Propidium monoazide (PMA) qPCR assay compared to the plate count method for quantifying the growth of Salmonella enterica serotypes in vacuum-packaged turkey breast combined with a mathematical modeling approach.

This study compares the plate count (PC) and the Propidium Monoazide-quantitative Polymerase Chain Reaction (PMA-qPCR) methods to assess the growth of a cocktail of three serotypes of Salmonella enterica (Heidelberg, Typhimurium, and Enteritidis) in cooked, sliced, and vacuum-packaged turkey breast (STB) under isothermal storage temperatures (8 °C-20 °C), using predictive models. Standard curves were developed for PMA-qPCR, demonstrating high efficiency (101%) and sensitivity, with quantification limits ranging from 1 to 2 log10 CFU/g for all temperatures studied. Comparative analysis revealed a significant correlation (R2 = 0.99; 95% CI) between the PC and PMA-qPCR methods; however, the agreement analysis indicated a mean difference (Bias) of -0.11 log10 CFU/g (p < 0.05), suggesting underestimation by the PC method. This indicates the presence of stressed or viable but nonculturable (VBNC) cells, detectable by PMA-qPCR but not by PC. The Baranyi and Roberts model showed a good ability to describe the behavior of S. enterica cocktail in STB for PC and PMA-qPCR data under all isothermal conditions. The exponential secondary model more accurately represented the temperature dependence of the maximum specific growth rate compared to the Ratkowsky square root model, with R2 values ≥ 0.984 and RMSE values ≤ 0.011 for both methods. These results suggest that combining PMA-qPCR with predictive modeling allows for a more accurate prediction of S. enterica growth, compared to PC method. In the event of cold chain disruptions of meat products, the use of PMA-qPCR method allow the quantification of VBNC cells, that can still pose a health risk to consumers, especially in ready-to-eat products.

Ligand-induced conformational changes in the ß1-adrenergic receptor revealed by hydrogen-deuterium exchange mass spectrometry.

G Protein Coupled Receptors (GPCRs) constitute the largest family of signalling proteins responsible for translating extracellular stimuli into intracellular functions. They play crucial roles in numerous physiological processes and are major targets for drug discovery. Dysregulation of GPCRs is implicated in various diseases, making understanding their structural dynamics critical for therapeutic development. Here, we use Hydrogen Deuterium Exchange Mass Spectrometry (HDX-MS) to explore the structural dynamics of the turkey ß1-adrenergic receptor (tß1AR) bound with nine different ligands, including agonists, partial agonists, and antagonists. We find that these ligands induce distinct dynamic patterns across the receptor, which can be grouped by compound modality. Notably, full agonist binding destabilises the intracellular loop 1 (ICL1), while antagonist binding stabilises it, highlighting ICL1's role in G protein recruitment. Our findings indicate that the conserved L72 residue in ICL1 is crucial for maintaining receptor structural integrity and stabilising the GDP-bound state. Overall, our results provide a platform for determining drug modality and highlight how HDX-MS can be used to dissect receptor ligand interaction properties and GPCR mechanism.

Immunological Response and Nutritional Effects of Lactobacillus spp.-fermented Garlic on Turkey Broilers.

In the era of free antibiotics used in animal production, the application of feed additives should be prioritized to improve poultry health and production.  The present study was conducted to evaluate the influences of garlic fermented by Lactobacillus spp. on the growth rate, intestinal microorganisms, and immune response of turkey broilers. A completely randomized design was used, involving 90 turkey broilers aged 1-56 days, with five treatments and three replicates per treatment. The birds were given feed and water ad libitum for the entire experiment period. The treatments included the supplementation of aqueous extract from fermented garlic (FG) to drinking water. The results showed that broilers supplemented with 0.8% FG exhibited the largest final body weight (1,158 g/bird), body weight gain (19.64 g/bird/day), and significantly improved feed conversion ratio (1.962) while decreasing the feed intake of turkey broilers from to 1-56 days. The immune organ indices, including the spleen, thymus, and bursa of Fabricius indices, were increased in the 0.6% FG treatment group (P<0.05), while antibody titers (at 28 and 42 days of age) were improved in the 0.6% and 0.8% FG treatments (P<0.05). Clostridium perfringens and Salmonella spp. were not detected in the intestines of these birds, while the amount of Escherichia coli was reduced (P<0.05) and Lactobacillus spp. increased (P>0.05) without a significant effect. It can be concluded that supplementation with 0.8% FG improved growth performance, and 0.6% FG may enhance the immunity of turkeys. Moreover, 0.6% and 0.8% FG could be widely used for poultry production.

