Research Note: The effect of selection for 16-week body weight on turkey serum metabolome.

The phenotype of modern commercial turkeys is substantially different than that of unselected, heritage turkey lines. These phenotypic changes have arisen from alterations in the genome/transcriptome, as well as the influence of many external factors on growth performance including nutrition, environment, and management. To investigate the phenotypic changes resulting from genetic selection for increased body weight, The Ohio State University maintains 2 unique genetic turkey lines: the randombred control (RBC2) line, which is comprised of genetics from 1960 era commercial turkeys and has been maintained without conscious selection for any trait; and the F line, which was originally selected from the RBC2 line and has been selected for increased 16 wk body weight for over 50 generations. This study used broad-spectrum mass-spectrometry profiling techniques to identify and quantify differences in the metabolome of the serum of F and RBC2 turkey lines. Serum samples from both F and RBC2 turkeys were subject to quantitative time of flight liquid chromatography tandem mass spectrometry analyses. Principle component analyses showed distinct populations of metabolites in the F vs. RBC2 serum, suggesting that increased body weight is associated with the accumulation of several metabolites. Comparing the spectral features to online databases resulted in the selection of 104 features with potentially identifiable chemical structures. Of these 104 features, 25 were found at higher levels in the serum of the RBC2 line turkeys, while 79 were found at a greater abundance in the F line turkeys. A more detailed analysis of these 104 features allowed for the putative identification of 49 compounds, which were clustered into 6 functional groups: 1) energy metabolism; 2) vitamins; 3) hormones and signaling molecules; 4) lipid derivatives, fatty acid metabolites, and membrane components; 5) amino acid/protein metabolism; and 6) microbial metabolites. Further validation and experimentation is needed to confirm the identity of these metabolites and understand their biological relevance and association with selection for increased body weight.

Effect of transportation and shackling on plasma concentrations of corticosterone and heterophil to lymphocyte ratios in market weight male turkeys in a commercial operation.

There is limited information on the effects of stress and/or physiological manipulation on either plasma concentrations of corticosterone (CORT) and/or heterophil: lymphocyte (H : L) ratios in turkeys. The present studies examine the effects of catching/transportation/lairage in a holding shed and shackling on plasma concentrations of CORT and H : L ratios in male market weight turkeys. Plasma concentrations of CORT were increased after transportation and lairage but not further elevated by shackling, irrespective of its duration up to 240 s. In one study, there were increased H : L ratios following catching/placing birds into transportation cages/transportation/lairage. In one study, H : L ratios declined following shackling. It is concluded that while moving turkeys from the farm to immediately before the shackling line is stressful, shackling for up to 4 min was not perceived as more stressful in turkeys. There were also differences between farms/houses for both plasma concentrations of CORT and H : L ratios.

Effects of putative stressors and adrenocorticotropic hormone on plasma concentrations of corticosterone in market-weight male turkeys.

There is limited information on the effects of stress and/or physiological manipulation on plasma concentrations of corticosterone (CORT) in turkeys. Under basal conditions, there was evidence for episodic release of CORT in turkeys. The present studies determine the effects of handling, herding, herding, the administration of Escherichia coli endotoxin, and challenge with turkey adrenocorticotropic hormone (ACTH) on plasma concentrations of CORT in market-weight male turkeys. Plasma concentrations of CORT were increased after challenge with turkey ACTH, handling together with saline injection or herding (moving birds from one pen to another). There were no effects on plasma concentrations of CORT of the following putative stressors: handling per se, endotoxin challenge, or of placing in an inverted position on simulated shackles.

Analysis of Toxoplasma gondii clonal type-specific antibody reactions in experimentally infected turkeys and chickens.

