Effects of dry period length and dietary energy source on inflammatory biomarkers and oxidative stress in dairy cows.

Negative energy balance in dairy cows in early lactation has been associated with increased inflammation and oxidative stress in these cows. The objective of this study was to evaluate the effects of dry period (DP) length and dietary energy source on inflammatory biomarkers and oxidative stress in dairy cows. Holstein-Friesian dairy cows (60 primiparous and 107 multiparous) were assigned randomly to a 3 × 2 factorial design with 3 DP length (0, 30, or 60 d) and 2 early lactation rations (glucogenic or lipogenic). Cows were fed a glucogenic or lipogenic ration from 10 d before the expected calving date. Blood was collected in wk -3, -2, -1, 1, 2, and 4 relative to calving. Dry period length affected inflammatory biomarkers and oxidative stress, especially in wk 1 and 2 after calving. Cows with a 0-d DP had higher levels of ceruloplasmin, cholesterol, and reactive oxygen metabolites, and they tended to have higher haptoglobin levels compared with cows with a 30- or 60-d DP. Cows with a 0-d DP had a lower plasma paraoxonase and bilirubin in the first 2 wk after calving and a lower liver functionality index compared with cows with a 60-d DP. Cows of parity >3 fed a glucogenic ration had higher cholesterol levels compared with cows of parity >3 fed a lipogenic ration. No interaction between DP length and ration was present for inflammatory biomarkers or oxidative stress variables. Plasma bilirubin levels for cows with a 0-d DP were negatively related to energy balance and metabolic status in these cows. Moreover, occurrence of clinical health problems (fever, mastitis, metritis, and retained placenta) was 41, 27, and 30% for cows with 0-, 30-, and 60-d DP, respectively. High levels of ceruloplasmin, cholesterol, and reactive oxygen metabolites in cows with 0-d DP were related to the occurrence of health problems in these cows. In conclusion, omitting the DP increased levels of ceruloplasmin, cholesterol, and reactive oxygen metabolites, and decreased levels of bilirubin and paraoxonase in plasma, independent of ration, compared with cows with a 60-d DP. These contrasting effects of DP length on inflammatory status could be explained in part by the improved energy balance and occurrence of health problems in these cows, but was not related to increased somatic cell count in cows with a 0-d DP. Cows with a 0-d DP had better energy balance, but also had higher levels of oxidative stress compared with cows with a 60-d DP. Moreover, occurrence of health problems did not differ between cows with different DP lengths.

Genetic parameters of IgM and IgG antibodies binding autoantigens in healthy chickens.

Levels of natural antibodies (NAb) binding keyhole limpet hemocyanin (KLH) in layers were shown to be heritable and to be potential indicative parameters for survival. A proportion of NAb are directed to self-molecules, or slightly changed self-molecules (neo-epitopes), labeled as natural autoantibodies (NAAb). It is unknown whether the levels of NAAb are heritable and genetically correlated. In this paper, we estimated genetic parameters in plasma of healthy layers for IgM and IgG antibodies binding ovalbumin (OVA), myosin (MYO), cardiolipin (CAR), lysozyme (LYS), and the model antigen keyhole limpet hemocyanin (KLH). A linear animal model was used to estimate (co)variance components, heritabilities, and correlations. The estimates of heritabilities ranged from 0.10 to 0.17 for IgM, and 0.02 to 0.11 for IgG, respectively. For both IgM and IgG, high genetic correlations were observed between levels of NAAb binding autoantigens and NAb binding KLH, except for IgG binding KLH and LYS, for which a low genetic correlation was found. Low to moderate phenotypic correlations were found between NAAb and NAb. In addition, significant maternal environmental effects of 0.03, 0.07, and 0.04 were observed for IgM binding OVA, LYS, and KLH, respectively. Results from this study indicated that NAAb or NAb levels in plasma were heritable and could provide tools to identify the health status of birds.

Phenotypic and genetic relationships of bovine natural antibodies binding keyhole limpet hemocyanin in plasma and milk.

