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

Veterinary Parasitology 178 (2011) 279–285

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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 Institut National de la Recherche Agronomique, Domaine expérimental de Gardel, 97160 Moule, Guadeloupe, France INRA UMR868, Systèmes d’Elevage Méditerranéens et Tropicaux (SELMET), Bâtiment 22, 12 Campus SupAgro-INRA, 2 Place Pierre Viala, 34060 Montpellier Cedex 1, France

b c

a r t i c l e

i n f o

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

a b s t r a c t 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 according 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 (L3 ) 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 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 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). 0304-4017/$ – see front matter © 2011 Elsevier B.V. All rights reserved. doi:10.1016/j.vetpar.2011.01.030

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, alternative control strategies are needed. Two main areas of research have been developed. First, as short-term strategies, is the reduction of host contact with infective larvae though different methods of grazing management, target-

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ing anthelmintic treatment of the most infected animals in the flock and the management of nutrition to increase host resistance and/or resilience (Hoste et al., 2008; TorresAcosta and Hoste, 2008; van Wyk and Bath, 2002); secondly is the improvement of the host response against GIN though the genetic selection of lines or breeds of resistant animals (Baker and Gray, 2003). In sheep, numerous studies suggest that an improved nutritional status could reduce the production losses and mortality rates due to GIN infection (Sykes and Coop, 2001; Walkden-Brown and Kahn, 2001). Although goats are markedly more susceptible to nematode infection than sheep, data on the relationships between dietary supplementation and parasitism in this model are still scarce (Hoste et al., 2008). A breeding scheme is under way for improved resistance to GIN infection within a local Creole goat breed in French West Indies. Indeed, it has been shown that naturally contaminated tropical pastures allow the genetic evaluation of Creole goat resistance to gastrointestinal strongyles (Mandonnet et al., 2001, 2006). Creole goats of the INRA-Gardel flock are indexed on their resistance under mixed natural infection conditions. However infection levels and proportions between Haemonchus contortus, Trichostrongylus colubriformis and Oesophagostomum columbianum populations vary across seasons and pasture managements. The aim of this study was to investigate the effect of dietary supplementation on the resilience and resistance of Creole goats towards an experimental infection with H. contortus. 2. Materials and methods 2.1. Animals and experimental design This experiment, replicated three times, involved a total of 154 female kids (10.1 ± 0.5 kg BW; 7 months old) that were chosen from three successive cohorts of the Creole flock of INRA-Gardel in 2007. The flock grazed all year on irrigated Digitaria decumbens pastures contaminated with H. contortus, T. colubriformis and O. columbianum. On average, the kids were mothered at pasture until weaning at 3 months of age. The pedigree of each animal was available from the foundation generation of 1979 and each 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 pedigree of each animal was available since the foundation generation was established in 1979. Faecal samples were collected regularly (during weeks 6 and 7 after drenching) at 7 and 11 months of age for genetic evaluation on the average of 2 FEC measures. Thus, the breeding value (BV) for FEC of each kid of the flock at 11 months old, under natural mixed infection on pasture conditions, was regularly estimated. Such estimation was made taking into account their own individual performances, the performances of its ascendants, and their pedigree (Bambou et al., 2009). At 7 months of age no differences of FEC means were observed between replicates when comparing resistant and susceptible kids. Then, animals were drenched with

Table 1 Average determined chemical composition (dry matter basis) of dietary components. Components

Dichantium hay

Commercial pelleta

Dry matter Organic matter Crude protein NDF ADF ADL

90.2 91.6 6.8 71.0 37.3 5.1

96.4 93.5 28.9 13.3 3.6 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 body weight) and housed indoors under worm-free conditions one month before the start of the experiment. They were placed into four groups according to the amount of commercial concentrate they received: G0 (no concentrate and 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 = 38 kids), G3 (300 g commercial concentrate d−1 and HAY, n = 39 kids) (Table 1). All groups were balanced according to live weight. The amount of metabolizable protein and net energy increased with the rate of concentrate supplementation, approximately at 17, 49, 86 and 127 g per day and 1.3, 2.5, 2.7 and 3.6 MJ per day for the experimental groups: G0, G1, G2 and G3, respectively. These estimates relate to healthy animals. For infected animals these values could be reduced by 20–30% (Dakkak, 1995). The animals were reared following European Union recommendations for animal welfare in accordance with the regulations of the Animal Care Committee of INRA H. contortus third stage larvae (L3 ) were obtained 42 days before the challenge from cultures of faeces taken from monospecifically infected Creole goats with isolates previously obtained from Creole goats reared on pasture. The G0–G3 groups were infected with a single dose of 10,000 H. contortus L3 at day 0 (D0). Each infected group was comprised of one half resistant and one half susceptible genetically indexed kids (Mandonnet et al., 2001, 2006; Bambou et al., 2009). The predicted 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. 2.2. Parasitological techniques, blood and serum samples Faecal samples were collected to determine the FEC using a modified McMaster method for rapid determination (Bambou et al., 2008). Blood samples from each animal were recovered once a week and centrifuged for 5 min at 5000 rpm. Serum was then frozen at −20 ◦ C until analysis. Blood samples were collected in EDTA coated tubes (Becton Dickinson, Plymouth, UK) to measure the number of circulating eosinophils according to the method of Dawkins et al. (1989). Eosinophils were counted using a Malassez cell counter. The packed cell volume was measured using the capillary microhaematocrit method.

