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This study was performed to investigate the effects of dietary threonine (Thr). level on performance, metabolizable energy, intestinal morphology, and immune system in coccidian–infected broiler chickens. The diets contained: 88%, 100% (Non challenged (NC) and challenged control (PC)), 112%, 124%, and 136% of Thr requirement according to Cobb 500 recommendation and fed during grower (pre challenged) and finisher (post challenged) periods. On d 23 (end of grower period), each bird received 0.5 mL of distilled water or received around 24000 sporulated oocysts. On d 23 and 31, one bird per replicate was slaughtered to measure the performance criteria. Mean dietary apparent metabolizable energy corrected for nitrogen (AMEn) and digestible energy were greater in NC birds than the challenged birds fed on 88% or 100% Thr diets. Feed intake and blood parameters were not significantly influenced by increasing levels of Thr in the diet. Compared with unchallenged birds with coccidia (NC), the growth performance, morphological parameters (not crypt depth), and immune responses decreased (P < 0.05) in the birds (PC) that were challenged with coccidia, and oocyte numbers were enhanced. Growth performance, jejunal morphology, and immune responses improved and oocyte count decreased in coccidian- challenged birds fed on the diets with greater levels of Thr (P < 0.05). Feeding the challenged birds with the diet containing greater levels of Thr improved (P < 0.05) their growth performance, morphology, and immune responses and decreased oocyte number. The birds fed on the diet with 124% Thr demonstrated a similar response as the NC birds. Increased diet Thr level linearly increased average daily gain and decreased feed conversion ratio in the grower and the whole experimentation periods. The AMEn and digestibility of crude protein were enhanced linearly Thr level increased in coccidian-challenged birds. It is concluded that diets containing 124% of Thr recommendation led to the greatest efficacy on the intestinal immuneresponse and normal growth maintenance of the birds contaminated with coccidia.

In ovo injection of camel lactoferrin (cLF36) as an antimicrobial peptide was applied in Ross 308 fertile eggs and tested in 320 post-hatched chickens challenged with Clostridium perfringens (Cp). In 8 treatments and five replicates of 8 birds each, performance, jejunum morphometry and ileal microbial counts of chickens were assayed. Feed intake and feed conversion ratio of the chickens affected by treatments. Together with the positive control group under the Cp (10 8 cfu/g) challenge and the negative control group under the antibiotic (AB) challenge, the highest villi length was observed. The highest crypt depth was related to the treatment with the Cp challenge and the lowest value was related to the in ovo injection of cLF36 group and combined Cp and AB challenges. The number of Clostridium spp. in the ileal contents increased in the chickens challenged with Cp (P < 0.05). The greatest change was observed in the treatment with injection of cLF36 during the embryonic period and challenge with Cp and the lowest value was related to negative control treatment. In addition, the difference between treatments with cLF36 in ovo injection during the embryonic period and challenge with or without Cp challenge was significantly increased. In the groups under the Cp challenge, the population of E. coli was numerically increased. Based on the obtained results, cLF36, derived from camel milk, could change some of the indices in performance. It caused morphological changes in the villi of ilium and caused a decrease the microbial counts of Clostridium spp., similar to the AB group in the chickens challenged with Cp. Our research attempts to create a new window for in ovo administration of cLF36, according to its beneficial effects in the present study, can be introduced as a candidate for growth-promoting antibiotics.

IntroductionIdentifying genomic regions under selection is the most challenging issue for improving important traits in animals. Few studies have focused on identifying genomic regions under selection in sheep. The aim of this study was to identify selective sweeps and to explore the relationship between these and quantitative trait loci (QTL) in both domestic and wild sheep species using single nucleotide polymorphism markers (SNPs).MethodsGenomic data were obtained from the NextGen project, which included genotyping 20 domestic and 14 wild sheep using the Illumina Ovine SNP50K BeadChip. The XP-EHH, iHS, and RSB methods were employed to detect signatures of positive selection.ResultsThe results of the iHS method indicated 405 and 275 selective sweeps in domestic and wild sheep, respectively. Additionally, RSB and XP-EHH analyses revealed approximately 398 and 479 selective sweeps in domestic and wild sheep, respectively. Some of the genes associated with important QTL traits in domestic sheep include ADGRB3, CADM1, CAPN2, GALNT10, MTR, RELN, and USP25, while in wild sheep, the relevant genes include ACAN, ACO1, GADL1, MGST3, and PRDM16. Selective sweeps identified in domestic sheep were associated with body weight, muscle weight, milk protein percentage, and milk yield. In contrast, selective sweeps found in wild sheep were linked to average daily gain, bone weight, carcass fat percentage, and dressing percentage.DiscussionThese results indicate that selection by humans and the environment have largely progressed in harmony, highlighting the importance of both economic and environmental traits for survival. Additionally, the identification of potential candidate genes associated with economic traits and genomic regions that have experienced selection can be utilized in sheep breeding programs. However, due to the incomplete information regarding the functional annotation of genes in sheep and the limited sample size, further research with a larger sample group is essential to gain a deeper understanding of the candidate genes linked to economic traits in both domestic and wild sheep. Advancing knowledge in this area can significantly enhance the effectiveness of breeding strategies. The quantitative trait loci identified in this study have the potential to be incorporated into breeding plans for both domestic and wild sheep.