To improve the health status (resilience) of dairy cows, levels of natural antibodies (NAb) might be useful. The objective of the present study was to compare levels and to estimate genetic parameters for NAb measured in milk and plasma samples. Titers of NAb IgM and IgG isotype-binding keyhole limpet hemocyanin of 2,919 cows, in both plasma and milk, were measured using ELISA. Analysis revealed that NAb levels in milk significantly increased with parity, whereas they remained constant in plasma. Moderate positive phenotypic correlations were found between NAb levels in milk and in plasma: 0.18 for IgG and 0.40 for IgM. This indicates that NAb from milk and plasma might reflect different aspects of dairy cow health status. However, high genetic correlations were found for NAb in milk and plasma: 0.81 for IgG and 0.79 for IgM. Heritabilities (SE in parentheses) for NAb measured in plasma [0.15 (0.05) for IgG and 0.25 (0.06) for IgM] were higher than heritabilities of NAb measured in milk [0.08 (0.03) for IgG and 0.23 (0.05) for IgM]. Our results indicate that NAb measured in milk and plasma are heritable and likely have a common genetic background, suggesting that NAb levels measured in milk might be useful for genetic improvement of disease resistance.

Effect of maternal dry period length on colostrum immunoglobulin content and on natural and specific antibody titers in calves.

The objective was to study the effect of dry period length in dairy cows on immunoglobulin content and natural antibodies (NAb) titers in colostrum, growth, and plasma natural and specific antibody titers in plasma of calves. Holstein-Friesian dairy cows (n=167) were randomly assigned to 3 dry period lengths (0, 30, or 60 d). Colostrum production, concentration of colostrum IgG and IgM, and titers of NAb (isotypes IgG and IgM) binding keyhole limpet hemocyanin (KLH) and human serum albumin (HuSA) in colostrum were measured. Female calves were immunized with both KLH and HuSA at wk 6 and 10 of life. Titers of NAb and specific antibody (SpAb) for isotypes IgG, IgM, and total immunoglobulin (IgT) binding KLH or HuSA were determined in plasma of female calves. Primary and secondary antibody responses to KLH or HuSA from wk 6 and 10 were expressed as the increase in antibody titers to wk 10 and 11 of life after primary and secondary challenges, respectively. Pregnancy length for cows with a 0-d dry period was 3d shorter compared with cows with a 30- or 60-d dry period. Birth weight of calves from cows with a 0-d dry period was lower compared with calves from cows with a 30-d dry period. Growth of calves until 12 wk of life was not affected by dry period length. Colostrum production and IgG and IgM concentration in colostrum were lower for cows with a 0-d dry period than a 60-d dry period. Natural IgG and IgM titers binding KLH or HuSA were lower in colostrum from cows with a 0-d dry period compared with cows with a 60-d dry period. Natural antibody titers (IgG, IgM, and IgT) binding KLH or HuSA in plasma were lower during the first 2 wk of life for calves from cows with a 0-d dry period compared with calves from cows with a 30- or 60-d dry period. After primary and secondary immunization of calves with KLH and HuSA, SpAb titers of calves were not affected by dry period length. After secondary immunization, the response of IgG and IgT binding KLH was higher in plasma of calves from cows with a 0-d dry period. The results of this study demonstrate that, although omission of the dry period of dairy cows leads to lower plasma NAb titers in calves during the first 2 wk of life, SpAb titers in calves were not affected and even the secondary antibody responses were enhanced compared with calves from cows with a 30- or 60-d dry period.

High natural antibody titers of indigenous chickens are related with increased hazard in confinement.

Natural antibody (NAb) levels and survival rates were evaluated in 4 breeds of laying hens in Ethiopia: indigenous, improved indigenous, exotic layer, and crossbred. Titers of NAb isotypes IgG and IgM binding keyhole limpet hemocyanin (KLH) in serum were measured at 20, 26, 35, and 45 wk age. Repeated-measure ANOVA showed that IgG and IgM levels vary with time within each breed (P < 0.05). Indigenous chickens had significantly (P < 0.05) higher NAb levels at all ages. The Cox proportional hazard analysis showed increased hazard with increased levels of NAbs in the exotic layers (P < 0.05). However, the reduced hazards with increased levels of NAbs were not significant in the improved indigenous and crossbred chickens. Indigenous chickens showed increased hazard with increasing levels of NAb (P > 0.05). We concluded that not only the NAb levels but also the effect of Nabs on survival vary between indigenous and improved breeds. The results indicate that NAb levels are associated with survival in elite (improved) breeds, but are associated with increased hazard in indigenous chickens.