Due to their ground-feeding behaviour, free-ranging chickens and turkeys are exposed to oocysts and are good indicators of the presence of Toxoplasma gondii in the environment. In addition, poultry may become infected by ingestion of tissues of infected intermediate hosts such as small rodents. Free-ranging poultry are considered an important source of T. gondii infection in humans, especially in developing countries. Knowledge on T. gondii genotypes in infected animals and humans is important for understanding the epidemiology of T. gondii infections. The aim of the present study was to analyse the ability of experimentally infected turkeys and chickens to develop a T. gondii clonal type-specific antibody response (IgY) after i.v. inoculation with tachyzoites of three T. gondii clonal lineages, types I, II and III. A peptide microarray displaying a panel of 101 different synthetic peptides was used for serotyping. Peptide sequences were derived from polymorphic regions of 16 T. gondii proteins (GRA1, GRA3-7, SAG1, SAG2A, SAG3, SAG4, SRS1, SRS2, ROP1, NTPase I and NTPase III and BSR4). The array was probed with 120 sera from experimentally infected chickens and turkeys inoculated with different doses of T. gondii tachyzoites (104, 103 and 102) collected from isolates representative for T. gondii clonal types I (RH), II (ME49) or III (NED) and uninfected controls. After screening of the peptides with reference sera from chickens and turkeys, and evaluation of data by Receiver Operating Characteristics analysis, 41 and 40 peptides were identified that appeared suitable to detect type-specific reactions with sera collected at 2, 5, 7 and 9 weeks p.i. Selected peptides allowed the identification of T. gondii clonal types, until 9 week p.i., which the chickens or turkeys had been inoculated with. At 9 weeks p.i., a high proportion of the experimentally infected chickens (67% (12/18)) and turkeys (61% (11/18)) no longer reacted with the selected peptides. Serotyping of the infection in individual chickens or turkeys was only possible when the whole peptide panel was applied. Clonal type-specific antibody responses were dynamic in both poultry species and depended on the individual animal and the time after infection.

Implementation of different feed withdrawal times and water temperatures in managing turkeys during heat stress.

This investigation studied the effects of different feed withdrawal times, water temperatures, and their interaction on growth performance, carcass traits, blood parameters, and health aspects of native turkeys (Egyptian local breed). We distributed native turkey poults (n = 180; 4 wk old) into 3 groups according to feed withdrawal time (ad libitum; feed withdrawal from 800 to 1,400 h, FW8 to 14; and feed withdrawal from 1,400 to 2,000 h, FW14 to 20). Each group was further divided into 2 subgroups corresponding to water temperature (ordinary and chilled water). Our results indicated that birds of FW14 to 20 exhibited the highest (P = 0.0001) body weight (BW) and average daily gain (ADG), whereas turkeys that drank chilled water displayed a higher BW than those that drank ordinary water. Turkeys that were kept under FW8 to 14 and drank chilled water displayed the highest BW at marketing and ADG from 4 to 16 wk of age. The lowest feed conversion ratio (FCR) was exhibited in turkeys kept off feed under the FW14 to 20 regime. Birds that were kept under FW14 to 20 and received chilled water showed the lowest FCR. The meat color scores of the birds that drank chilled water were higher than those that drank ordinary water, whereas birds kept off feed under the FW14 to 20 regime displayed the highest tenderness and juiciness, followed by those fed ad libitum. Turkeys reared under FW14 to 20 exhibited the highest albumin/globulin ratio and glucose content but the lowest globulin and aspartate transaminase (AST) values. Birds that were kept under FW14 to 20 and drank chilled water displayed the lowest corticosterone concentration. Turkeys reared under FW14 to 20 presented the lowest body temperature. Birds that drank chilled water exhibited a lower body temperature than those that drank ordinary water. Turkeys that were fed ad libitum and drank chilled water displayed the lowest body temperature. Conclusively, applying feed withdrawal and cold water had benefits in turkeys during heat stress. Applying these treatments as managerial alternatives for raising native turkeys during the summer season is highly recommended.

Comparative Toxicokinetics and Plasma Protein Binding of Ochratoxin A in Four Avian Species.

Ochratoxin A (OTA, 0.25 mg/kg body weight) was absorbed rapidly ( Tmax = 0.31-1.88 h) in all avian species (broiler chickens, laying hens, turkeys, and Muscovy ducks) but more slowly in broiler chickens ( Tmax = 1.43-4.63 h). The absolute oral bioavailability was complete in these bird species (88.0-109.6%). Ducks have a significantly higher volume of distribution ( Vd) and turkeys a lower Vd compared to chickens and layers (broiler chickens, 0.27 ± 0.12 L/kg; layers, 0.23 ± 0.08 L/kg; turkeys, 0.18 ± 0.04 L/kg; ducks, 0.76 ± 0.44 L/kg). This difference in Vd can be attributed to the species-dependent differences in plasma protein binding of OTA, namely ranging between 82.2 and 88.9% in ducks and between 96.5 and 98.8% in turkeys. No significant gender differences were found in toxicokinetics or plasma protein binding.