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

12000

Every morning, the kids were offered the concentrate first and, once finished the consumption, then the Dichantium hay was immediately distributed ad libitum. The concentrate was distributed individually with the help of yoke traps during the consumption lapses’ time. Feeding stalls were long enough to avoid competition for hay between the kids. Offered and refused feed were recorded weekly for each experimental group in order to estimate average voluntary dry matter intake (DMI). However, data are expressed individually by metabolic body weight (g/BW0.75 ). Daily mean individual intake was estimated by dividing total intake of each group by the number of kids being measured. Such values were then divided by the mean metabolic weight of the same animals.

10000

2.4. Antibody detection in serum by indirect ELISA The ELISA analyses were performed on sera from 60 kids, representative of the three cohorts. 2.4.1. Worm antigen preparation Prior to experimentation five donor goats infected with 10,000 L3 of H. contortus were sacrificed at 42 days postinfection (d.p.i.) and adult worms were harvested from the abomasum. These worms were thoroughly washed in PBS (pH 7.4) containing penicillin (100 IU/ml) and streptomycin (1 mg/ml). Fifty adult worms per millilitre of the same buffer were maintained in a 5% CO2 atmosphere at 37 ◦ C overnight. Next, the supernatant containing excretory/secretory products (ESP) was collected, filtered (0.2 ␮m) and stored at −70 ◦ C until further use. A crude extract of H. contortus L3 was prepared after three cycles of freezing and thawing (−70 ◦ C, +25 ◦ C), homogenization at 4 ◦ C and centrifugation at 30,000 × g for 30 min at 4 ◦ C. The supernatant was used as the crude extract of the L3 antigen. The protein concentration of both antigenic preparations was determined using the method of Bradford. 2.4.2. Serum specific IgA Crude extracts of H. contortus L3 and ESP were diluted at 2 ␮g/ml in carbonate buffer (pH 9.6), distributed in 96-well plates (Nunclon surface, Nunc, Denmark) and incubated overnight at 4 ◦ C. The wells were washed three times with PBST (0.01 M phosphate, 0.15 M sodium chloride, pH 7.2 and 0.1% Tween 20). Non-specific binding sites were blocked by 3 h incubation with PBS-1% bovine serum albumin (BSA, Sigma, St. Louis, USA) and 5% sucrose. Duplicate serum samples diluted 1:2 in PBST were incubated for 2 h at room temperature (RT). The plates were washed three times with PBST before the addition of a horseradish peroxidase-conjugated Rabbit anti-goat IgA (Alpha Diagnostic) diluted 1:1000 in carbonate buffer (60 min of incubation at RT). Three final washes with PBST were carried out before the addition of 100 ␮l per well of the chromogen (2, 2 -azino-bis, 3-ethylbenzthiazoline6-sulfonic acid), and incubation at RT. After 20 min, the optical densities were determined with a spectrophotometer by measuring the absorbance at 405 nm. In order to compare results between the assays, a positive control

FEC (eggs/g of faeces)

2.3. Feed intake measurements

281

G0 G1 G2 G3

8000

* * *

6000 4000 2000 0 0

7

14 21 28 Days post-infection

35

42

Fig. 1. Geometric means of faecal egg counts (FEC) according to the 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 commercial concentrate d−1 , n = 38) and × G3, Group 3 (300 g commercial concentrate d−1 , n = 39). Significant differences (P < 0.05) between supplemented (G1, G2 and G3) and non-supplemented groups (G0) are indicated by asterisk marks.