Genetic relations between natural antibodies binding keyhole limpet hemocyanin and production traits in a purebred layer chicken line.

Natural antibodies (NAb) are an important component of the first line of immune defense. Selective breeding for enhanced NAb levels in chickens may improve general disease resistance. It is unknown what the consequences of selection for NAb will be on the productive performance of laying hens. In this paper we describe the genetic relations between NAb titers binding keyhole limpet hemocyanin at 19 wk age and production traits in a white purebred leghorn chicken line observed in several time periods. A linear animal model was used to estimate (co)variance components, heritabilities, and correlations. Negative genetic correlations were found between egg weight and NAb titers, and between egg breaking strength and NAb titers. Positive genetic correlations were found between the feed conversion ratio (consumed feed/egg mass produced) and NAb titers, and egg production and NAb titers. Negative phenotypic correlations were found between body weight and NAb titers, between egg weight and NAb titers, and between egg breaking strength and NAb titers. Positive phenotypic correlations were found between egg production and NAb titers, and feed conversion ratio and NAb titers. In general, phenotypic correlations were more often significant, but less pronounced than genetic correlations. Other production traits were not found to be significant related to NAb titers. These findings suggest that there is a genetic tradeoff between levels of immunity and some production traits, although the underlying mechanism(s) remain(s) unclear. The results suggest possible consequences for production efficiency as a result of selective breeding for improved general disease resistance by natural antibodies.

Modelling of feather pecking behavior in beak-trimmed and non-beak-trimmed crossbred laying hens: variance component and trait-based approach.

Because of a ban on the use of beak trimming in some European countries, feather pecking is becoming a substantial problem in the layer industry, both from animal welfare and economic points of view. The feather condition score (FCS) as a measure of feather damage has been shown to be closely related to feather pecking behavior in laying hens housed in groups. To obtain a better understanding of genetic and other biological mechanisms underlying feather pecking behavior, data on FCS of a population of 2,724 female offspring from crossing 50 male W1 and 907 female WB purebred lines were used. The offspring of 25 sires were beak-trimmed, and the offspring of another 25 sires were non-beak-trimmed. Titers of plasma natural antibody (NAb) isotypes IgM and IgG binding keyhole limpet hemocyanin at 24 wk of age were measured. Feather condition was scored at 53 wk of age. In the first part of the present study, we estimated genetic parameters for FCS with 2 variance components models: a traditional linear animal model and a model combining direct and associative genetic effects. In the second part of the present study, a trait-based analysis for FCS was conducted to investigate whether NAb isotype titers can explain variation in FCS among individuals, by fitting a linear mixed model. Though the estimated associative genetic variance was substantial, associative effects for FCS were not statistically significant in both populations (P = 0.09 in beak-trimmed birds, and P = 0.08 in non-beak-trimmed birds). This suggests an insufficient number of records on FCS. Individual's NAb isotypes titers did not show direct effect for FCS of itself, but individual's IgG titers showed a suggestive effect on the FCS of cage mates (associative effect) in beak-trimmed laying hens, which need further confirmation.

Genetic parameters and across-line SNP associations differ for natural antibody isotypes IgM and IgG in laying hens.

In an earlier study, serum levels of natural antibody isotypes IgM- and IgG-binding keyhole limpet hemocyanin were found to be indicative for survival through the laying period of hens and therefore considered as promising traits for future implementation in breeding programs for higher survival of layers. In the present study, we first estimated the genetic parameters for the two isotypes at 20, 40 and 65 weeks of age (IgM20, IgM40 and IgM65; IgG20, IgG40 and IgG65). Pooled genetic parameters were estimated from the total population of 2504 hens from nine purebred layer lines, with line included in the model to account for admixture. Moderate heritabilities (0.14-0.44) indicated that selection for isotype titers is feasible, especially for IgM. Secondly, associations between 1022 SNP markers and the above-mentioned six immunological traits were estimated in 650 genotyped hens from the nine lines. The association study was performed across lines to detect markers that are closer to the QTL and have the same phase of association in the entire population. Forty-three significant associations between SNPs and isotype titers were detected. The SNPs of interleukins IL10 and IL19 were associated with both isotypes; SNPs of tripartite motif containing 33 (TRIM33) and IL6 showed significant association with IgG20 and IgM20 respectively; SNPs of heat shock protein 90kDa alpha (cytosolic), class B member 1 (HSP90AB1) were associated with IgG titers at older ages. Some detected SNPs were also reported associated with other immune and behavioral traits.

