Effect of IL8 haplotype on immunological traits in periparturient dairy cows.

Interleukin 8 (IL8) is a major mediator of the innate immune response. Polymorphisms in this gene are associated with susceptibility to inflammatory disease in humans. Two major promoter polymorphic haplotypes (IL8-h1 and IL8-h2) segregating in cattle populations have shown a significant effect on the immune response profile in calves but their implications for transition cow immunity have not been established. The aims of this study were to assess functional relevance of the IL8 haplotypes on the immunological traits of periparturient cows (n = 32) belonging to three genetic groups: Holstein (HO), Simmental (SI) and their crosses (CR) and to evaluate the frequency of IL8 haplotypes in the HO (dairy) and SI (dual purpose) pure breeds. IL8 haplotypes showed a significant effect on circulating number of both T helper lymphocytes (P = 0.0133) and T cytotoxic lymphocytes (P = 0.0024). Differences in percentage of CD14+ monocytes and T lymphocyte subsets were found between haplotype groups at different time points. Plasma concentrations of Serum Amyloid A (SAA) and Haptoglobin (Hp) were enhanced at calving in IL8-h2 (P = 0.0019, P = 0.0029) and IL8-het (P = 0.050 and P = 0.052) respectively, compared with IL8-h1 cows. In contrast, significantly lower levels of reactive oxygen metabolites (d-ROMs) activation were identified in IL8-h2 and IL8-het cows after calving compared with IL8-h1 cows. Furthermore, genotyping results showed that SI cows have a high frequency of the homozygous IL8-h2 haplotype compared to the HO cows (87.5 % vs 40 %) which reflects the different selective pressure between the two pure breeds. In conclusion, our preliminary data suggests that IL8 promoter haplotype is associated with significant and dynamic changes in immunological traits during peripartum and early lactation period. Future work will focus on a more comprehensive assessment of immune changes in additional cows.

Simmental × Holstein crossbred: comparison of immunological traits with parental breeds during peripartum and early lactation period.

The experiment described in this research communication aimed to compare the immunological traits of Simmental (sire) × Holstein (dam) crossbred cows with the two parental breeds in the peripartum and early lactation period and to estimate the effects of heterosis for these traits. Flow cytometric evaluation of leukocyte subpopulations was assessed in 16 Crossbred (CR), 8 Holstein (HO) and 8 Simmental (SI) cows. Estimated average values of innate and adaptive immune cells showed statistically significant differences between the crossbred cows and parental breeds. Interestingly, the most relevant differences between the three groups related to adaptive immune cells. In particular, the CR cows showed a lower percentage of CD3+ T lymphocytes compared with the SI group (P < 0.0001) and the highest proportions of CD21+ B lymphocytes among the three groups (P < 0.0001). Furthermore, we found the highest positive value of heterosis for the CD21+ B lymphocytes (7.0) and the lowest negative value for CD3+ T lymphocytes (-4.8) in F1 derived population. It seems reasonable to believe that these differences could affect immune function of crossbred cows.

Peripartum issues in the inflammatory arthritis patient: A survey of the RAPPORT registry.

Childbearing women with rheumatoid (RA) and psoriatic arthritis (PsA) have significant peripartum issues. A retrospective anonymous RedCAP survey of peripartum period in females with RA/PsA in the RAPPORT registry was performed. Completed analyses included descriptive statistics, Chi-square and Fisher's exact test. 162 patients (133 RA/29 PsA) completed the survey (103 women having 234 pregnancies), 164 pregnancies occurring before and 70 pregnancies occurring after diagnosis. Pregnancy outcomes from 103 patients included: 96% live births, 1.9% stillbirths, 23% miscarriages, and 15% therapeutic abortions. A third of patients had fewer children than desired due to disease activity, medications and other reasons. For 63 pregnancies after diagnosis: (1) 49% of pregnancies received pre-conception counseling; (2) 65% described good disease control during pregnancy but 74% flared in the first 3 months postpartum; (3) 79% of pregnancies discontinued IA medications; (4) 35% of pregnancies occurred on biologic therapy at or prior to conception. Gestational age at time of delivery was 37-40 weeks in 58% (33/57) post-arthritis vs 66% (83/126) pre-arthritis pregnancies. No statistically significant differences occurred between pregnancies before or after RA/PsA diagnosis for: pregnancy planning, fertility treatment, pregnancy and labour/delivery complications, birth defect frequency or neonatal complications. Neonatal ICU admissions were significantly lower in pre- compared to post-arthritis pregnancies (3.2% vs 14.5%). No pregnancy complications were noted in 24/54 pregnancies on medications compared to 6/9 pregnancies not on medications. The impact of RA/PsA before, during and after pregnancy varied considerably in this cohort emphasizing the importance of informed-decision making at all stages.

