	 the	 most	 nutritionally	 appropri-
ate	 strains.	 Such	 algae	 as	 Spirulina,	 Chlorella,	
Haematococcus,	and	Dunaliella	are	easily	mass-
produced	 because	 they	 can	 be	 cultivated	
in	open	ponds	at	low	cost,	but	they	all	lack	
the	omega-3	PUFAs	EPA	and	DHA	that	are	
essential	 for	 production	 of	 live	 feeds	 that	
provide	 adequate	 nutrition	 to	 marine	 fish.	
High-PUFA	algae	in	wide	use	include	strains	
of	 Nannochloropsis	 (Eustigmatophyceae),	
favoured	 for	 rotifer	 production	 and	 green-
water;	Tetraselmis	(Prasinophyceae);	Isochrysis	
and	Pavlova	(Prymnesiophyceae);	Thalassiosira,	
Chaetoceros,	 and	
Skeletonema	 (diatoms);	
and	 Rhodomonas	
(Cryptophyceae).
Although	 the	 PUFA	
content	of	many	strains	
has	by	now	been	well-
documented,	 sterol	
profiles	 have	 been	
more	 challenging	 to	
characterise	 because	
there	is	far	more	strain-
to-strain	variation,	even	
among	 strains	 suppos-
edly	 of	 the	 same	 spe-
cies,	 as	 revealed	 in	 a	
recent	 investigation	 of	
over	100	diatom	strains	
(Rampen	 et	 al.	 2010).	
Protein	 content	 is	 less	
variable,	with	a	study	of	
40	strains	of	microalgae	
in	 seven	 algal	 classes	
finding	consistently	high	
contents	 of	 essential	
amino	 acids	 (Brown	
et	 al.	 1997).	 Vitamin	
contents	 of	 microalgae	
also	appear	to	be	con-
sistently	 high	 (Brown	
&	 Miller	 1992,	 Brown	
et	al.	1999,	De	Roeck-
Holtzhauer	et	al.	1991).
Although	 various	 nutritional	 components	
have	been	well-documented	in	many	strains,	
it	 remains	 difficult	 to	 assemble	 complete	
nutritional	profiles	of	many	strains	so	that	the	
optimal	combination	of	strains	can	be	selected	
for	 a	 particular	 application.	 It	 is	 unfortunate	
that	 so	 many	 studies	 of	 the	 nutritional	 per-
formance	 of	 microalgae	 have	 tested	 single	
strains	 as	 the	 only	 feed,	 when	 it	 should	 be	
obvious	that	no	single	strain	is	likely	to	provide	
an	 optimal	 nutritional	 profile	 comparable	 to	
that	 provided	 by	 a	 natural	 phytoplankton	
assemblage.	
In	 practice,	 microalgae	 have	 repeatedly	
been	shown	to	dramatically	improve	the	PUFA	
content	of	rotifers	and	Artemia (Chakraborty	
et	 al.	 2007,	 Ferreira	 et	 al.	 2008,	 Kjell	 et	 al.	
1993,	Lie	et	al.	1997,	Øie	et	al.	1994,	Reitan	et	
al.	1997),	which	frequently	results	in	improved	
larval	 performance.	 But	 it	 is	 important	 to	
recognise	 that	 the	 high	 nutritional	 quality	 of	
enriched	 live	 feeds	 can	 be	 maintained	 after	
delivery	to	the	larval	tank	only	by	application	
of	‘greenwater’	techniques.	Unless	microalgae	
are	 added	 to	 the	 larval	 tank	 water,	 the	 live	
feed	 organisms	 quickly	 begin	 to	 starve,	 and	
can	 metabolize	 a	 significant	 fraction	 of	 their	
biomass	before	they	are	eaten	by	the	larvae.	
The	 algal	 cells	 themselves	 can	 also	 function	
as	live	feeds,	since	they	have	been	shown	to	
be	eaten	and	digested	by	larvae	(Reitan	et	al.	
1997,	Van	Der	Meeren	et	al.	2007),	and	may	
Nauplius stage of copepod
Parvocalanus crassirostris
14 | InternatIonal AquAFeed | May-June 2013
FEATURE
They are what they eat - Enhancing the nutritional value of live feeds with microalgae
also	 stimulate	 digestive	 enzyme	 production	
(Cahu	et	al.	1998).