Transgenerational epigenetic effects on innate immunity in broilers: an underestimated field to be explored?

Transgenerational epigenetics is becoming more and more important for understanding the variation of physiological responses of individuals to the environment and the inheritance of these responses based on all mechanisms other than the actual DNA nucleotide sequence. Transgenerational epigenetics is the phenomenon that the information of the environment of (usually) a female animal is translated into memory-like responses preparing the offspring. As a consequence, individuals of the next generation may show different phenotypic traits depending whether their mothers were kept under different environmental conditions. This may result in either positive or negative effects on the next-generation individuals, which is different from individuals from mothers that have been kept in a different environment. Transgenerational epigenetic effects have been proposed and indicated for specific immune (T cell and antibody) responses (especially in mammals, but also in birds) and innate immunity (nonvertebrates), but surprisingly very little is known of transgenerational effects on innate immunity in chickens. Given the short lifespan of the chicken and therefore the likely dependence of chicken on innate immune mechanisms, more attention should be given to this arm of immunity and mechanisms of inheritance including transgenerational effects that can be initiated in the breeder generation. In addition, it is becoming evident that innate immunity also underlies metabolic disorders in broilers. In the current paper, we will argue that although very little is known of transgenerational effects of innate immunity in poultry, more attention should be given to this type of study. We will illustrate examples of transgenerational epigenetics, and finally propose strategies that should reveal the presence of transgenerational epigenetic effects on innate immunity in chickens and strategies to modulate breeder birds such that these effects positively affect innate immunity of broilers. It is suggested that a mismatch between breeder environment and broiler environment may account for unwanted effects of innate immunity in the broiler.

Genetic parameters of natural antibody isotypes and survival analysis in beak-trimmed and non-beak-trimmed crossbred laying hens.

Natural antibodies (NAb) are important humoral components of innate immunity. As the first line of defense, NAb provide protection against infection and support adaptive immunity. An earlier study indicated that serum levels of NAb isotypes IgM and IgG at a young age were predictive for survival in non-beak-trimmed purebred laying hens during the laying period. In the present study, genetic parameters of NAb isotypes were estimated and relationships between survival and NAb isotypes levels in crossbred laying hens were investigated. In total, 1,555 beak-trimmed and 1,169 non-beak-trimmed crossbred laying hens were used. Genetic parameters of IgM and IgG titers binding keyhole limpet hemocyanin at 24 wk of age were estimated with a linear animal model. The heritabilities of NAb isotypes IgG and IgM were 0.21 (SE = 0.04) and 0.26 (SE = 0.04), respectively. The genetic correlation between IgG and IgM isotypes was 0.43 (SE = 0.11). These results indicated that NAb isotype titers were heritable traits in the crossbred laying hens. Both NAb isotypes can be selected for simultaneously because the detected positive genetic correlation (0.43, SE = 0.11) between them is positive. Both row and level of the cage were indicated to be associated environmental factors for NAb isotype titers. Different from an earlier study with purebred hens, survival analysis showed no significant associations of survival with NAb isotype titers in beak-trimmed or non-beak-trimmed crossbred hens. Non-health-related causes of mortality, especially in birds with intact beaks, overruled the anticipated relationships between NAb isotype titers and survival.

Localization and (semi-)quantification of fluorescent beads of 2 sizes in chickens over time after simultaneous intratracheal and cloacal administration.

