Methionine sources and genotype affect embryonic intestinal development, antioxidants, tight junctions, and growth-related gene expression in chickens.

Methionine (Met) is an essential and first limiting amino acid in the poultry diet that plays a significant role in chicken embryonic development and growth. The present study examined the effect of in ovo injection of DL-Met and L-Met sources and genotypes on chicken embryonic-intestinal development and health. Fertilized eggs of the two genotypes, TETRA-SL layer hybrid (TSL) - commercial layer hybrid and Hungarian Partridge colored hen breed (HPC) - a native genotype, were randomly distributed into four treatments for each genotype. The treatment groups include the following: 1) control non-injected eggs (NoIn); 2) saline-injected (SaIn); 3) DL-Met injected (DLM); and 4) L-Met injected (LM). The in ovo injection was carried out on 17.5 d of embryonic development; after hatching, eight chicks per group were sacrificed, and the jejunum was extracted for analysis. The results showed that both DLM and LM groups had enhanced intestinal development as evidenced by increased villus width, villus height, and villus area (P < 0.05) compared to the control. The DLM group had significantly reduced crypt depth, glutathione content (GSH), glutathione S-transferase 3 alpha (GST3), occludin (OCLN) gene expression and increased villus height to crypt depth ratio in the TSL genotype than the LM group (P < 0.05). The HPC genotype has overexpressed insulin-like growth factor 1 (IGF1) gene, tricellulin (MD2), occludin (OCLN), superoxide dismutase 1 (SOD1), and GST3 genes than the TSL genotype (P < 0.05). In conclusion, these findings showed that in ovo injection of Met enhanced intestinal development, and function, with genotypes responding differently under normal conditions. Genotypes also influenced the expression of intestinal antioxidants, tight junction, and growth-related genes.

In ovo feeding of methionine affects antioxidant status and growth-related gene expression of TETRA SL and Hungarian indigenous chicks.

Methionine (Met) plays a substantial role in poultry due to its involvement in several pathways, including enhancing antioxidant status and improving growth performance and health status. This study examined how in ovo feeding of Met affects hatching performance, antioxidant status, and hepatic gene expression related to growth and immunity in the TETRA-SL LL hybrid (TSL) commercial layer and Hungarian partridge colored hen (HPC) indigenous genotypes. The eggs were injected with saline, DL-Met, and L-Met on 17.5 days of embryonic development. The results showed that the in ovo feeding of DL-Met significantly increased the hatching weight and ferric reducing the ability of the plasma (FRAP) compared with L-Met. The in ovo feeding of either Met source enhanced the liver health and function and hepatic antioxidant status of the chicks. The genotype's differences were significant; the TSL genotype had better hatching weight, an antioxidant defense system, and downregulated growth-related gene expression than the HPC genotype. In ovo feeding of either Met source enhanced the chicks' health status and antioxidant status, and DL-Met improved the hatching weight of the chicks more than L-Met. Genotype differences were significantly evident in the responses of growth performance, antioxidant status, blood biochemical parameters, and gene expression to Met sources.

Monitoring physiological processes of fast-growing broilers during the whole life cycle: Changes of redox-homeostasis effected to trassulfuration pathway predicting the development of non-alcoholic fatty liver disease.

In the broiler industry, the average daily gain and feed conversion ratio are extremely favorable, but the birds are beginning to approach the maximum of their genetic capacity. However, as a consequence of strong genetic selection, the occurrence of certain metabolic diseases, such as myopathies, ascites, sudden cardiac death and tibial dyschondroplasia, is increasing. These metabolic diseases can greatly affect the health status and welfare of birds, as well as the quality of meat. The main goal of this study was to investigate the changes in the main parameters of redox homeostasis during the rearing (1-42 days of age) of broilers with high genetic capacity, such as the concentrations of malondialdehyde, vitamin C, vitamin E, and reduced glutathione, the activities of glutathione peroxidase and glutathione reductase, and the inhibition rate of superoxide dismutase. Damage to the transsulfuration pathway during growth and the reason for changes in the level of homocysteine were investigated. Further, the parameters that can characterize the biochemical changes occurring in the birds were examined. Our study is the first characterize plasma albumin saturation. A method was developed to measure the levels of other small molecule thiol components of plasma. Changes in redox homeostasis induce increases in the concentrations of tumor necrosis factor alpha and inflammatory interleukins interleukin 2, interleukin 6 and interleukin 8 in broilers reared according to current large-scale husbandry technology and feeding protocols. A significant difference in all parameters tested was observed on the 21st day. The concentrations of cytokines and homocysteine increased, while the concentrations of glutathione and cysteine in the plasma decreased. Our findings suggest that observed changes in the abovementioned biochemical indices have a negative effect on poultry health.

