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Veterinary Parasitology 178 (2011) 279–285 Contents lists available at ScienceDirect Veterinary Parasitology journal homepage: www.elsevier.com/locate/vetpar Effect of dietary supplementation on resistance to experimental infection with Haemonchus contortus in Creole kids J.C. Bambou a , H. Archimède a , R. Arquet b , M. Mahieu a , G. Alexandre a , E. González-Garcia a,c , N. Mandonnet a,a Institut National de la Recherche Agronomique, Unité de Recherches Zootechniques, Domaine Duclos, 97170 Petit-Bourg (French West Indies), France b Institut National de la Recherche Agronomique, Domaine expérimental de Gardel, 97160 Moule, Guadeloupe, France c INRA UMR868, Systèmes d’Elevage Méditerranéens et Tropicaux (SELMET), Bâtiment 22, 12 Campus SupAgro-INRA, 2 Place Pierre Viala, 34060 Montpellier Cedex 1, France article info Article history: Received 16 June 2010 Received in revised form 11 January 2011 Accepted 17 January 2011 Keywords: Haemonchus contortus Goats Genetic resistance Dietary supplementation abstract The aim of the present study was to test the effect of dietary supplementation on resistance to experimental infection with Haemonchus contortus in Creole kids. One trial with three replicates involved a total of 154 female kids that were chosen from three successive cohorts of the Creole flock of INRA-Gardel in 2007. The kids were placed into four treatments accord- ing to the amount of concentrate they received: G0 (no concentrate and a quality Dichantium spp. hay ad libitum, HAY), G1 (HAY + 100 g commercial concentrate d 1 ), G2 (HAY + 200 g commercial concentrate d 1 ), G3 (HAY + 300 g commercial concentrate d 1 ). The G0–G3 groups were infected with a single dose of 10,000 H. contortus third stage larvae (L 3 ) at Day 0 (D0). Each infected group was comprised of one half resistant and one half susceptible genetically indexed kids. The average breeding values on egg excretion at 11 months of age were distant of 0.70, 0.65, 0.61 and 0.61 genetic standard deviations in G0, G1, G2 and G3, respectively. The faecal egg count (FEC), packed cell volume (PCV), eosinophilia (EOSI) and dry matter intake (DMI) indices were monitored weekly until 42 days post-infection. Enzyme-linked immunosorbent assay was carried out on serum samples to determine the level of IgA anti-H. contortus L 3 crude extracts and adult excretion/secretion products (ESP). The 10,000 L 3 dose received by the kids induced a severe infection: 8000 eggs per gram at the FEC peak, a PCV less than 15% and mortality. Interestingly, the supplemented animals in G3 showed a higher level of EOSI but a lower level of IgA anti-L3 and IgA anti-ESP than non-supplemented animals (G0). Resistant and susceptible kids had significantly different FEC variations within the groups. Susceptible kids had a 1.6 times higher egg output than resistant kids in G0. This difference was not found in the supplemented groups. The results of this study showed that supplementary feeding improved resistance of Creole kids to H. contortus experimental infection. © 2011 Elsevier B.V. All rights reserved. 1. Introduction Gastrointestinal nematode (GIN) infection remains a major constraint on small ruminant production through- Corresponding author at: INRA-URZ, Domaine Duclos, Prise d’eau, 97170 Petit-Bourg, France. Tel.: +33 590 25 54 08; fax: +33 590 25 59 36. E-mail address: [email protected] (N. Mandonnet). out the world. These parasitic diseases cause significant production loss, particularly in young animals. Due to the emergence of anthelmintic resistance (Jackson and Coop, 2000; Papadopoulos, 2008; Waller, 2005) and public concern about chemical residues in animal products, alter- native control strategies are needed. Two main areas of research have been developed. First, as short-term strate- gies, is the reduction of host contact with infective larvae though different methods of grazing management, target- 0304-4017/$ – see front matter © 2011 Elsevier B.V. All rights reserved. doi:10.1016/j.vetpar.2011.01.030

Effect of dietary supplementation on resistance to experimental infection with Haemonchus contortus in Creole kids

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Page 1: Effect of dietary supplementation on resistance to experimental infection with Haemonchus contortus in Creole kids

