robiotics are well known androutinely used additives inthe main livestock species.They claim to improve guthealth by stabilising gut flora beingtheir effect reflected in a betteroverall health status, welfare and per-formance of the animals.
However, their use in fish production isstill scarce, being nowadays only one addi-tive registered to be used in the EuropeanUnion.Different factorsmight be behindthe lack of this typeof products foraquaculture:1) Gut microbiota and physiology of fishes cultured arestill being studiedand seems to differin a high extentfrom one speciesto another2) Probioticsused in monogastricand ruminants avail-able nowadays aremainly based on bac-teria or yeast thatneed temperatureenough to developin the animals’ gut.It can be difficultto reach taking intoaccount that fishare poiquilotermsand in some specificproductions watertemperature isextremely low3) Up today, it is not well known if the microorganisms from probiotics candevelop multiply as well as modify fishgut flora in these environmental and gutconditions4) Feed fish processing is extremely hardin terms of temperature and pressure so,how to apply these alive microorganismsto fish pellets is still being studied. Theirinclusion by coating after pelleting can bethe solution, although the stability of thismicroorganism in this oily solutions as wellas once reach water in tanks or sea needsstill to be demonstratedDespite all this, and taking into accountthe increasing importance of fish productionall over the world, Rubinum SA is investinga lot of effort in this field. In this regard, itrecently ran a trial in collaboration withIRTA to study the effect of the probiotic
cereus var. toyoi on rainbow trout(Oncorhynchus mykiss) fingerlings.In the trial, fingerlings of rainbow trout(4.2 ± 0.1g) were fed two diets, a commer-cial diet (Aller FuturaTM from AllerAqua,Denmark) and the same diet containingthe probiotic B. cereus var. toyoi at thefinal concentration of 2*104 UFC/g, during93 days.Each treatment was tested in triplicate(400-L tank, 125 fish/tank, initial density:1.3kg/m3). During the trial, water tempera-ture, conductivity, pH and dissolved oxygenwere 13.2 ± 0.2ºC, 1800 ± 200 µS/cm, 7.5± 0.01 and 8.0 ± 0.3 mg/L (mean ± S.D.),respectively.Tanks were connected to a recircula-tion system (IRTAMAR®) which maintainedadequate waterquality param-eters. Fish werefed at appar-ent satiation(3.3 percent)with automaticfeeders (ARVO-TEC T-Drum-2000TM,Finland). Theproximatebiochemicalcompositionof diets was64 percentprotein, 12percent fat and11 percent ash(2.0 mm pelletsize).All fish fromeach tank were
Use of probiotics in aquaculture:
can these additives be useful?
by E Gisbert, DVM, PhD, Researcher, Irta Sant Carles de la Ràpita, Spain and M Castillo, DVM, PhD, R&Dand Customer Support Manager, Rubinum SA, Spain
8 | IntnatInal
| november-December 2011-
measured for their body size, and 45specimens per condition (15 per replicate)sacrificed for histology (size of intestinal villiand number of goblet cells), assessment of digestive system functionality and quantifi-cation of the intestinal microbiota by RFLP.
At the end of the trial, fish fed thediet containing the probiotic were slightlysignificantly heavier and longer (43.9 ± 9.1g, 14.4 ± 1.1cm) than those fed the controldiet (42.5 ± 7.6g, 14.1 ± 1.1; n = 276).Distribution of size classes in bodyweight were also affected by the incorpora-tion of the probiotic in the diet.The frequency of fish belonging to the51-70g size class was higher in the group fedthe probiotic (53.6 ± 1.1 vs. 47.1± 2.2%; t-test,P < 0.05; n = 3; see figure 1 and 2), whereasthose trouts fed the control diet showed ahigher frequency of smaller individuals (6.9 ±0.5 vs 3.6 ± 0.7%; t-test, P < 0.05; n = 3).The percentage of fish with intermediateweights (41-50 g) was also significantly higheramong those fish fed the probiotic (16.2 ± 0.5vs. 13.0 ± 1.5; t-test, P < 0.05; n = 3).The above-mentioned changes in growthand size classes did not affect the proximalcomposition of fish fed both diets (protein:40.0 ± 2.2%, lipids: 20.5 ± 2.0%, ash: 1.8 ± 0.5%).The inclusion of the probiotic into thecontrol diet did not affect the functional-ity of the digestive system, as indicatedby the absence of significant differences inthe specific activity of pancreatic (trypsin,chymotrypsin, total protease) and intes-tinal brush border (alkaline phosphatase,aminopeptidase-N, maltase) enzymes.However, the number of goblet cells(1.6 ± 0.1 vs. 1.3 ± 0.2 cells/100 µm; n =15) and height of villi (928.5 ± 137 vs. 527± 130 µm; t-test, P < 0.001; n = 15; seefigure 3 and 4) in the intestinal mucosa wassignificantly higher in those fish fed the dietcontaining the probiotic. Goblet cells, orso-called mucous cells, reside throughoutthe intestine and are the main source of mucins production in the gut.Mucins are considered to play importantroles in host defense by forming a physicalbarrier between the host and the contentsof the intestinal lumen.Thus, these results indicated that theinclusion of the probiotic in the diet pro-moted goblet cell proliferation and pos-sibly the immune response in the intestinalmucosa.In addition, theintestinal micro-biota was alsoaffected by thediet, showing dif-ferent RFLP results(clades) dependingon the tested die-tary group. Theseresults indicatethat the inclusionof the probiotic inthe diet was ableto modulatehost micro-biota, although themolecular tech-niques used in thisstudy did not allowthe identificationof the bacterialgenus or species.
In conclusion,the inclusion of B. cereus var.toyoi at the finalconcentration of 2*10
UFC/g in acommercial dietpromoted growthin rainbow troutfingerlings, as well as the organisation of theintestinal mucosa (number of goblet cellsand villi height), whereas did not affect thespecific activity of selected pancreatic andintestinal digestive enzymes.Therefore,the inclusion of this probiotic introut feeds could be beneficial and advantageousin terms of the fish host,as well as for the inten-sive production of the species,although morestudies are needed to study mode of action of Gram positive bacteria in the gut as well as thecorrect dosage to administer.
-november-December 2011 | IntnatInal