Production of microalgae
Despite	the	many	advantages	of	microalgae,	
their	 wider	 use	 is	 hampered	 by	 difficulties	 in	
culturing,	 storage,	 and	 high	 costs.	 Microalgae	
culture	can	consume	a	significant	fraction	of	the	
resources	 of	 a	 hatchery,	 and	 requires	 special	
equipment,	 skilled	 labour,	 and	 a	 large	 alloca-
tion	of	space	that	is	unproductive	during	the	
seasons	when	live	feeds	are	not	needed.	
Low-cost	 open-pond	 culture	 methods	
carry	high	risks	of	contamination	and	culture	
failure	due	to	the	impossiblity	of	tightly	con-
trolling	culture	conditions,	and	the	most	highly	
prized	high-PUFA	strains	such	as	Isochrysis	and	
Pavlova	require	indoor	culture.	
It	is	very	difficult	to	synchronize	microalgal	
production	 with	 live	 feed	 requirements	 to	
prevent	feed	shortages	or	wasteful	overpro-
duction,	 and	 it	 is	 difficult	 to	 accurately	 dose	
algae	cultures	directly	into	live	feed	cultures.	
If	the	algae	are	harvested	and	concentrated,	
the	tightly-packed	cells	can	deteriorate	rapidly	
in	refrigerated	storage.	Some	microalgae	have	
been	 freeze-	 or	 spray-dried,	 but	 dried	 cells	
are	subject	to	protein	denaturation,	and	when	
they	 are	 rehydrated	 the	 leaching	 of	 water-
soluble	 substances	 can	 rapidly	 deplete	 their	
nutritional	value,	as	with	other	dry	feeds.	
Microalgae concentrates
The	 best	 solution	 to	 these	 problems	
can	 be	 the	 use	 of	 commercially-available	
refrigerated	or	frozen	algae	concentrates	
or	 ‘pastes’	 (Guedes	 &	 Malcata	 2012,	
Shields	&	Lupatsch	2012).	These	products,	
which	 are	 actually	 viscous	 liquids,	 have	
proven	 to	 be	 effective	 feeds	 for	 rotifers,	
Artemia,	shellfish	and	other	filter-feeders,	
as	well	as	for	greenwater	applications.	
In	 products	 formulated	 to	 provide	 a	
long	shelf-life,	the	concentrated	microalgae	
are	 suspended	 in	 buffer	 media	 that	 pre-
serve	cellular	integrity	and	nutritional	value,	
although	 the	 cells	 are	 non-viable.	 When	
concentrates	 with	 well-defined	 biomass	
densities	 are	 employed,	 the	 algae	 can	 be	
accurately	 dosed	 into	 live	 feed	 cultures	
with	 a	 metering	 pump,	 and	 non-viability	
confers	 the	 advantage	 that	 the	 products	
pose	 no	 risk	 of	 introducing	 exotic	 algal	
strains.	The	best	refrigerated	products	typi-
cally	 have	 a	 shelf-life	 of	 3-6	 months,	 and	
frozen	products	several	years.	This	means	
that	a	reliable	supply	of	algae	can	be	kept	
on	hand,	available	for	use	in	any	season	or	
if	 an	 unexpected	 need	 arises.	 Algae	 costs	
become	 predictable,	 and	 often	 prove	 to	
be	less	than	on-site	production	when	total	
production	 costs	 and	 inefficiencies	 are	
accounted	for.
Although	 costs	 of	 liquid	 algae	 concen-
trates	 are	 higher	 than	 for	 dried	 algae	 or	
formulated	feeds,	they	offer	all	the	nutritional	
advantages	 of	 live	 cultures.	 The	 nutritional	
quality	 of	 live	 feeds	 can	 be	 no	 better	 than	
the	 food	 sources	 used	 to	 produce	 them.	
Success	 of	 early	 larvae	 is	 so	 critical	 to	 the	
success	of	a	hatchery	that	even	a	relatively	
small	improvement	in	survival	or	growth	rate	
can	yield	great	benefits.
Outlook
Live	 feeds	 remain	 indispensable	 for	
larviculture	 of	 many	 fish.	 Although	 micro-
algae	are	among	the	costliest	food	sources	
used	 to	 produce	 live	 feeds,	 their	 many	
advantages	 justify	 the	 cost	 for	 hatcheries	
producing	high-value	fish.	Research	contin-
ues	 to	 better	 characterise	 the	 nutritional	
properties	 of	 various	 algae	 strains	 and	 to	
optimise	 algae	 production	 technologies.	
