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China is the world’s largest producer of aquaculture, currently producing more than ten times more than India, and over 30 different species of aquaculture products. Now that financial incomes in developing nations such as China have increased, the demand for meat and dairy has also risen as more people can afford to consume these higher protein foods.
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July | August 2010
Feature title: Effect of Orego-Stim on the growth performance and
intestinal bacterial populations of channel catfish (Ictalurus punctatus)
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The International magazine for the aquaculture feed industry
F: Growth performance performance
F: Growth Aqua News
AwF launch
Optimisation of range of
Lallemand feed of the diet science-based, field-approved
nutritional composition
intake and
redesigned website
helps to ensure for a catfish
solutionsthat channel sustainable aquaculture
Effect of Orego-Stim on the A
are extracting the maximum possible quaculture without
growth from what they consume. Frontiers (AwF) is
L
Ingestion of Animal Nutrition Further applications being under guarantying its purity and con-
allemand aquatic feed inevita- pleased to announce
the launch of its completely
growth performance and
causes drinking develop- investigation to cover other major sistent quality. Agrimos contains
bly is dedicated to theof the localised
aquatic environment and mar- aquaculture species (tilapia, carp, one of the highest guaran-
ment, production, putting the gut redesigned website.
keting of profitable, naturaldirect chal- Performance trials have teed levels of MOS on the
interface of catfish into and etc). This fulfills the commitment
to raise the profile of AwF and
intestinal bacterial populations
lenge with yeast and bacteria- proven beneficial effects in terms market.
differentiated the indigenous pathogenic
microbial community.
based solutions for animal nutri- of growth enhancement, feed uti- Independent trials in various improve fundraising activities
The gastrointestinal tract lisation, as well as improved resist- salmonid and tropical fish species
tion and health. is that Dr Barry Costa-Pierce and
The company hassensitive to aa wide under specific challenging have shown the benefits of
responsive and developed ance Dr Mudnakudu Nandeesha,
international co-chairs, made at
of channel catfish (Ictalurus punctatus)
range microbial-based nutri- conditions.
range ofof stressors. yeast mannan-oligosaccharides
tional Various phytogenic ingredients
solutions to support per- on gut health, intestinal micro- the AwF Board of Directors
have been health of aquatic Overcoming intestinal
formance andshown to facilitate ben- flora and global performances meeting on September
species in effects on gut environment
eficial a natural way. disturbance under various challenging condi- 26, 2009 during the World
Channel Catfish are an important aquaculture species originally from USA but introduced into China Lallemand’s aquaculture inno- plant
and microflora, though some Levucell® SB, a probiotic yeast tions. Aquaculture Society confer-
in the mid 1980s and now widely cultured in China. Last year Chinese channel catfish production vative solutions stronger impact than
species have a are based on strain (Saccharomyces cerevi- Laltide® is a combination of ence in Veracruz, Mexico.
was estimated at over 500 thousand MT in both small scale and larger commercial monoculture and others.
specific yeast and bacteria strains siae var. boulardii CNCM 1079). available nucleotide specifically The AwF Board of Directors
polyculture systems. grown in the company’s effects S.of
The antimicrobial own find the
boulardii Figures 1-4. Growth performance for aquaculture total weight hopes visitors willgrowth new
yeast is well-known formulated measured by appli- gain, specific
phenolic compounds which are the cations. It is recommended as a
production plants. Each product’s in human and animal health for ratio and protein efficiency ratio. website informative and provide
ratio, feed conversion
origin, safety and concentration feedpositive effects against intes- well-balanced source of nucle-
active ingredient of phytogenic its a clear and comprehensive view
by Z. L. Zheng of China and Matt Pearce of the UK
additives, target pathogenic bacte- disturbance. Its modes of otides adapted to the needs of
are guaranteed. tinal of work to date.
