Acute phase proteins and stress markers in the immediate response to a combined vaccination against Newcastle disease and infectious bronchitis viruses in specific pathogen free (SPF) layer chicks

Vaccination is an important tool in poultry health, but is itself a stressor often resulting in a reduction in feed intake, body weight gain, and nutrient digestibility. In other species, vaccination is associated with an immediate acute-phase response. As an important immune parameter, the circulating heterophil/lymphocyte (H/L) ratio is a well-recognized parameter of stress in poultry. In this study, the effects of a routinely used commercial poultry vaccine on the acute phase response (APR) and H/L ratios in specific pathogen-free (SPF) layer chicks was examined to determine if post vaccination (PV) stress and an APR occur. A combined Newcastle disease and infectious bronchitis vaccine (Nobalis Ma5+Clone 30) was administered to SPF chicks by the intraocular route at age 7 d. Acute phase proteins (APP), alpha-1 acid glycoprotein (AGP) and serum amyloid A (SAA) were measured by enzyme-linked immunosorbent assays at d 0 (pre-vaccination) and d 0.5, 1, 2, 3, 4, 5, 6, and 21 PV. Stress was determined in the chicks by measurement of the H/L ratio. The immune response to the vaccine was estimated by measurement of the antibody (IgY) response to the vaccine at d 21. The antibody titer was significantly (P < 0.05) higher in the vaccinated group at 21 d PV, confirming stimulation of the immune system. The H/L ratio was also significantly higher in the vaccinated group at 1 to 2 d (P < 0.01) and at 3 d (P < 0.05) PV. The concentration of SAA increased by 2.8-fold, from 63.7 μg/mL in controls to 181 μg/mL in the vaccinated group, (P < 0.05) at 1 d PV. AGP increased 1.6-fold at 2 d PV, (from 0.75 g/mL in the control group to 1.24 g/mL in the vaccinated group, P < 0.05). In conclusion an immediate but mild APR occurred in the chicks following intraocular vaccination, whereas the stress response as measured by H/L ratio seemed to be more specific and sensitive. Measurement of these biomarkers of the host response could be a tool in vaccine development

Acute phase proteins as a biomarker of health and disease in chickens

Animals undergoing a challenge to their state of health mount a vigorous response which involves both the innate and acquired immune systems. The varied non-specific responses of an animal to infection, inflammation or trauma are collectively referred to as the acute phase response (APR). The APR is a very complex reaction involving both local and systemic effects. Acute-phase proteins (APP) are a group of blood proteins primarily synthesised in the liver in response to pro-inflammatory cytokines being released primarily from leukocyte activation during an APR. The use of APP for diagnostic purposes in both human and veterinary medicine has increased greatly within the last decade. The aim of this thesis was to investigate the APR in chickens in response to i) vaccination using a Newcastle disease and Infectious Bronchitis (N/B) Live, freeze-dried virus vaccine; ii) an experimental challenge with Poultry red Mite (PRM); (iii) an LPS challenge experiment with an E. coli LPS. Four APP namely serum amyloid A (SAA), alpha 1 acid glycoproteins (AGP), ovotransferrin (OVT) and ceruloplasmin (CP) were studied in detail along with other blood component including heterophile / lymphocyte (H/L) ratios and corticosterone. For SAA, AGP and CP, species specific ELISA kits are now commercially available, but these had not previously been validated. The results from chapter 2 demonstrated that all three ELISAs gave good accuracy, had a suitably low detection limit and allowed discrimination between different levels of APP in chicken samples. For OVT a lab based radial immunodiffusion assay (RID) was used. The results of the current study have shown that a mild response of APP (SAA and AGP) in SPF chick post vaccination at day one and two post vaccination. H/L ratios also increased, and this measure was deemed more sensitive and consistent in terms of measuring the mild stress response under the conditions employed than the APP though of these the SAA was the most promising. In the other experimental challenge with PRM in laying hens, the serum levels of SAA were also significantly increased, and this was subsequently found to be positively correlated with the level of PRM infestation which was monitored for several months, further serum protein profile alterations were described in this study. Whereas, LPS challenge induced significant with high magnitude of APP (SAA, AGP and OVT). In addition, local expression of APP has been investigated and significant changes of other blood components were described. Looking for the APR by non-invasive way in chickens challenged with LPS, there were interesting sensitive detection of the body surface temperature. Also, a significant correlation of the cloacal with the body surface temperature changes in development of pyrexia. In summary, the research presented in this thesis has demonstrated that SAA is the most sensitive APP and therefore potentially the most useful biomarker in chickens (layers and broilers): this APP increased more rapidly and by a greater magnitude than the other APP following a range of stimuli. AGP and OVT moderately increased and usually peaked later than SAA, while CP rarely changed. Further research is needed to investigate the relationship between the local and systemic APR and to determine the significance of extrahepatic versus hepatic production of APP in the chicken and, the associated H/L ratio and thermal imaging in early disease recognition as both of these measurements proved to be useful in such studies