Cochlosoma anatis in Poultry and its Industry Impact.

Cochlosoma anatis is a flagellated protozoan parasite classified in the Trichomonadidae family and is the causative agent of cochlosomiasis, an enteric disease of turkeys, waterfowl, and other wild birds. Cochlosomiasis symptoms largely consist of watery diarrhea, lethargic birds, depressed weight gain, and widespread flock morbidity causing flock nonuniformity. The known distribution of C. anatis is centered around areas of turkey production farms in the southeast United States, e.g., North Carolina, Missouri and Arkansas, but has been reported in other states and some other countries. Diagnosis is confirmed through examination of enteric mucosal scrapings using light microscopy. Following the withdrawal of approval of effective antiprotozoal medications for use in commercial animal production, cochlosomiasis has become a greater concern for commercial turkey industry professionals. Transmission of C. anatis occurs via the fecal-oral route, but the organism is fragile outside the host, suggesting the implication of a vector in the introduction of disease to susceptible farms. Research regarding C. anatis pathogenicity, transmission, and environmental involvement has been limited, creating a gap in cochlosomiasis knowledge. Future research is needed to further explore ways to prevent and treat cochlosomiasis, with needs centered on disease pathogenesis, transmission patterns, and prophylaxis and treatment methods.

Estudio recapitulativo- Cochlosoma anatis en la avicultura comercial y su impacto en esta industria. Cochlosoma anatis es un parásito protozoario flagelado clasificado dentro de la familia Trichomonadidae y es el agente causante de la coclosomiasis, que es una enfermedad entérica de pavos, aves acuáticas y otras aves silvestres. Los signos de la coclosomiasis consisten principalmente en diarrea acuosa, aves con letargo, disminución de la ganancia de peso y morbilidad generalizada de las parvadas, lo que provoca su falta de uniformidad. La distribución conocida de C. anatis gira en torno a zonas de granjas de producción de pavos en el sudeste de los Estados Unidos, principalmente en Misuri y Arkansas, pero se ha notificado su presencia en otros estados y en algunos otros pa'ises. El diagnóstico se confirma mediante el examen de raspados de mucosa entérica con microscop'ia óptica. Tras el retiro de la aprobación de medicamentos antiprotozoarios eficaces para su uso en la producción animal comercial, la coclosomiasis se ha convertido en una preocupación importante para los profesionales de la industria de pavos comerciales. La transmisión de C. anatis se produce por v'ia fecal-oral, pero es frágil fuera del hospedador, lo que sugiere la participación de un vector en la introducción de la enfermedad en granjas susceptibles. La investigación sobre la patogenicidad, la transmisión y el efecto ambiental de C. anatis ha sido limitada, lo que crea un vac'io en el conocimiento de la coclosomiasis. En el futuro será necesario investigar más a fondo las formas de prevenir y tratar la coclosomiasis, con la necesidad de enfocarse en la patogenia de la enfermedad, los patrones de transmisión y los métodos de profilaxis y tratamiento.

Comparison of magnetic seed and RFID methods in the localization of non-palpable breast lesions.

BACKGROUND: Many methods have been developed for localizing non-palpable breast lesions. This study investigated the success rate and surgical results of the magnetic seed (Magseed) and radiofrequency identification (RFID) method, which are relatively new compared to standard wire-guided localizations. MATERIALS AND METHODS: 20 simulation (10 Magseed, 10 RFID) models were created using turkey breasts and raisins. Raisins containing magnetic seed and RFID tags were placed on the turkey breast. Sentimag® probe was used for the Magseed group, and Faxitron LOCalizer™ System device was used in the RFID group. Both methods were evaluated in terms of accuracy in detecting breast lesion localization, operation times, excised tissue weights, total resection volume, surgical margin negativity, and re-excision rates. RESULTS: Lesion localization success in both techniques was 100%. While procedure times were statistically significantly shorter in the Magseed group, incision lengths were shorter in the RFID group (P = 0.013, P = 0.007, respectively). No statistically significant difference was found between the groups for the weight of the removed parts, total resection volume, and surgical margin distance (P > 0.05). CONCLUSION: In this feasibility study, it was concluded that neither the RFID nor Magseed methods had a significant advantage over each other, in terms of localization detection and surgical margin negativity, and both methods could be used successfully for localization.