Visit were used for
(Chongqing, China) the website atthe
C
catfish aimed to fish the response of
The development and valida- with in the gut are well under- analyseand crustacean species, par-
rial cell walls by interaction action
hina is the world’s largest duced into China in tion of cytoplasmic membrane andstood, creating a real populations whenduring periods of intense
the our ‘field led and science changing bacterial ‘shield ticularly exposed to www.aquaculturewithout
experiment. Experiments were conducted
Table 1: Formulation (g kg-1) and proximate composition of frontiers.org
producer of aquaculture, the mid 1980s and now the diet
supported’ products iscations, on H+ and K+. Orego-Stim®.
its permeability for based like effect’, reinforcing the microbial metabolism needs. in a recirculation aquaculture system at
currently producing more widely cultured in China. scientific research and perform- ecosystem and protecting the Both aero-
than ten times more than Last year Chinese Ingredients Chemical composition Inhibitory effects on bic and anaero-
ance trials conducted in par t- animal’s digestive tract. Levucell
India, and over 30 different species of channel catfish produc- microorganisms bic
nership with leading interna- SB is also a source ofbeneficial beneficial
aquaculture products. Now that finan- tion was estimated and pathogenic
tional Orego-Stim® contains essential oilsparticularly useful to
research centres, universi- nutrients
cial incomes in developing nations
such as China have increased, the
at over 500 thousand
MT in both small scale
Soybean meal
Rapeseed meal
300.00
260.00
DE (kJ kg-1)
CP
12.51
345.00
ties and commercial firms around promote the growthuof certain
which include precise natural composi-
of
the world: a-terpinene, p-cymene, aquaculture species. bacteria
tions of carvacrol
pop lations
FIGAP/ VIV
demand for meat and dairy has also and larger commercial
Corn gluten meal 50.00 EE 38.50
and thymol. These components Alkosel®, yeast isolated
Bactocell® (live bacteria strain are found a were product América Latina 2010
risen as more people can afford to monoculture and poly- Pediococcus inhibitory CNCM on micro- elevatedquantified
to have acidilactici effects containing and levels of
consume these higher protein foods. culture systems. Fish meal 100.00 ASH 62.40 organisms especially in spore forming
MA18/5M), the first probiotic selenomethionine, the highest from differ- Your natural partner for aquaculture
Channel catfish is Wheat shorts 220.00 Ca 8.50 organisms. ent regions of
strain authorised for use in aqua- bioavailable form of selenium.
Between 1995 and 2009 soybean imports tolerant to poorer It has European Union, is Alkosel is the intestine
culture in thebeen clearly demonstrated that recommended as a We offer a range of innovative microbial-based
Rapeseed oil 20.00 TP 12.20
increased from 150 thousand tonnes to 41 water quality conditions chronic of its measured by source of of
a guaranteestress safety, quality prolonged natural selenium for channel solutions for sustainable aquaculture:
million tonnes to help sustain this increased with dissolved oxygen Ca(H2PO4)2 16.00 AP 8.00 and efficacy blood plasma cortisol concentra-
elevated in aquaculture. fish and shrimpcatfish. Various
production. October 21-23, 2010
Vitamin premixa
Guadalajara
demand in animal feed. (DO) of less than 1ppm, 1.70 Lys 17.50 Bactocell is one of the best dose dependent s ® ,other c ogrowth
tion in trout results in sci- Agrimo a ncen-
During the same time period broiler though when produced increased mortality probiotic trated sourcep a r ayeastt e r s
entifically-documented from common aquatic of m e cell Probiotics
Choline chloride 2.00 Met + Cys 13.20
production doubled to over 12 million in aquaculture DO is bacterial and fungal diseases.
strains in aquatic species with wall mannan-oligosaccharidessuch as weight
Sodium chloride 1.00
tonnes and pork production increased by maintained at or above many This finding isinnot just confined to sal-
applications salmonids, (MOS) obtained through again, FCR, SGR,
over 50 percent. 4ppm in order to opti- Mineral premixb 5.00 shrimps and other marinenormal physiological Protein
monid species but is a fishes. specific production process effi-
China is undeniably the world’s most mise production response in all fish species. The hypotha- ciency ratio and
Selenium Enriched Yeast
Bentonite 24.30
powerful nation in its appetite for agricul- efficiency. lamic-pituitary-interrenal axis in fish which mortality were Register now
Note: Crude protein, crude lipid, ash, crude fibre are expressed
ture and aquaculture commodity use and can be stimulated by both environmental also quantified. for free entr
has consistently consumed over
on a dry matter basis and given as means (n=2)
and stress events affects the production of ance
a Vitamin mix provided the following vitamins (mg kg-1 diet Yeast Mannan-Oligosaccharides (MOS)
20 percent of the world’s
unless otherwise stated): vitamin A, 4000 IU; vitamin D3, 2000
lymphocytes and antibody response as well Fish and www.viv.net
production of fishmeal IU; vitamin K, 10; vitamin E, 50; thiamine, 10; riboflavin, FES Consultants (UK) Ltd culture
as the reproductive capacity.