Introduction :Ascites syndrome, as a metabolic disorder, is one of the most non-infectious causes of death in the old age of breeding in the poultry industry, which causes annual losses of over one hundred million dollars and is an important economic concern. The growth rate of broiler and their high need for oxygen causes an increase in pumping, followed by heart failure, and heart failure is often associated with other diseases. Due to the relationship between the heart and the kidney, chronic heart failure leads to a decrease in filtered blood volume and a decrease in kidney function, which over time causes permanent damage to the kidneys. lncRNAs play important roles in a variety of different mechanisms related to cellular homeostasis and in a wide range of pathophysiological processes and pathogenesis of many diseases, including cardiovascular disorders, respiratory and kidney diseases, preliminary studies in human samples show shows that lncRNAs are strongly involved in the development and disease of the kidney. For this reason, the lncRNAs obtained from the RNA-seq data of the kidney tissue were investigated in the occurrence of ascites.Materials and Methods :700 one-day-old chicks from one of the paternal lines of a commercial strain were kept under standard conditions until 21 days old. On the 21st day of the breeding period, cold stress (24 degrees Celsius) began and continued until the age of 48 days. After applying cold stress, the birds with ascites symptoms were grouped in the Ascites group and the rest in the Healthy group. On the 39th day of the breeding period, 70 slaughtered birds and kidney tissue samples from each bird were immediately transferred to the liquid nitrogen tank after separation. 16 ascites birds and 16 healthy birds were used for RNA extraction. Total RNA extraction was done individually using trizol (YTzol) solution, and then an equal amount of RNA from four tissue samples was combined and then cDNA was prepared from 4 ascites tissue samples and 4 healthy tissue samples. All small RNAs, such as rRNAs, tRNAs, etc., were removed by dt oligo beads, and finally, all mRNAs were used to prepare the library. Novagen company was used for sequencing using Illumine hiseq 2500 technology and paired-end reads. Several software such as hisat2, cufflinks, stringtie, cuffmerge and cuffdiff were used for mapping, aligning and analysis of gene expression differences. In order to extract coding and non-coding genes, FEELnc software was used with default settings. Potential target genes of lncRNAs were investigated by searching for protein coding genes located within 100 kb upstream and downstream of each lncRNA. In the following, a positive and negative correlation of 90% was obtained between two groups of coding and non-coding genes based on the degree of expression change. In order to investigate the metabolic, structural and functional pathways of significant genes, the Enrichr database, which is connected with other databases such as KEGG, was used.Results and Discussion:In the current research, a total of eight samples produced 187640642 million paired reads with a size of 150 nucleotides and after quality control, 185258819 uncontaminated reads were obtained. The number of 1421 lncRNA transcripts related to 921 gene loci and 154 related target genes were identified in the comparison between ascites and healthy birds. Then, this number of genes were identified (154 genes) in order to check their functional characteristics using the Enrichr database, five functional pathways Glycine, serine and threonine metabolism, Rap1 signaling pathway, Sphingolipid metabolism, Phosphatidylinositolsignaling system, Mucin type O-glycan biosynthesis and related genes were significant. Out of 13 significant lncRNAs in these biological pathways, 12 are located on the antisense direction and one is located on the sense direction. In addition, 9 lncRNAs are exonic, 3 intronic and 1 in downstream position. These pathways are activated as damage modifiers in hypoxic conditions caused by ascites and provide the required energy and maintain kidney homeostasis in response to oxygen tension caused by ascites. On the other hand, they act as cell survival, increase proliferation and anti-apoptosis, which on the one hand reduce kidney damage and on the other hand make it function better, and in this way, reduce complications caused by ascites kidney damage.Conclusion: Therefore, by targeting the important pathways of biology obtained and the genes affecting them for the prevention and treatment of ascites disease, it will provide a new insight for breeding.

Introduction[1]: Over the years, animals have been exposed to various factors such as natural selection, genetic drift, and multiple mutations, so such factors have caused changes between and within species. The genetic mutations that occur in the populations of domestic animals, will be added to the merits of animals who contain these genetic mutations and they will have more breeds. These mutations are also repeated in their breeds. If a new SNP in a population increases the competence of its carriers compared to other members of society, this choice will make the more deserving individual more involved in shaping the next generation. The most important statistical tests based on demographic differentiation are the FST statistics, which identify distinct positions under positive selection, which are of particular importance for economic characteristics. One of the best ways to understand physiological processes is to analyze gene regulation networks. Identification of genes involved in economic traits as molecular markers in breeding is of special importance. Gene regulation networks enable the researcher to study all of the genes together. The aim of this study was to identify selection signature regions and candidate genes related to economic traits.Materials and Methods: The necessary data for this research were acquired from two sources, namely NEXTGEN and HAPMAP. The dataset encompassed breeds such as Afshari (41 individuals), Ghezel (35 individuals), Moghani (35 individuals), and eight wild sheep. The initial objective was to assess data quality and perform filtration on raw data. For the remaining single nucleotide polymorphisms (SNPs), those not conforming to Hardy-Weinberg equilibrium were considered indicative of genotyping errors. A stringent probability level of 10⁻⁶, determined through Bonferroni correction, was applied. Various stages of quality control were meticulously executed using PLINK v1.9. Additionally, the study involved identifying animals positioned outside their respective groups, contributing to a comprehensive understanding of the population structure within the two groups. Principal component analysis (PCA) were done in R software. The FST index was proposed to study the distinction between subpopulations and identification of selection signature. the population structure of wild and domestic sheep breeds was analyzed. PCA analysis was performed using genotype information of the samples to investigate how the animals were grouped Investigation of identified genes using SNPs in the upper 1% range of FST were identified by Plink v1.9 software. In addition, the DAVID database (http://david.abcc.ncifcrf.gov) was used to determine biological routes. At this stage, it is assumed that genes that belong to a functional class can be considered as a group of genes that have some specific and common characteristics. GeneCards (http://www.genecards.org) and UniProtKB (http://www.uniprot.org) databases were also used to interpret the function of the obtained genes.Results and Discussion: The results showed that adjacent SNPs are highly dispersed in several genomic regions. From 34556 SNPs after filtration above 1%, SNPs with higher FST stabilization index (340 SNP) with FST range from 0.304 to 0.472 were selected. Selected SNPs consisted of 95 genomic regions on 23 chromosomes between domestic and wild sheep. Most regions were located on chromosomes 13 and 7 had 14 and 9 gene regions, respectively. Examination of the relationship between QTLs and important genes in selected areas showed that 95 genes related to economic traits were identified. QTLs with important economic characteristics including quality and quantity of meat, milk, fat, bone, immune system and parasite resistance were reported. Most QTLs were located on chromosomes 2, 3, 5, 6, and 7, indicating that the most positive mutations occurred on these chromosomes. Most of the identified biological pathways related to ion channels through cell membranes are neuromuscular processes, Brain and cerebellum growth, metencephalon growth, membrane ion membrane transport, and pathways involved in regulating ion transport in cell membranes. Genes identified in different genomic regions can be considered as selective candidates. A number of genes studied as selection signatures reported were consistent with previous studies. Important genes were included: GABRB1, GRM3, HERC1, HERC3 and KCND2.Conclusion: The study of genomic regions showed that these regions are directly and indirectly related to the quality and quantity of meat, milk, fat, bone, immune system and parasite resistance. Identifying important economic traits and locating parts of the genome that have changed as a result of selection could be used in sheep breeding programs. However, in this research we had limitations such as the incompleteness of information related to functional annotation of genes in sheep species and also the small sample size of this study. Therefore, in subsequent studies with more samples and more breeds of domestic and wild sheep in Iran, a better understanding of candidate genes for important economic traits in domestic and wild species would be achieved.