Pedigree-Based Description of Three Traditional Hungarian Horse Breeds.

The Mezohegyes Stud was founded in 1784 where three different horse breeds were developed: the Furioso-North Star, the Gidran, and the Nonius. These breeds were based on the same mare population, but each breed had different utilization purposes. Our aim was to analyze the pedigree information of these three indigenous breeds. The genealogical information was traced back from the actual breeding population back to the founder animals, and the final database contained more than 47,000 horses. The reference populations were defined as the registered breeding animals in 2019. The complete generation equivalent was 16.45 for the Gidran breed, 15.18 for Furioso-North Star, and 12.64 for Nonius, respectively. Due to the utilization of English Thoroughbred during the breeding history, the average maximum generations were close to 36 generations for each breed. The average relatedness was approximately 4%. The average Wright's inbreeding coefficient was the highest for the Nonius breed (5.59%). Kalinowski's decomposition of inbreeding showed that inbreeding is originated mainly from the past; the current fixation of alleles was higher for the Nonius horse breed. There was a reasonable bottleneck effect for each breed. The estimated effective population sizes suggest that there is no problem with the maintaining of Mezohegyes horse breeds.

Effects of DL and L-Methionine on Growth Rate, Feather Growth, and Hematological Parameters of Tetra-SL Layers from 1-28 Days of Age.

The study was carried out to determine whether sources or levels of methionine (Met) affect the health status of Tetra-SL (TSL) chicks by examining growth performance, feather growth, and hematological parameters. A total of ninety-six (96) day-old (44.2 ± 0.18 g lw) TSL chicks were randomly allotted to six treatment groups (three levels for each DL and L-Met source) with four replicates of four chicks each. Chicks were fed ad libitum diets supplemented with 90, 100, and 110% of methionine requirements for four weeks after hatch. The parameters examined are bodyweight (BW), average daily gain (ADG), feather length (FL), and hematological parameters, including: red blood cell (RBC) count, hemoglobin (Hb) concentration in the blood, hematocrit (Ht; %), number of white blood cells (WBC), platelet count, mean corpuscular volume of red blood cells (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), lymphocyte (LYM), mid-range (MID), and granulocyte (GRAN). There was no significant effect of Met sources and levels on BW, ADG, and FL of TSL chicks for the first four weeks of rearing. The RBC, Hb, Ht, WBC, LYM, MID, and GRAN values of TSL chicks were statistically influenced by dietary Met sources and Met levels (p < 0.05). Among the treatment groups, the number of white blood cells (WBC) on 90% DL−Met was the lowest. WBC, RBC, Hb, and Ht were higher from chicks that received 100% DL-Met than all other levels, regardless of the Met sources. The MCV, MCH, and MCHC values were not affected by either Met source or levels or their interactions. Met source and level interactively affected the Ht, WBC, LYM %, and GRAN values of TSL chicks (p < 0.05). The platelet number was affected by Met source only (p < 0.05) with chicks receiving L-Met source having more than twofold higher platelet values than DL-Met source. In conclusion, varying Met levels by ±10% of the requirement does not adversely affect the growth performance, feather growth, and hematological parameters of TSL chicks reared for up to four weeks of age. DL-Met increased the body weight and improved the white blood cells, red blood cells, and hematocrit at 28 days after hatch. DL-Met showed similar biological efficacy as L-Met for body weight and feather growth but not for the hematological parameters.

Comparison of VividTrac, King Vision and Macintosh laryngoscopes in normal and difficult airways during simulated cardiopulmonary resuscitation among novices.