in the past 10 years 12; pyridoxine, 10; panthothenic acid, 32; nicotinic acid, Gut Associated Lymphoid Tissue (GALT) allconditions
• Wear parts for single and
to help support its 80; folic acid, 2; biotin, 0.2; vitamin B12, 0.01; L-ascorbyl-2- is the interface between thetwin screw extruders h a n n e l
diet, host C Purified Nucleotides
Guadalajara, Jalisco - Mexico
expanding aquaculture polyphosphate (250 g kg-1 vitamin C activity), 60 • GALT activ-
physiology and gut microflora. The supply and refurbishment of
catfish (Ictalurus
b Trace mineral mix provided the following minerals (mg kg-1 used extruders for the pet, aqua
industry. the diet by
ity can be modified through and human food industryt a t u s )
punc
Channel Catfish are an diet): zinc (as ZnSO4·7H2O), 150; iron (as FeSO4·7H2O), three principles – competitive bacterial propagated It is time to explore a wide range of
important aquaculture 40; manganese (as MnSO4·H2O), 25; copper (as CuCl2), 3; www.fes-ltd.com by
exclusion, bacterial antagonism and immune and reared challenging opportunities
iodine (as KI), 5; cobalt (as CoCl2·6H2O), 0.05; selenium (as Unit 11, Barnwell Manor Estate, Lallemand Animal Nutrition- 19, rue des Briquetiers BP 59 31702 Blagnac Cedex − France
species originally the health
modulation which in turn affect Barnwell, Peterborough, PE8 5PL
the Research e-mail: aqua@lallemand.com Tel.: +33 (0)5 62 74 5555 Fax:+33 (0)5 62 74 5500
Na2SeO3), 0.09 Tel: +44 (0)1832 270831 Email:
from USA but and commercial productive status of fish. Institute
welcome@fes-ltd.com for www.lallemandanimalnutrition.com
intro- latest experimental trial in Channel
This extrusion equipment we can saveFyou h e r i e s
If you use i s money! www.lallemand.com
adv viv americalatina 132x190a.indd 1 25-06-2010 14:09:21
10 30 | InternatIonal AquAFeed | July-august 2010
| InternatIonal AquAFeed | July-august 2010 July-august 2010 | InternatIonal AquAFeed |
July-august 2010 | InternatIonal AquAFeed | 11 31
F: Growth performance
GROWiNG iS
At each feeding, an excess amount of The formulating method of FWA as fol-
feed was fed to the fish and uneaten feed lows: MgSO4 (0.05g), beef extract (2.5g),
was collected one hour after feeding, dried NaCl (500g), K2HPO4 (0.2g), glucose (1g),
at 70°C and reweighed. Leaching of uneaten peptone (5.0g), yeast extract (2.5g), agar
feed was estimated by placing weighed powder (15.0g), add water to 1000 ml, pH
beautiful
samples of each diet into a tank without fish 7.2-7.4, and was sterilised for 30 min at
for one hour and then recovered, dried and 121℃.
reweighed. 18 FWA plates were spread with
The average leaching value homogenates of the intestinal sections in
was used to correct the amount each group. Nine plates were incubated
Southwest U n i ve r s i t y, of uneaten feed. at 30℃ for 24-48 hours in an incubator,
Chongqing, China. Channel catfish with and nine plates were incubated at 30℃
an average initial weight of 50 g were Intestine bacteriological for 48-72 hours in an anaerobic incubator.
held in 200 L fibreglass tanks. Fish in sampling Plates with 20-200 bacterial colonies were
randomly assigned duplicate aquaria (40 Six channel catfish were sampled from counted and bacterial numbers in plates
fish per aquaria) were fed one of the two each diet group after one-day starvation. were converted to the number per unit
experimental diets to apparent satiation The fish were killed by physical destruc- weight of intestine.
six times daily. Water temperature and pH tion of the brain, and the number of inci-
were constant (23-24°C; pH 8.5) during dental organisms was reduced by washing Identification of bacteria
the experimental period. Water flow was the fish skin with 700 ml L-1 ethanol before Approximately 10 colonies were ran-
maintained at 4.5 L/minute. Orego-Stim® opening the ventral surface with sterile domly picked from plates containing 20 to
(50g kg-1) was a commercial product of scissors to expose the body cavity. 200 bacteria. A total of 100 isolates were
Meriden Animal Health Ltd., UK. The ventral belly surface of the fish inoculated with a method of slope culture,
The dietary formulation of each of the was opened to expose the peritoneal cav- respectively. 60 tubes were incubated at
diets was as follows: ity, and then the spleen, gallbladder and 30℃ for eight to 12 hours in an incubator,
Control: without feed additives liver removed. The intestinal sections were and placed at 4℃ after incubation.