Introduction: Among different sheep breeds in the world, the Texel breed is known as a meaty and muscular breed. Skeletal muscle growth is a step-by-step and exponential process from differentiation, development and maturation, which is regulated by gene networks and cell signaling pathways, and several genes and factors are involved in the process of muscle fiber formation and their growth and hypertrophy (Badday Betti et al. 2022). The study of gene expression is done with several methods, and this gene expression information is used in breeding programs as a tool to improve phenotypic choices. Databases are a large source of expression data that can be used by bioinformatics methods to integrate heterogeneous data from different studies and platforms. In this study, by integrating the microarray and RNA-Seq data available in the database belonging to the muscle tissue of Texel breed sheep, the transcriptomic profile of the muscle was compared at two ages of embryonic and adult.Materials and Methods: Microarray data related to longissimus dorsi muscle tissue with three replicates d-70 embryos from GEO database with accession number GSE23563 and RNA-Seq data related to muscle tissue from six samples with two replicates from adult individuals from ArrayExpress database were selected. Limma, Biobase and GEOquery software packages were used to calculate the expression values of the microarray data related to the embryonic age in the R environment, and Tuxedo, HTSeq and DESeq2 packages were used in the Linux and R environment to calculate the expression values of the RNA-Seq data (Kamali et al. 2022; Sahraei et al. 2019). Then two types of expression values were integrated and to eliminate non-biological effects, the batch effects were also removed. Next, differential genes were identified with the limma software package. In order to identify the relationship between the identified differential genes, the gene network was drawn between them by software of Cytoscape version 3.7.1 and String 1.5.1 program. next, due to the vastness of the gene network, each network was clustered with MCODE 1.6.1 and CytoCluster 2.1.0 programs (ClusterOne algorithm) and significant clusters (P-value < 0.05) were identified (Saedi et al. 2022). In order to better understand the ontology and function of the identified differential genes, the Gene Ontology of the genes was investigated using software of Cytoscape version 3.7.1 and ClueGO 2.5.9 and CluePedia 1.5.9 programs. After receiving the Gene Ontology results, significant Gene Ontology terms (P-Value < 0.05) related to functional groups were identified. Finally, the selected genes (Adj P-Value < 0.05) were identified and introduced in these two age groups.Results and Discussion: After quality control, correcting and normalizing the microarray data, the GPL10778 platform annotation file with 1042520 Probe ID was used to calculate their expression values. After relevant analyzes of 9289 Probe ID identified related to the data of this study, 7918 Gene Symbol was identified finally. After quality control, trimming and normalizing the RNA-Seq data in total, the number of Ensembl_Genes based on which the reading values were calculated by HTSeq was 27056. After removing IDs that had zero readings in all 6 samples, 10855 IDs remained. Then, these 10855 Ensembl ID were merged with the annotation file to obtain Gene Symbol, and finally 9417 common genes were identified between the six samples of adult age. The results of differential expression analysis showed that there were significant differences in the expression of 62 genes (37 increased and 25 decreased) in the muscle tissue between adult and embryonic age. By creating a gene network between differential genes, 15 selected genes were identified, including MYH1, ACTN3, CASQ1, TMOD4, FBP2, SLC2A4, MX1, COX4I1, SOD2, MFN2, UQCRB, UCP3, PRKAB2, PHKG2, PPP1R3C. The function of these genes has been proven in cell proliferation, protein synthesis, myofibril formation, and lipid metabolism. Differential gene enrichment analysis revealed some biological processes such as Vasculogenesis, positive regulation of ossification, positive regulation of muscle tissue development, regulation of muscle contraction, contractile fiber part, calcium signaling, calcineurin-NFAT signaling cascade and regulation of receptor signaling pathway via JAK-STAT, the molecular function of regulating cation channel activity and the cellular components of the contractile fiber.Conclusion: This study in addition to confirming the accuracy of the integration method of two types of heterogeneous data, provided a general view of the transcriptomic differences of Texel sheep muscle tissue at two important age points to be a useful source for biological investigations of genes related to muscle growth and development in sheep.

