H.K. Parmentier

Effects of housing system (outdoor vs cages) and age of laying hens on egg characteristics

1. Effects of two housing systems (cages vs outdoor) on external and internal egg characteristics were investigated. 2. In total 785 eggs from three different lines in cages and 268 eggs from outdoor-housed layers were examined for egg weight, albumen, yolk and shell content, albumen height and pH, and albumen and yolk dry matter content. 3. Interactions between layer age and housing systems were found for egg weight, eggshell content, albumen height, albumen pH, and dry matter content of the albumen and yolk. This was mainly due to the greater variation with age in the outdoor layers, compared to the caged layers. 4. Irrespective of age eggs from outdoor layers were relatively broader than eggs from the caged layers. Yolk colour was considerably darker in the outdoor group (11.0 vs 9.3). 5. We concluded that it is more difficult to maintain constant external and internal egg quality in an outdoor housing system than in a battery cage system. Factors that determine the greater fluctuations in internal egg quality need to be investigated.

Divergent Antibody Responses to Vaccines and Divergent Body Weights of Chicken Lines Selected for High and Low Humoral Responsiveness to Sheep Red Blood Cells

Primary and secondary antibody responses to intramuscularly administered proteins of Eschericia coli (F11), Newcastle disease virus (NCD), infectious bronchitis virus (IB), and infectious bursal disease virus (IBD), respectively, were measured at weekly intervals in two chicken lines. The latter had been divergently selected for high and low antibody responses to sheep red blood cells (SRBC), and in a random-bred control line. An oil-based adjuvant was required to induce primary and secondary antibody responses to NCD, IB, and IBD. With respect to F11, elevated antibody responses were found in birds sensitized and boosted to F11 with and without adjuvant. The humoral response to F11 and to all viral antigens was significantly higher in the high (H) line than in the low (L) line, whereas the control (C) line showed intermediate titers. At 5 and 17 weeks of age, L line birds were significantly heavier than birds of the H and the C lines. A negative phenotypic correlation within lines between body weight at 17 weeks of age and antibody titers at 1 week after sensitization was found, but no further correlations between humoral responses and body weight or growth could be established. The present results suggest that selection for enhanced humoral responsiveness to SRBC resulted in enhanced responsiveness to components of several vaccines. Mechanisms underlying the relationship between divergent selection for immune responsiveness and body weight are discussed.

Kinetic and immunohistochemical characteristics of mitogen-induced cutaneous hypersensitivity in chickens selected for antibody responsiveness

Mitogen-induced cutaneous hypersensitivity was evaluated in chickens selected for high and low antibody responses to SRBC, and in a random bred control line. Wing web swelling responses were found after subcutaneous administration of phytohemagglutinin (PHA), concanavalin A (Con A), pokeweed mitogen (PWM), and Escherichia coli lipopolysaccharide (LPS), respectively, in all three lines. All mitogens induced significant acute 4 h wing web swelling responses, followed by a significant (classical) late 24 h wing web swelling response. The 4 h responses were significantly lower in the L line, whereas a tendency for lower responses at 24 h in the L line was found as well. Immunohistochemical evaluation of the early and late wing web swelling responses revealed extravascular localisation of leukocytes at 24 h after sensitization with mitogens, which consisted of CD4+ cells, CD8+ cells, TCR-1+ cells, and heterophils, but no B cells, whereas the 4 h swelling response was primarily characterized by oedema. Cutaneous hypersensitivity either initiated by T-cell mitogens as well as B-cell mitogens may depend for an important part on the rapid induction of local homing of lymphocytes towards the sensitizing agent, which may be mediated by an acute local expression of molecules with chemo-attractive capacities. Interpretation of cellular immunity responses in vivo such as delayed-type hypersensitivity should therefore incorporate oedema-initiating characteristics of sensitizing agents. The relationship between the magnitude of cutaneous hypersensitivity to mitogens and selection for antibody responsiveness is discussed.

Genetic parameters for levels of natural antibodies in chicken lines divergently selected for specific antibody response

The present study estimated the heritability of natural antibody (NAb) levels binding rabbit red blood cells at the day of immunization with SRBC (NAb0) and 5 d postimmunization (NAb5) in 2 chicken lines divergently selected for specific antibody levels (SpAb) against SRBC and a randombred control line. In addition, genetic correlations between the levels of NAb binding rabbit red blood cells and levels of SpAb binding SRBC were estimated. The heritability of the SpAb level against SRBC was estimated based on data from 23 generations of selection and in total 21,842 chickens. The heritability of NAb levels against rabbit red blood cells was estimated on 1,764 chickens in generations 22 and 23 using a bivariate analysis including the trait under selection (i.e., SpAb responses to SRBC). Maternal environmental effects were accounted for in the analyses. The heritability for SpAb was 0.17, the heritability for NAb0 levels was 0.23, and the heritability for NAb5 was 0.09. The genetic correlation between SpAb and NAb0 was 0.15 and the genetic correlation between SpAb and NAb5 was 0.50. The weak, though positive, genetic correlation between SpAb responses and the NAb0 levels in nonimmunized chickens, as well as the moderate genetic correlation between SpAb responses and the NAb levels 5 d postimmunization with SRBC, suggest that selection for enhanced adaptive immune system results in a positive correlated response of the innate immunity of chickens.