Orego-Stim®: 50g kg-1 Orego-Stim® emptied and thoroughly rinsed three times Another 40 tubes were incubated at
in 2ml sterile 9g/kg-1 saline to remove non- 30℃ for 24 to 36h in an anaerobic incuba-
Experimental design adherent bacteria. tor, and were identified after 4 hours.
and diets Anterior intestine (AI), mid intestine The identification process of aerobic bac-
Both diets were formulated to be isoca- (MI), and posterior intestine (PI) of the cor- teria was formulated according to Bergey’s
loric (12.51kJ kg-1 diet) and isonitrogenous responding sections were treated separately, Manual of Determinative Bacteriology and
(3450g/kg-1 crude protein). All ingredients and were then transferred to sterile plastic classified according to genera or groups on
were finely ground, mixed in a Hobart mixer bags and homogenised in a Stomacher the basis of cell morphology, motility, gram
and pelleted through a 2.4mm diameter die (Seward Laboratory, London, UK). reaction, catalase and oxidase and glucose
in a Hobart meat grinder. The pellets were fermentation, O/F test, salt tolerance for
air-dried at room temperature, fragmented Bacterial plates growth.
and stored in a freezer until use. spreading and count Anaerobic bacteria were identified with
Ingredients and proximate composition Homogenates of the intestinal sections an automatic microorganism identifica-
of the experimental diets are presented in were diluted in sterile 9g/kg-1 saline and tion instrument (VITEK-�) by the French
Table 1. appropriate dilutions were spread on the BioMerieux Company and identification
The feeding trial lasted eight weeks. surface of fresh water agar (FWA) plates. plate (VITEK-ANA) for anaerobic bacteria.
Results - Growth
Table 2: WG, SGR, FCR, PER and survival of Channel catfish fed different diets with feed additives performance
Diets WG1 (g) SGR2 (%/d) FCR3 PER4 Survival Survival rate of the experi-
ment was high (>95%) and unre-
lated to dietary treatment (see
Control 102.63±2.22a 1.95±0.04a 1.94±0.092a 1.49±0.08a 95.83±2.89a Table 2). Data on the growth
Car+Thy 116.77±2.49b 2.15±0.05b 1.84±0.03a 1.58±0.02a 95.00±2.50a performance of channel catfish,
OS 127.53±6.40c 2.24±0.12b 1.64±0.04b 1.77±0.07b 98.33±1.44a including weight gain (WG), spe-
cific growth ratio (SGR), feed
Data were presented as mean±SD (n=2). Value in the same column having different superscripts is
conversion ratio (FCR) and pro-
FRANCE / 00 33 2 97 40 42 09
significant (P<0.05).
1 WG (Weight gain, g) = final body weight − initial bodyweight
tein efficiency ratio (PER) are
shown in Table 2.
2 SGR (Specific growth ratio, /d) = (loge average final weight - loge average initial weight) / no days) x
WG of fish fed the OS diet Our hydrolysates have a superior concentration of Natural Active NutrientsTM
100 was significantly higher than
3 FCR (Feed conversion ratio) = feed consumed (g, dry weight) / weight gain (g) those of fish fed the Con diet Thanks to their biological action on fish and shrimp growth mechanisms,
4 PER (Protein efficiency ratio) = weight gain (g) / protein intake (g) (P<0.05). they boost your feed performance and help you substituting fish meal.
SGR of fish fed the OS diet
contact@aquativ-diana.com - www.aquativ-diana.com
32 | InternatIonal AquAFeed | July-august 2010
F: Growth performance
Numbers of Aer. also order to maximise growth performance
declined with the and immune defence.
Orego-Stim® diet This experimental trial isolated the bac-
and this result was terial species, compositional proportion and
significantly different the region of the intestine that different
from the control bacteria species colonised.