Dynamics of pro- and anti-inflammatory cytokine changes in serum and assessment of their diagnostic utility during lactation impairment in pigs.

Postpartum dysgalactia syndrome (PDS) in sows is a frequent and important clinical problem in the field. Currently, the diagnosis is based on physical examination performed during first days after the farrowing. The present study aimed at evaluation the dynamics of pro- and anti-inflammatory cytokine (IL-4, IL-6, IL-8, IL-10 and TNF-α) changes in serum of sows during peripartum period (day - 28 to + 28) and assessment of their diagnostic utility during lactation impairment in pigs. The study was done on 139 sows divided into 3 groups: clinically healthy sows, sows with lactation disorders, sows which had experienced difficult parturitions, lameness, etc. In order to measure the level of serum cytokines, the quantitative species-specific ELISA assays were used. The investigation demonstrated a different kinetics of changes of studied cytokines in sows from various groups. IL-6 and TNF-alfa shown high dynamic changes after farrowing in in sows. The levels of IL-8 and IL-10 were relatively stable in healthy sows, while in sows with peripartum disorders usually increased during lactation. However, the detailed examination revealed that investigated cytokines cannot be a useful early diagnostic markers of lactation impairments in sow. They do not allow to detect with high probability which sows are susceptible to lactation disorders before the parturition.

Transcriptomic analysis of circulating neutrophils in metabolically stressed peripartal grazing dairy cows.

The high metabolic demand during the transition into lactation places cows at greater risk of metabolic and infectious disease than at any other time in their lactation cycle. Additionally, a change occurs in the innate immune response during this period, which contributes to increased risk of disease. In the current study, we compared the transcriptomes of neutrophils from dairy cows divergent in their metabolic health post-calving. Cows (n = 5 per risk group) were selected from a parent experiment (n = 45 cows). Those with high or low concentrations of plasma nonesterified fatty acids, plasma ß-hydroxybutyrate, and liver triacylglycerol in both wk 1 and 2 were deemed to be at "high risk" (HR) or "low risk" (LR) of metabolic dysfunction, respectively. Circulating neutrophils were isolated at 3 time points during the transition period (d 0 and wk 1 and 4 post-calving), and gene expression was analyzed using RNA sequencing. Differential gene expression between the risk groups was determined using edgeR (http://bioconductor.org), and pathway analysis was conducted using Ingenuity Pathway Analysis (Ingenuity Systems, Qiagen, Valencia, CA). Statistical analysis indicated no interaction between risk and week. Therefore, the overall effect of risk was analyzed across all time points. In total, 3,500 genes were differentially expressed between the HR and LR cows (false discovery rate < 0.05). Of these, 2,897 genes were identified by Ingenuity Pathway Analysis and used for pathway analysis. Of the relevant pathways identified, neutrophils isolated from HR cows showed downregulation of genes involved in the recruitment of granulocytes, interferon signaling, and apoptosis, and upregulation of genes involved in cell survival. The results indicate that metabolically stressed cows had reduced neutrophil function during the peripartum period, highlighting a potential relationship between subclinical metabolic disease and innate immune function that suggests that metabolic health negatively affects the innate immune system and may contribute to the state of immunosuppression during the peripartum period. In this way, the metabolic stress among the HR cows may reduce their ability to combat infection during the transition period.

Evaluation of the immune status of peripheral blood monocytes from dairy cows during the periparturition period.

Calving is a critical but stressful event required for milk production in dairy cows. In the present study, we investigated the immune status of peripheral blood mononuclear cells (PBMCs) isolated from periparturient cows to better understand and, thus, possibly prevent stress during the periparturient period. To evaluate the immune response of PBMCs, we assessed their proliferation with or without a mitogen (concanavalin A, ConA). Blood samples were collected 24 h before and after calving and 1 week after calving. The proliferation of non-treated cells remained unchanged throughout the examination period. The immune response of PBMCs isolated from the cows before calving was relatively low, even after ConA stimulation; however, the immune response of PBMCs collected at both time points after calving was significantly higher than those of non-stimulated controls. Next, we examined the expression patterns of T cell related and inflammatory cytokine genes in PBMCs. We found that the mRNA expression levels of both CD4 and CD8 showed decreasing trends after calving. The expression of the Th1 cell marker gene IFNG also decreased after calving. The mRNA expression level of the inflammatory cytokine gene TNFA increased after parturition. Overall, our results suggest that the PBMC immune response was weakened in cows before delivery and part of the expression of the immune cell-related genes in these cells is altered 24 h before and after calving.