INTRODUCTION: Early endotracheal intubation improves neurological outcomes in cardiopulmonary resuscitation, although cardiopulmonary resuscitation is initially carried out by personnel with limited experience in a significant proportion of cases. Videolaryngoscopes might decrease the number of attempts and time needed, especially among novices. We sought to compare videolaryngoscopes with direct laryngoscopes in simulated cardiopulmonary resuscitation scenarios. MATERIALS AND METHODS: Forty-four medical students were recruited to serve as novice users. Following brief, standardized training, students executed endotracheal intubation with the King Vision®, Macintosh and VividTrac® laryngoscopes, on a cardiopulmonary resuscitation trainer in normal and difficult airway scenarios. We evaluated the time to and proportion of successful intubation, the best view of the glottis, esophageal intubation, dental trauma and user satisfaction. RESULTS: In the normal airway scenario, significantly shorter intubation times were achieved using the King Vision® than the Macintosh laryngoscope. In the difficult airway scenario, we found that the VividTrac® was superior to the King Vision® and Macintosh laryngoscopes in the laryngoscopy time. In both scenarios, we noted no difference in the first-attempt success rate, but the best view of the glottis and dental trauma, esophageal intubation and bougie use were more frequent with the Macintosh laryngoscope than with the videolaryngoscopes. The shortest tube insertion times were achieved using the King Vision® in both scenarios. CONCLUSION: All providers achieved successful intubation within three attempts, but we found no device superior in any of our scenarios regarding the first-attempt success rate. The King Vision® was superior to the Macintosh laryngoscope in the intubation time in the normal airway scenario and noninferior in the difficult airway scenario for novice users. We noted significantly less esophageal intubation using the videolaryngoscopes than using the Macintosh laryngoscope in both scenarios. Based on our results, the KingVision® might be recommended over the VividTrac® and Macintosh laryngoscopes for further evaluation.

Ollieopteryx, a new genus of caddisflies from Madagascar (Trichoptera: Dipseudopsidae).

New taxa of Dipseudopsidae from Madagascar are described, including a new genus, Ollieopteryx, and three new species, Ollieopteryx fianarana, O. inopinata, and O. dakshan. The hypothetical phylogenetic tree of the Dipseudopsidae (Weaver Malicky 1994) is revised to include Ollieopteryx, an isolated relic. Scenarios on the historical biogeography of the family are hypothesized.

Newly Grown Wool Mineral Content Response to Dietary Supplementation in Sheep.

Determination of wool mineral content to assess the animal' mineral status has been extensively used, but the results are controversial. One of the possible contributing factors is that the sampling material in previous studies was collected from a long staple, a fact that could mask the response to recent differences in mineral intake. Therefore, the aim of the present study was to test the sensitiveness of newly grown wool to different dietary mineral intake. Twenty Tsigai ewes were allocated into five dietary treatments with similar hay and concentrate intake but different premix inclusion rates in the concentrate (3, 4, 5, 6, and 7%). Wool was sampled on the left side from a 5 × 5 cm area using bent scissors at the beginning of the trial and from the very same area 28 days later. Samples after cleaning and mineralization were analyzed with ICP-OES (Perkin-Elmer, Optima 3300 DV) for calcium, phosphorus, magnesium, sodium, selenium, zinc, copper, and sulfur content. Long fleeces had significantly lower Ca and Se content compared to the newly grown wool samples of the group at the premix manufacturer's suggested level of supplementation (5%). Macrominerals in fresh wool did not respond to increased dietary supplementation. Se and Zn content of wool had a strong relationship with the daily intake (R2 = 0.95 and R2 = 0.97, respectively.) In conclusion, the mineral content of long fleeces can be different compared to recently developed wool fiber. This indicates that, in some cases, analyzing long staples for mineral status can be misleading. Our results showed that wool could be a sensitive indicator of low selenium and high zinc intake. Mineral interactions can significantly affect the actual availability of trace minerals; therefore, a more careful design of premixes is needed. The described method seems to be applicable in livestock farming, but the mineral interactions that may alter the results need to be further explored.

Effect of Carotenoids, Oligosaccharides and Anthocyanins on Growth Performance, Immunological Parameters and Intestinal Morphology in Broiler Chickens Challenged with Escherichia coli Lipopolysaccharide.