Modification of the lipid profile and antioxidant status of the blood plasma of turkey hens fed mixtures with raw or extruded linseed.

The aim of the study was to determine the most beneficial proportion of raw linseed in complete feed mixtures for turkey hens on the basis of lipid and redox indicators in the blood. In experiment 1, the turkey hens received the complete mixture with 2%, 4% or 6% linseed. On the basis of the results obtained in experiment 1, we selected the most effective proportion of linseed, which was given to the birds in the group receiving a 4% linseed additive. In experiment 2, the birds were fed mixtures with a 4% addition of raw or extruded linseed. The use of 4% raw linseed was found to improve production effects (improvement of weight gain, and lower feed conversion ratios), while extruded linseed in the diet of turkey hens did not affect growth performance. The use of linseed (4% and 6%) as a feed component for turkey hens led to an increase in indicators of antioxidant potential, that is the total antioxidant potential of the plasma, vitamins E and C, bilirubin and creatinine. A benefit resulting from the use of linseed, particularly in the amounts of 2% and 4% was a marked improvement in lipid indicators in the blood. The reduced percentage of unsaturated fatty acids (n-3) following the use of extruded linseed resulted in a decrease in lipid peroxidation (lower content of malondialdehyde, superoxide and vitamins C and E in the blood). The most effective dose and form of linseed in the diet of turkey hens is 4% raw linseed.

Determination of tigecycline in turkey plasma by LC-MS/MS: validation and application in a pharmacokinetic study.

Tigecycline (TIG), a novel glycylcycline antibiotic, plays an important role in the management of complicated skin and intra-abdominal infections. The available data lack any description of a method for determination of TIG in avian plasma. In our study, a selective, accurate and reversed-phase high performance liquid chromatography-tandem mass spectrometry method was developed for the determination of TIG in turkey plasma. Sample preparation was based on protein precipitation and liquid-liquid extraction using 1,2-dichloroethane. Chromatographic separation of TIG and minocycline (internal standard, IS) was achieved on an Atlantis T3 column (150 mm × 3.0 mm, 3.0 µm) using gradient elution. The selected reaction monitoring transitions were performed at 293.60 m/z → 257.10 m/z for TIG and 458.00 m/z → 441.20 m/z for IS. The developed method was validated in terms of specificity, selectivity, linearity, lowest limit of quantification, limit of detection, precision, accuracy, matrix effect, carry-over effect, extraction recovery and stability. All parameters of the method submitted to validation met the acceptance criteria. The assay was linear over the concentration range of 0.01-100 µg/ml. This validated method was successfully applied to a TIG pharmacokinetic study in turkey after intravenous and oral administration at a dose of 10 mg/kg at various time-points.

Models describing metronidazole pharmacokinetics in relation to hemodynamics in turkeys.

Linking haemodynamic (HD) and pharmacokinetic (PK) parameters provides much insight into interrelations between circulatory system and drug disposition. This effect is particularly pronounced in rapidly growing animals. Heart rate (HR), cardiac output (CO) and stroke volume (SV) are tightly linked with the animal's age and correlate with the increasing body weight (BW). The aim of this study was to establish and validate the relations between BW, HD and chosen PK parameters of metronidazole (MTZ) and its metabolite - hydroxymetronidazole (MTZ-OH) in growing turkeys. The study was carried out on broiler turkeys (BUT-9, n=26). All individuals were subjected to single dose PK studies four times, that is when the mean BW in the group reached: 1.4 (group A); 2.7 (group B); 5.5 (group C); 10.7kg (group D). Some PK parameters normalized with regard to HR were found to take constant values in all the age groups under investigation. CO↔1/MRT, SV↔1/MRT and SV↔MRT model was validated with regard to both metabolite and drug PK. This study proposed a model for the analysis of the relations HD↔BW and HD↔PK. Models developed in this study provide empirical evidence that HD affect the PK of MTZ and MTZ-OH in a different fashion.