Sequencing and characterization of complete mitogenome DNA of worldwide turkey (Meleagris gallopavo) populations.

The history of turkey (Meleagris gallopavo) domestication can be traced back to the period between 700 and 200 BC in Mexico. This process involved multiple contributors and resulted in the development of modern local turkey breeds. This research investigates the complete mitochondrial diversity across a diverse range of local turkeys. Seventy-three turkeys were sampled from various populations, including autochthonous Italian breeds, an American breed (Narragansett), as well as wild turkeys from the USA and Mexico. The mitochondrial DNA (mtDNA) was employed as a powerful tool for biodiversity and breed phylogeny investigation. An analysis of the entire mtDNA was conducted to identify breed-specific unique traits, mitochondrial-specific characteristics, and the phylogenetic relationship among turkey populations. A total of 44 polymorphic sites were identified. Brianzolo and Narragansett birds were characterized as genetically homogeneous populations. Thirty-two different haplotypes were identified when our samples were compared with mtDNA D-loop of 96 online available turkeys from various geographical countries. H1 and H2, differing for one mutation, were the most abundant, comprising 132 of the 185 sequences. H1 included samples coming from every region, while H2 was predominantly characterized by Italian samples. USA and Mexican samples appear to be more variable in their mtDNA than the other populations.

A Comparison of White and Yellow Seminal Plasma Phosphoproteomes Obtained from Turkey (Meleagris gallopavo) Semen.

Seminal plasma is rich in proteins originating from various male reproductive organs. The phosphorylation of these proteins can significantly impact sperm motility, capacitation, and acrosome reaction. Phosphoproteomics identifies, catalogues, and characterizes phosphorylated proteins. The phosphoproteomic profiling of seminal plasma offers valuable insights into the molecular mechanisms that influence semen quality and male fertility. Thus, the aim of this study was a phosphoproteomic analysis of white and yellow turkey seminal plasma. The experimental material consisted of 100 ejaculates from BIG-6 turkeys between 39 and 42 weeks of age. The collected white and yellow turkey seminal plasmas were analyzed for total protein content; the activity of selected enzymes, i.e., alkaline phosphatase (ALP), acid phosphatase (ACP), superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT); and the content of reduced glutathione (GSH) and malondialdehyde (MDA). Phosphoproteins were isolated from white and yellow seminal fluids, and the resulting protein fractions were separated by SDS-PAGE and Western blotting. Phosphorylated residues were immunodetected, and the isolated phosphoproteins were identified (nano LC-MS/MS). Yellow seminal plasmas were characterized by higher levels of total protein, GSH, and MDA, as well as higher levels of ALP, ACP, and GPx activity. There were no significant differences in the activity of SOD and CAT. A total of 113 phosphoproteins were identified in turkey seminal fluids. The functional analysis demonstrated that these phosphoproteins were mainly involved in oocyte fertilization, organization and metabolism of the actin cytoskeleton, amplification of the intracellular signal transduction pathway, general regulation of transport, vesicular transport, proteome composition of individual cellular compartments, and the organization and localization of selected cellular components and macromolecules. Increased phosphorylation of the fractions containing proteins encoded by SPARC, PPIB, TRFE, QSOX1, PRDX1, PRDX6, and FASN genes in white plasmas and the proteins encoded by CKB, ORM2, APOA1, SSC5D, RAP1B, CDC42, FTH, and TTH genes in yellow plasmas was observed based on differences in the optical density of selected bands. The obtained results indicate that the phosphorylation profiles of turkey seminal plasma proteins vary depending on the type of ejaculate.

Salmonella associated with agricultural animals exhibit diverse evolutionary rates and show evidence of recent clonal expansion.