Innate immune activation and depressive and anxious symptoms across the peripartum: An exploratory study.

BACKGROUND: There are complex associations between immune function and mental illness, yet studies in the perinatal period focus primarily on individual inflammatory markers and depressive symptoms only, cross-sectionally. We sought to examine associations between both depressive and anxious symptoms and immune activation longitudinally across the peripartum. METHODS: We measured mood (Beck Depression Inventory, BDI-1 A) and anxiety (State-Trait Anxiety Inventory, STATE) and levels of 23 cytokines at 5 points in pregnancy and postpartum in 51 women. Within subject cytokine trajectories over time by depressive and anxious symptom grouping were assessed using linear mixed effects models with random intercept and slope. We also undertook an exploratory cluster analysis based on third trimester cytokine values. RESULTS: Based on categorical BDI scores, IL-6 (p < 0.001), IL-15 (p = 0.047), GCSF (p = 0.003), and CCL3 (p < .001) were significantly different across time, with IL-6 (p < 0.001), IL-15 (p = 0.003), and CCL3 (p < 0.001) higher at the third trimester visit in more depressed subjects. Based on categorical STATE scores, GM-CSF significantly decreased across pregnancy for the less anxious group (p = 0.016), but not for the more anxious, and CCL3 (p = 0.017), CXCL8 (p = 0.011), and IL-6 (p < 0.001) were higher at the third trimester visit for more anxious subjects. In exploratory cluster analysis based on cytokine level, there were no differences in mood or anxiety scores, but significant differences by race/ethnicity and overweight/obesity status. Women with higher pro-inflammatory cytokine values are more likely to be Hispanics (69.2% vs. 21.4%, p = 0.015), but less likely to be African American (23.1% vs. 60.7%, p = 0.015) or overweight/obese (25% vs. 69.2%, p = 0.016) compared to women with lower pro-inflammatory cytokine values. CONCLUSION: We identified a pro-inflammatory burst at the third trimester, indicative of innate immune activation, in women with higher levels of both depressive and anxious symptoms, as well as differences in pro-inflammatory changes across time. We also found significant differences in cytokine levels by race, ethnicity, and overweight/obesity status. These results point the way toward future longitudinal work that considers race/ethnicity, timing, and weight status, and evaluates perinatal mood and anxiety disorders in the context of changing immune functioning across the peripartum.

Distinct neutrophil C5a receptor inflammatory events in cows initiated by chemoattractant C5a and lipopolysaccharide around parturition and in mid lactation.

In neutrophils, toll-like receptor and complement component 5a (C5a) signaling are critical pathways regulating innate immunity. In cows, not much is known about the second C5a receptor, complement component 5a receptor 2 (C5AR2). It is an interesting player in sepsis treatment because it is considered to have an anti-inflammatory effect during normal inflammation. Periparturient cows are prone to severe infections, and the objectives of this study were to investigate the expression and functionality of C5AR2 during peripartum. We investigated the effect of 2 major inflammatory stimuli, C5a and lipopolysaccharide (LPS), on the expression of a selected number of genes (C5AR1, C5AR2, TLR4, ITGAM, COX2, and CXCL8) and functions linked to these receptors. Overall, TLR4, ITGAM, and C5AR2, all of which are involved in early inflammation, showed a lower expression in periparturient cows. However, an overall lower expression seems not to be the only explanation for the increased risk of sepsis in periparturient cows. Normally, in response to inflammation and as seen in the mid-lactation group, the expression of these genes increases after stimulation with LPS. However, in periparturient cows, stimulation with LPS led to a decrease in expression of these receptors, indicating a different response of neutrophils in response to LPS during this period. A decrease in ITGAM (coding for CD11b) expression complicates correct neutrophil localization and phagocytosis. Its downregulation upon stimulation might be detrimental for adequate eradication of the pathogen and might increase the risk of an imbalanced inflammation; C5AR2 seems to play a central role in this altered response. In addition, myeloperoxidase (MPO) activity in periparturient cows is lower in response to C5a stimulation. It has been suggested that MPO plays an important role in neutrophil shutdown and, thereby, timely resolution of inflammation. A decreased MPO activity might thus prolong the inflammatory reaction of the neutrophils. This finding was supported by the increased viability of the neutrophils obtained from periparturient cows. Even after stimulation, we found a lower caspase-3 activity in this group, indicating that they might be activated for a longer time compared with the neutrophils from mid-lactation cows. Accordingly, these alterations might contribute to a temporal mismatch in inflammatory responses, as often seen in severe periparturient infections.