consisting of a pool of sera containing IgA antibodies was included on each plate, and the OD405 of unknown samples were altered in proportion to the changes of this standard. 2.5. Statistical analysis The FEC and EOSI variables were log transformed in order to normalize the variances. The kinetics of each variable was modelled using the Mixed procedure of SAS v. 8.6 (SAS Institute Inc., 1999). The model included the fixed effects of experiment replicate (n = 3), the combined effect 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 data were analysed using an autoregressive structure with kid, within dietary treatment and replicate as the subjects. The peaks of the different variables were also localised using the mixed procedure of SAS software, including time as discrete variation factor. The results are presented after back-transformation. Pearson’s rank correlations were calculated in order to determine associations between the data also using SAS (1999). Significant results were considered for P < 0.05. 3. Results 3.1. Parasitological and zootechnical measures The FEC remained at zero until 21 d.p.i. in all dietary treatments (Fig. 1). A peak was observed between 28 and 35 d.p.i. in all groups. When compared to the supplemented kids (G1, G2 and G3), the overall FEC mean was 2-fold higher (P < 0.05) in kids without supplementation (G0), independently of the level of supplementation. No significant difference was observed between the supplemented groups. Susceptible kids had a 1.6 times higher FEC than the resistant kids in G0 (Fig. 2). However, this difference was not found in the supplemented groups. The PCV values significantly decreased during the experiment in all groups until 28 d.p.i. (P < 0.001; Fig. 3).

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Eosinophils (106×cells/ml)

6 5

*

4

*

3 2

G0 G100 G200 G300

1 0 0

7

14

21

28

35

42

Days post-infection Fig. 2. Differences between faecal egg counts (FEC) in resistant and susceptible kids according to the experimental dietary treatments: G0, Group 0 (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 supplemented groups. However, a significant difference was observed between supplemented (G1, G2 and G3) and and nonsupplemented groups (G0) from 14 to 42 d.p.i. (P < 0.001). The PCV increased significantly (P < 0.0001) in all groups from 28 to 42 d.p.i. No difference was observed between resistant and susceptible animals. Eosinophil counts in blood significantly increased after infection and showed a peak between 7 and 14 d.p.i. in all groups except for G0 which remained constant throughout the experiment (P < 0.001; Fig. 4). Then, eosinophilia decreased significantly in all groups (P < 0.001). The overall level of eosinophilia was significantly higher in G3 compared to G0 (P < 0.05). Time variation within groups was not significant. All groups gained weight throughout the experiment and no BW losses were observed, which support the idea that undernourishment was effectively avoided when planning experimental diets. Average daily gains (ADG) during the experimental period were different (P < 0.01) among dietary treatments and increased with protein allowance (Fig. 5). Nonetheless, a significant difference was

Fig. 4. Geometric means of number of blood eosinophils/ml according to the 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 commercial concentrate d−1 , n = 38) and × G3, Group 3 (300 g commercial concentrate d−1 , n = 39). Significant differences (P < 0.05) between supplemented G3 and and non-supplemented G0 are indicated by asterisk marks.

observed between G0 and G1 compared to G2 and G3 (Fig. 5, P < 0.05). Hay intake increased with the level of concentrate when the animals consumed the mixed diets. However, regardless of the mixed ration, the hay intake was always lower than that observed in the control diet (G0, ad libitum hay) (Table 2; P < 0.05). Whatever the diet considered, the concentrate contributed less than 30% of the total DMI. 3.2. Serum antibody responses The OD’s levels of sera from parasite-free kids used as negative controls were not significantly different from the background levels (data not shown). Following infection with H. contortus, the levels of the IgA anti-L3 response increased in all groups to peak at 21 d.p.i. and then decreased to reach a baseline level at the end of the infection (Fig. 6a). The levels of IgA specific antibody response to H. contortus adult excretion/secretion products (IgA anti-ESP) increased in all groups to peak at 42 d.p.i.

40 35 30 PCV (%)

25

*

20 15

G0 G1 G2 G3

10 5

* *

* *

0 0

7

14 21 28 Days post-infection

35

42

Fig. 3. Means of packed cell volume (PCV) and standard error of the mean according to the 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 commercial concentrate d−1 , n = 38) and × G3, Group 3 (300 g commercial concentrate d−1 , n = 39). Significant differences (P < 0.001) between supplemented (G1, G2 and G3) and non-supplemented groups (G0) are indicated by asterisk marks.

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283

Table 2 Means of dry matter intake (DMI) of Creole kids during experimental infection with Haemonchus contortus according to the experimental groups: G0, Group 0 (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 commercial concentrate d−1 ). Means identified as significantly different (P < 0.05) have different letters listed with the respective values.