Environmental particles enter the chicken via several routes. Entry via the respiratory and cloacal routes likely activates immune responses. We studied the localization of simultaneous intratracheally and cloacally applied beads of 2 sizes in the chicken body in time, and when possible, semiquantified the amount of beads. Ten broiler hens, 3.5 wk of age, received 1.25 × 10(9) 1.0-µm beads and 1.05 × 10(7) 10-µm fluorescein isothiocyanate (green) labeled cloacally, and simultaneously the same number and same sizes of tetramethylrhodamine isothiocyanate (red) labeled beads intratracheally. The bursa of Fabricius, lung, liver, kidney, gallbladder, spleen, thymus, small intestine (upper ileum), cecum, intestinal luminal contents, aerated bones, feces, and blood, from 2 chickens per moment were sampled at 1 h, 6 h, 24 h, 48 h, and 1 wk after challenge and studied for the presence of beads using fluorescence microscopy. The highest amount of beads was found in organs closest to the application site after 1 h (i.e., the lungs for red beads, and the bursa for green beads). All tissue samples showed all 4 types of beads at all time moments, most of them within 1 h. Lower levels of beads were found in lungs and bursa after 6 h and in all other organs after 24 h, except for the kidneys where levels declined after 48 h. Surprisingly, beads were found in thymus tissue and only relatively few beads were found in the spleen. At 1 h, 1-µm intratracheally applied red beads were also found in the cecal luminal content and cecal tissue, but not in the small intestinal luminal content, suggesting that ceca are capable of excreting small particles entering the body via the respiratory route. The presence of nondegradable and nonimmunogenic beads of different sizes in all sampled organs throughout the whole chicken body for 7 d suggested potentially negative chronic health and welfare risks for the chicken of environmental particles.

Effects of 2 size classes of intratracheally administered airborne dust particles on primary and secondary specific antibody responses and body weight gain of broilers: a pilot study on the effects of naturally occurring dust.

We studied the effects of a concurrent challenge on slow-growing broilers with 1) airborne particles of 2 sizes: fine dust (smaller than 2.5 microns) and coarse dust (between 2.5 and 10 microns) that were directly collected from a broiler house and 2) lipopolysaccharide on intratracheal immunizations with the specific antigen human serum albumin (HuSA) and measured primary and secondary systemic (total) antibody responses and (isotype-specific) IgM, IgG, and IgA responses at 3 and 7 wk of age. All treatments affected immune responses at several ages, heart morphology, and BW gain, albeit the latter only temporarily. Dust particles significantly decreased primary antibody (IgT and IgG) responses to HuSA at 3 wk of age but enhanced IgM responses to HuSA at 7 wk of age. Dust particles decreased secondary antibody responses to HuSA, albeit not significantly. All of the birds that were challenged with dust particles showed decreased BW gain after the primary but not after the secondary challenge. Relative heart weight was significantly decreased in birds challenged with coarse dust, fine dust, lipopolysaccharide, and HuSA at 3 wk of age, but not in birds challenged at 7 wk of age. Morphology (weight, width, and length) of hearts were also affected by the dust challenge at 3 wk of age. The present results indicate that airborne dust particles obtained from a broiler house when intratracheally administered at an early age affect specific humoral immune responsiveness and BW gain of broilers to simultaneously administered antigens differently than when administered at a later age. The hygienic status of broiler houses at a young age may be of importance for growth and immune responsiveness, and consequently, for vaccine efficacy and disease resistance in broilers. The consequences of our findings are discussed.

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 repeated intratracheally administered lipopolysaccharide on primary and secondary specific antibody responses and on body weight gain of broilers.