IntroductionSex differences in prenatal growth may contribute to sex-dependent programming effects on postnatal phenotype. MethodsWe integrated for the first time phenotypic, histomorphological, clinico-chemical, endocrine and gene expression analyses in a single species, the bovine conceptus at mid-gestation. ResultsWe demonstrate that by mid-gestation, before the onset of accelerated growth, the female conceptus displays asymmetric lower growth compared to males. Female fetuses were smaller with lower ponderal index and organ weights than males. However, their brain:body weight, brain:liver weight and heart:body weight ratios were higher than in males, indicating brain and heart ‘sparing’. The female placenta weighed less and had lower volumes of trophoblast and fetal connective tissue than the male placenta. Female umbilical cord vessel diameters were smaller, and female-specific relationships of body weight and brain:liver weight ratios with cord vessel diameters indicated that the umbilico-placental vascular system creates a growth-limiting environment where blood flow is redistributed to protect brain and heart growth. Clinico-chemical indicators of liver perfusion support this female-specific growth-limiting phenotype, while lower insulin-like growth factor 2 (IGF2) gene expression in brain and heart, and lower circulating IGF2, implicate female-specific modulation of key endocrine mediators by nutrient supply. ConclusionThis mode of female development may increase resilience to environmental perturbations in utero and contribute to sex-bias in programming outcomes including susceptibility to non-communicable diseases.

Introduction: Lactoferrin is secreted in the apo-form from epithelial cells in most exocrine fluids, such as saliva, bile, pancreatic and gastric fluids, tears and, particularly, in milk. Lactoferrin is the most dominant protein in milk after casein. This protein plays a crucial role in many biological processes including the regulation of iron metabolism, induction and modulation of the immune system, the primary defense against microorganisms, inhibiting lipid peroxidation and presenting antimicrobial activity against various pathogens such as parasites, fungi, bacteria, and viruses. The major antimicrobial effect of lactoferrin is related to its N-terminal tail where different peptides for instance lactoferricin and lactoferrampin which are important for their antimicrobial abilities are present. cLF chimera (CLF36 peptide ) was derived from camel lactoferrin (cLF) consisting of 42 amino acids and has primary sequence of DLIWKLLVKAQEKFGRGKPSKRVKKMRRQWQACKSSHHHHHH. In addition, the results of previous study showed that cLFchimera had anti-inflammatory and regulatory activity of the immune system. Nuclear factor kappa B (NF-kB) transcription factors regulate several important physiological processes, including inflammation and immune responses, cell growth, apoptosis, and the expression of certain viral genes. NF-kB dimers are located in the cytoplasm in an inactive form through association with any of several IkB inhibitor proteins. The phosphorylation and degradation of IkB have received great attention as key steps for the regulation of Nuclear factor kappa B (NF-kB) complexes. Phosphorylation of the IkB by IKK signals it for ubiquitination at specific lysine residues. The aim of this study was to investigate the inhibitory effects of the recombinant camel Lactoferrampin-Lactoferricin on nuclear factor kappa B (NF-кB) pathway. Materials and Methods: CLF36 peptide was prepared through a previous study in Biotechnology Laboratory of Animal Science Department, Faculty of Agriculture, Ferdowsi University of Mashhad (GenBank accession number: MH327768.1). Protein data for inhibitory combinations of the Nuclear factor kappa B (NF-kB) pathway was collected from the Protein Data Bank (PDB) And UniProt.Physico-chemical properties analysis (atomic state, isoelectric point, half-life, hydrophobicity hydrophilicity , barometric and pH) was done using Expasy Prot Param online server. The inhibitory effects of this peptide was assessed by Molecular Docking Simulation (In silico). the simulation of the interaction (Docking) of CLF36 peptide with Nuclear factor kappa B (NF-kB) signaling pathway in upstream, pro-inflammatory cytokines (e.g., TNF-α and IL-6) and in downstream, cytoplasmic IKKB and NF-κBp65 was done using ClusPro 2.0 software online.interaction diagram created by LigPlot+ showing the hydrogen bond network and the hydrophobic interactions of CLF36 peptid with the upstream and downstream Nuclear factor kappa B (NF-kB) pathways. Results and Discussion: Bioinformatics analysis on The molecular interaction of this peptide with the active site of NF-κB proteins suggested that in may have role of the modulators and Anti-Inflammatory of immune processes by inhibitory effect on the TNF-α and IL-6 cytokines in upstream and IKK-β and NF-κB-p65 in downstream of the NF-κB signaling pathway. This results are similar to the inhibitory effects of Infliximab, Camelid Fab , NEMO and GILZ on the Nuclear factor kappa B (NF-kB) signaling pathway. Infliximab (Remicade) is a chimeric mAband its use is not very well tolerated in the majority of patients, infliximab therapy leads to the production of antibodies to infliximab in a small subset of patients. the Camelid Fab antibody with the highest (femtomolar) potency, displays a very large surface of interaction with IL-6. the regulatory role of NEMO in the IKK complex, since a number of genetic and biochemical studies clearly demonstrate that proinflammatory IKK activation is absolutely dependent upon the presence of functional NEMO. Glucocorticoid-induced leucine zipper (GILZ) is a glucocorticoid responsive protein that links the nuclear factor-kappa B (NFκB) and the glucocorticoid signaling pathways. Func­tional and binding studies suggest that the proline-rich region at the carboxy terminus of Glucocorticoid-induced leucine zipper (GILZ) binds the p65 subunit of Nuclear factor kappa B (NF-kB) and suppresses the immunoinflammatory response. Conclusion: The results of the interaction (Docking) of CLF36 peptide with Nuclear factor kappa B (NF-kB) signaling pathway indicates that, in upstream, will be inhibitory effect of CLF36 peptide in active site of pro-inflammatory cytokines (e.g., TNF-α and IL-6) and in downstream, cFL36 peptide will bind to protein receptors of cytoplasmic IKKB and NF-κB p65. Therefore, these findings may provide a theoretical basis for therapeutic mechanisms of CLF36 peptide.