.75 b

c

DMI (g/BW ) (forage) DMI (g/BW.75 ) (forage + concentrate)d

G2

G3

SEMa

75.9a 75.9a

53.2b 66.0b

54.4b 79.6a

69.3c 99.9 c

1.7 1.7

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

(Fig. 6b). No difference between resistant and susceptible animals was observed within groups for the IgA response (IgA anti-L3 and IgA anti-ESP). The G0 group had an IgA response more pronounced than the supplemented groups (P < 0.002; Fig. 6a and b). The values of PCV negatively correlated with IgA anti-L3 and IgA anti-ESP values (r = 0.530 and r = 0.560, respectively; P < 0.001; data not shown). The values of FEC positively correlated with IgA anti-L3 and IgA anti-ESP values (r = 0.432 and r = 0.229, respectively; P < 0.05, data not shown). 4. Discussion It has been postulated that protein metabolism is more disturbed by GIN infection than any other component of the diet, including energy (Vanhoutert et al., 1995a). This was well discussed in a previous study in our conditions evaluating effect of GIN parasitism on feed intake and digestibility of Creole kids (Bambou et al., 2009). However, some recent studies have underlined the fact that energy and protein should both be considered (Hoste et al., 2005). The aim of the present study was to investigate the effect of supplementary feeding on the resistance of Creole kids genetically resistant and susceptible to GIN infection, in an experimental infection with H. contortus. We showed that supplementary feeding in Creole kids was associated with increased resilience and resistance to GIN infection. This was shown by increased growth rate (ADG), decreased excretion of GIN eggs in the faeces (FEC) and absence of

acute anaemia in the supplemented groups compared to those not supplemented. Similar findings showing a significant effect of supplementation on resilience in browsing kids and in pen trials with goats have been reported (Blackburn et al., 1991; Torres-Acosta et al., 2004), as well as in field trials with grazing sheep (Vanhoutert et al., 1995a). It has been suggested that the positive impact of supplementation on GIN infection is due to the compensation of endogenous protein loss induced in part by the maintenance of gastrointestinal tract integrity, increased mucus secretion and the immune response (Blackburn et 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-

a Mean of Optical Density

c d

G1

0.25

G0

* *

G1

0.20

*

G2 G3

0.15

*

0.10 0.05 0.00 0

7

14

21

28

35

42

Days post-infection

b

0.16

Mean of Optical Density

a b

G0

0.14

*

G0 G1

0.12

G2

0.1

G3

*

0.08

*

0.06 0.04 0.02 0 0

7

14

21

28

35

42

Days post-infection

Fig. 5. Means of average daily gain (ADG) of Creole kids during experimental infection with Haemonchus contortus according to the 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 commercial concentrate d−1 , n = 38) and G3, Group 3 (300 g commercial concentrate d−1 , n = 39). Means identified as significantly different (P < 0.05) have different letters listed above the respective columns.

Fig. 6. (a) Systemic IgA response against H. contortus crude extract of L3 antigen (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 䊉 G3, Group 3 (300 g commercial concentrate d−1 , n = 15). Significant differences (P < 0.05) between supplemented (G1, G2 and G3) and non-supplemented groups (G0) are indicated by asterisk marks and (b) Systemic IgA response against adult H. contortus excretion secretion products (anti-ESP) 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 䊉 G3, Group 3 (300 g commercial concentrate d−1 , n = 15). Significant differences (P < 0.05) between supplemented (G1, G2 and G3) and non-supplemented groups (G0) are indicated by asterisk marks.