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Veterinary Parasitology 178 (2011) 279–285

Contents lists available at ScienceDirect

Veterinary Parasitology

journa l homepage: www.e lsev ier .com/ locate /vetpar

ffect of dietary supplementation on resistance to experimentalnfection with Haemonchus contortus in Creole kids

.C. Bamboua, H. Archimèdea, R. Arquetb, M. Mahieua,. Alexandrea, E. González-Garciaa,c, N. Mandonneta,∗

Institut National de la Recherche Agronomique, Unité de Recherches Zootechniques, Domaine Duclos, 97170 Petit-Bourg (French West Indies), FranceInstitut National de la Recherche Agronomique, Domaine expérimental de Gardel, 97160 Moule, Guadeloupe, FranceINRA UMR868, Systèmes d’Elevage Méditerranéens et Tropicaux (SELMET), Bâtiment 22, 12 Campus SupAgro-INRA,Place Pierre Viala, 34060 Montpellier Cedex 1, France

r t i c l e i n f o

rticle history:eceived 16 June 2010eceived in revised form 11 January 2011ccepted 17 January 2011

eywords:aemonchus contortusoatsenetic resistanceietary supplementation

a b s t r a c t

The aim of the present study was to test the effect of dietary supplementation on resistanceto experimental infection with Haemonchus contortus in Creole kids. One trial with threereplicates involved a total of 154 female kids that were chosen from three successive cohortsof the Creole flock of INRA-Gardel in 2007. The kids were placed into four treatments accord-ing to the amount of concentrate they received: G0 (no concentrate and a quality Dichantiumspp. hay ad libitum, HAY), G1 (HAY + 100 g commercial concentrate d−1), G2 (HAY + 200 gcommercial concentrate d−1), G3 (HAY + 300 g commercial concentrate d−1). The G0–G3groups were infected with a single dose of 10,000 H. contortus third stage larvae (L3) at Day0 (D0). Each infected group was comprised of one half resistant and one half susceptiblegenetically indexed kids. The average breeding values on egg excretion at 11 months ofage were distant of 0.70, 0.65, 0.61 and 0.61 genetic standard deviations in G0, G1, G2 andG3, respectively. The faecal egg count (FEC), packed cell volume (PCV), eosinophilia (EOSI)and dry matter intake (DMI) indices were monitored weekly until 42 days post-infection.Enzyme-linked immunosorbent assay was carried out on serum samples to determine thelevel of IgA anti-H. contortus L3 crude extracts and adult excretion/secretion products (ESP).The 10,000 L3 dose received by the kids induced a severe infection: 8000 eggs per gram atthe FEC peak, a PCV less than 15% and mortality. Interestingly, the supplemented animals

in G3 showed a higher level of EOSI but a lower level of IgA anti-L3 and IgA anti-ESP thannon-supplemented animals (G0). Resistant and susceptible kids had significantly differentFEC variations within the groups. Susceptible kids had a 1.6 times higher egg output thanresistant kids in G0. This difference was not found in the supplemented groups. The results

ed thatntal in

of this study showcontortus experime

. Introduction

Gastrointestinal nematode (GIN) infection remains aajor constraint on small ruminant production through-

∗ Corresponding author at: INRA-URZ, Domaine Duclos, Prise d’eau,7170 Petit-Bourg, France. Tel.: +33 590 25 54 08; fax: +33 590 25 59 36.

E-mail address: [email protected] (N. Mandonnet).

304-4017/$ – see front matter © 2011 Elsevier B.V. All rights reserved.oi:10.1016/j.vetpar.2011.01.030

supplementary feeding improved resistance of Creole kids to H.fection.

© 2011 Elsevier B.V. All rights reserved.

out the world. These parasitic diseases cause significantproduction loss, particularly in young animals. Due tothe emergence of anthelmintic resistance (Jackson andCoop, 2000; Papadopoulos, 2008; Waller, 2005) and public

concern about chemical residues in animal products, alter-native control strategies are needed. Two main areas ofresearch have been developed. First, as short-term strate-gies, is the reduction of host contact with infective larvaethough different methods of grazing management, target-
Page 2: Effect of dietary supplementation on resistance to experimental infection with Haemonchus contortus in Creole kids

ry Parasitology 178 (2011) 279–285

Table 1Average determined chemical composition (dry matter basis) of dietarycomponents.