Most foodborne salmonellosis outbreaks are linked to agricultural animal products with a few serovars accounting for most Salmonella isolated from specific animal products, suggesting an adaptation to the corresponding animal hosts and their respective environments. Here, we utilized whole-genome sequence (WGS) data to analyze the evolution and population genetics of seven serovars frequently isolated from ground beef (Montevideo, Cerro, and Dublin), chicken (Kentucky, Infantis, and Enteritidis), and turkey (Reading) in the United States. In addition, publicly available metadata were used to characterize major clades within each serovar with regard to public health significance. Except for Dublin, all serovars were polyphyletic, comprising 2-6 phylogenetic groups. Further partitioning of the phylogenies identified 25 major clades, including 12 associated with animal or environmental niches. These 12 clades differed in evolutionary parameters (e.g., substitution rates) as well as public health relevant characteristics (e.g., association with human illness, antimicrobial resistance). Overall, our results highlight several critical trends: (i) the Salmonella generation time appears to be more dependent on source than serovar and (ii) all serovars contain clades and sub-clades that are estimated to have emerged after the year 1940 and that are enriched for isolates associated with humans, agricultural animals, antimicrobial resistance (AMR), and/or specific geographical regions. These findings suggest that serotyping alone does not provide enough resolution to differentiate isolates that may have evolved independently, present distinct geographic distribution and host association, and possibly have distinct public health significance. IMPORTANCE: Non-typhoidal Salmonella are major foodborne bacterial pathogens estimated to cause more than one million illnesses, thousands of hospitalizations, and hundreds of deaths annually in the United States. More than 70% of Salmonella outbreaks in the United States have been associated with agricultural animals. Certain serovars include persistent strains that have repeatedly contaminated beef, chicken, and turkey, causing outbreaks and sporadic cases over many years. These persistent strains represent a particular challenge to public health, as they are genetically clonal and widespread, making it difficult to differentiate distinct outbreak and contamination events using whole-genome sequence (WGS)-based subtyping methods (e.g., core genome allelic typing). Our results indicate that a phylogenetic approach is needed to investigate persistent strains and suggest that the association between a Salmonella serovar and an agricultural animal is driven by the expansion of clonal subtypes that likely became adapted to specific animals and associated environments.

The effect of early administration of antibiotics or feeding a diet containing coccidiostats on the level of their accumulation in liver and the redox status of turkeys.

Early administration of antibiotics may worsen the functioning of the turkeys' antioxidant system. It was also assumed that the longer the time of administration of an antibiotic, e.g. a coccidiostat, the greater the risk of its accumulation in the liver. The study aimed to determine whether early administration of antibiotics or feeding a diet containing coccidiostats causes accumulation in the liver and whether it affects the deterioration of the antioxidant system, and whether preventive vaccinations can intensify it. A total of 3 080 female turkeys were randomly allocated to eight groups. The experiment had a two-factorial design, with four treatments (C, M, E, D) and two groups of birds (vaccinated +, unvaccinated -). The C group did not receive the coccidiostat or antibiotics. Group M was administered monensin at 90 mg/kg feed for 56 days of life. Group E received enrofloxacin at 10 mg/kg BW, and group D received doxycycline at 50 mg/kg BW, added to drinking water, for the first 5 days of life. One-day-old turkeys from groups C+, M+, E+, and D+ were administered live-attenuated vaccines against turkey rhinotracheitis and Newcastle disease by coarse spray; 28-day-old birds were administered a subcutaneously injected inactivated vaccine against Ornithobacterium rhinotracheale. Turkeys from groups C-, M-, E-, and D- were not vaccinated. It was determined that as a result of administration of enrofloxacin or doxycycline until the 5th day of life, biotransformation of these antibiotics occurred in the liver until the 56th day of life of the turkeys, which was confirmed by their lower level than the Maximum Residue Level. Because the concentration of monensin in the liver of turkeys gradually increased with the extension of the time of its administration in the diet, it is probable that discontinuing its addition a day before the slaughter of birds will result in the presence of this coccidiostat in the liver of turkeys. Despite the accumulation of monensin in the liver of turkeys, this coccidiostat did not increase oxidative reactions in the organism of turkeys. Vaccination of turkeys can reduce oxidative reactions and apoptosis in the body. However, the effect of the redox system reaction is different immediately after vaccination, which is due to the mechanism of action of the immune system. If it is necessary to administer an antibiotic in the early rearing period, the effects of doxycycline on the organism's immunity including antioxidant defence will be less severe than those of enrofloxacin.

Amino acid and energy digestibility of conventional corn dried distillers grains with solubles, high protein distillers dried grains, and corn fermented protein, and impacts on turkey poult growth performance and intestinal characteristics.