We	 can	 anticipate	 that	 introduction	 of	
novel	 algae	 strains	 and	 nutritionally-opti-
mised	 combinations	 of	 strains,	 along	 with	
improved	 feeding	 protocols,	 will	 ensure	
that	microalgae	remain	the	food	of	choice	
for	 production	 of	 the	 highest-quality	 live	
feeds.
References
www.aquafeed.co.uk/referencesIAF1303
May-June 2013 | InternatIonal AquAFeed | 15
FEATURE
Naturally ahead
MYC OFIX
Myco
toxin Ri
sk
M
a
n a g e M
ent
Mycofix®
More protective.
Mycotoxins decrease performance and interfere
with the health status of your animals.
Mycofix®
is the solution for mycotoxin risk management.
mycofix.biomin.net
www.aquafeed.co.uk
LINKS
•	 See the full issue
•	 Visit the International Aquafeed website
•	 Contact the International Aquafeed Team
•	 Subscribe to International Aquafeed
They are what they eat
Enhancing the nutritional value of live feeds
with microalgae
Controlling mycotoxins with
binders
Ultraviolet
water disinfection for fish
farms and hatcheries
Niacin
– one of the key B vitamins for sustaining
healthy fish growth and production
Volume 16 Issue 3 2013 - mAY | Ju Ne
INCORPORATING
fIsh fARmING TeChNOlOGy
This digital re-print is part of the May | June 2013 edition of International
Aquafeed magazine. 	
Content from the magazine is available to view free-of-charge, both as a full
online magazine on our website, and as an archive of individual features on
the docstoc website.
Please click here to view our other publications on www.docstoc.com.
To purchase a paper copy of the magazine, or to subscribe to the paper
edition please contact our Circulation and Subscriptions Manager on the link
above.  
INFORMATION FOR ADVERTISERS - CLICK HERE

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They are what they eat - Enhancing the nutritional value of live feeds with microalgae

  • 1. May | June 2013 They are what they eat - Enhancing the nutritional value of live feeds with microalgae The International magazine for the aquaculture feed industry International Aquafeed is published six times a year by Perendale Publishers Ltd of the United Kingdom. All data is published in good faith, based on information received, and while every care is taken to prevent inaccuracies, the publishers accept no liability for any errors or omissions or for the consequences of action taken on the basis of information published. ©Copyright 2013 Perendale Publishers Ltd.All rights reserved.No part of this publication may be reproduced in any form or by any means without prior permission of the copyright owner. Printed by Perendale Publishers Ltd. ISSN: 1464-0058 INCORPORATING f ish farming technolog y
  • 2. www.oj-hojtryk.dk Die and roll re-working machines O&J Højtryk A/S Ørnevej 1, DK-6705 Esbjerg Ø CVR.: 73 66 86 11 Phone: +45 75 14 22 55 Fax: +45 82 28 91 41 mail: info@oj-hojtryk.dk O&J HØJTRYK A/S endeavours to be a powerful, vigorous and energetic company. With our customers needs and the immediate environment as our point of departure, we want to be a trendsetter in our core area of business - Mechanical Engineering - Re-working of Dies and Rollers for the manufacture of Feedstuffs and Biopellets, as well as the sale and delivery of wearing parts in connection with Roller Re-working - as well as to remain open to new initiatives and business opportunities. O&J HØJTRYK A/S has drawn up a strategy plan setting out guidelines for how we as an organisation must conduct and develop ourselves in order to make our mark, not just in the present, but also in the future. O&J HØJTRYK A/S must achieve success by adapting to the conditions of the market more quickly and efficiently than our competitors. As a service organisation we must, in relation to our customers, attach great importance to our own vitality and efficiency by constantly living up to the ever-growing demands made on the services we provide.