Integrated dataset on acute phase protein response in chicken challenged with Escherichia coli lipopolysaccharide endotoxin

Data herein describe the quantitative changes in the plasma proteome in chickens challenged with lipopolysaccharide (LPS), a bacterial endotoxin known to stimulate the host innate immune system obtained by shotgun quantitative proteomic tandem mass tags approach using high-resolution Orbitrap technology. Statistical and bioinformatic analyses were performed to specify the effect of bacterial endotoxin. Plasma from chicken (N=6) challenged with Escherichia coli (LPS) (2 mg/kg body weight) was collected pre (0 h) and at 12, 24, 48, and 72 h post injection along with plasma from a control group (N=6) challenged with sterile saline. Protein identification and relative quantification were performed using Proteome Discoverer, and data were analysed using R. Gene Ontology terms were analysed by the Cytoscape application ClueGO based on Gallus gallus GO Biological Process database, and refined by REVIGO. Absolute quantification of several acute phase proteins, e.g. alpha-1-acid glycoprotein (AGP), serum amyloid A (SAA) and ovotrensferrin (OVT) was performed by immunoassays to validate the LC-MS results. The data contained within this article are directly related to our research article”Quantitative proteomics using tandem mass tags in relation to the acute phase protein response in chicken challenged with Escherichia coli lipopolysaccharide endotoxin” [1]. The raw mass spectrometric data generated in this study were deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD009399 (http://proteomecentral.proteomexchange.org/cgi/GetDataset?ID=PXD009399)

Serum and acute phase protein changes in laying hens, infested with poultry red mite

The poultry red mite (PRM) is one of the most economically important ectoparasites of laying hens globally. This mite can have significant deleterious effects on its fowl host including distress, anemia, reduced egg production, and reduced egg quality. This study was conducted to evaluate the influence of PRM on the serum protein profile in laying hens and its effect on the acute phase proteins (APPs) to assess their potential as biomarkers for mite infestation. Three APPs: alpha-1 acid glycoprotein (AGP), serum amyloid-A (SAA), and ceruloplasmin (CP) were measured in serum samples collected from laying hens at 12 and 17 wk of age, and then for up to 4 mo after a challenge with PRM (starting at 18.5 wk of age). The serum protein profile (SDS-PAGE/nanoflow HPLC electrospray tandem mass spectrometry) and concentration of individual serum proteins (SDS-PAGE-band densitometry) were also compared. Post challenge there was a positive correlation (r = 0.489; P < 0.004) between the levels of SAA and the PRM numbers. The levels of SAA steadily increased after the PRM challenge and were significantly different than the pre-challenge levels at 28, 32, and 36 wk of age (P < 0.01). The PRM numbers also peaked around 31-33 wk of age. The results for AGP and CP in comparison were inconsistent. Proteomics revealed the presence of 2 high molecular weight proteins in the serum between 12 and 17 wk of age. These were identified as Apolipoprotein-B and Vitellogenin-2, and their increase was commensurate with the onset of lay. No other major differences were detected in the protein profiles of blood sera collected pre and post challenge. We conclude that SAA could be used as a useful biomarker to monitor PRM infestation in commercial poultry flocks and that PRM infestation does not disrupt the production of the major proteins in the serum that are associated with egg formation

Evaluation of AGP, SSA concentration with the relationship of antibody titer in broiler chicken vaccinated with IBD virus

Gumboro or Infectious bursal disease (IBD) is one of the most important diseases in poultry industry. IBD causes economic losses as a result to mortality and reduction in performance and medicine expenses(de S. B. Fróes et al., 2018). IBD is Severe acute disease of 3-6-week-old birds is associated with high mortality, but a less acute or subclinical disease is common in 0-3-week-old birds (Sali, 2019). IBD virus causes lymphoid depletion of the bursa and if this occurs in the first 2 weeks of life, significant depression of the humoral antibody response may result. The current study aims to measurement of antibody titer and acute phase proteins (AGP and SSA) in broiler chickens subjected to the Gumboro vaccine to explore the intensity of the vaccine strain and the response of both the early innate immune response and late humoral immune responses (antibody titers) raised against the vaccine. A handed Ross 308 chicks were divided into three groups, control, intermediate and intermediate plus vaccine. At 10 days old, the chicks were vaccinated and then blood samples collected at 12, 24 hours, and 10 and 15 days.&nbsp