Amino acids (AA) are an expensive nutritional components of poultry diets. Distillers dried grains with solubles (DDGS) is the primary co-product produced by the dry grind bioethanol industry, although new technologies are being implemented to produce high protein distillers dried grains (HP-DDG) and corn fermented protein (CFP), but data on their nutritive value in poultry are lacking. Two experiments (EXP) were conducted to determine the energy and AA digestibility of DDGS, HP-DDG, and CFP in poults in addition to a feeding trial to evaluate increasing dietary levels of HP-DDG and CFP on growth performance and intestinal characteristics. In EXP 1, 6 different DDGS sources were evaluated using poults to determine their nitrogen-corrected apparent metabolizable energy (AMEn) concentrations, and cecectomized roosters were used to determine their standardized ileal (SID) AA digestibility (SID-AA). In EXP 2, AMEn and SID-AA for HP-DDG and CFP were determined in young poults, and a feeding trial was conducted to evaluate growth performance and intestinal morphology and permeability of poults fed diets containing 7.5 and 15% HP-DDG or CFP from 1 to 32 d of age. In EXP 1, the AMEn concentration among the DDGS samples ranged from 2,530 to 3,573 kcal/kg DM but was not different (P = 0.57) among the samples, with an average SID for LYS of 66.6%. In EXP 2, different (P = 0.001) AMEn concentrations for HP-DDG and CFP were observed (3,114 and 3,760 kcal/kg DM, respectively), with the SID for LYS being 66.55 and 77.00% for HP-DDG and CFP, respectively. Including HP-DDG or CFP into the diet at 7.5 and 15% had no effect (P > 0.05) on growth, feed intake, or feed conversion. Neither co-product nor its inclusion rate affected intestinal morphology and permeability (P > 0.05). Overall, DDGS, HP-DDG, and CFP are excellent sources of AMEn and digestible AA, with dietary inclusion rates of up to 15% of HP-DDG or CFP having no impact on growth or intestinal characteristics.

The effect of avian influenza outbreaks on retail price premiums in the United States poultry market.

This study analyzes the effect of avian influenza outbreaks on retail price premiums in the US poultry market. We estimate hedonic price models for eggs, chickens, and turkeys, controlling for quality characteristics, unobserved time, and regional factors. To measure the impact of avian influenza outbreaks we use 2 proxies. The first proxy is a measure of the number of new bird infections at the production level. The second proxy measures online search queries related to the outbreak. The results show that, on average, prices increase across product categories, that is, egg, broiler, and turkey markets, during avian influenza outbreaks. Furthermore, we observe price convergence and reduced dispersion within product categories, which is consistent with the economic theory of asymmetric substitutability between conventional and premium products. Our analysis finds that the HPAI outbreak caused a reduction of the price gap between conventional and premium products.

Dietary concoction of formic acid and thymol and its effects on zoo-technical performance, immunity, jejunal architecture and gut health in Turkey.

With increasing demand for improved protein-rich food, diverse poultry birds like turkey are gaining popularity in India. However, with the rising concerns of antibiotic residues and drug resistance, safe alternatives like formic acid (FA) and thymol (TH) have attracted the attention of researchers as effective replacer of antibiotic growth promoters (AGP). This experiment investigated the effects of combinations of FA and TH on growth performance, immunity, gut microflora and jejunal microstructures in turkey. A total of 240 turkey poults were reared in 6 treatment groups for a period of 16 weeks with standard management and feeding: T1 (basal diet only); T2 (AGP); T3 (FA@2.5 ml + TH@240 mg/kg); T4 (FA@2.5 ml + TH@360 mg/kg); T5 (FA@7.5 ml + TH@240 mg/kg) and T6 (FA@7.5 ml + TH@360 mg/kg). The results revealed that T5 group had the highest final body weight and best FCR while the feed consumption across the groups was comparable. The immune status of the turkey improved (p ≤ 0.05) in treatment groups compared to control with T4 and T5 group showcasing the best CBH response, antibody titres and relative immune organ weight index. A significant improvement (p ≤ 0.05) in jejunal microstructures was observed at 16th week in T5 group over control and AGP fed group. There was significant (p ≤ 0.001) reduction in total plate and coliform counts, but a positive shift was seen in Lactobacillus population in T5 group as compared to control and AGP fed group. In conclusion, the concoction of formic acid (7.5 ml/kg) + thymol (240 mg/kg) in-feed supplement improved the performance, immune status and gut health in turkey as an efficient alternative to AGPs.