Earlier, we reported that pathogen-associated molecular patterns such as lipopolysaccharide (LPS), when administered intratracheally (i.t.), affected primary and secondary specific antibody responses to antigens administered concurrently, either i.t. or systemically, and also affected BW gain (BWG) of layers and broilers. In the present study, we evaluated the effects of repeated i.t. challenge with LPS concurrently with or before i.t. immunizations with the specific antigens human serum albumin (HuSA) and rabbit gamma globulin (RGG) on primary (HuSA, RGG) and secondary (HuSA) systemic antibody responses and (isotype) IgM and IgG responses at 2 different ages. Broilers were challenged via the trachea at 3 and 7 wk of age with various combinations of LPS, HuSA, and RGG. All treatments affected immune responses at several time points and also affected BWG, albeit temporarily for the latter. Lipopolysaccharide enhanced primary antibody responses to HuSA and to RGG, when challenged concurrently, but birds challenged solely with LPS at 3 wk of age also showed enhanced primary antibody responses to HuSA and RGG given at 7 wk of age. This was true for IgM as well as IgG isotype responses. Lipopolysaccharide challenge negatively affected BWG at 3 wk of age, whereas the negative effects of LPS after a secondary LPS challenge at 7 wk of age were most pronounced in the birds challenged with LPS at 3 wk of age. The present results indicated that LPS, when administered i.t. at a young age, may affect specific humoral immune responsiveness to antigens administered simultaneously and to BWG of broilers, but also when challenged 4 wk later with specific antigens, suggesting an enhanced status of immune reactivity or sensitivity. The hygienic status of broiler houses at a young age may thus influence BWG, immune responsiveness, and, consequently, the vaccine efficacy and disease resistance in broilers at later ages. The consequences of our findings are discussed.

Effects of early nutrition and sanitary conditions on antibody levels in early and later life of broiler chickens.

Immune maturation of broiler chickens may be affected by management, such as early life feeding strategy (early versus delayed nutrition) or by low or high sanitary conditions (LSC versus HSC). We compared systemic maternal (MAb), natural (NAb), natural auto- (NAAb), and antigen specific antibody (SpAb) levels (IgM, IgY) between broilers (n = 48 per treatment) that received early (EN) or delayed nutrition for 72 h (DN) housed in either low (LSC) or high sanitary conditions (HSC) between 7 and 35 d of age. We found minimal interactions between feeding strategy and sanitary conditions. At 7 d of age, broilers receiving EN compared with DN, had elevated levels of IgM binding keyhole limpet hemocyanin (KLH), phosphoryl-conjugated ovalbumin (PC-OVA), and muramyl dipeptide (MDP), whereas effects of feeding strategy diminished at later ages. In LSC compared with HSC broilers, levels of NAb agglutinating RRBC and sheep red blood cells (SRBC) were already elevated from 14 d of age onwards. At 33 d of age, antibody levels (NAb, NAAb, anti-LPS, anti-MDP) were all elevated in LSC, compared with HSC broilers, for both IgM and IgY, but not IgM against KLH. Western blotting revealed different binding patterns of NAAb against chicken liver homogenate, which may indicate that the NAAb repertoire is affected by antigenic pressure. Our data suggest that antibody levels are affected for an important part by environmental conditions (feeding strategy and sanitary conditions), but minimally by their interaction. However, it remains to be further studied whether the enhanced levels of antibodies as initiated by EN and LSC contribute to enhanced resistance to infectious diseases.

Adhesion of human B cells to follicular dendritic cells involves both the lymphocyte function-associated antigen 1/intercellular adhesion molecule 1 and very late antigen 4/vascular cell adhesion molecule 1 pathways.

Presentation of antigen in the form of immune complexes to B lymphocytes by follicular dendritic cells (FDC) is considered to be a central step in the generation of memory B cells. During this process, which takes place in the microenvironment of the germinal center, B cells and FDC are in close physical contact. In the present study, we have explored the molecular basis of FDC-B cell interaction by using FDC and B cells derived from human tonsils. We found that FDC express high levels of the adhesion receptors intercellular adhesion molecule 1 (ICAM-1 [CD54]) and vascular cell adhesion molecule 1 (VCAM-1), while the B lymphocytes express lymphocyte function-associated antigen 1 (LFA-1 [CD11a/18]), very late antigen 4 (VLA-4 [CD49d], and CD44. Furthermore, we established that both the LFA-1/ICAM-1 and VLA-4/VCAM-1 adhesion pathways are involved in FDC-B lymphocyte binding, and therefore, these pathways might be essential in affinity selection of B cells and in the formation of B memory cells.

Across-line SNP association study of innate and adaptive immune response in laying hens.