This study was conducted to investigate the effect of carotenoid, oligosaccharide and anthocyanin supplementation in broiler diets under Escherichia coli lipopolysaccharide (LPS) challenge. Ross 308 chickens were fed 5 diets: basal diet (control diet), diet supplemented with ß-glucan in 0.05% (positive control) and diets with 0.5% carotenoid-, oligosaccharide- or anthocyanin contents. On the 26th days of age, chickens were challenged intraperitoneally 2 mg LPS per kg of body weight. 12 h after injection, birds were euthanized, then spleen and ileum samples were collected. LPS induced increased relative mRNA expression of splenic (p = 0.0445) and ileal (p = 0.0435) interleukin-1ß (IL-1ß), which was lower in the spleen in carotenoid (p = 0.0114), oligosaccharide (p = 0.0497) and anthocyanin (p = 0.0303)-treated chickens compared to LPS-injected control birds. Dietary supplementation of carotenoids also decreased relative gene expression of splenic interleukin-6 (IL-6) (p = 0.0325). In the ileum, ß-glucan supplementation showed lower relative mRNA expression of toll-like receptor 5 (TLR-5) (p = 0.0387) compared to anthocyanin treatment. Gene expression of both splenic and ileal interferon-α (IFN-α), interferon-γ (IFN-γ), toll-like receptor 4 (TLR-4) and toll-like receptor 5 (TLR-5) were not influenced by dietary supplements. In conclusion, carotenoids, oligosaccharides and anthocyanins could partially mitigate the immune stress caused by LPS challenge. All of the compounds impacted longer villus height (p < 0.0001), villus height:crypt depth ratios were higher after ß-glucan (p < 0.0001) and anthocyanin (p = 0.0063) supplementations and thickened mucosa was observed in ß-glucan (p < 0.0001), oligosaccharide (p < 0.0001) and anthocyanin (p = 0.048) treatments. All of these findings could represent a more effective absorption of nutrients.

Proteomics alterations in chicken jejunum caused by 24 h fasting.

The small intestine is the longest part of the chicken (Gallus gallus) gastrointestinal system that is specialized for nutrient absorption. It is known that decrease in intestinal villi area or height in early age can cause a reduction in essential nutrient intake, which may lead to delayed growth and consequently poorer performance of broiler chickens. The small intestinal absorptive surface is known to be affected by various factors, among others things the nutritional state. In our experiment, we aimed to investigate the possible protein expression alterations that lie behind the villus area and height decrease caused by feed deprivation. A total of 24 chickens were divided into three groups, namely ad libitum fed, fasted for 24 h, fasted for 24 h then refed for 2 h. The morphometric parameters were also measured in the duodenum, jejunum and ileum tissue sections using image analysis. Differential proteome analyses from jejunum samples were performed using two-dimensional difference gel electrophoresis followed by tryptic digestion and protein identification by matrix-assisted laser desorption/ionization mass spectrometry. Overall 541 protein spots were detected after 2D. Among them, eleven showed 1.5-fold or higher significant difference in expression and were successfully identified. In response to 24 h fasting, the expression of nine proteins was higher and that of two proteins was lower compared to the ad libitum fed group. The functions of the differentially expressed proteins indicate that the 24 h fasting mainly affects the expression of structural proteins, and proteins involved in lipid transport, general stress response, and intestinal defense.

Feeding state and age dependent changes in melanin-concentrating hormone expression in the hypothalamus of broiler chickens.

We aimed to quantify the gene expression changes of the potent orexigenic melanin-concentrating hormone (MCH) in chicken (Gallus gallus) hypothalamus with quantitative real-time polymerase chain reaction (qPCR), and for the first time determine peptide concentrations with a novel radioimmunoassay (RIA) under different feeding status. Three different experimental conditions, namely ad libitum feeding; fasting for 24 h; fasting for 24 h and then refeeding for 2 h, were applied to study changes of the aforementioned target and its receptor (MCHR4) gene expression under different nutritional status. The relative changes of MCH and MCHR4 were also studied from 7 to 35 days of age. Expression of PMCH and MCHR4 along the gastrointestinal tract (GIT) was also investigated. We found that expression of both targets was significant in the hypothalamus, while only weak expression was detected along the GIT. Different nutritional states did not affect the PMCH and MCHR4 mRNA levels. However, fasting for 24 h had significantly increased the MCH-like immunoreactivity by 25.65%. Fasting for 24 h and then refeeding for 2 h had further significantly increased the MCH peptide concentration by 32.51%, as compared to the ad libitum state. A decreasing trend with age was observable for both, the PMCH and MCHR4 mRNA levels, and also for the MCH-like immunoreactivity. Correlation analysis did not result in a significant correlation between MCH peptide concentration and abdominal fat mass in ad libitum fed birds. In conclusion, MCH peptide concentration altered in response to 24 h fasting, which indicated that this peptide may take part in feed intake regulation of broiler chickens.

X- and Y-chromosome-specific variants of the amelogenin gene allow non-invasive sex diagnosis for the detection of pseudohermaphrodite goats.