Ultrasound Shear Wave Elastography for Noninvasive Diagnosis of Acute Compartment Syndrome Using a Novel In Vivo Turkey Model.

INTRODUCTION: Acute Compartment Syndrome (ACS) is a severe trauma caused by elevated intra-muscle-compartment pressure (ICP). The current standard method for diagnosis is to insert a needle into the muscle sterilely under anesthesia. However, to secure the environment is sometimes not easy and leads to delays in diagnosis. Recently, we have focused on shear wave ultrasound elastography (SWE) as an alternative, which can be done concisely in unclean environment and without anesthesia. We would like to report the usefulness of SWE for ACS diagnosis using 2-pedal walking turkey model recently developed in our lab. MATERIALS AND METHODS: A total of 32 1-year-old Bourbon turkeys were used. 5% solution of chicken albumin was infused continuously into the tibialis cranialis (TC) muscle using IV pump. The ICP was increased stepwise from 0 to 50 mmHg. During the rising of ICP, the correlation between values of SWE (kPa) and ICP (mmHg) was measured. After the ICP reached 50 mmHg, half of the turkeys were maintained at this pressure for 2 hours and the rest for 6 hours. After infusion, a fasciotomy was performed on the half turkey. Half of the turkeys were euthanized after 2 weeks and the rest after 6 weeks. SWE of TC muscle and walking gait data on turkeys using a portable walkway system were measured weekly until euthanasia. At euthanasia, isometric tetanic muscle force (ITF) tests to TC muscle and histological evaluations were performed. RESULTS: SWE value (kPa) was highly significantly correlated to the actual ICP (mmHg) (R2 = 0.91). Stance of ACS side leg were significantly extended, and swing of the control side shortened from the second to the third week after ACS in the 6 hours infusion-no-fasciotomy group (P < 0.05*). ITF was significantly reduced mainly in the 6 hours infusion group (P < 0.05*). Histological evaluation revealed that in the 6 hours infusion and 6 weeks survival group, both the muscle fiber and intercellular distances were significantly expanded (P < 0.05). CONCLUSION: SWE seems to be a substitute measure of ICP in diagnosing ACS. With regard to our in vivo ACS model using turkey, survival at 50 mmHg ICP for 6 hours and 6 weeks post ACS would be an appropriate situation.

Impact of Thyme Essential Oil on the Aroma Profile and Shelf Life of Vacuum-Packed Minced Turkey Meat.

There is considerable interest in the use of essential oils for food preservation, but their effect on the aroma profile of a product is poorly understood. This study investigated the effect of thyme essential oil (EO) addition at increasing concentrations (0.005, 0.01, 0.02, and 0.03% v/w) on the volatile compound composition of vacuum-packed minced turkey meat after storage for 8 days at 1-2 °C. The aroma profile of the meat was determined using the HS-SPME/GCMS (headspace solid-phase microextraction/gas chromatography-mass spectrometry) method. The results were also analysed by PCA (principal component analysis). The addition of thyme EO had a modifying effect on the aroma profile of meat-derived components, e.g., the formation of benzeneacetaldehyde, benzyl alcohol, 4,7-dimethylbenzofuran, hexathiane, hexanal, and 1-hexanol was reduced and the appearance of 9-hexadecenoic acid was observed in the stored samples. The increase in EO concentration affected the levels of its individual components in the meat headspace in different ways. In terms of fat rancidity indices, even a 0.005% addition of this essential oil significantly reduced the peroxide value. Quantitative descriptive analysis (QDA) showed that the addition of thyme EO reduced or masked the intensity of unpleasant odours associated with meat spoilage. In the aroma analysis, the turkey with 0.02% v/w EO scored highest, and pleasant citrus notes were found.

Characterizing the splice map of Turkey Hemorrhagic Enteritis Virus.