Components Dichantiumhay

Commercialpelleta

Dry matter 90.2 96.4Organic matter 91.6 93.5Crude protein 6.8 28.9NDF 71.0 13.3ADF 37.3 3.6

280 J.C. Bambou et al. / Veterina

ing anthelmintic treatment of the most infected animalsin the flock and the management of nutrition to increasehost resistance and/or resilience (Hoste et al., 2008; Torres-Acosta and Hoste, 2008; van Wyk and Bath, 2002); secondlyis the improvement of the host response against GINthough the genetic selection of lines or breeds of resistantanimals (Baker and Gray, 2003).

In sheep, numerous studies suggest that an improvednutritional status could reduce the production losses andmortality rates due to GIN infection (Sykes and Coop,2001; Walkden-Brown and Kahn, 2001). Although goatsare markedly more susceptible to nematode infection thansheep, data on the relationships between dietary supple-mentation and parasitism in this model are still scarce(Hoste et al., 2008).

A breeding scheme is under way for improved resis-tance to GIN infection within a local Creole goat breed inFrench West Indies. Indeed, it has been shown that nat-urally contaminated tropical pastures allow the geneticevaluation of Creole goat resistance to gastrointestinalstrongyles (Mandonnet et al., 2001, 2006). Creole goatsof the INRA-Gardel flock are indexed on their resis-tance under mixed natural infection conditions. Howeverinfection levels and proportions between Haemonchuscontortus, Trichostrongylus colubriformis and Oesophagos-tomum columbianum populations vary across seasons andpasture managements. The aim of this study was to investi-gate the effect of dietary supplementation on the resilienceand resistance of Creole goats towards an experimentalinfection with H. contortus.

2. Materials and methods

2.1. Animals and experimental design

This experiment, replicated three times, involved a totalof 154 female kids (10.1 ± 0.5 kg BW; 7 months old) thatwere chosen from three successive cohorts of the Creoleflock of INRA-Gardel in 2007. The flock grazed all yearon irrigated Digitaria decumbens pastures contaminatedwith H. contortus, T. colubriformis and O. columbianum. Onaverage, the kids were mothered at pasture until wean-ing at 3 months of age. The pedigree of each animal wasavailable from the foundation generation of 1979 andeach animal was genetically indexed for faecal egg counts(FEC) at 11 months of age (Mandonnet et al., 2001, 2006).Briefly, in the flock of INRA Domaine Gardel, the pedi-gree of each animal was available since the foundationgeneration was established in 1979. Faecal samples werecollected regularly (during weeks 6 and 7 after drenching)at 7 and 11 months of age for genetic evaluation on theaverage of 2 FEC measures. Thus, the breeding value (BV)for FEC of each kid of the flock at 11 months old, undernatural mixed infection on pasture conditions, was reg-ularly estimated. Such estimation was made taking intoaccount their own individual performances, the perfor-

mances of its ascendants, and their pedigree (Bambou et al.,2009).

At 7 months of age no differences of FEC means wereobserved between replicates when comparing resistantand susceptible kids. Then, animals were drenched with

ADL 5.1 0.3

a Consisting of maize (68%), soybean cake (15%), wheat bran (11%), urea(1%) and vitamin and mineral supplements (5%).

Ivermectin (Oramec®, Merial, Lyon, France, 300 �g/kg bodyweight) and housed indoors under worm-free conditionsone month before the start of the experiment. They wereplaced into four groups according to the amount of com-mercial concentrate they received: G0 (no concentrateand Dichantium spp. hay ad libitum, HAY, n = 39 kids), G1(100 g commercial concentrate d−1 and HAY, n = 38 kids),G2 (200 g commercial concentrate d−1 and HAY, n = 38kids), G3 (300 g commercial concentrate d−1 and HAY,n = 39 kids) (Table 1). All groups were balanced accordingto live weight. The amount of metabolizable protein andnet energy increased with the rate of concentrate supple-mentation, approximately at 17, 49, 86 and 127 g per dayand 1.3, 2.5, 2.7 and 3.6 MJ per day for the experimentalgroups: G0, G1, G2 and G3, respectively. These estimatesrelate to healthy animals. For infected animals these valuescould be reduced by 20–30% (Dakkak, 1995). The animalswere reared following European Union recommendationsfor animal welfare in accordance with the regulations ofthe Animal Care Committee of INRA