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etation in tropical Mexico and in sheep grazing dry or semi-arid environments. Positive digestive interactions (better intake and digestion of the roughage) would be expected from the intake of concentrate, representing less than 30% of the total DMI. The reduction of hay intake from the first level of concentrate would indicate that this was not the case in the present trial. This can be explained by the relatively good quality of the hay used. Generally, positive digestive interactions are explained by the improved ruminal environment classically observed with poor roughage (Archimede et al., 1999). We hypothesized that the increase of intake observed with the amount of concentrate could be explained by the improvement of the protein allowance to the kids, as previously suggested by Egan (1980). However, the method used in our study to measure intake (e.g. weekly intervals and animals in communal pens with yoke traps) could probably not be sufficiently fine for short term trials and not adapted to monitor digestibility. In further research we aim to investigate nutrition-parasitism interactions in this experimental model with a more adapted method to measure intake and digestibility: individual feeding and daily individual feed consumption measures. In this study, we showed that the physiopathological effects of H. contortus experimental infection were pronounced in the non-supplemented groups as revealed by the clinical signs typical of haemonchosis, such as anaemia, submandibular oedema, prostration, apathy and, in some cases, death. In contrast, the supplemented kids were resistant to the infection. The advantage of supplementation was already obvious at 100 g of concentrate per day. In sheep, numerous studies have suggested that the benefits of supplementation on the deleterious effects of GIN parasitism are more pronounced in susceptible genotypes compared to resistant ones (Coop and Kyriazakis, 1999). In this study, susceptible kids were more responsive to the influence of increased supplementation, resulting in the absence of difference in resistance to infection between resistant and susceptible animals in the supplemented groups. In contrast, it has been reported that increased protein supplementation resulted in increased resistance to H. contortus infection in the native, more resistant Santa Ines lambs compared with the more susceptible Ile de France breed (Bricarello et al., 2005). Coop and Kyriazakis suggested that when the benefits of a more resistant genotype are not decreased by a lower protein diet, this would indicate that such animals, as Creole goats, would be better able to survive in areas of the world where forage quality is poor (Coop and Kyriazakis, 1999). Many studies suggested that eosinophils play a role in resistance to helminth infection since significant correlations between resistance/susceptibility to endoparasite infection and the magnitude of the peripheral eosinophil response have been shown (Meeusen et al., 2005). Here, we showed that supplementation enabled blood eosinophilia of Creole kids to increase after infection whereas no variation was observed in non-supplemented groups. These results are in agreement with previous studies in Criollo kids from tropical Mexico under natural infection conditions and in sheep artificially infected with H. contortus, T. colubriformis or Teladorsagia circumcincta (Datta et al., 1998; Valderrabano et al., 2002; Vanhoutert

et al., 1995b). Thus, these results suggest that the immune response against GIN infection of supplemented animals was enhanced compared to that of lambs kept on a restricted diet and that eosinophils may play a role in this mechanism. Regardless of the experimental diet, no significant difference was found between resistant and susceptible kids in the levels of IgA anti-L3 and IgA anti-ESP (IgA response) throughout the study. In contrast, a significant higher level of the IgA response was found in non-supplemented animals compared to supplemented ones. These results are not consistent with a previous study in sheep which suggested that the plane of nutrition may be positively correlated with the antibody response against GIN (Martinez-Valladares et al., 2005). Our data would suggest that the IgA response is better correlated with the nematode burden, as reflected by the FEC and the PCV. 5. Conclusion Numerous studies have already reported the benefit of supplementation on resistance and resilience in small ruminants to parasitic infection (Jackson and Miller, 2006; Knox et al., 2006). The present study establishes that about 75 g of the commercial concentrate per day prevented the deleterious effect of H. contortus infection in growing Creole kids when considering resistance parameters. This effect must be confirmed on growth parameters and on trickle infections. Acknowledgements This study was supported by ‘La Région Guadeloupe’ and the European Community (FEOGA). The authors thank L. Abinne-Molza and H. Varo for their technical assistance in parasitological measurements in the laboratory. They are also 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 fellowship from Le Conseil Régional de la Guadeloupe. References Archimede, H., Aumont, G., Saminadin, G., Depres, E., Despois, P., Xande, A., 1999. Effects of urea and saccharose on intake and digestion of a Digitaria decumbens hay by black belly sheep. Animal Science 69, 403–410. Baker, R.L., Gray, G.D., 2003. Worm control for small ruminants in tropical Asia. Australian Centre for International Agricultural Research (ACIAR). Monograph 113, 63–95. Bambou, J.C., Arquet, R., Archimede, H., Alexandre, G., Mandonnet, N., Gonzalez-Garcia, E., 2009. Intake and digestibility of naive kids differing in genetic resistance and experimentally parasitized (indoors) with Haemonchus contortus in two successive challenges. Journal of Animal Science 87, 2367–2375. Bambou, J.C., de la Chevrotiere, C., Varo, H., Arquet, R., Kooyman, F.N.J., Mandonnet, N., 2008. Serum antibody responses in Creole kids experimentally infected with Haemonchus contortus. Veterinary Parasitology 158, 311–318. Blackburn, H.D., Rocha, J.L., Figueiredo, E.P., Berne, M.E., Vieira, L.S., Cavalcante, A.R., Rosa, J.S., 1991. Interaction of parasitism and nutrition and their effects on production and clinical-parameters in goats. Veterinary Parasitology 40, 99–112. Bricarello, P.A., Arnarante, A.F.T., Rocha, R.A., Cabral, S.L., Huntley, J.F., Houdijk, J.G.M., Abdalla, A.L., Gennari, S.M., 2005. Influence of dietary protein supply on resistance to experimental infections with

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