(P<0.05). Conversely, Orego-Stim® demonstrated the ability
Bac. numbers to selectively reduce or increase different
increased signifi- species of bacteria. Numbers of key patho-
cantly upon addition genic bacteria such as Ent. Aer. and Vib were
of OS (P<0.05). reduced in the Orego-Stim® group of
Addition of Orego- catfish, where as beneficial bacteria which
Stim® had no effect helped to promote gut health such as Lac.
on the numbers and Bif had higher numbers of bacteria.
of Staphylococcus This study shows that Orego-Stim®
(Stap). acts in a similar way as probiotics using
In MI, the num- competitive exclusion, but unlike probiot-
bers of Ent. sig- ics Orego-Stim® has much more potent
nificantly decreased antimicrobial properties.
upon addition of
OS (P<0.05). The Orego-Stim® is a
feed additives had phytogenic feed additive.
no effect on the This specific discipline of aquaculture
numbers of Aer., feed science is still an innovative and pio-
Stap. and Vib.. The neering technology.
numbers of Bac. At the time of writing the exact bio-
also increased chemical mechanism by which beneficial
significantly in the bacteria are allowed to grow in the intes-
Orego-Stim® diets tines and pathogenic bacteria are killed, is
compared to con- still not fully understood. The theory is that
trol (P<0.05). there must be a structural difference in
In PI, the numbers the cell wall of the two classes of bacteria
of Ent., Aer., Stap. and possibly relating to epitopes and virulence
Vib. decreased with factors which selectively promotes or pre-
Figures 5-7: Composition of Aerobic and Anaerobic the Orego-Stim® vents cell lysis upon exposure to Orego-
bacteria species in the Anterior, Mid and posterior diet. Stim®, but this needs further research to
intestine of Channel Catfish Anaerobic get the required scientific evidence.
bacteria What is known is that both the holistic
The three intes- and generic effects of Orego-Stim® work
were significantly higher than those of fish tinal anaerobic bacteria identified with in a way that alters intestinal bacterial
fed the control diet. FCR and PER were also the identification plate (VITEK-ANA) of populations and benefits the host fish by
affected by OS. anaerobic bacteria were Bifidobacterium increasing aquaculture productivity, both
Lowest FCR and highest PER values (Bif.), Lactobacillus (Lac.) and Bacteroides experimentally and commercially.
were observed for catfish fed the OS diet (Bact.), respectively. The predominant References are available on request.
compared to catfish fed the control diet intestinal anaerobic bacteria were Bact. Bif.
Dry with 20-50% less energy
(P<0.05). and Lac. (Figure 7).
In the AI, MI and PI numbers of Bif. and
Composition of bacteria Lac in the Orego-Stim® group significantly About the authors:
flora - Aerobic Bacteria increased compared to that of the control Z. L. Zheng of China - School
Bacterial numbers are shown in tables (P<0.05). Numbers of Bact. significantly of Veterinary Medicine of Sichuan
5-7. decreased in the Orego-Stim® group com- Agriculture University, Sichuan 625014,
Intestinal aerobic bacteria of 4 families pared to that of control (P<0.05). PR China and the Fisheries Breeding
(genera) of channel catfish were identi- and Healthy Cultivation Research
fied. The intestinal predominant aerobic Discussions and conclusion Centre, South-west University,
bacteria were Enterobacteriaceae Ent.）, As there is evidence that intestinal com- Chongqing 402460, PR China
Aeromonas (Aer.), Bacillus (Bac.) and Vibrio petitive bacterial exclusion and bacterial Matt Pearce - Meriden Animal
(Vib.) (Figure 5-6). antagonism are the unique modes of action Health Limited, Cranfield Innovation
In AI, the numbers of Enterobacteriaceae that help the immune response of fish, it is Centre, University Way, Cranfield Geelen Counterflow / T +31-475-592315
in fish fed Orego-Stim® decreased sig- pertinent to understand how Orego-Stim® Technology Park MK43 0BT, United Geelen Counterflow USA Inc. / T +1-772-559-4338
nificantly compared to control (P<0.05). can beneficially balance the gut flora in Kingdom Geelen Counterflow América Latina / T +54-9-2362-418899
E info@geelencounterflow.com / I www.geelencounterflow.com
34 | InternatIonal AquAFeed | July-august 2010
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