H. contortus third stage larvae (L3) were obtained 42days before the challenge from cultures of faeces takenfrom monospecifically infected Creole goats with isolatespreviously obtained from Creole goats reared on pasture.The G0–G3 groups were infected with a single dose of10,000 H. contortus L3 at day 0 (D0). Each infected groupwas comprised of one half resistant and one half suscepti-ble genetically indexed kids (Mandonnet et al., 2001, 2006;Bambou et al., 2009). The predicted average breeding val-ues on egg excretion at 11 months of age were distant of0.70, 0.65, 0.61 and 0.61 genetic standard deviations in G0,G1, G2 and G3, respectively.

2.2. Parasitological techniques, blood and serum samples

Faecal samples were collected to determine the FECusing a modified McMaster method for rapid determi-nation (Bambou et al., 2008). Blood samples from eachanimal were recovered once a week and centrifuged for5 min at 5000 rpm. Serum was then frozen at −20 ◦Cuntil analysis. Blood samples were collected in EDTAcoated tubes (Becton Dickinson, Plymouth, UK) to mea-

sure the number of circulating eosinophils according tothe method of Dawkins et al. (1989). Eosinophils werecounted using a Malassez cell counter. The packed cell vol-ume was measured using the capillary microhaematocritmethod.
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y Parasitology 178 (2011) 279–285 281

2

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2

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2

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0

2000

4000

6000

8000

10000

12000

0 7 14 21 28 35 42

FE

C (

eggs

/g o

f fae

ces)

Days post-infection

G0G1G2G3

*

*

*

Fig. 1. Geometric means of faecal egg counts (FEC) according to the exper-imental groups: � G0, Group 0 (no concentrate, n = 39); � G1, Group 1(100 g commercial concentrate d−1, n = 38); � G2, Group 2 (200 g com-

J.C. Bambou et al. / Veterinar

.3. Feed intake measurements

Every morning, the kids were offered the concentraterst and, once finished the consumption, then the Dichan-

ium hay was immediately distributed ad libitum. Theoncentrate was distributed individually with the help ofoke traps during the consumption lapses’ time. Feed-ng stalls were long enough to avoid competition for hayetween the kids. Offered and refused feed were recordedeekly for each experimental group in order to esti-ate average voluntary dry matter intake (DMI). However,

ata are expressed individually by metabolic body weightg/BW0.75). Daily mean individual intake was estimatedy dividing total intake of each group by the number ofids being measured. Such values were then divided by theean metabolic weight of the same animals.

.4. Antibody detection in serum by indirect ELISA

The ELISA analyses were performed on sera from 60ids, representative of the three cohorts.

.4.1. Worm antigen preparationPrior to experimentation five donor goats infected with

0,000 L3 of H. contortus were sacrificed at 42 days post-nfection (d.p.i.) and adult worms were harvested fromhe abomasum. These worms were thoroughly washed inBS (pH 7.4) containing penicillin (100 IU/ml) and strep-omycin (1 mg/ml). Fifty adult worms per millilitre ofhe same buffer were maintained in a 5% CO2 atmo-phere at 37 ◦C overnight. Next, the supernatant containingxcretory/secretory products (ESP) was collected, filtered0.2 �m) and stored at −70 ◦C until further use. A crudextract of H. contortus L3 was prepared after three cycles ofreezing and thawing (−70 ◦C, +25 ◦C), homogenization at◦C and centrifugation at 30,000 × g for 30 min at 4 ◦C. The

upernatant was used as the crude extract of the L3 antigen.he protein concentration of both antigenic preparationsas determined using the method of Bradford.