Changes of leukocyte counts and expression of pro- and anti-inflammatory cytokines in peripheral leukocytes in periparturient dairy cows with retained fetal membranes.

In dairy cows, retained fetal membranes (RFM) affect reproductive performance. The aim of this study was to examine the leukocyte counts and the gene expression of tumour necrosis factor α (TNFα), interleukin 1ß (IL-1ß), IL-8, and IL-10 in polymorphonuclear leukocytes (PMNs) and peripheral blood mononuclear cells (PBMCs) in cows with (n = 5) or without (n = 5) RFM during the peripartum period. The lymphocyte counts in RFM cows were higher than those in control cows throughout the experiment (p < .05). The expression of IL-8 in PMNs of control cows was higher (p < .05) compared with that of RFM cows postpartum. In cows with RFM, IL-1ß expression was higher (p < .05) in PMNs at 6 weeks postpartum whereas the expression of IL-1ß was lower (p < .05) in PBMCs at 4 weeks postpartum. The expression of IL-10 in PBMCs of control cows was higher (p < .05) than that of RFM cows at 2 weeks prepartum and 4 weeks postpartum. Taken together, our data indicate that changes of gene expression of pro- and anti-inflammatory cytokines in RFM cows might be associated with the delayed placental separation and development of uterine inflammation in RFM cows.

Characterisation of peripheral blood mononuclear cell populations in periparturient dairy cows that develop metritis.

Bacterial contamination of the uterus following calving is ubiquitous in the dairy cow, 40% of cows develop postpartum uterine infection, including metritis. While predisposing factors like twinning and dystocia are associated with metritis, it is unclear why some cows remain healthy following calving and others develop uterine infection, negatively impacting animal health, milk production and economic return. Here, we profiled peripheral blood mononuclear cells of cows before calving and during postpartum metritis. We hypothesized that peripheral blood mononuclear cell function and proportions would be altered during the prepartum period in cows that develop postpartum metritis. Using flow cytometry we observed reduced proportions of peripheral CD3+/CD4+, CD4+/CD62L+, FOXP3+ and CD21+ populations from -10 to 40 days relative to calving associated with metritis, while the proportion of peripheral CD3+/CD4+ lymphocytes were specifically reduced in the prepartum period before the onset of metritis. Peripheral blood mononuclear cells from cows with metritis had a perturbed capacity to secrete IL-1ß or IFNγ in response to in vitro stimulus; cells collected during the prepartum period from cows that would go on to develop metritis failed to increase IL-1ß secretion in response to stimulation, while IFNγ secretion was altered at calving and postpartum in cows with metritis compared to healthy herd mates. No effect of metritis was observed in the capacity of cows to mount a humoral immune response to antigen administered on the day of calving. The studies discussed here suggest that while minor changes to the prepartum immune system are observed in cows that develop metritis, changes observed in the postpartum period are more prevalent and likely a consequences of disease and not causative. Future studies to modulate the prepartum immune system may help to limit postpartum metritis.

Type I and type II cytokine production of CD4+ T-cells in immune response biased dairy cattle around calving.