Abstract The lack of cost-effective methods for producing antimicrobial peptides has made it impossible to use their high potential as a new and powerful class of antimicrobial agents. In recent years, extensive research has been conducted to decrease the cost of recombinant proteins production through microorganisms, transgenic animals, and plants. Well-known genetic and physiological characteristics, short-term proliferation, and ease of manipulation make E. coli expression system a valuable host for recombinant proteins production. Expression in periplasmic space is recommended to reduce the inherently destructive behavior of antimicrobial peptides against the expressing microorganism and to decline susceptibility to proteolytic degradation. In this study, a pET-based expression system was used to express buforin I at E. coli periplasmic space, and its antimicrobial, hemolytic, and cell toxicity activities as well as structural stability were evaluated. The hemolysis activity and cytotoxicity of His-tagged buforin I were negligible and its antimicrobial activity did not show a significant difference compared to synthetic buforin I. In addition, in silico investigating of stability of native and His-tagged buforin I showed that RMSF, RMSD and Rg curves had followed a similar trend during 150 ns simulation. Furthermore, evaluating the modelled structures, FTIR and X-ray methods of both peptides indicated an insignificant structural difference. It was concluded that the recombinant buforin I could be a viable alternative to some currently used antibiotics by successfully expressing it in the pET-based expression system.

Habitat eradication and loss of animal species have created a new international hazard for wildlife conservation. National parks are considered as suitable places that can serve dual functions of biodiversity conservation and ecotourism. As recommended by the Food and Agriculture Organization (FAO), microsatellites have been used for animal biodiversity assessment. For this reason, Iranian urials population genetic diversity was studied by analyzing of 10 microsatellite markers in 75 skeletal muscle samples that were collected from Tandooreh National Park, Northeastern of Iran. Species of samples validated by sequencing of the control region from mtDNA. Allelic frequencies for each locus in the population and different measurements of within-breed genetic variations were computed by the POPGENE32 software. The number of alleles per locus counted from 5 to 8, with an average of 6.1. The polymorphism information content was calculated between 0.66-0.74 with the average of 0.7. Observed heterozygosity ranged from 0.223 (MaF214) to 0.776 (OarFCB128) with an average about 0.584 while the average expected heterozygosity for all studied loci was 0.785 ranging from 0.765 (BM8125) to 0.807 (MaF36). High levels of expected heterozygosity can be attributed to some factors such as low level of inbreeding, low selection pressure, and high allele number. However, findings of the present study of the high variability of the Iranian urials showed the presence of a possible ‘hot spot’ genetic diversity for wild urial population in the Northeast of Iran. In conclusion, values of genetic diversity revealed that the Iranian urial population harbor unique and appreciable reservoirs of diversity.

[1]Introduction: Poultry and meat products are the largest sources of non-typhoid salmonella infections in most countries. Studies have shown that raw foods of animal origin, especially poultry and its products, are the main source of contamination of kitchens and restaurants. In terms of growth conditions, these microorganisms are resilient bacteria and easily adapt to their environmental conditions. Salmonella has been known to cause intestinal disease for many years and has been reported as the most important cause of food poisoning. According to Iranian and international standards, there should be no S. enteritidis or S. typhimurium in 25 grams of food. DNA-based methods for the identification and differentiation of Salmonella serovars have been designed and applied using specific primers at the genus and serovar levels. Therefore, they can be used as useful and rapid screening tests, as well as to supplement or replace conventional biochemical and serological tests. Real-time PCR, with the most accurate and reliable results using a fluorescence probe, which of course has a high cost. In this method, sequence specific fluorescence probes are used, and as a result, in the target molecule, screening and determination the presence or even the concentration of specific sequences is possible. Therefore, even in the presence of other types of nucleic acid molecules, the results are obtained quickly and have a high level of specificity. Under these conditions, if specific probes with different florescence dyes are used, even multiple targets can be detected in a single PCR reaction. The aim of this study was to identify S. enteritidis or S. typhimurium by PCR and Salmonella spp. by real time PCR method in poultry products. Material and Method: In total, 45 samples of poultry products, including chicken breast, liver and gizzard (15 samples each) were purchased from different regions of Mashhad and from various companies and transferred to the laboratory in accordance with hygienic standards. For each sample, 25 g of tissue was isolated and homogenized under sterile conditions and DNA extraction was then performed using a DNA extraction kit. The extracted DNA was evaluated by agarose gel electrophoresis. The purity and quantity of DNA extracted from each sample was examined by spectrophotometry method. In the next step, in order to identify the genus Salmonella, the samples were examined by real time PCR. In this method we used an internal control to ensure that negative results are not false negative due to inhibitors. The results of real time PCR showed that out of 45 samples, nine samples were infected with Salmonella. Then, these nine samples were evaluated for Salmonella typhimurium and Salmonella enteritidis infection by conventional PCR method. Result and Discussion: The results showed that out of nine samples that were positive in real time PCR test, seven samples were contaminated with Salmonella typhimurium, of which five samples were related to chicken breast and two to liver. Regarding Salmonella enteritidis infection, out of nine samples, only one sample was contaminated, which was related to chicken breast. Conventional methods have been traditionally used to enumerate target bacteria in food. However, these methods have some limitations and require considerable time and labor. Previous studies have already shown that real time PCR is more effective than conventional bacteriological methods for the detection of Salmonella spp. In a study by Whyte et al. (2002) The presence of Salmonella was assessed by traditional culture methods and by a Salmonella-specific polymerase chain reaction (PCR) test. Salmonella was recovered from 16% of samples using traditional culture methods. In contrast, the PCR assay proved to be more sensitive and detected Salmonella DNA in 19% of the examined samples (Whyte et al. 2002). Results of PCR with specific primers showed that reactions in real time PCR with general primers of Salmonella spp. were done correctly. Despite of accuracy and speed of real time PCR to detect DNA of microorganisms, further studies are developed to have more advantages. Loop-mediated isothermal amplification (LAMP) showed a higher sensitivity of Salmonella detection in compare to qPCR (Vichaibun & Kanchanaphum, 2020). Although LAMP could detect trace amount of Salmonella DNA but primer design for this reaction is very difficult. However, it is important to highlight that non-viable cells can be detected by real time PCR or other DNA-based methods, which does not occur in traditional methods of culture and isolation that require viable cells for quantification (Zeng et al., 2016).