.4.2. Serum specific IgACrude extracts of H. contortus L3 and ESP were diluted

t 2 �g/ml in carbonate buffer (pH 9.6), distributed in6-well plates (Nunclon surface, Nunc, Denmark) and incu-ated overnight at 4 ◦C. The wells were washed threeimes with PBST (0.01 M phosphate, 0.15 M sodium chlo-ide, pH 7.2 and 0.1% Tween 20). Non-specific bindingites were blocked by 3 h incubation with PBS-1% bovineerum albumin (BSA, Sigma, St. Louis, USA) and 5% sucrose.uplicate serum samples diluted 1:2 in PBST were incu-ated for 2 h at room temperature (RT). The plates wereashed three times with PBST before the addition of aorseradish peroxidase-conjugated Rabbit anti-goat IgAAlpha Diagnostic) diluted 1:1000 in carbonate buffer60 min of incubation at RT). Three final washes with PBSTere carried out before the addition of 100 �l per well

f the chromogen (2, 2′-azino-bis, 3-ethylbenzthiazoline--sulfonic acid), and incubation at RT. After 20 min, theptical densities were determined with a spectrophotome-er by measuring the absorbance at 405 nm. In order toompare results between the assays, a positive control

mercial concentrate d−1, n = 38) and × G3, Group 3 (300 g commercialconcentrate d−1, n = 39). Significant differences (P < 0.05) between supple-mented (G1, G2 and G3) and non-supplemented groups (G0) are indicatedby asterisk marks.

consisting of a pool of sera containing IgA antibodies wasincluded on each plate, and the OD405 of unknown sampleswere altered in proportion to the changes of this standard.

2.5. Statistical analysis

The FEC and EOSI variables were log transformed inorder to normalize the variances. The kinetics of eachvariable was modelled using the Mixed procedure of SASv. 8.6 (SAS Institute Inc., 1999). The model included thefixed effects of experiment replicate (n = 3), the combinedeffect of birth-rearing rank (n = 4), the dietary treatment(n = 4) and genetic predisposition to GIN within groups(n = 2; resistant and susceptible). Repeated measures datawere analysed using an autoregressive structure with kid,within dietary treatment and replicate as the subjects. Thepeaks of the different variables were also localised usingthe mixed procedure of SAS software, including time asdiscrete variation factor. The results are presented afterback-transformation. Pearson’s rank correlations were cal-culated in order to determine associations between thedata also using SAS (1999). Significant results were con-sidered for P < 0.05.

3. Results

3.1. Parasitological and zootechnical measures

The FEC remained at zero until 21 d.p.i. in all dietarytreatments (Fig. 1). A peak was observed between 28 and35 d.p.i. in all groups. When compared to the supplementedkids (G1, G2 and G3), the overall FEC mean was 2-foldhigher (P < 0.05) in kids without supplementation (G0),independently of the level of supplementation. No signif-icant difference was observed between the supplemented

groups. Susceptible kids had a 1.6 times higher FEC thanthe resistant kids in G0 (Fig. 2). However, this differencewas not found in the supplemented groups.

The PCV values significantly decreased during theexperiment in all groups until 28 d.p.i. (P < 0.001; Fig. 3).

Page 4: Effect of dietary supplementation on resistance to experimental infection with Haemonchus contortus in Creole kids

282 J.C. Bambou et al. / Veterinary Parasitology 178 (2011) 279–285

Fig. 2. Differences between faecal egg counts (FEC) in resistant and sus-

0

1

2

3

4

5

6

0 7 14 21 28 35 42

Eos

inop

hils

(10

6 ×ce

lls/m

l)

Days post-infection

G0

G100

G200

G300

* *

Fig. 4. Geometric means of number of blood eosinophils/ml according tothe experimental groups: � G0, Group 0 (no concentrate, n = 39); � G1,Group 1 (100 g commercial concentrate d−1, n = 38); � G2, Group 2 (200 g

ceptible kids according to the experimental dietary treatments: G0, Group0 (no concentrate, n = 39); G1, Group 1 (100 g commercial concentrate d−1,n = 38); G2, Group 2 (200 g commercial concentrate d−1, n = 38) and G3,Group 3 (300 g commercial concentrate d−1, 39).