Effect of Single or Combined Climatic and Hygienic Stress in Four Layer Lines: 1. Performance

Effects of long-term climatic stress (heat exposure), short-term hygienic stress [lipopolysaccharide (LPS)], or a combination of both challenges on performance of 4 layer lines were investigated. The lines were earlier characterized by natural humoral immune competence and survival rate. At 22 wk of age, 80 hens per line were randomly divided over 2 identical climate chambers and exposed to a constant high temperature (32 degrees C) or a control temperature (21 degrees C) for 23 d. Half of the hens housed in each chamber were i.v. injected with LPS at d 1 after the start of the heat stress period. The effect of heat, LPS, or a combined challenge on feed intake, BW, hen-day egg production, egg weight, and egg shell thickness were investigated. Feed intake, BW, hen-day egg production, egg weight, and egg shell thickness were significantly reduced by heat stress. Administration of LPS significantly reduced feed intake, BW (LPS x time interaction), hen-day egg production, and egg weight (LPS x time interaction). Hens were able to recover from LPS administration but did not completely adapt to heat stress. Hens still lost weight, had a lower feed intake and hen-day egg production after 23 d of continuous exposure to heat stress. These data suggest a different nature of short-term LPS exposure versus long-term heat exposure affecting performance parameters of laying hens, and different adaptation mechanisms of hens toward these stressors. Neither natural humoral immune competence nor survival rate, for which the lines had been earlier characterized, were indicative of the response to different stressors. However, significant line x heat interactions were found for feed intake and hen-day egg production, and a line x heat x time interaction for BW, whereas a line x LPS interaction was found for hen-day egg production and a line x LPS x time interaction for BW. The lines had similar response patterns, but differed in response levels, suggesting that some lines were better able to adapt to stressors than others.

Natural antibody isotypes as predictors of survival in laying hens

To identify possible relationships between survival and titers of natural antibody (NAb) isotypes in serum of laying hens, birds from 12 purebred layer lines of 2 commercial breeds, Rhode Island Red (n = 524) and White Leghorn (n = 538), were monitored for survival during one laying period (from 20 until 70 wk of age). Titers of NAb isotype IgM- and IgG-binding keyhole limpet hemocyanin (KLH) in serum were measured at 20, 40, and 65 wk of age, respectively. Overall, the titers of IgM and IgG binding KLH decreased with aging. At the same age, lines within breed showed significantly different titers of isotypes (P < 0.0001). Multivariable logistic regression analysis showed that NAb isotype IgM and IgG titers at 20 wk of age were associated with survival at 20 to 40 wk of age. In the R breed, odds ratios of 0.56 (P < 0.0001) for IgM and 0.72 (P = 0.02) for IgG were estimated; in the W breed, these were 0.74 (P < 0.01) and 0.99 (P = 0.95) for IgM and IgG, respectively. We conclude that titers of Nab isotypes, especially the IgM-binding KLH at 20 wk of age, are indicative for survival during the laying period. The higher the titers of NAb isotypes, the higher the probability of layers to survive.

Effects of dust and airborne dust components on antibody responses, body weight gain, and heart morphology of broilers

Pathogen-associated molecular patterns (PAMP) such as lipopolysaccharide (LPS), lipoteichoic acid, beta-glucans (BGL), and possibly many others are important parts of (fine) dust in animal houses. When intratracheally (i.t.) administered, PAMP affected specific primary and secondary humoral immune responses to concurrently i.t. or systemically administered antigens and BW gain (BWG) of layer chickens. In the present study, we evaluated the effects of i.t. challenge with various PAMP known to be present in dust: LPS, lipoteichoic acid, zymosan-A (containing 1,3 BGL), next to heat-inactivated dust particles as a representative of mechanical stress, a combination of the former components, and NH3 as a chemical component of dust on primary and secondary (total) systemic antibody (Ab) responses and (isotype) IgM and IgG responses to concurrently i.t.-administered human serum albumin (HuSA) in broilers. Birds were challenged via the trachea for 2 consecutive days at 3 and 7 wk of age, respectively. All treatments affected immune responses at several moments, BWG, and heart morphology. beta-Glucans and LPS affected the birds most pronounced and for a prolonged period. Intratracheally administered LPS and BGL significantly enhanced primary and secondary total Ab, IgM Ab, and IgG Ab responses to HuSA. All birds that were challenged with dust, PAMP, or NH3 concurrently with HuSA showed a decreased BWG especially after primary, but also after secondary challenge. Weight, width, and length of hearts were enhanced in dust and PAMP-treated birds as well when these birds were challenged with HuSA. The present results indicated that components of dust such as PAMP when i.t. administered affect humoral immune responsiveness of broilers, which may lead to an enhanced status of immune reactivity. Furthermore, our results suggest that the hygienic status of the environment influences BWG and may affect heart morphology, and as a consequence physiology in broilers. The consequences of our findings with respect to dust, (airborne) PAMP, hygienic conditions in the barn, and immune responsiveness of broilers are discussed.