Ameliorative Effects of Neutral Electrolyzed Water on Growth Performance, Biochemical Constituents, and Histopathological Changes in Turkey Poults during Aflatoxicosis.

Different in vitro and in silico approaches from our research group have demonstrated that neutral electrolyzed water (NEW) can be used to detoxify aflatoxins. The objective of this investigation was to evaluate the ability of NEW to detoxify B-aflatoxins (AFB1 and AFB2) in contaminated maize and to confirm detoxification in an in vivo experimental model. Batches of aflatoxin-contaminated maize were detoxified with NEW and mixed in commercial feed. A total of 240 6-day-old female large white Nicholas-700 turkey poults were randomly divided into four treatments of six replicates each (10 turkeys per replicate), which were fed ad libitum for two weeks with the following dietary treatments: (1) control feed containing aflatoxin-free maize (CONTROL); (2) feed containing the aflatoxin-contaminated maize (AF); (3) feed containing the aflatoxin-contaminated maize detoxified with NEW (AF + NEW); and (4) control feed containing aflatoxin-free maize treated with NEW (NEW). Compared to the control groups, turkey poults of the AF group significantly reduced body weight gain and increased feed conversion ratio and mortality rate; whereas turkey poults of the AF + NEW group did not present significant differences on productive parameters. In addition, alterations in serum biochemical constituents, enzyme activities, relative organ weight, gross morphological changes and histopathological studies were significantly mitigated by the aflatoxin-detoxification procedure. From these results, it is concluded that the treatment of aflatoxin-contaminated maize with NEW provided reasonable protection against the effects caused by aflatoxins in young turkey poults.

Intensive trapping of blood-fed Anopheles darlingi in Amazonian Peru reveals unexpectedly high proportions of avian blood-meals.

Anopheles darlingi, the main malaria vector in the Neotropics, has been considered to be highly anthropophilic. However, many behavioral aspects of this species remain unknown, such as the range of blood-meal sources. Barrier screens were used to collect resting Anopheles darlingi mosquitoes from 2013 to 2015 in three riverine localities (Lupuna, Cahuide and Santa Emilia) in Amazonian Peru. Overall, the Human Blood Index (HBI) ranged from 0.58-0.87, with no significant variation among years or sites. Blood-meal analysis revealed that humans are the most common blood source, followed by avian hosts (Galliformes-chickens and turkeys), and human/Galliforme mixed-meals. The Forage Ratio and Selection Index both show a strong preference for Galliformes over humans in blood-fed mosquitoes. Our data show that 30% of An. darlingi fed on more than one host, including combinations of dogs, pigs, goats and rats. There appears to be a pattern of host choice in An. darlingi, with varying proportions of mosquitoes feeding only on humans, only on Galliformes and some taking mixed-meals of blood (human plus Galliforme), which was detected in the three sites in different years, indicating that there could be a structure to these populations based on blood-feeding preferences. Mosquito age, estimated in two localities, Lupuna and Cahuide, ranged widely between sites and years. This variation may reflect the range of local environmental factors that influence longevity or possibly potential changes in the ability of the mosquito to transmit the parasite. Of 6,204 resting An. darlingi tested for Plasmodium infection, 0.42% were infected with P. vivax. This study provides evidence for the first time of the usefulness of barrier screens for the collection of blood-fed resting mosquitoes to calculate the Human Blood Index (HBI) and other blood-meal sources in a neotropical malaria endemic setting.

Serological and haematological values in geriatric broad-breasted white turkeys.

Blood samples from 1-, 5- and 10-year-old broad-breasted white turkeys were used to determine haematological and serum values. There were 8 turkeys in each age group. Mean haematocrit, haemoglobin and creatinine values were higher in the 1-year-old than in 10-year-old turkeys. Mean alanine aminotransferase was greater in 1 year old than in 10 year olds and greater in 5 year olds than in 10 year olds. Aspartate aminotransferase values were significantly different between all age groups. Mean alkaline phosphatase was less in 1-year-old than in 10-year-old turkeys. There were no significant differences between the three groups for cholesterol, glucose, total protein, albumin, globulin, calcium and bilirubin. All male turkeys had higher haemoglobin and haematocrit values than age-matched females. These data will help establish basic background references values in geriatric turkeys used in research.