No difference was observed among the supplementedgroups. However, a significant difference was observedbetween supplemented (G1, G2 and G3) and and non-supplemented groups (G0) from 14 to 42 d.p.i. (P < 0.001).The PCV increased significantly (P < 0.0001) in all groupsfrom 28 to 42 d.p.i. No difference was observed betweenresistant and susceptible animals.

Eosinophil counts in blood significantly increased afterinfection and showed a peak between 7 and 14 d.p.i. in allgroups except for G0 which remained constant through-out the experiment (P < 0.001; Fig. 4). Then, eosinophiliadecreased significantly in all groups (P < 0.001). The overalllevel of eosinophilia was significantly higher in G3 com-pared to G0 (P < 0.05). Time variation within groups wasnot significant.

All groups gained weight throughout the experimentand no BW losses were observed, which support the

idea that undernourishment was effectively avoided whenplanning experimental diets. Average daily gains (ADG)during the experimental period were different (P < 0.01)among dietary treatments and increased with proteinallowance (Fig. 5). Nonetheless, a significant difference was

0

5

10

15

20

25

30

35

40

0 7 14 2

PC

V (

%)

Days post-in

G0G1G2G3

*

*

Fig. 3. Means of packed cell volume (PCV) and standard error of the mean accor� G1, Group 1 (100 g commercial concentrate d−1, n = 38); � G2, Group 2 (200 g cconcentrate d−1, n = 39). Significant differences (P < 0.001) between supplementeasterisk marks.

commercial concentrate d−1, n = 38) and × G3, Group 3 (300 g commercialconcentrate d−1, n = 39). Significant differences (P < 0.05) between sup-plemented G3 and and non-supplemented G0 are indicated by asteriskmarks.

observed between G0 and G1 compared to G2 and G3 (Fig. 5,P < 0.05).

Hay intake increased with the level of concentrate whenthe animals consumed the mixed diets. However, regard-less of the mixed ration, the hay intake was always lowerthan that observed in the control diet (G0, ad libitum hay)(Table 2; P < 0.05). Whatever the diet considered, the con-centrate contributed less than 30% of the total DMI.

3.2. Serum antibody responses

The OD’s levels of sera from parasite-free kids used asnegative controls were not significantly different from thebackground levels (data not shown). Following infectionwith H. contortus, the levels of the IgA anti-L3 response

increased in all groups to peak at 21 d.p.i. and thendecreased to reach a baseline level at the end of theinfection (Fig. 6a). The levels of IgA specific antibodyresponse to H. contortus adult excretion/secretion products(IgA anti-ESP) increased in all groups to peak at 42 d.p.i.

1 28 35 42fection

*

**

ding to the experimental groups: � G0, Group 0 (no concentrate, n = 39);ommercial concentrate d−1, n = 38) and × G3, Group 3 (300 g commerciald (G1, G2 and G3) and non-supplemented groups (G0) are indicated by

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J.C. Bambou et al. / Veterinary Parasitology 178 (2011) 279–285 283

Table 2Means of dry matter intake (DMI) of Creole kids during experimental infection with Haemonchus contortus according to the experimental groups: G0, Group0 (no concentrate); G1, Group 1 (100 g commercial concentrate d−1); G2, Group 2 (200 g commercial concentrate d−1) and G3, Group 3 (300 g commercialconcentrate d−1). Means identified as significantly different (P < 0.05) have different letters listed with the respective values.

G0 G1 G2 G3 SEMa

DMI (g/BW.75)b (forage)c 75.9a 53.2b 54.4b 69.3c 1.7DMI (g/BW.75) (forage + concentrate)d 75.9a 66.0b 79.6a 99.9 c 1.7

(a(r(ravIP

4

dtwedsaTogatwTe

Fee1cc(

mucus secretion and the immune response (Blackburnet al., 1991; Coop and Kyriazakis, 2001). In this study,supplementation significantly increased the growth rate(ADG), as was shown in Criollo kids browsing native veg-

0.00

0.05

0.10

0.15

0.20

0.25a

Mea

n of

Opt

ical

Den

sity

G0

G1

G2

G3

*

*

**

a Standard error mean.b DMI per kg of metabolic weight (BW.75).c DMI of forage.d DMI of forage + concentrate.