Immune responses of piglets to Pasteurella multocida toxin and toxoid

Experimental atrophic rhinitis (AR), serum antibody titres and in vitro lymphoproliferation to Pasteurella multocida derived toxin (Pm-T) were studied in piglets. Specific immune responses to Pm-T and Pm-T induced conchae atrophy were compared with AR immunity. This immunity was initiated by the Nobi-VAC AR-T vaccine administered at various times with respect to Pm-T challenge. Animals challenged with Pm-T developed conchae atrophy, but no antibodies nor cellular immune responses to Pm-T were detected. Vaccination 3 weeks before Pm-T challenge protected pigs against breakdown of nasal bony tissues. This protection was accompanied by an increase of serum antibodies and in vitro lymphoproliferation to Pm-T. Animals vaccinated 10 days before or after Pm-T challenge also had antibodies and cellular immune responses. However, these animals developed AR. In vitro, Pm-T appeared mitogenic for quiescent (non-immune) peripheral lymphocytes and Concanavalin A stimulated lymphocytes from some pigs. These in vitro lymphoproliferative responses could be partly abrogated by the addition of monomorphic anti-swine major histocompatibility complex class II DQ and DR specific monoclonal antibodies. We conclude that Pm-T is poorly immunogenic in vivo and does not initiate a protective Pm-T specific immune response. Pigs were protected from AR by vaccination, but protection was dependent on the timing of vaccine administration. We speculate that Pm-T modifies the immune response such that the response is not directed towards the toxin but to an unidentified component in the nose of piglets.

Natural antibodies related to metabolic and mammary health in dairy cows

Natural antibodies (NAb) are defined as antibodies that circulate in normal healthy individuals under the absence of deliberate antigenic stimulation. Two types of NAb are distinguished: NAb towards exogenous antigens and NAb towards autoantigens (N(A)Ab). The objectives of the current study were threefold. First, we studied the relation between metabolic health and concentrations of NAb binding keyhole limpet hemocyanin (KLH) or lipopolysaccharide (LPS) in milk and plasma of dairy cows in early lactation. Second, we determined the presence of N(A)Ab binding transferrin, myosin and thyroglobulin in bovine milk. Third, we studied the relation between N(A)Ab in bovine milk and mammary health. For the first objective, dairy cows were either fed a control (C) (n=8) or a diet where 2 kg of concentrates were replaced by an iso-energetic concentrate containing marine algae (ALG) from week -3 till 8 postpartum (experiment 1). Plasma and milk samples were analyzed weekly for NAb binding either KLH or LPS. Plasma was analyzed for glucose, non-esterified fatty acids (NEFA) and β-hydroxybutyrate (BHBA). For the second and third objective, milk samples were collected weekly from 96 dairy cows from week 2 till 9 postpartum and analyzed for milk composition and N(A)Ab binding myosin, transferrin and thyroglobulin (experiment 2). For both datasets, N(A)Ab titers are expressed as (2)log values of the highest dilution giving a positive reaction. Data are expressed as means ± SEM. Repeated observations were analyzed in a mixed model. In experiment 1, no diet effect (P>0.05) was observed on NAb binding LPS in plasma or milk, NAb binding KLH in milk was greater (P=0.05) for cows fed the control diet. Concentration of NAb binding KLH and LPS in plasma was negatively related to plasma NEFA concentration (P<0.05). In experiment 2, NAb binding myosin (5.66 ± 0.06), thyroglobulin (4.85 ± 0.06), and transferrin (5.76 ± 0.07) were identified in milk. Clinical mastitis incidence (9%) tended to be positively related to concentration of NAb binding myosin (P=0.06) and negatively related to Nab binding transferrin (P=0.08). In conclusion, NAb binding KLH and LPS in plasma and milk are related to metabolic health, as indicated by plasma NEFA concentration. Furthermore, this study demonstrates the presence of N(A)Ab in bovine milk and shows trends for a relation between N(A)Ab binding auto-antigens and mastitis. Future studies should confirm these trends and shed light on the predictive value of N(A)Ab in bovine milk for mammary health.

Does enhancement of specific immune responses predispose laying hens for feather pecking

To mimic airborne immune challenges, layer hens were intratracheally and concurrently challenged with various doses of the protein antigen human serum albumin (HuSA) and the pathogen-associated molecular pattern lipopolysaccharide (LPS) at 7 and 13 wk of age. All groups received 1 similar dose of HuSA plus LPS at 11 mo of age. Evaluation of plumage and body condition at 12 mo of age revealed that birds that had undergone intratracheal immunization with a high dosage of HuSA, irrespective of the concurrent dose of LPS, had significantly more feather damage but less wounds to the vent region, as opposed to birds not receiving HuSA. On the other hand, a high dosage of LPS was related to comb damage. These results suggest that stimulation of specific (humoral) immune responses (to HuSA) rather than innate responses (to LPS) at a young age may predispose layers for feather pecking (FP) behavior at later ages. Involvement of immune mechanisms in FP or vent damage may differ. Predisposal for FP behavior by specific immunity can have consequences for health and vaccine management.