A comparison of Mott cell morphology of three avian species. II. - Bad behavior by plasmacytes?

Mott cells are atypical plasmacytes recognized microscopically by endoplasmic reticulum (ER) distensions (Russell bodies) a result of retained secretory product (antibody). Originally associated with parasitism, they are observed in a broad spectrum of immunopathology, sometimes involving hypergammaglobulinemia. Few descriptions of Mott cells appear in avian literature. The purpose of the manuscript is to provide examples identified by light microscopy in three poultry species. Transmission electron micrographs (TEM) of plasmacytes from the turkey oviduct mucosa are included for comparison with Mott cell light microscopic images. Wright's stained blood and bone marrow from commercial and specific pathogen free (SPF) chickens, ducks, and turkeys are the sources. Mott cell positive samples commonly occurred with leukocytosis or leukemoid reactions, polymicrobial bacteremia, and fungemia. Atypical granulocytes and leukocytes regularly accompanied Mott cells. It is proposed that circulating Mott cells are "sentinels" indicative of stress, dyscrasia, and pathology. Moreover, Mott cells, like other atypia, complicate the interpretation of simple heterophil/lymphocyte (H/L) ratios. As Mott cells are defective plasmacytes these observations address hematology, immunology, pathology, and welfare issues.

Effect of dietary supplementation with clay-based binders on biochemical and histopathological changes in organs of turkey fed with aflatoxin-contaminated diets.

This study was carried out to investigate the effect of dietary supplementation with molecular or nano-clay binders on biochemical and histopathological examination of organs of turkeys fed diets contaminated with aflatoxin B1. Two hundred and sixteen unsexed 1-day-old British United Turkeys were randomly allotted to nine diets in a 3 × 3 factorial arrangement of diets supplemented with no toxin binder, molecular toxin binder (MTB) and nano-clay toxin binder, each contaminated with 0, 60 and 110 ppb aflatoxin B1 respectively. There were three replicates per treatment with eight turkeys per replicate. Biochemical analyses, organ weights and histopathological changes of some organs were examined at the end of the study which lasted for 84 days. Turkeys fed diets supplemented with molecular and nano-binders showed higher (p < 0.001) total serum protein, reduced (p < 0.001) serum uric acid and GGT concentration values when compared with those fed aflatoxin-contaminated diets supplemented with no binder. Turkeys fed aflatoxin-contaminated diets supplemented with no binder had increased (p < 0.001) AST and ALT concentration when compared with other treatments. The heaviest (p < 0.001) liver and intestinal weight was noticed with turkeys fed diets supplemented with no binder and contaminated with 110 ppb aflatoxin B1 . Pathologically, there was no visible morphological alteration noticed in all turkeys fed uncontaminated diets and nano-clay-supplemented group. Hepatic paleness, hepatomegaly and yellowish discolouration of the liver were observed with turkeys fed diets containing no binder but contaminated with 60 and 110 ppb aflatoxin B1. Intestinal histopathological changes such as goblet cell hyperplasia, villous atrophy and diffuse lymphocytic enteritis were more prominent in turkeys fed diets containing no toxin binder and MTB. In conclusion, there were improved biochemical parameters and reduced deleterious effects of aflatoxin B1 in turkeys fed diet supplemented with clay binders. However, the improvement was more conspicuous in the nano-clay-supplemented group than molecular clay group.

Effect of protein source and protease addition on performance, blood metabolites and nutrient digestibility of turkeys fed on low-protein diets from 28 to 55 d post hatch.