The aim of the present study was to detect quantitative trait loci (QTL) for innate and adaptive immunity in laying hens. For this purpose, the associations between 1022 single nucleotide polymorphism (SNP) markers and immune traits were studied in 583 hens from nine different layer lines. Immune traits were natural antibodies for keyhole limpet haemocyanin (KLH) and lipopolysaccharide (LPS) at 20, 40 and 65 weeks, acquired antibodies to the vaccinal virus of Newcastle disease at 20 weeks, and complement activity measured on sheep and bovine red blood cells at 20, 40 and 65 weeks. We adopted a novel approach based on across-line analysis and testing of the SNP-by-line interaction. Among lines, linkage disequilibrium is conserved at shorter distances than in individual lines; therefore, SNPs significantly associated with immune traits across lines are expected to be near the functional mutations. In the analysis, the SNPs that had a significant across-line effect but did not show significant SNP-by-line interaction were identified to test whether the association was consistent in the individual lines. Ultimately, 59 significant associations between SNPs and immune traits were detected. Our results confirmed some previously identified QTL and identified new QTL potentially involved in the immune function. We found evidence for a role of IL17A (chromosome 3) in natural and acquired antibody titres and in the classical and alternative pathways of complement activation. The major histocompatibility genes on chromosome 16 showed significant association with natural and acquired antibody titres and classical complement activity. The IL12B gene on chromosome 13 was associated with natural antibody titres.

Genetic variation of natural antibodies in milk of Dutch Holstein-Friesian cows.

Defense mechanisms of dairy cows against diseases partly rest on their naturally present disease resistance capacity. Natural antibodies (NAb) form a soluble part of the innate immune system, being defined as antibodies circulating in animals without prior intentional antigenic stimulation. Genetic selection on NAb titers in milk, therefore, might improve disease resistance. We estimated genetic parameters of NAb titers binding lipopolysaccharide, lipoteichoic acid (LTA), peptidoglycan, and keyhole limpet hemocyanin, and titers of the NAb isotypes IgG1, IgM, and IgA binding LTA in milk of Dutch Holstein-Friesian heifers. Natural antibody titers were measured in 1 milk sample from each of 1,939 Holstein-Friesian heifers and used for estimating genetic parameters of NAb titers. The data show that phenotypic variation exists among heifers in NAb titers binding lipopolysaccharide, LTA, peptidoglycan, and keyhole limpet hemocyanin, and the NAb isotypes IgG1, IgM, and IgA binding LTA in milk. High genetic correlations among NAb (ranging from 0.45 to 0.99) indicated a common genetic basis for the levels of different NAb in bovine milk. Intra-herd heritability estimates for NAb ranged from 0.10 to 0.53. The results indicated that NAb levels have potential for genetic selection.

Effect of aging and repeated intratracheal challenge on levels of cryptic and overt natural antibodies in poultry.

Natural antibodies (NAb) have been divided in 2 classes: overt and cryptic. Overt NAb can be detected in unfractionated normal sera of nonimmunized mammals and chickens. Cryptic NAb as described in mammals need an in vitro physical or biochemical treatment to be detected, which may reflect their biochemical modification in situ during inflammation or infection. We studied the effect of concurrent primary, secondary, and tertiary intratracheal (i.t.) challenges with lipopolysaccharide (LPS) and human serum albumin on levels of 2 cryptic NAb [i.e., NAb binding actin (ACT), or thyroglobulin (THYRO)] and levels of an overt NAb binding keyhole limpet hemocyanin (KLH). In addition, effect of aging of the birds on levels of NAb was taken into account. Presence and changes on the level of the cryptic antibodies directed to ACT and THYRO were expected after the i.t. challenges with LPS (and human serum albumin). However, levels of NAb binding ACT were only significantly enhanced by LPS after primary challenge, whereas levels of NAb binding THYRO were not affected by primary nor secondary challenges but were significantly decreased by LPS after the third immunization. On the other hand, no changes in the levels of overt NAb binding KLH were expected, but levels of NAb binding KLH were significantly enhanced after the primary and secondary challenges with LPS. Levels of all three NAb increased with aging, but the different challenges performed at 3 moments during aging did not significantly or consistently affect levels of the overt nor cryptic NAb. Our results suggest that chickens might react by overt NAb as well as cryptic NAb to an infection or inflammation rather than by cryptic NAb only. The relation between various types of NAb and i.t. immunization with antigen and LPS and aging 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.