The peripartum period is a period of high stress, transition and management changes for dairy cows. It is associated with higher incidence of both metabolic and pathogenic disease. Both antibody-(AMIR) and cell-(CMIR) mediated immune responses play a key role in the maintenance of health in mammals protecting against extracellular and intracellular pathogens, respectively. Generally, interferon gamma (IFN-γ) has been associated with CMIR, whereas interleukin 4 (IL-4) has been associated with AMIR bias, and interleukin 17 (IL-17A) is associated with pro-inflammatory. It has been previously demonstrated that cows can be classified as high (H), average (A), and low (L) immune responders based upon their AMIR and CMIR to test antigens, and that this classification is associated with disease occurrence throughout lactation. The mechanisms behind these differences in phenotype and the effects of the peripartum period have not been fully investigated. The aim of this study was to determine the effects of the peripartum period on cytokine production of CD4+ T-cells or T helper (Th) cells, key mediators of the adaptive immune response. Immune response phenotyped cows were selected based on H-AMIR/L-CMIR (H-AMIR cows; n = 10) and H-CMIR/L-AMIR (H-CMIR cows; n = 11) response to test antigens. Isolated CD4+ T-cells collected at 28 days before calving (prepartum samples), 4 days after calving (early postpartum samples), and 21 days after calving (late postpartum samples) from these groups were stimulated with Concanavalin-A (ConA) with unstimulated controls. Subsequently, IL-4, IFN-γ, and IL-17A concentrations were quantified by ELISA. Overall, there was no obvious decline in IL-4, IFN-γ or IL-17A close to calving observed from CD4+ T-cells from each of these phenotypically distinct groups of cows. However, CD4+ T-cells isolated from H-CMIR secreted higher amounts of IL-4 (746.43 ±â€¯428 pg/mL), IL-17A (446IL ±â€¯62 pg/mL), and IFN-γ (7755.79 ±â€¯4449 pg/mL) than H-AMIR cows (IL-4 (212.15 ±â€¯121 pg/mL), IL-17A (163.15 ±â€¯87 pg/mL), and IFN-γ (2909.771 ±â€¯1671 pg/mL)) on day 21 after calving, late postpartum. This study indicates a genetic predisposition based on immune response phenotype of cytokine production from CD4+ T-cells around calving.

Assessment of the innate immune response in the periparturient cow.

The transition period is the most critical phase in the life of high yielding dairy cows. Within a few weeks, cows are submitted to many challenges (physiological, nutritional, psychological, management) that require prompt and effective adaptive responses. The immune system is involved in this process, and many changes of the cow's immune system components have been observed around calving. Cows are considered to be immunosuppressed in late lactation, and available data suggest that the immune system is dysregulated around parturition. Significant attention has been focused on modification of cellular functions (e.g. the reduction of phagocytosis and diapedesis), but growing interest concerns the components of the innate immune system, which often exhibits increased responses such as susceptibility to inflammatory events and the related acute phase response (APR). Systemic inflammation plays a significant role in early lactation, affects many liver functions and has been associated with the impairment of cow performance (i.e. reduced feed intake, milk yield, fertility, welfare). The assessment of variations in immune-metabolic indices offers opportunities to predict the onset of the health troubles and to anticipate the proper therapies needed to guarantee health, good welfare and fertility in the following lactation. The frequency of diseases (metabolic and infectious) before calving is rare, but several clues suggest that various metabolic and immune variations can begin during the dry period. Interesting preliminary results encourage this perspective and possible candidates are suggested.

Expression profiles of immunity and reproductive genes during transition period in Holstein cattle.

The transition period is a critical time for dairy cows as the animal is subjected to the physiological stress accompanying parturition. Immunosuppression and health status were examined during this period in 80 Holstein cows. Blood samples were taken from each cow 3, 2 and 1 week before and after calving, and at calving (0 day). RNA was extracted and subjected to real-time PCR to determine mRNA levels for the immune-related genes TLR 2, 4, 6, 7 and ß-defensin 5 in addition to the reproduction-related genes prolactin and IGF-I. Results showed significant up-regulation of pro-inflammatory-selected genes, TLR 4, 6 7 and ß-defensin 5 at the third-week post-calving; however, earlier periods had lower expression of such genes. In contrast, the immunosuppression biomarker TLR2 gene was up-regulated at calving and 1 week after parturition and then down-regulated again at second and third week. Prolactin and IGF-I genes expression levels were significantly and gradually increased mainly post-partum. This research highlights that the expression patterns of TLRs, BNBD5, PRL and IGF-I could be biomarkers to follow up immune and reproductive status of dairy cow at peri-parturient period to predict the most susceptible risk time for disease incidence and to build up management protocol.

Depressed polymorphonuclear cell functions in periparturient cows that develop postpartum reproductive diseases.