  • 3. “Today the most costly and perhaps least understood live food are the unicellular algae” - Dhert & Sorgeloos 1995 L ive feeds are often essential for larval fish. Live feeds are proven to be essential first-feed for many larval fish, essentially all those that hatch from small eggs with limited yolk reserves and often immature feeding and digestive functions. Live feeds provide larval fish with essential nutrients that are naturally ‘microencapsulated’ in bite-sized packages. They include a high proportion of easily- assimilated free amino acids and free fatty acids, as well as digestive enzymes and beneficial bacterial microfloras in the gut contents of the prey. The swimming activity of live prey also stimulates feeding responses in larval fish, a vital concern because small larvae with very limited metabolic reserves can quickly starve if they do not promptly begin feeding actively. The natural live foods of such larvae are of course microplankton, both zooplankton and (although often not appreciated) phytoplank- ton. Natural zooplankton assemblages are often highly diverse and may include protozoa, rotifers, arroworms, microcrustaceans such as copepods, and eggs and larvae of nearly every group of marine animals including sponges, coelenterates, polychaetes, various crusta- ceans, molluscs, echinoderms, and even fish. This diverse array of prey organisms supplies multiple sources of essential nutrients. But it can be very difficult to obtain sufficient natural plankton to supply the needs of a hatchery, and natural plankton can introduce predators, parasites and pathogens. Hatchery-cultured live feeds are therefore the only practical and safe feed for many larval fish. Use of live feeds in aquaculture By far the most commonly-used live feeds in hatcheries are rotifers (Brachionus spp.) and brine shrimp (Artemia) (Conceição et al. 2010), with some use of copepods such as species of Acartia, Calanus, Tisbe, and Parvocalanus. Although copepods gener- ally provide better nutritional value, their culture presents so many difficulties that they are not commonly used in hatcheries (Drillet et al. 2006, 2011). Rotifers can read- ily be mass-cultured at high densities and can double their numbers in a day. Rotifers are smaller than newly-hatched Artemia, which can be too large for some larvae. Artemia are most convenient because their resting eggs (cysts) can be purchased and hatched when needed, but newly-hatched Artemia nauplii do not begin to feed until after the first molt, so their nutritional value depends entirely on the nutritional environ- ment of the previous wild generation that produced the eggs. One study found that the content of the important omega-3 Poly-Unsaturated Fatty Acid (PUFA) EPA in Artemia cysts from the same source can vary as much as 44-fold (Dhert & Sorgeloos 1995). Such variations mean that the nutri- tional content of newly hatched Artemia may be largely unknown, and only after the first molt can their nutritional value be improved by feeding. It is important to understand that nei- ther Brachionus rotifers nor Artemia are truly marine organisms. Rather they are found in ‘saline’ habitats, which are mostly inland environments with often extreme seasonal variations in temperature, salinity, and even availability of water. Adaptation to such extreme conditions has endowed these species with characteristics that are very useful in aquaculture, such as tolerance of a wide range of culture conditions, rapid asexual reproduction by parthenogenesis (Brachionus), and formation of resistant rest- ing cysts (Brachionus and Artemia). They are also relatively omnivorous and do not have stringent nutritional requirements, and so can be fed on low-cost feeds such as yeast, starch, rice bran, and dried Spirulina (cyano- bacteria). It may be no surprise that feeding larvae only one or two species of hatchery-pro- duced live feeds might not provide adequate nutrition. But the underlying cause of such nutritional inadequacy is often the low quality of the low-cost food sources used to pro- duce the live feeds. It is therefore necessary to choose carefully the food sources used for hatchery-produced live feeds if they are to provide adequate nutritional support for larval fish. They are what they eat Enhancing the nutritional value of live feeds with microalgae by Eric C Henry PhD, research scientist, Reed Mariculture Inc., USA 12 | InternatIonal AquAFeed | May-June 2013 FEATURE
  • 4. Innovations for a better world. Bühler AG, Feed & Biomass, CH-9240 Uzwil, Switzerland, T +41 71 955 11 11, F +41 71 955 28 96 fu.buz@buhlergroup.com, www.buhlergroup.com Fatten up your bottom line. Bühler high-performance animal and aqua feed production systems are used by leading companies around the world. These producers know they can rely not just on the technology itself, but also on the support that accompanies it. A service combining local presence with global expertise both lowers feed mill operating costs and increases capacity utilization. To find out more, visit www.buhlergroup.com