BACKGROUND: Hemorrhagic enteritis, caused by Turkey Hemorrhagic Enteritis Virus (THEV), is a disease affecting turkey poults characterized by immunosuppression and bloody diarrhea. An avirulent THEV strain that retains the immunosuppressive ability is used as a live vaccine. Characterizing the splice map of THEV is an essential step that would allow studies of individual genes mediating its immunosuppressive functions. We used RNA sequencing to characterize the splice map of THEV for the first time, providing key insights into the THEV gene expression and mRNA structures. METHODS: After infecting a turkey B-cell line with the vaccine strain, samples in triplicates were collected at 4-, 12-, 24-, and 72-hours post-infection. Total RNA was extracted, and poly-A-tailed mRNA sequenced. Reads were mapped to the THEV genome after trimming and transcripts assembled with StringTie. We performed PCR of THEV cDNA, cloned the PCR products, and used Sanger sequencing to validate all identified splice junctions. RESULTS: Researchers previously annotated the THEV genome as encoding 23 open reading frames (ORFs). We identified 29 spliced transcripts from our RNA sequencing data, all containing novel exons although some exons matched some previously annotated ORFs. The three annotated splice junctions were also corroborated by our data. During validation we identified five additional unique transcripts, a subset of which were further validated by 3' rapid amplification of cDNA ends (3' RACE). Thus, we report that the genome of THEV contains 34 transcripts with the coding capacity for all annotated ORFs. However, we found six of the previously annotated ORFs to be truncated ORFs on the basis of the identification of an in-frame upstream start codon or the detection of additional coding exons. We also identified three of the annotated ORFs with longer or shorter isoforms, and seven novel unannotated ORFs that could potentially be translated; although it is beyond the scope of this manuscript to investigate whether they are translated. CONCLUSIONS: Similar to human adenoviruses, all THEV transcripts are spliced and organized into five transcription units under the control of their cognate promoters. The genes are expressed under temporal regulation and THEV also produces multiple distinctly spliced transcripts that code for the same protein. Studies of the newly identified potential proteins should be urgently performed as these proteins may have roles in THEV-induced immunosuppression. Also, knowing the splicing of THEV genes should be invaluable to future research focusing on studying THEV genes, as this will allow accurate cloning of the mRNAs.

A Novel Training Model for Superficial Temporal Artery- Middle Cerebral Artery Anastomosis Using Microsurgical Techniques.

OBJECTIVE: To create a reusable and inexpensive training model with technological tools that simulates cerebral bypass surgery and a sensor system that provides tactile feedback to the surgeon. Furthermore, we aimed to evaluate the anastomotic stability and contribution to the surgeon's learning curve. METHODS: We created a superficial temporal artery-middle cerebral artery bypass simulation model using chicken and turkey brachial arteries. A cranium model was printed with a three-dimensional printer for craniotomy and cerebral parenchyma was created by pouring silicone into the cranial mold. A blood flow simulation system was also prepared. Pressure-sensitive sensors were placed on parenchyma and tactile conditioning was performed via audible warning from the sensors. Twenty-four anastomosis were performed with different sutures and hand tools. Anastomosis completion times and durability and the number of touches and pressures applied to the parenchyma were recorded. The stability of the anastomoses was evaluated by increasing the pressure in the blood flow simulation system, so usefulness of the training model was evaluated. RESULTS: The time required for anastomosis completion decreased as the number of practices increased (P < 0.05). As the number of practices increased, the number of parenchymal touches decreased (P < 0.05). CONCLUSIONS: With practice, the time required for anastomosis completion and number of parenchymal touches decreased. Thus, the model is useful, inexpensive, reusable, easily accessible, and contributes to the surgeon's learning curve. Our model with pressure-sensitive sensors can be used for microsurgery practice, enabling the surgeons to gain tactile conditioning and evaluate anastomotic stability and leakage.

Reliability, practicability and farmers' acceptance of an animal welfare assessment protocol for broiler chickens and turkeys.