Abstract Background Lactoferrin is a versatile protein with important modulatory functions in inflammation and immune response. This glycoprotein can bind and sequester iron and LPS, thereby intervening in certain signaling pathways and biological processes. In the present meta‐analysis, we aimed to pool experimental data regarding the immunomodulatory effects of lactoferrin and its derived peptides on the NF‐κB signaling pathway. Materials We searched PubMed, Google Scholar, and Web of Science databases and obtained all related articles published before April 2022. Finally, 25 eligible studies were selected, and their reports were analyzed. Methods We used Review Manager Version 5.2 to compute the standardized mean difference (SMD) and its 95% confidence interval. In addition, the source of heterogeneity was explored using meta‐regression and sensitivity analysis. The symmetry of the funnel plot and Egger's test were also used to evaluate publication bias utilizing Comprehensive Meta‐Analysis Version 2. Results Comparing the group of cells and animals exposed to lipopolysaccharide alone with the group that received pretreatment with lactoferrin and its derivatives, we observed significant reductions in TNF‐α, IL‐1 beta, and IL‐6 levels by 8.73 pg/mL, 2.21 pg/mL, and 3.24 pg/mL, respectively, in the second group. Additionally, IKK‐β, p‐IκB, and NF‐κB (p65) levels were significantly lower by 7.37‐fold, 15.02‐fold, and 3.88‐fold, respectively, in various cells and tissues. Conclusion Based on the results of this meta‐analysis, lactoferrin and its derived peptides can be considered potent prophylactic and therapeutic candidates against inflammation‐associated diseases by targeting the NF‐kB pathway.

The effects of curcumin and its nano-micelle form on body weight, insulin resistance, adiponectin, and blood biochemical parameters of streptozotocin-induced diabetic rats were studied. Diabetes was induced in fifty male Wistar rats which were divided into five groups treated with 1) no dietary supplements, 2 and 3) 40 and 80 mg curcumin/kg of feed, and 4 and 5) 40 and 80 mg nano-micelle curcumin/kg of feed. A group of ten untreated male Wistar rats was also considered a healthy control group. The serum concentrations of AST, ALT, glucose, insulin, triglycerides, cholesterol, HDL-C, LDL-C, and adiponectin, as well as insulin resistance, were assessed. Body weight and weight of liver, heart, and pancreas were also evaluated. Induction of diabetes increased the serum concentrations of AST, ALT, glucose, triglycerides, cholesterol, LDL-C, and insulin resistance and decreased the serum levels of insulin, adiponectin, and HDL-C, as well as body weight and weight of the heart and pancreas (p < 0.05). Nano-micelle form of curcumin alleviated the negative effects of glucose, lipid profile, and liver enzymes in diabetic rats (p < 0.05). In conclusion, the nano-micelle form of curcumin showed better efficiency compared to curcumin for improving the adverse effects of diabetes. It can be suggested that the nano-micelle form of curcumin at specific doses might be useful for diabetes treatment.

Objective(s): Early, specific, and sensitive detection methods of COVID-19 are essential for force stopping its worldwide infection. Although CT images of the lung and/or viral RNA extraction followed by real-time reverse-transcriptase-polymerase chain reaction (rRT-PCR) are widely used; they have some limitations. Here, we developed a highly sensitive magnetic bead-based viral RNA extraction assay followed by rRT-PCR. Materials and Methods: Case group included oropharyngeal/nasopharyngeal and blood samples from 30 patients diagnosed positive by PCR test for COVID-19 and control group included 30 same samples from COVID-19 negative PCR test individuals. RNA was extracted, using viral RNA extraction kit as well as using our hand-made capture bead-based technique. A one-step cDNA synthesis and Real Time PCR was conducted. A two-step comparison of the different viral RNA extraction methods for oropharyngeal/nasopharyngeal and blood samples was performed. Student t-test was applied with a P