  • 5. Limitations of formulated feeds for live feed production Formulated feeds offer low cost and con- venience, but they have fundamental short- comings. Zooplankton, including rotifers and Artemia, can feed only on micro particles of appropriate size (from bacteria to 10 µm for Brachionus [Baer et al. 2008, Vadstein et al. 1993], and from bacteria to 28 µm, with the optimum about 8-16 µm for Artemia [Makridis and Vadstein 1999, Fernández 2001]). It is difficult to produce dry feeds that provide uniform particle sizes, and even when uniform dry particles can be produced they can be subject to clumping when dispersed into water for feeding. But probably the most criti- cal shortcoming of dry feeds is rapid leaching of water-soluble nutrients; the smaller the par- ticle, the faster nutrients are leached out. Not only are leached nutrients unavailable to the live feeds, they can cause fouling of the water. Rotifer Brachionus plicatilis Algae concentrate (Reed Mariculture Tetraselmis 3600) May-June 2013 | InternatIonal AquAFeed | 13 FEATURE
  • 6. Lipid emulsions of high-PUFA oils may be used to improve the fatty acid profile of live feeds. Although their contents are not subject to leaching, lipid droplets are prone to stick to surfaces, including the walls of the culture tank and the live feed organisms themselves. Lipid enrichment protocols therefore often must include a rinsing step to clean the rotifers or Artemia of adhering lipid droplets, which would otherwise foul the larval tank. Short-term feeding of oil emulsions results in lipid-enriched rotifers with high EPA and DHA levels, but, they are prone to rapid loss of their gut contents and acquire an extreme lipid:protein ratio (Dhert et al. 2001). Moreover, it has been shown that when the rotifers are collected on screens, as they are for rinsing, this mechanical stress can cause ejection of the nutritious gut contents that were ingested during enrichment feeding (Romero-Romero & Yúfera 2012), defeating the purpose of the enrichment. Advantages of microalgae Microalgae are the base of the plankton food web, and their great biochemical diver- sity is the source of the high nutritional value of natural zooplankton. As the natural food of zooplankton, microalgae offer a number of advantages over formulated feeds. They are natural ‘microencapsulation’ particles bounded by a cell membrane that retains the nutri- tious contents. They naturally contain a wide spectrum of nutritional components, such as essential amino acids, PUFAs, sterols, vitamins, and phytopigments. Different species provide a wide range of cell sizes and nutritional factors, as well as components that enhance digestion and immune functions (Guedes & Malcata 2012). Some strains have been found to have antibacterial effects (Austin & Day 1990, Kokou et al. 2012, Regunathan & Wesley 2004). Selecting the right microalgae Although hundreds of microalgae strains have been tested as feeds for aquaculture, fewer than 20 are in widespread use (Guedes & Malcata 2012). Because these strains vary so greatly in their nutritional profiles, careful consideration is necessary in order to select the most nutritionally appropri- ate strains. Such algae as Spirulina, Chlorella, Haematococcus, and Dunaliella are easily mass- produced because they can be cultivated in open ponds at low cost, but they all lack the omega-3 PUFAs EPA and DHA that are essential for production of live feeds that provide adequate nutrition to marine fish. High-PUFA algae in wide use include strains of Nannochloropsis (Eustigmatophyceae), favoured for rotifer production and green- water; Tetraselmis (Prasinophyceae); Isochrysis and Pavlova (Prymnesiophyceae); Thalassiosira, Chaetoceros, and Skeletonema (diatoms); and Rhodomonas (Cryptophyceae). Although the PUFA content of many strains has by now been well- documented, sterol profiles have been more challenging to characterise because there is far more strain- to-strain variation, even among strains suppos- edly of the same spe- cies, as revealed in a recent investigation of over 100 diatom strains (Rampen et al. 2010). Protein content is less variable, with a study of 40 strains of microalgae in seven algal classes finding consistently high contents of essential amino acids (Brown et al. 1997). Vitamin contents of microalgae also appear to be con- sistently high (Brown & Miller 1992, Brown et al. 1999, De Roeck- Holtzhauer et al. 1991). Although various nutritional components have been well-documented in many strains, it remains difficult to assemble complete nutritional profiles of many strains so that the optimal combination of strains can be selected for a particular application. It is unfortunate that so many studies of the nutritional per- formance of microalgae have tested single strains as the only feed, when it should be obvious that no single strain is likely to provide an optimal nutritional profile comparable to that provided by a natural phytoplankton assemblage. In practice, microalgae have repeatedly been shown to dramatically improve the PUFA content of rotifers and Artemia (Chakraborty et al. 2007, Ferreira et al. 2008, Kjell et al. 1993, Lie et al. 1997, Øie et al. 1994, Reitan et al. 1997), which frequently results in improved larval performance. But it is important to recognise that the high nutritional quality of enriched live feeds can be maintained after delivery to the larval tank only by application of ‘greenwater’ techniques. Unless microalgae are added to the larval tank water, the live feed organisms quickly begin to starve, and can metabolize a significant fraction of their biomass before they are eaten by the larvae. The algal cells themselves can also function as live feeds, since they have been shown to be eaten and digested by larvae (Reitan et al. 1997, Van Der Meeren et al. 2007), and may Nauplius stage of copepod Parvocalanus crassirostris 14 | InternatIonal AquAFeed | May-June 2013 FEATURE