The study was planned to see if there is any important and significant changes in the PMN function in cows suffering from postpartum reproductive diseases (PRD). Blood sampling was done from 41 pregnant cows on 15 days prepartum (-15d), calving day (0d), 15 days (15d) and 30 days (30d) postpartum and thorough gynaecological examination was performed on 0d, 15d, 30d and 45d for diagnosis of PRD like retained placenta (RP), clinical metritis (CM), clinical endometritis (CE) and delayed involution of uterus (DIU). The heparinised blood was used for isolation of PMN leukocytes for estimation of superoxide (SO), hydrogen peroxide (H2O2) and enzyme myeloperoxidase (MPO) activity in each group of cows. The SO production (ΔOD) was greater for normal (0.19 ± 0.05) than cows suffering from RP (-0.12 ± 0.09), CM (-0.15 ± 0.13) and CE (-0.07 ± 0.05) at -15d. The mean value was greater for normal cows (0.12) than the cows with PRD (0.05 to 0.9) at 30d. The H2O2 production was greater for normal than cows with PRD at all sampling days and significantly greater than cows with RP and CE at 15d (p < 0.01) and 30d (P < 0.05). The MPO activity (µmol/1 × 107) was greater for normal (18.77 ± 1.27) than for RP (12.52 ± 2.57) and CM (11.31 ± 3.30) cows on 0d. The depressed capability of the PMN from the cows with PRD to produce SO, H2O2 and MPO during the periparturient period indicated their association with the development of RP, CM and CE.

Perfil sérico proteico de vacas Holandesas no período de transição / Serum protein profile of Holstein cows during the transition period

O objetivo deste trabalho foi avaliar, de forma longitudinal, o perfil sérico proteico de 13 vacas Holandesas durante o período de transição. Amostras de sangue (n=78) foram coletadas semanalmente, da segunda semana pré-parto (M-2) até a terceira semana pós-parto (M3), para determinação do perfil sérico proteico, por meio de teste bioquímico (proteínas séricas totais - PT) e eletroforese em gel de poliacrilamida SDS-PAGE, para as outras proteínas analisadas. Os valores de PT diminuíram de forma gradativa (P=0,000) de M-2 (6,4g/dL) a M0 (6,2g/dL), aumentando nos momentos subsequentes (M3=7,3g/dL). As concentrações da IgG de cadeia pesada (M-2=919,4; M-1=1074,5mg/dL) e de cadeia leve (M-2=393,9; M-1=466,7mg/dL) foram menores no pré-parto em relação ao pós-parto (M1=1283,3; M2=1374,2 e M3=1630,3 mg/dL para IgG pesada e M1=463,4; M2=573,7; M3=651,8mg/dL para IgG leve). Para a IgA, houve diminuição nos valores (P=0,001), de M-2 (51,9mg/dL) a M1 (34,0mg/dL), e aumento em M2 (45,4g/dL) e M3 (62,6g/dL). Os valores de haptoglobina (Hp) e ceruloplasmina (Cp) aumentaram (P=0,000) de M-2 (Hp=16,6mg/dL; Cp=8,6mg/dL) a M3 (Hp=60,9mg/dL; Cp=127,1mg/dL). A albumina apresentou ligeiras variações durante o período de transição (P=0,000), enquanto a transferrina sérica (P=0,101) e a glicoproteína ácida (P=0,105) foram estáveis. O escore de condição corporal (ECC) também foi analisado durante o período de transição, verificando-se diferença (P=0,003) entre M-2 (ECC=4,0) e M1 (ECC=3,0). Foi relatada ainda a ocorrência de distocias (4/13), retenção de placenta (1/13) e hipocalcemia (1/13) no dia da parição (M0) e infecções uterinas (5/13) e cetose (1/13) ocorridas no pós-parto. Concluiu-se, portanto, que houve aumento nas concentrações séricas de Hp e Cp e diminuição nos valores de imunoglobulina e transferrina em vacas Holandesas no período de transição, relacionados às doenças ocorridas nesse período e ao elevado ECC, promovendo modificações metabólicas e imunossupressão.(AU)