The effect of different levels of Scrophularia striata extract in normal and challenged conditions with E. coli on performance, carcass characteristics, cellular immune response, blood antioxidant status, intestinal histo-morphometry, and microbial population (E.coli and Lactobacillus) of Cobb 500 broilers were evaluated. The experiment was performed in a complete block design with a 2 × 4 factorial arrangement with five replications of 10 birds each for 35 days in two separated halls [include challenge (C) and non-challenge (N)] with similar experimental conditions. Experimental treatments in two breeding halls were: 1) basal diet or control (CONT), 2) basal diet + 0.1 g/kg of herbal extract (EXTR 1), 3) basal diet +0.2 g/kg of herbal extract (EXTR 2), 4) Basal diet +0.1 g/kg an antibiotic (Oxytetracycline) (ANTB). On the 16th and 24th days of the experiment, one dose of 1×107 CFU K99 E. Coli was gavaged to chickens in a challenged conditioned hall. The results showed that average body weight and daily weight gain in the whole period (days 35 and 1-35) in EXTR 2 was better than the control treatment (P < 0.05). The percentage of breast weight in the ANTB was significantly different from the control treatment (P < 0.05). Glutathione peroxidase (GPX) in EXTR 2 was better than the control treatment (P < 0.05). Malondialdehyde (MDA) in EXTR 1 was significantly lower than the control group (P < 0.05). The values of E. coli in the treatment ANTB were less than control treatment (P < 0.05). Lactobacillus value in treatment EXTR 2 was higher than control treatment (P < 0.05). The value of total immunoglobulin in 28 days in ANTB, EXTR 1, and EXTR 2 were significantly higher than that of control treatment (P < 0.05). The length of villi was affected by treatments (P < 0.05). In conclusion, dietary inclusion of 0.2 g/kg of Scrophularia striata extract may improve the health status of the birds during E. coli challenge.

Abstract There is a continuing need to prevent the increasing use of common antibiotic and find the replacement to combat the drug/antibiotic resistant bacteria such as antimicrobial peptides (AMPs) such as thanatin peptide. In this study, recombinant thanatin peptide was expressed in the HEK293 cell line. Then the antimicrobial properties of this peptide on some poultry and farm animal’s pathogen strains were assessed. The thermal-stability of thanatin was predicted in various temperatures through in silico analysis. Afterwards, according to Minimum Inhibitory Concentration (MIC) results, Escherichia coli and Pseudomonas aeruginosa were chosen to test the hypothesis of LptA/LptD–thanatin interaction, computationally. Relative amino acid sequences and crystallography structures were retrieved and missed tertiary structures were predicted. The interaction of thanatin with LptA and LptD of Escherichia coli and Pseudomonas aeruginosa were analyzed subsequently. The antibacterial activity of thanatin peptide was evaluated between 6.25 and 100 μg/mL using minimum inhibitory concentration. Also, the amounts of minimum bactericidal concentrations (MBC) were between 12.5 and 200 μg/mL. The bioinformatics analysis followed by the in vitro assessment, demonstrated that thanatin would be thermally stable in the body temperature of poultry and farm animals. Thanatin could penetrate to the outer membrane domain of LptD in Escherichia coli and it could block the transition path of this protein while the entrance of LptD in Pseudomonas aeruginosa was blocked for thanatin by extra residues in comparison with Escherichia coli LptD. In addition, the quality of interaction, with regard to the number and distance of interactions which leads to higher binding energy for thanatin and LptD of Escherichia coli was much better than Pseudomonas aeruginosa. But the site and quality of interaction for thanatin and LptA was almost the same for Escherichia coli and Pseudomonas aeruginosa. Accordingly, thanatin can prevent the assembly of LptA periplasmic bridge in both pathogens. The antibacterial and thermal stability of the thanatin peptide suggested that thanatin peptide might serve as a natural alternative instead of common antibiotics in the veterinary medicine. The outcome of this in silico study supports the MIC results. Therefore, a probable reason for different level of activity of thanatin against Escherichia coli and Pseudomonas aeruginosa might be the quality of LptA/LptD–thanatin interaction.

In ovo injection of camel lactoferrin (cLF36) as an antimicrobial peptide was applied in Ross 308 fertile eggs and tested in 320 post-hatched chickens challenged with Clostridium perfringens (Cp). In 8 treatments and five replicates of 8 birds each, performance, jejunum morphometry and ileal microbial counts of chickens were assayed. Feed intake and feed conversion ratio of the chickens affected by treatments. Together with the positive control group under the Cp (108 cfu/g) challenge and the negative control group under the antibiotic (AB) challenge, the highest villi length was observed. The highest crypt depth was related to the treatment with the Cp challenge and the lowest value was related to the in ovo injection of cLF36 group and combined Cp and AB challenges. The number of Clostridium spp. in the ileal contents increased in the chickens challenged with Cp (P < 0.05). The greatest change was observed in the treatment with injection of cLF36 during the embryonic period and challenge with Cp and the lowest value was related to negative control treatment. In addition, the difference between treatments with cLF36 in ovo injection during the embryonic period and challenge with or without Cp challenge was significantly increased. In the groups under the Cp challenge, the population of E. coli was numerically increased. Based on the obtained results, cLF36, derived from camel milk, could change some of the indices in performance. It caused morphological changes in the villi of ilium and caused a decrease the microbial counts of Clostridium spp., similar to the AB group in the chickens challenged with Cp. Our research attempts to create a new window for in ovo administration of cLF36, according to its beneficial effects in the present study, can be introduced as a candidate for growth-promoting antibiotics. [ABSTRACT FROM AUTHOR]