The Polled Intersex Syndrome (PIS) is responsible for the absence of horns in homozygous and heterozygous goats causing a female-to-male sex reversal in the homozygous polled genotypic female (XX) goats. A simple and efficient non-invasive method was elaborated to detect the genotypic sex from hair and faecal samples using a pair of primers to amplify the X- and Y-linked alleles of the amelogenin gene. The PCR products were easily distinguishable using agarose gel electrophoresis: we detected an X-specific single band in samples originating from healthy phenotypic females and double (X- and Y-) bands in samples from males. The new PCR method is applicable for diagnosing the sex of PIS-affected animals already as newborn kids, in contrast with the phenotypic findings appearing only after puberty, and thus it may replace the cumbersome chromosome investigations.

Changes in expression of neuropeptides and their receptors in the hypothalamus and gastrointestinal tract of calorie restricted hens.

The list of orexigenic and anorexigenic peptides, those are known to alter feed intake, is continuously growing. However, most of them are studied in mammalian species. We aimed to investigate plasma level and mRNA expression of the pituitary adenylate cyclase-activating polypeptide (PACAP), gene expression of its receptor (PAC1), furthermore the gene expression of galanin (GAL), neuromedin U (NMU), and its two receptors (NMUR1 and NMUR2) in the hypothalamus, proventriculus, and jejunum of hens exposed to 40% calorie restriction. Feed restriction resulted in a 88% increase in mRNA and a 27% increase in peptide level of PACAP in proventriculus measured with qPCR and RIA, respectively. Increases were found in the gene expression of PAC1 (49%) and NMUR1 (63%) in the hypothalamus. Higher expressions of peptide encoding genes (76% for PACAP, 41% for NMU, 301% for NMUR1 and 308% for GAL, P < 0.05) were recorded in the jejunum of hens exposed to restricted nutrition. The results indicate that PACAP level responds to calorie restriction in the proventriculus and jejunum, but not in the hypothalamus and plasma.

Description of two new filtering carnivore Drusus species (Limnephilidae, Drusinae) from the Western Balkans.

Two new species of the genus Drusus (Trichoptera, Limnephilidae, Drusinae) from the Western Balkans are described. Additionally, observations on the biodiversity and threats to the region's endemic aquatic fauna are discussed. Drususkrpachi sp. n. is a micro-endemic of the Korab Mountains, Macedonia, and Drususmalickyi sp. n. is a micro-endemic of the Prokletije Mountains, Albania. Both new species are most similar to Drususmacedonicus but differ from the latter in the shape of segment IX, the shape of the tips of the intermediate appendages in lateral view, the shape of the inferior appendages, and the form and shape of the parameres. In addition, males of the European species of filtering carnivore Drusinae are diagnosed and illustrated, including Cryptothrixnebulicola McLachlan, Drususchrysotus Rambur, Drususdiscolor Rambur, Drususmacedonicus Schmid, Drususmeridionalis Kumanski, Drususmuelleri McLachlan, Drususromanicus Murgoci and Botosaneanu, and Drusussiveci Malicky. These additions to the Western Balkan fauna demonstrate the significance of this region for European biodiversity and further highlight the importance of faunistic studies in Europe.

A hairy case: The evolution of filtering carnivorous Drusinae (Limnephilidae, Trichoptera).

The caddisfly subfamily Drusinae BANKS comprises roughly 100 species inhabiting mountain ranges in Europe, Asia Minor and the Caucasus. A 3-gene phylogeny of the subfamily previously identified three major clades that were corroborated by larval morphology and feeding ecologies: scraping grazers, omnivorous shredders and filtering carnivores. Larvae of filtering carnivores exhibit unique head capsule complexities, unknown from other caddisfly larvae. Here we assess the species-level relationships within filtering carnivores, hypothesizing that head capsule complexity is derived from simple shapes observed in the other feeding groups. We summarize the current systematics and taxonomy of the group, clarify the systematic position of Cryptothrix nebulicola, and present a larval key to filtering carnivorous Drusinae. We infer relationships of all known filtering carnivorous Drusinae and 34 additional Drusinae species using Bayesian species tree analysis and concatenated Bayesian phylogenetic analysis of 3805bp of sequence data from six gene regions (mtCOI5-P, mtCOI3-P, 16S mrDNA, CADH, WG, 28S nrDNA), morphological cladistics from 308 characters, and a total evidence analysis. All analyses support monophyly of the three feeding ecology groups but fail to fully resolve internal relationships. Within filtering carnivores, variation in head setation and frontoclypeus structure may be associated with progressive niche adaptation, with less complex species recovered at a basal position. We propose that diversification of complex setation and frontoclypeus shape represents a recent evolutionary development, hypothetically enforcing speciation and niche specificity within filtering carnivorous Drusinae.