  • 8. also stimulate digestive enzyme production (Cahu et al. 1998). Production of microalgae Despite the many advantages of microalgae, their wider use is hampered by difficulties in culturing, storage, and high costs. Microalgae culture can consume a significant fraction of the resources of a hatchery, and requires special equipment, skilled labour, and a large alloca- tion of space that is unproductive during the seasons when live feeds are not needed. Low-cost open-pond culture methods carry high risks of contamination and culture failure due to the impossiblity of tightly con- trolling culture conditions, and the most highly prized high-PUFA strains such as Isochrysis and Pavlova require indoor culture. It is very difficult to synchronize microalgal production with live feed requirements to prevent feed shortages or wasteful overpro- duction, and it is difficult to accurately dose algae cultures directly into live feed cultures. If the algae are harvested and concentrated, the tightly-packed cells can deteriorate rapidly in refrigerated storage. Some microalgae have been freeze- or spray-dried, but dried cells are subject to protein denaturation, and when they are rehydrated the leaching of water- soluble substances can rapidly deplete their nutritional value, as with other dry feeds. Microalgae concentrates The best solution to these problems can be the use of commercially-available refrigerated or frozen algae concentrates or ‘pastes’ (Guedes & Malcata 2012, Shields & Lupatsch 2012). These products, which are actually viscous liquids, have proven to be effective feeds for rotifers, Artemia, shellfish and other filter-feeders, as well as for greenwater applications. In products formulated to provide a long shelf-life, the concentrated microalgae are suspended in buffer media that pre- serve cellular integrity and nutritional value, although the cells are non-viable. When concentrates with well-defined biomass densities are employed, the algae can be accurately dosed into live feed cultures with a metering pump, and non-viability confers the advantage that the products pose no risk of introducing exotic algal strains. The best refrigerated products typi- cally have a shelf-life of 3-6 months, and frozen products several years. This means that a reliable supply of algae can be kept on hand, available for use in any season or if an unexpected need arises. Algae costs become predictable, and often prove to be less than on-site production when total production costs and inefficiencies are accounted for. Although costs of liquid algae concen- trates are higher than for dried algae or formulated feeds, they offer all the nutritional advantages of live cultures. The nutritional quality of live feeds can be no better than the food sources used to produce them. Success of early larvae is so critical to the success of a hatchery that even a relatively small improvement in survival or growth rate can yield great benefits. Outlook Live feeds remain indispensable for larviculture of many fish. Although micro- algae are among the costliest food sources used to produce live feeds, their many advantages justify the cost for hatcheries producing high-value fish. Research contin- ues to better characterise the nutritional properties of various algae strains and to optimise algae production technologies. We can anticipate that introduction of novel algae strains and nutritionally-opti- mised combinations of strains, along with improved feeding protocols, will ensure that microalgae remain the food of choice for production of the highest-quality live feeds. References www.aquafeed.co.uk/referencesIAF1303 May-June 2013 | InternatIonal AquAFeed | 15 FEATURE Naturally ahead MYC OFIX Myco toxin Ri sk M a n a g e M ent Mycofix® More protective. Mycotoxins decrease performance and interfere with the health status of your animals. Mycofix® is the solution for mycotoxin risk management. mycofix.biomin.net
  • 9. www.aquafeed.co.uk LINKS • See the full issue • Visit the International Aquafeed website • Contact the International Aquafeed Team • Subscribe to International Aquafeed They are what they eat Enhancing the nutritional value of live feeds with microalgae Controlling mycotoxins with binders Ultraviolet water disinfection for fish farms and hatcheries Niacin – one of the key B vitamins for sustaining healthy fish growth and production Volume 16 Issue 3 2013 - mAY | Ju Ne INCORPORATING fIsh fARmING TeChNOlOGy This digital re-print is part of the May | June 2013 edition of International Aquafeed magazine. Content from the magazine is available to view free-of-charge, both as a full online magazine on our website, and as an archive of individual features on the docstoc website. Please click here to view our other publications on www.docstoc.com. To purchase a paper copy of the magazine, or to subscribe to the paper edition please contact our Circulation and Subscriptions Manager on the link above. INFORMATION FOR ADVERTISERS - CLICK HERE