This study presents an evaluation of on-farm self-assessment using animal-based indicators to support fattening poultry farmers in managing the welfare of their animals. Self-assessment guidelines elaborated by a German expert group were evaluated together with 11 trained broiler and 11 trained turkey farmers. The participating farmers tested a protocol with 18 indicators for broilers and a protocol with 20 indicators for turkeys on their farms for 1 y. The reliability of individual animal scoring, the practicability of the protocols, their implementation, and acceptance were then evaluated. Reliability was tested during 2 farm visits by the accompanying scientists, using the scientist as a silver standard. On average, the farmers achieved very good reliability (mean PABAK - prevalence-adjusted and bias-adjusted kappa, broilers: 0.90; turkeys: 0.86), with no detectable influence of the previous training method (online versus in-person), the first versus the second visit of the scientists, the fattening stage of the animals scored, number of animals on the farm or the farmers' professional background. The assessment took longer at the end of the fattening period for both animal species. On farms with more personnel and for farmers with a higher position on farm, the assessment was easier to integrate into their work. Most farmers did not fully document their self-assessment and almost no farmer processed and evaluated the data properly, even though it seemed interesting to them. Overall, there was only moderate acceptance of the welfare self-assessment approach, with mixed responses as to whether it provided early warning information or benefits to animal management. Farmers often pointed to the increased cost associated with carrying out self-assessments in terms of the additional working time required, which is likely to be an important barrier to continuing. Digital applications for data recording, processing and evaluation may help to overcome barriers. Overall, it appears unlikely that the welfare self-assessment approach will be widely implemented in the poultry fattening sector without further incentives.

An affordable detection system based on RT-LAMP and DNA-nanoprobes for avian metapneumovirus.

Airborne animal viral pathogens can rapidly spread and become a global threat, resulting in substantial socioeconomic and health consequences. To prevent and control potential epidemic outbreaks, accurate, fast, and affordable point-of-care (POC) tests are essential. As a proof-of-concept, we have developed a molecular system based on the loop-mediated isothermal amplification (LAMP) technique for avian metapneumovirus (aMPV) detection, an airborne communicable agent mainly infecting turkeys and chickens. For this purpose, a colorimetric system was obtained by coupling the LAMP technique with specific DNA-functionalized AuNPs (gold nanoparticles). The system was validated using 50 different samples (pharyngeal swabs and tracheal tissue) collected from aMPV-infected and non-infected chickens and turkeys. Viral detection can be achieved in about 60 min with the naked eye, with 100% specificity and 87.88% sensitivity for aMPV. In summary, this novel molecular detection system allows suitable virus testing in the field, with accuracy and limit of detection (LOD) values highly close to qRT-PCR-based diagnosis. Furthermore, this system can be easily scalable to a platform for the detection of other viruses, addressing the current gap in the availability of POC tests for viral detection in poultry farming. KEY POINTS: •aMPV diagnosis using RT-LAMP is achieved with high sensitivity and specificity. •Fifty field samples have been visualized using DNA-nanoprobe validation. •The developed system is a reliable, fast, and cost-effective option for POCT.

Exploring the Effects of an Alfalfa Leaf-Derived Adsorbent on Microbial Community, Ileal Morphology, Barrier Function, and Immunity in Turkey Poults during Chronic Aflatoxin B1 Exposure.

This article follows-up on our recently published work, which evaluated the impact of the addition of an alfalfa leaf-derived adsorbent in the aflatoxin B1 (AFB1)-contaminated diet in regard to the production parameters, blood cell count, serum biochemistry, liver enzymes, and liver histology of turkey poults. This paper presents complementary results on microbial community, ileal morphology, barrier function, and immunity. For this purpose, 350 1-day-old female turkey poults were randomly distributed into five groups: (1) Control, AFB1-free diet; (2) AF, AFB1-contaminated diet at 250 ng/g; (3) alfalfa, AFB1-free diet + 0.5% (w/w) adsorbent; (4) alfalfa + AF, AFB1-contaminated diet at 250 ng/g + 0.5% (w/w) adsorbent; and (5) YCW + AF, AFB1-contaminated diet at 250 ng/g + 0.5% (w/w) commercial yeast cell wall-based adsorbent (reference group). In general, in the AF group, the growth of opportunistic pathogens was promoted, which lead to gut dysbacteriosis, mainly influenced by Streptococcus lutetiensis. Conversely, a significant increase in beneficial bacteria (Faecalibacterium and Coprococcus catus) was promoted by the addition of the plant-based adsorbent. Moreover, the AF group had the lowest villus height and a compromised barrier function, as evidenced by a significant (p < 0.05) increase in fluorescein isothiocyanate dextran (FITC-d), but these negative effects were almost reversed by the addition of the alfalfa adsorbent. Furthermore, the AF + YCW and alfalfa + AF groups exhibited a significant increase in the cutaneous basophil hypersensitivity response compared to the rest of the experimental groups. Taken together, these results pointed out that the alfalfa counteracts the adverse effects of AFB1 in poults, facilitating the colonization of beneficial bacteria and improving the barrier function of the turkey poults.