Fig. 6b). No difference between resistant and susceptiblenimals was observed within groups for the IgA responseIgA anti-L3 and IgA anti-ESP). The G0 group had an IgAesponse more pronounced than the supplemented groupsP < 0.002; Fig. 6a and b). The values of PCV negatively cor-elated with IgA anti-L3 and IgA anti-ESP values (r = 0.530nd r = 0.560, respectively; P < 0.001; data not shown). Thealues of FEC positively correlated with IgA anti-L3 andgA anti-ESP values (r = 0.432 and r = 0.229, respectively;< 0.05, data not shown).

. Discussion

It has been postulated that protein metabolism is moreisturbed by GIN infection than any other component ofhe diet, including energy (Vanhoutert et al., 1995a). Thisas well discussed in a previous study in our conditions

valuating effect of GIN parasitism on feed intake andigestibility of Creole kids (Bambou et al., 2009). However,ome recent studies have underlined the fact that energynd protein should both be considered (Hoste et al., 2005).he aim of the present study was to investigate the effectf supplementary feeding on the resistance of Creole kidsenetically resistant and susceptible to GIN infection, in

n experimental infection with H. contortus. We showedhat supplementary feeding in Creole kids was associatedith increased resilience and resistance to GIN infection.

his was shown by increased growth rate (ADG), decreasedxcretion of GIN eggs in the faeces (FEC) and absence of

ig. 5. Means of average daily gain (ADG) of Creole kids duringxperimental infection with Haemonchus contortus according to thexperimental groups: G0, Group 0 (no concentrate, n = 39); G1, Group

(100 g commercial concentrate d−1, n = 38); G2, Group 2 (200 gommercial concentrate d−1, n = 38) and G3, Group 3 (300 g commer-ial concentrate d−1, n = 39). Means identified as significantly differentP < 0.05) have different letters listed above the respective columns.

acute anaemia in the supplemented groups compared tothose not supplemented. Similar findings showing a signif-icant effect of supplementation on resilience in browsingkids and in pen trials with goats have been reported(Blackburn et al., 1991; Torres-Acosta et al., 2004), as wellas in field trials with grazing sheep (Vanhoutert et al.,1995a). It has been suggested that the positive impact ofsupplementation on GIN infection is due to the compen-sation of endogenous protein loss induced in part by themaintenance of gastrointestinal tract integrity, increased

b

423528211470

Days post-infection

0

0.02

0.04

0.06

0.08

0.1

0.12

0.14

0.16

423528211470

Mea

n of

Opt

ical

Den

sity

Days post-infection

G0

G1

G2

G3*

*

*

Fig. 6. (a) Systemic IgA response against H. contortus crude extract of L3antigen (anti-L3) according to the experimental groups: � G0, Group 0(no concentrate, n = 15); � G1, Group 1 (100 g commercial concentrate d−1,n = 15); � G2, Group 2 (200 g commercial concentrate d−1, n = 15) and � G3,Group 3 (300 g commercial concentrate d−1, n = 15). Significant differences(P < 0.05) between supplemented (G1, G2 and G3) and non-supplementedgroups (G0) are indicated by asterisk marks and (b) Systemic IgA responseagainst adult H. contortus excretion secretion products (anti-ESP) accord-ing to the experimental groups: � G0, Group 0 (no concentrate, n = 15);� G1, Group 1 (100 g commercial concentrate d−1, n = 15); � G2, Group 2(200 g commercial concentrate d−1, n = 15) and � G3, Group 3 (300 g com-mercial concentrate d−1, n = 15). Significant differences (P < 0.05) betweensupplemented (G1, G2 and G3) and non-supplemented groups (G0) areindicated by asterisk marks.

