The Probiotic approach to elevating health of fish

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July | August 2010

Feature title: The Probiotic approach to elevating health o fsh

The International magazine or the aquaculture eed industry

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Probiotics and their applica-

tions in fish culture have

been studied and discussed

for much of the past decade.

However, we must not continue to use the

umbrella term too loosely and be mindful that

not all probiotics display the same properties

or yield the same subsequent benefits.

Probiotics are inclusive of a wide range

of microbes: Gram-positive bacteria, Gram-negative bacteria, yeasts and cyanobacteria

have all shown some varying degree of

success when applied to fish. They provide

benefits to the host fish, in part at least,

by improving the microbial balance and

functionality of the host gastrointestinal

microbial communities.

Improving fish healthFrom current literature, it is clear that

probiotic applications can improve fish

health, disease resistance, growth perform-

ance, feed utilisation, carcass composition,

intestinal morphology and reduce mal-

formations; however, we are yet to find a

specific probiotic candidate that can achieve

all of these effects.

Some candidates are able to mediate

more benefits than others, and perhaps

one of the most well studied, which we

have been working on at the Fish Nutrition

and Health Research Aquarium, at The

University of Plymouth, is Pediococcus aci-

dilactici (Bactocell, Lallemand).

P. acidilactici strains produce a range of

antimicrobial proteins (pediocins) and organic

acids (such as lactic and acetic acid) which

gives them antagonistic properties against a

range of Gram-positive and Gram-negative

bacteria, notably pathogenic Vibrio spp..

A clear effort to assess the use of this

probiotic in fish culture has been under-

taken; information regarding the benefits of

P. acidilacticiis available for pollock Pollachius

pollachius (Gatesoupe 2002), channel catfish

Ictalurus punctatus

(Shelby et al. 2007),

tilapia Oreochromis

niloticus (Shelby et

al. 2006; Ferguson et

al. 2010) and shrimp

(Castex et al. 2008,2010) but the bene-

fits to rainbow trout

O n c o r h y n c h u s

mykiss health and

production is most

well documented

(Quentel et al. 2004;

Aubin et al. 2005a,b;

Merrifield et al.

2010a,b).

We have previ-

ously shown at

Plymouth that the

provision of dietary

P. acidilacticican lead

to potentially resi-

dent populations

in close association

with the intestinal

epithelium of rain-

bow trout, even

though other com-

mercially available

probiotics could

not (Merrifield

et al. 2010a,b).

Subsequently, the

application of P.

acidilacticienhanced

the gut ultrastruc-

ture and improved

microvilli morphology of the enterocytes

in the anterior intestine of rainbow trout

(Figure 1; Merrifield et al. 2010b).

Not only could such intestinal improve-

Figure 1. TEM micrographs reveal that althoughmicrovilli appear healthy in both groups they aresignificantly longer in trout fed P. acidilactici (B)than the control (A). Scale bar = 1 m. (Merrifield etal. 2010b)

36 | InernaIona AquAFeed | July-august 2010 -

F: Probiotic

The Probiotic approachto elevating health of fishMolecular profiling of gastro-intestinal biota and effects on gut ultra-structural characteristics

by Dr Daniel L Merrifield and Matthew Emery, Senior microbiology technician, The School of Biomedical and BiologicalSciences, The University of Plymouth, Plymouth, UK, Email: [email protected]

ments provide a foundation for enhanced

digestive function but maintaining epithelial

integrity is paramount to defend against

enteric infections.

This is particularly relevant to salmo-

nid culture as the gastrointestinal tract is

a potential port of entry for many fish

pathogens and both Vibrio anguillarum and

Aeromonas salmonicida (the causative agents

of vibriosis and furunculosis, respectively)

can infect and cause severe tissue damage to

the anterior intestine of salmonid fish. From

these findings we hypothesise that dietary

applications of P. acidilacticimay be able to

prevent or reduce pathogen damage and

translocation in the anterior gut of salmonids

and we are presently using novel approaches

at The University of Plymouth to test this

hypothesis.

Indeed, a study supported by the

OFIMER (Office National Interprofessionnel

des Produits de la Mer et de l'Aquaculture),

demonstrated that P. acidilactici could

improve rainbow trout resistance to yers-

iniosis, as demonstrated by reduced accu-

mulated mortalities after an intraperitoneal

injection challenge.

As well as benefits to intestinal morphol-

ogy and disease resistance, P. acidilacticican

also alleviate rainbow trout vertebral col-

umn compression syndrome (VCCS) and

improve aesthetic quality by lowering con-

dition factor (Aubin et al. 2005a; Merrifield

et al. 2010a). These benefits which improve

the quality of the final product could effec-

tively boost production and elevate trout

farming profit margins.