The objective of this study was to investigate the effect of a monocomponent protease and dietary inclusion of canola meal (CM) and poultry by-product meal (PBM) on growth performance, carcass characteristics and blood metabolites of turkeys fed on low crude protein (CP) diets from 28 to 55 d post hatch. Experimental treatments included control, maize-soybean meal diet including 258.3 g/kg CP; negative control 1 (NC1), maize-soybean meal diet with reduced CP (232.4 g/kg); NC2, control diet (CP, 258.3 g/kg) including CM (80 g/kg) and PBM (80 g/kg); NC3, maize-soybean meal diet with reduced CP (232.4 g/kg) including CM (80 g/kg) and PBM (80 g/kg). Also, the NC1 + P and NC3 + P diets were created by addition of protease enzyme (30 000 units/kg of diet) to the NC1 and NC3 diets, respectively. The NC3 group had lower body weight gain (BWG) compared to those fed on the control diet, and no improvement with enzyme addition (NC3 + P) was achieved. The protease addition to the NC1 diet (NC1 + P) improved BWG to the level of the control diet. The NC1 group had higher feed conversion ratio (FCR) compared to the control and NC3 + P, but protease addition to the NC1 diet improved FCR. Protease addition to the low CP diets resulted in higher nitrogen (N) retention than in the control and NC2 groups. Also, the NC1 + P and NC3 + P diets increased apparent ileal digestibility (AID) of CP compared to the control group. It was concluded that addition of CM (up to 80 g/kg) and PBM (up to 80 g/kg) to turkey diets had no negative effect on growth performance from 28 to 55 d of age. The NC1 + P group achieved the BWG of the control group which was partially due to increases in N retention and AID of CP, but the NC3 + P group failed to recover the growth losses. This difference implies that the efficacy of the protease may depend upon the protein source in the ration.

Antioxidant status of blood and liver of turkeys fed diets enriched with polyunsaturated fatty acids and fruit pomaces as a source of polyphenols.

It was hypothesized that dietary polyphenol-rich fruit pomaces can improve the antioxidant status of both diets and the tissues of turkeys fed such diets. Turkeys were fed diets containing a cellulose preparation (C) or 5% dried apple pomace (AP), blackcurrant pomace (BCP), strawberry pomace (SP) and seedless strawberry pomace (SSP). Blood and liver biochemical parameters were determined in 7 birds from each experimental group slaughtered at 15 weeks of age, after 5 weeks of feeding diets containing soybean oil and linseed oil (approx. 1:1 ratio). Dietary linseed oil added to diets at 2.5% lowered the n-6/n-3 PUFA ratio from approx. 7:1 to below 2:1, thus reducing the antioxidant properties of diets measured using DPPH, ABTS and photo-chemiluminescence assays, compared with diets containing only soybean oil and administered to birds in the first phase of feeding. Fruit pomaces, in particular SSP with the highest polyphenol content (32.81 g/kg) and the highest antioxidant activity (256.4 µM Trolox/g), increased the antioxidant capacity of turkey diets. In comparison with the control group, the dietary treatments with fruit pomaces improved blood antioxidant parameters, including catalase activity (groups AP and BCP), the total antioxidant capacity of hydrophilic (group AP) and lipophilic (groups AP, SP, and SSP) compounds, peroxide levels (groups AP and SSP) and antioxidant capacity measured by the FRAP (ferric reducing antioxidant power of plasma) assay (groups AP, BCP and SSP). Significantly lower concentrations of both vitamin E and thiobarbituric acid reactive substances (TBARS) were noted in the livers of turkeys fed all diets with dried fruit pomaces.

The effect of dietary methionine levels on fattening performance and selected blood and tissue parameters of turkeys.

A total of 490 eight-week-old female Hybrid Converter turkeys (body weight 4.11 ± 0.03 kg) were divided into 5 groups with 7 replicates of 14 birds each. For 8 weeks, basal diets were supplemented with methionine (Met) at following levels (weeks 9-12/weeks 13-16 of age): Group 1 - 0.34/0.29%, Group 2 - 0.39/0.34%, Groups 3 and 4 - 0.45/0.38% and 0.51/0.41%, respectively, Group 5 - 0.58/0.47%. Only in the first feeding phase the body weight gain (BWG) was affected by Met levels with the significantly highest BWG in Group 3. No treatment effects were found for feed conversion ratio, carcass yield, carcass composition and meat colour. The blood superoxide dismutase activity was significantly highest in Groups 2 and 3. The concentrations of reduced glutathione in the liver were linearly increased (p = 0.018), whereas the ratio of reduced glutathione to oxidised glutathione was highest in Group 3 (quadratic contrast, p = 0.004). It can be concluded that turkeys from Group 3 (Met levels age depending 15% and 10% above recommendations by NRC) were characterised by a well-balanced physiological response. Attention should be paid to the immune response of birds to higher dietary Met levels: plasma IgA concentrations decreased, whereas IL-6 and TNF-α levels increased in turkeys fed diets with the highest Met content.