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ry Parasi

284 J.C. Bambou et al. / Veterina

etation in tropical Mexico and in sheep grazing dry orsemi-arid environments. Positive digestive interactions(better intake and digestion of the roughage) would beexpected from the intake of concentrate, representing lessthan 30% of the total DMI. The reduction of hay intake fromthe first level of concentrate would indicate that this wasnot the case in the present trial. This can be explained by therelatively good quality of the hay used. Generally, positivedigestive interactions are explained by the improved rumi-nal environment classically observed with poor roughage(Archimede et al., 1999). We hypothesized that the increaseof intake observed with the amount of concentrate couldbe explained by the improvement of the protein allowanceto the kids, as previously suggested by Egan (1980). How-ever, the method used in our study to measure intake (e.g.weekly intervals and animals in communal pens with yoketraps) could probably not be sufficiently fine for short termtrials and not adapted to monitor digestibility. In furtherresearch we aim to investigate nutrition-parasitism inter-actions in this experimental model with a more adaptedmethod to measure intake and digestibility: individualfeeding and daily individual feed consumption measures.

In this study, we showed that the physiopathologicaleffects of H. contortus experimental infection were pro-nounced in the non-supplemented groups as revealed bythe clinical signs typical of haemonchosis, such as anaemia,submandibular oedema, prostration, apathy and, in somecases, death. In contrast, the supplemented kids were resis-tant to the infection. The advantage of supplementationwas already obvious at 100 g of concentrate per day. Insheep, numerous studies have suggested that the bene-fits of supplementation on the deleterious effects of GINparasitism are more pronounced in susceptible genotypescompared to resistant ones (Coop and Kyriazakis, 1999). Inthis study, susceptible kids were more responsive to theinfluence of increased supplementation, resulting in theabsence of difference in resistance to infection betweenresistant and susceptible animals in the supplementedgroups. In contrast, it has been reported that increased pro-tein supplementation resulted in increased resistance to H.contortus infection in the native, more resistant Santa Ineslambs compared with the more susceptible Ile de Francebreed (Bricarello et al., 2005). Coop and Kyriazakis sug-gested that when the benefits of a more resistant genotypeare not decreased by a lower protein diet, this would indi-cate that such animals, as Creole goats, would be better ableto survive in areas of the world where forage quality is poor(Coop and Kyriazakis, 1999).

Many studies suggested that eosinophils play a role inresistance to helminth infection since significant corre-lations between resistance/susceptibility to endoparasiteinfection and the magnitude of the peripheral eosinophilresponse have been shown (Meeusen et al., 2005). Here, weshowed that supplementation enabled blood eosinophiliaof Creole kids to increase after infection whereas novariation was observed in non-supplemented groups.

These results are in agreement with previous studies inCriollo kids from tropical Mexico under natural infec-tion conditions and in sheep artificially infected withH. contortus, T. colubriformis or Teladorsagia circumcincta(Datta et al., 1998; Valderrabano et al., 2002; Vanhoutert

tology 178 (2011) 279–285

et al., 1995b). Thus, these results suggest that the immuneresponse against GIN infection of supplemented animalswas enhanced compared to that of lambs kept on arestricted diet and that eosinophils may play a role in thismechanism.

Regardless of the experimental diet, no significantdifference was found between resistant and suscepti-ble kids in the levels of IgA anti-L3 and IgA anti-ESP(IgA response) throughout the study. In contrast, a sig-nificant higher level of the IgA response was found innon-supplemented animals compared to supplementedones. These results are not consistent with a previous studyin sheep which suggested that the plane of nutrition maybe positively correlated with the antibody response againstGIN (Martinez-Valladares et al., 2005). Our data would sug-gest that the IgA response is better correlated with thenematode burden, as reflected by the FEC and the PCV.

5. Conclusion

Numerous studies have already reported the benefitof supplementation on resistance and resilience in smallruminants to parasitic infection (Jackson and Miller, 2006;Knox et al., 2006). The present study establishes that about75 g of the commercial concentrate per day prevented thedeleterious effect of H. contortus infection in growing Creolekids when considering resistance parameters. This effectmust be confirmed on growth parameters and on trickleinfections.

Acknowledgements

This study was supported by ‘La Région Guadeloupe’ andthe European Community (FEOGA). The authors thank L.Abinne-Molza and H. Varo for their technical assistance inparasitological measurements in the laboratory. They arealso grateful to the Gardel team in charge of the goat flock:T. Kandassamy, W. Troupé, J. Gobardhan and S.-A. Matou.J.-C. Bambou was supported by a post-doctoral fellowshipfrom Le Conseil Régional de la Guadeloupe.

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