The financial benefitsof this probiotic

We have recently also begun to assess

the efficacy of this probiotic in tilapia at

Plymouth (Ferguson et al. 2010).

We fed triplicate tanks of genetically male

tilapia (12 x 175g fish per tank) a basal diet

with or without the addition ofP. acidilactici.

After 14 days feeding on the experimen-

Figure 2. Probiotic P.acidilactici

significantlyelevated the circulatingleucocyte levels and serumlysozyme activity of tilapia

after 14 days feeding(Ferguson et al. 2010)

- July-august 2010 | InernaIona AquAFeed | 37

F: Probiotic

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mykiss Walbaum). Aqua. Nutr. in press. DOI:10.1111/j.1365-2095.2009.00712.x.

Merrifield, D. L., Harper, G., Baker, R. T. M., Ring, E.,Davies, S. J. (2010b). Possible influence of probioticadhesion to intestinal mucosa on the activity andmorphology of rainbow trout (Oncorhynchusmykiss) enterocytes. Aquacult. Res. in press DOI:10.1111/j.1365-2109.2009.02397.x.

Shelby, R.A., Lim, C., Yildirim-Aksoy, M. & Delaney,M.A. (2006) Effects of probiotic supplements ondisease resistance and immune response of youngNile tilapia, Oreochromis niloticus. J. Appl. Aqua.,18, 2234.

Shelby, R.A., Lim, C., Yildirim-Aksoy, M. & Klesius,P.H. (2007) Effects of probiotic bacteria as dietarysupplements on growth and disease resistancein young Channel catfish, Ictalurus punctatus(Rafinesque). J. Appl. Aqua., 19, 8191.

Quentel, C., Gatesoupe, F.J., Lamour, F., Abiven,A., Baud, M., Aubin, J. (2004) Effects of oraladministration of probiotics on the resistanceof rainbow trout, Oncorhynchus mykiss, againstYersinia ruckeri: asymptomatic carriers andhumoral immune parameters. 6th Symposium onFish Immunology, 2629 May 2004. bo/Turku,Finland, p. 60. book of abstracts.

sal interface which help

mediate the reported

host benefits in order to

maximise the benefits of

future applications.

References

Aubin, J., Gatesoupe,F.-J., Labbe, L., Lebrun, L.(2005a). Trial of probiotics

to prevent the vertebralcolumn compressionsyndrome in rainbow trout(Oncorhynchus mykissWalbaum). Aquacult. Res. 36,758767.

Aubin, J., Gatesoupe, F.J.,Quentel, C., Forraz, M.,Facqueur, J.M., Rochet, B.,Usache, V. (2005b). tudede l'amlioration de larentabilit et de la scuritsanitaire des filires truitearc-en-ciel et bar parl'utilisation de probiotiquespar voie alimentaire. RapportFinal, Convention N008-03C, Contract OFIMERNA01959. 67 pp.

Castex, M., Chim, L., Pham,D., Lemaire, P., Wabete, N.,Nicolas, J.-L., Schmidely, P.& Mariojouls, C. (2008)Probiotic P. acidilacticiapplication in shrimpLitopenaeus stylirostrisculture subject to vibriosis in New Caledonia.Aquaculture, 275, 182193.

Castex, M., Lemaire, P., Wabete, N., Chim, L.(2010) Effect of probiotic Pediococcus acidilacticion antioxidant defences and oxidative stressstatus of Litopenaeus stylirostris. Under Vibrionigripulchritudo challenge. Fish and ShellfishImmunology 28, 622-631.

Ferguson R. M. W., Merrifield, D. L., Harper,G. M., Rawling, M. D., Mustafa, S., Picchietti, S.,Balczar, J. L. &. Davies S. J. (2010) The effect ofPediococcus acidilactici on the gut microbiotaand immune status of on-growing red t ilapia(Oreochromis niloticus). J. Appl. Microbiol., earlyview doi:10.1111/j.1365-2672.2010.04713.x

Gatesoupe, F.-J. (2002) Probiotic andformaldehyde treatments of artemia naupliias food for lar val pollack, Pollachius pollachius.Aquaculture, 212, 347360.

Merrifield, D.L., Bradley, G., Harper, G.M.,Baker, R.T.M., Munn, C.B., Davies, S. J. (2010a).Assessment of the effects of vegetative andlyophilised Pediococcus acidilactici on growth,feed utilisation, intestinal colonisation and healthparameters of rainbow trout (Oncorhynchus

tal diets serum lysozyme activity and cir-

culating leucocyte levels were significantly

higher in the P. acidilacticigroup than the

control group (Figure 2).

At day 32, PCR-DGGE demonstrated

clear modulations of gastrointestinal micro-

bial profiles and the presence ofP. acidilactici

(OTU Pa) in the probiotic fed fish (Figure

3). The level of lactic acid bacteria in

the intestine significantly increased from

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