Circulating extracellular miR-22, miR-155, and miR-365 as candidate biomarkers to assess transport-related stress in turkeys.

MicroRNA (miRNA) have been identified in circulating blood and might have the potential to be used as biomarkers for several pathophysiological conditions. To identify miRNA that are altered following stress events, turkeys (Meleagris gallopavo) were subjected to 2 h of road transportation. The expression levels of five circulating miRNA, namely miR-22, miR-155-5p, miR-181a-3p, miR-204 and miR-365-3p, were detected and assessed by quantitative polymerase chain reaction using TaqMan® probes, as potential biomarkers of stress. The areas under the receiver operating characteristic curves were then used to evaluate the diagnostic performance of miRNA. A panel of three stress-responsive miRNA, miR-22, miR-155 and miR-365 were identified; their expression levels were significantly higher after road transportation and the area under the curve (AUC) were 0.763, 0.71 and 0.704, respectively. Combining the three miRNA a specificity similar to the one found for the three miRNA separately was found. The AUC of the weighted average of the three miRNA was 0.763. This preliminary study suggests that the expression levels of circulating miR-22, miR-155 and miR-365 are increased during transport-related stress and that they may have diagnostic value to discriminate between stressed- and unstressed animals.

Antioxidant status of turkey breast meat and blood after feeding a diet enriched with histidine.

The objective of this study was to investigate the effects of 1) spray dried blood cells rich in histidine and 2) pure histidine added to feed on the antioxidant status and concentration of carnosine related components in the blood and breast meat of female turkeys. The experiment was performed on 168 Big7 turkey females randomly assigned to 3 dietary treatments: control; control with the addition of 0.18% L-histidine (His); and control with the addition of spray dried blood cells (SDBC). Birds were raised for 103 d on a floor with sawdust litter, with drinking water and feed ad libitum. The antioxidant status of blood plasma and breast muscle was analyzed by ferric reducing ability (FRAP) and by 2,2-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radicals scavenging ability. The activity of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) was analyzed in the blood and breast meat, with the content of carnosine and anserine quantified by HPLC. Proximate analysis as well as amino acid profiling were carried out for the feed and breast muscles. Growth performance parameters also were calculated. Histidine supplementation of the turkey diet resulted in increased DPPH radical scavenging capacity in the breast muscles and blood, but did not result in higher histidine dipeptide concentrations. The enzymatic antioxidant system of turkey blood was affected by the diet with SDBC. In the plasma, the SDBC addition increased both SOD and GPx activity, and decreased GPx activity in the erythrocytes. Feeding turkeys with an SDBC containing diet increased BW and the content of isoleucine and valine in breast muscles.

DIAGNOSING LYMPHOPROLIFERATIVE DISEASE VIRUS IN LIVE WILD TURKEYS (MELEAGRIS GALLOPAVO) USING WHOLE BLOOD.

Lymphoproliferative disease virus (LPDV) is a retrovirus that infects wild and domestic turkeys ( Meleagris gallopavo ). The first cases of LPDV in the United States were diagnosed in 2009, and subsequent surveillance has revealed the virus to be widespread in wild turkey populations throughout the eastern half of the country. More research is needed to determine whether LPDV is having a negative effect on turkey populations, but progress has been impeded by the lack of a simple method for diagnosing the virus in living birds. Infected animals may appear asymptomatic, and diagnostics currently rely on tissue or bone marrow, which can be difficult to obtain. This study investigated the reliability of polymerase chain reaction (PCR) to detect LPDV in whole blood, compared with previous methods using buffy coat (concentrated white blood cells) and bone marrow. Paired samples of whole blood and buffy coat were collected from 137 live turkeys and paired samples of whole blood and bone marrow were collected from 32 turkeys postmortem. Compared with buffy coat, whole blood had 97% sensitivity and 100% specificity. When compared with bone marrow, whole blood had 100% sensitivity and 89% specificity. Both comparisons had a high degree of agreement using Cohen's kappa statistic. Based on these results, PCR of whole blood provides detection of LPDV in living birds that is on par with both buffy coat and bone marrow.