The aim of this study was to evaluate longitudinally the serum protein profile of 13 Holstein cows during the transition period. Blood samples (n=78) were taken weekly, from the second week before parturition (M-2) to the third week after parturition (M3) for determination of serum protein profile by biochemical tests (total serum protein - PT) and SDS-PAGE electrophoresis, for the other proteins analyzed. PT values decreased gradually (P = 0.000) from M-2 (6.4g/dL) to M0 (6.2g/dL), increasing in subsequent moments (M3 = 7.3g/dL). The concentrations of the heavy chain (M-2=919.4; M-1=1074.5mg/dL) and the light chain of IgG (M-2=393.9; M-1=466.7mg/dL) were lower in pre-calving compared to post calving (M1= 1,283.3; M2=1,374.2 and M3=1,630.3 mg/dL for the heavy chain, and M1=463.4; M2=573.7 and M3=651,8 mg/dL for the light chain of IgG). For IgA there was a decrease in the values (P=0.001), from M-2 (51.9mg/dL) to M1 (34.0mg/dL), and increase in M2 (45,4g/dL) and M3 (62,6g/dL). The values of haptoglobin (Hp) and ceruloplasmin (Cp) increased (P=0,000) from M-2 (Hp=16.6mg/dL; Cp=8.6 mg/dL) to M3 (Hp=60.9mg/dL; Cp=127.1mg/dl). Albumin showed slight variations during the transition period (P=0.000), while the serum transferrin (P=0.101) and acid glycoprotein (P=0.105) were stable. Body condition score (BCS) was also analyzed during the transition period, checking the difference (P=0.003) between M-2 (ECC= 4.0) and M1 (ECC=3.0). It was also reported the occurrence of dystocia (4/13), retained placenta (1/13) and hypocalcemia (1/13) on the day of calving (M0) and uterine infections (5/13) and ketosis (1/13) occurring in the post calving. In conclusion, there was an increase in serum Hp and Cp, and a decrease in the immunoglobulins and transferrin amounts in Holstein cows during the transition period, related to diseases occurring during this period and the high BCS, promoting metabolic changes and immunosuppression.(AU)

Long-term impact of puerperal metritis on the profiles of peripheral blood leukocytes in peripartum dairy cows.

To determine the effects of puerperal metritis on the immune response, changes in the differential peripheral blood leukocyte counts were analyzed during the peripartum period in cows with or without metritis. Multiparous Holstein cows were examined for uterine health disorders and classified into two groups: healthy (n = 11) or metritis (n = 5) cows. The lymphocyte and monocyte counts and the proportion of CD8(+) lymphocytes were higher in cows with metritis compared to healthy cows. Moreover, the effects of puerperal metritis on the lymphocyte counts and CD4(+)/CD8(+) ratio persisted weeks after the uterine inflammation had self-resolved. Taken together, the findings of the present study indicate the possible long-term alterations of systemic immune responses in cows with puerperal uterine inflammation.

Maternal and Infant Secretory Immunoglobulin A across the Peripartum Period.

BACKGROUND: Salivary secretory immunoglobulin A (sIgA) concentrations change over early infancy. The primary immunoglobulin in breast milk is sIgA, however, no study has examined the role of maternal sIgA in relation to infant salivary sIgA. OBJECTIVES: This study aimed to examine within-source associations and mean level changes of maternal and infant sIgA across the first 6 months of life, to examine the interrelations between maternal and infant sIgA across the first 6 months of life, and to determine the association between breastfeeding and infant sIgA. METHODS: Participants were a convenience sample of 51 mother-infant dyads. Salivary sIgA was collected from the mother in the third trimester. Infant and maternal salivary and maternal breast milk sIgA was collected at approximately 1, 3, and 6 months postpartum. RESULTS: Maternal salivary sIgA showed no mean level change across the visits, and levels were moderately associated over time. Breast milk sIgA was moderately associated over time; infant salivary sIgA was weakly associated over time. Both breast milk and infant sIgA levels decreased from 1 to 3 months postpartum. Maternal salivary sIgA was not related to infant or breast milk sIgA. Breastfed infants had lower levels of salivary sIgA. Likewise, higher concentrations of breast milk sIgA were related to lower concentrations of infant sIgA. CONCLUSION: Maternal salivary sIgA is highly stable over the peripartum period, whereas breast milk and infant salivary sIgA was variable. Infant secretory IgA development does not depend positively on maternal salivary or breast milk sIgA.

Periparturient stress and immune suppression as a potential cause of retained placenta in highly productive dairy cows: examples of prevention.