Effect of Betaine on some productive traits of turkey under heat stress

This study was conducted at the poultry farm, collage of veterinary, of Kufa included field experimentation which was conducted during the period from 24th of December 2010 to 23th April 2011. The experiment aimed to investigate the effect of betaine supplemented to the diet to alleviate the negative effects of heat stress through study of some productive traits of turkey were being exposed to heat stress.    Sixty unsexed one day old chick turkey(Nicholas 300) average weight (58gram). The chicks were reared on a ground throughout two stages: the first stage from one day old to 12 weeks old with no treatment whereas second stage was from 13-16 weeks of age. They were randomly allocated 3 treatment groups, each comprising of 2 replicates of 10  chicks, the treatment containing three betaine levels at 0,1000,1500 mg/kg diet T1,T2 and T3 respectively, birds reared under cyclic high temperature 27-38-27 cilices and relative humidity average 55-60%.1. Birds in T2 have a higher (P<0.05) body weight in average at 16 weeks of age. And the average of  weight gain during the period of 13-16 weeks of age is also higher as compared to other treatments2.  During the period of 13-16 weeks of age, food conversion rate for T2 was best at significant level (P<0.05) compared with other treatment.3. Birds in T2 have a higher (P<0.05)food intake average during the period 13-16 weeks of age as compared with the other treatments.4.Abdomnal fat significantly decreased (p<0.05) while breast muscle ratio and breast meat yield were increased (p<0.01) in T3 as compared with T1 .5. birds in T2 and T3 resulted in significant decrease( P<0.05) in mortality as compared with control treatment.6. Production index (PI) and economic figure (EF) were significantly improved for T2 and T3 compared with the control

Quantitative proteomics using tandem mass tags in relation to the acute phase protein response in chicken challenged with Escherichia coli lipopolysaccharide endotoxin

The inflammatory response in chickens (Gallus Gallus domesticus) is an integral part of the bird's response to infection. Detailing proteomic changes occurring during infection would be beneficial to the poultry industry, offering opportunities for comparative pathophysiological analysis. The objective of this study was to quantify the changes in the plasma proteome in chickens challenged with lipopolysaccharide (LPS), a bacterial endotoxin known to stimulate the host innate immune system. Plasma from chicken (N = 6) challenged with Escherichia coli (LPS) (2 mg/kg body weight) was collected pre (0 h) and at 12, 24, 48, and 72 h post-injection along with plasma from a control group (N = 6) challenged with sterile saline. Samples were analysed by a quantitative Tandem Mass Tags approach using a Q-Exactive-Plus mass-spectrometer. Identification and relative quantification were performed using Proteome Discoverer, and data were analysed using R. Gene Ontology terms were analysed by Cytoscape based on the Gallus gallus database. Finally, 87 significantly regulated proteins were found, including serum-amyloid-A, ovotransferrin and alpha-1-acid-glycoprotein, showing a significant effect of time post-injection in the LPS-treated group. Different pathways related with protein activation cascade and heterotopic cell-cell adhesion were affected by LPS-challenge. LPS-challenged chickens demonstrate significant changes to the plasma proteome with both increases and decreases of individual proteins within 12 h of challenge. Significance: The injection of chicken with bacterial lipopolysaccharide followed by sequential plasma and clinical analysis of the bird, is a long established and a widely used model for inflammation and infection studies. This study, utilising and combining proteomic and immunoassay analysis with bioinformatic analysis, revealed that several biological pathways are modulated during this early period of inflammation. In addition, proteins with biomarker potential were identified and successfully validated. This experimental model also demonstrated potential for pathophysiological mechanism investigation and as an inflammatory model for biomedical research. There is, despite plasma being an easily accessible biological matrix which is representative of the health status of the bird, scarce data on the chicken plasma proteome. This research makes a positive contribution to the current field, generating significant data for continuing comparative analysis

Increased mechanically-induced ectopy in the hypertrophied heart

IntroductionLeft ventricular hypertrophy (LVH) predisposes patients to arrhythmias, but the mechanism of these arrhythmias is unclear. Here we show that hearts from spontaneously hypertensive rats (SHR) have a lower threshold for induction mechanically induced arrhythmias compared to age matched Wistar-Kyoto (WKY).MethodsRecordings were made from isolated hearts from nine month old SHR (n = 18) and WKY (n = 17) rats. A water filled balloon in the left ventricle had its volume controlled by a servo-driven syringe. LVEDP was abruptly increased in increments until an ectopic beat was detected by an epicardial MAP electrode. Alternatively, LVEDP was abruptly reduced back to 5 mm Hg from an elevated pressure.ResultsSHR hearts had a lower threshold for stretch induced ectopics (29.87 ± 2.79 mm Hg vs. 42.23 ± 2.33 mm Hg, p ConclusionsHearts from SHR are more susceptible to both stretch-induced and release-induced arrhythmia.Do Yeong Kim, Ed White, David A. Sain