The immune system during the periparturient period is impaired. At this time the most important factor causing immune-suppression in highly productive cows is metabolic stress resulting from hormonal and metabolic fluctuations, a negative energy balance, shortage of proteins, minerals and vitamins which are required to meet the demands of the fetus as well as the onset of lactation. This stress can activate the hypothalamic-pituitary-adrenocortical axis (HPA), which results in increase plasma corticosteroids. As a result, the cortisol concentration during the periparturient period increases by several folds particularly on the day of calving. Cortisol is a powerful immune-suppressive agent. During stress, this hormone causes depression of the leukocyte proliferation and their functions. Decreased phagocytosis of neutrophils, decreased cytotoxic ability of lymphocytes, as well as depressed activity of their cytokines, make it impossible for the normal, efficient maternal immune recognition and rejection of fetal membranes (as a foreign, allogeneic tissue expressed fetal antigens-MHC class I proteins by trophoblast cells) and finally results in their retention in cows. The metabolic periparturient stress also activates production of catecholamines, especially adrenalin. Adrenalin activates adrenoreceptors of the myometrium and then causes hypotony or atony of the uterus. Thus, cortisol and adrenalin inhibit rejection and expulsion of fetal membranes and cause their retention. These mechanisms of retained placenta (RP) often have a metabolic etiology and occur in herds, where important infectious diseases causing placentitis are absent or prevented. The aim of this article is to show the fundamental mechanisms occurring during periparturient stress and the accompanied immune-suppression in cows, as well as their consequences in relation to RP. The paper also gives examples of the symptomatic prevention of RP in cows caused by metabolic and immune suppressive factors. The prevention of RP was carried out using drugs which inhibit the activity of cortisol or adrenalin in dairy cows during calving.

Factors contributing to immunosuppression in the dairy cow during the periparturient period.

The transition from late gestation to early lactation results in dramatic physiological changes including metabolic changes and immunosuppression in the dairy cow. As a result, cows are at a high risk for disease during this time. Evidence supporting a link between metabolic status and naturally occurring immunosuppression is growing. This review focuses on the impacts of metabolic status, and the metabolites that characterize it, on the immune response of cows during the transition period. Glucose is the preferred fuel for immune cells and its low concentration during the transition period may partly explain the naturally occurring immunosuppression at this time. To our knowledge, ketones are not utilized by immune cells and primarily have been shown to inhibit the immune response when concentration is relatively high. The effect of fatty acids on the immune system response remains unclear. Evidence suggests that the type of fatty acid can either stimulate (i.e. saturated fatty acids) or inhibit (i.e. unsaturated fatty acids) the immune response. We have suggested that an index for physiological imbalance (PI), based on circulating metabolites that characterize metabolic status, directly relates to mechanisms associated with the development of disease and is superior to calculated energy balance and therefore is a better predictor of risk of disease. The usefulness of the PI index as a predictor of risk of disease and the mechanisms associated with the links between degree of PI and immunosuppression for dairy cows during the transition period warrants further investigation.

Modulation of innate immune function and phenotype in bred dairy heifers during the periparturient period induced by feeding an immunostimulant for 60 days prior to delivery.

The purpose of this study was to evaluate the effect of a feed additive (OmniGen-AF(®), reported to have immune modulating activity) on innate immunity and health events during the periparturient period in dairy heifers when immunity is suppressed. From 60 days prepartum through calving, supplemented heifers (n=20) received OmniGen-AF(®) daily and were compared with unsupplemented controls (n=20). Blood leukocyte innate immune activity (phenotype markers, phagocytic activity, and reactive oxygen species--ROS production) was measured prior to feeding (60 days prepartum), 30 days later, and on days 1, 7, 14, and 30 postpartum. Adverse health events (udder edema, ketosis, displaced abomasum, and death) and milk production were measured at calving and into early lactation. The fraction of leukocytes with measurable CD62L (L-selectin) on their surface from supplemented heifers tended to be greater during the periparturient period in treated heifers than controls (p=0.100). Likewise, leukocyte phagocytosis of Escherichia coli and Staphylococcus aureus during this time period tended to be greater in heifers supplemented with OmniGen-AF(®) (p=0.100). Conversely, ROS production in response to phorbol myristate acetate or when leukocytes were stimulated with killed S. aureus lysate tended to be greater among control heifers compared with supplemented animals (p=0.100). Supplemented heifers exhibited fewer incidents of udder edema than controls (p=0.030) and tended to exhibit a lower rate of new cases of mastitis (p=0.098); however, no differences were observed in milk somatic cell counts or level of milk production. Results demonstrate a positive role of OmniGen-AF(®) in amplifying leukocyte function consistent with antibacterial activity during the periparturient period, and support the continued study of dietary supplementation to enhance mammary gland health in dairy cows.