Due to the increasing global demand for Gamma-aminobutyric acid (GABA) in the food and pharmaceutical industries, the expression of the recombinant Glutamate decarboxylase (GAD) and its industrial production is currently requested. Culture conditions were optimized to increase the expression level of the recombinant enzyme in different pH, temperature, incubation time, aeration levels, inoculation concentrations, concentrations of IPTG, and several carbon sources using the RSM based on a central composite design. According to the results of the quadratic regression equation, recombinant Escherichia coli BL21 (DE3) had the highest GAD expression at pH=7.2, aeration at about 120 rpm, inoculation concentrations of about 3% v/v, 2.25 mM IPTG, 37 °C, and 6 h of incubation time in the presence of 0.2% glucose. Using pure glucose as a carbon source on an industrial scale is not cost-effective for producing recombinant proteins. Therefore, using low-cost carbon sources such as corn syrup and molasses with concentrations of 1.5 and 5.65% (w/v) is an efficient method for the industrial production of recombinant GAD. The concentration of purified recombinant GAD in carbon sources of 0.2% glucose, 1.5 corn syrup and 5.65% molasses was 2.155, 2.07 and 1.96 mg/mL, respectively. In this way, the global need for GABA can be met by the industrial production of GAD. [ABSTRACT FROM AUTHOR]

Objective Due to high cytotoxicity effect, ribonucleases (RNases) are known as pharmacological agents with therapeutic potential. The high potential of these enzymes in the destruction of RNA strand and their other substrates and subsequently, cell destruction or cessation of cell division causes a very high cytotoxic effect of this enzyme in various cancer cells. Hence, this review investigates the therapeutic applications of ribonucleases, cell pathways and their mode of action as cytotoxic agents in order to pay attention to them in researches related to the health and therapy fields. Materials and methods In this study the keywords such as cancer, ribonuclease, ribonuclease inhibitor and RNA were used to search in databases including Scopus, SID, IranDoc, PubMed, Google Scholar, Web of Science and IranMedex. In order to select the documents used, all articles published in non-English and Persian languages, duplicate articles, articles that could not be accessed to the full text, as well as articles that were presented as abstracts were removed. Finally, the selected cases were thoroughly studied and summarized in order to prepare the current review. Results The study and review of research conducted showed that ribonucleases have the ability to kill tumor cells and that these enzymes have antiviral properties. Bovine pancreatic ribonuclease (RNase A), bovine seminal ribonuclease (BS-RNase), Onconase, and angiogenin are known as RNases with high antitumor activity which exert cytotoxic activity on cancer cells selectively by involving different cellular pathways and/ or enhance the cytotoxicity by mutation. Also, the study of articles related to the function of ribonucleases showed that the investigation of genetic pathways of synthesis and mechanisms of cytotoxicity in these enzymes will provide the development of new pharmaceutical products in the future. Utilization of engineering processes and chemical changes in ligand / receptor structure may increase the toxicity of ribonuclease molecules in order to select cytotoxic pathways on malignant cells. Therefore, it is necessary to discover the therapeutic potential and to study in detail the therapeutic values of ribonucleases. Conclusions Ribonucleases are potentially bioavailable drug candidates, and in regard to their toxicity, which is based on the selective hydrolysis of intracellular RNA molecules and specific cell membrane recognition processes, these enzymes can be used as anti-tumor drugs or therapeutic agents.

Objective Immunotoxins are one of the most promising ways in therapeutic fields, specially cancer therapy which have a unique toxin-antibody structure, and kill the cancer cell by passing through the cell membrane and entering the target cell. The aim of this study was to engineer and design bovine pancreatic enzyme (RNase A) for to escape from ribonuclease inhibitor (RI) as a toxin segment in designing of immunotoxins used in therapeutic fields. Materials and Methods After bioinformatics investigation using PyMol software, engineering and substitution of target amino acid in wild gene coding RNase A sequence (K91A) were performed. The wild type and recombinant proteins in E.coli (BL21(D3) strain) were expressed and refolded, and then approved using SDS-PAGE. The purification of produced proteins was conducted using an immobilized-metal affinity chromatography for His-tagged proteins. Results The results showed that the protein concentrations were 3 and 2.2 g/L for wild RNase A and recombinant enzyme. The hydrolysis activity measurement of the produced enzymes, in presence of different concentrations of RI, indicated that recombinant variant had lower sensitivity and higher catalytic activity in comparison with the wild enzyme. The results obtained from enzyme activity of recombinant enzyme compared to the wild enzyme, in presence or absence of RI, demonstrated that the interaction effect between RNase A and RI can be disrupted by manipulation of amino acids involved in this interaction. Conclusion In respect to the current results, it was concluded that RNase A enzyme can be used as a promising drug in engineering and production of immunotoxins.

Sheep satellite cells more than satellite cells of the rat and mouse are similar to human satellite cells. These cells are widely used in the modeling and treatment of diseases like heart insufficiency, neurological diseases, muscular dystrophy, cerebral cell transplantation for the treatment of migraines, screening, and the production of new drugs. This study was aimed to isolate and culture primary satellite cells (PSCs) obtained from sheep fetus, and perform clonal expansion of transfected PSCs. Skeletal muscle tissues of hind limbs were collected from sheep fetuses obtained from a local abattoir. After enzymatic digestion, flasks were replaced after 3 hours to isolate non-myogenic cells, such as fibroblasts. After six days, the cells were differentiated to myoblasts. Using a differentiation medium containing the horse serum, myotube cells were observed in the flask, indicating that the cultured cells were satellite cells. The mRNA expression of the PAX7 gene was used to confirm the presence of satellite cells. In addition, the results showed that satellite cells grow in a culture medium containing 5% FBS without differentiation, while 10% FBS initiates their differentiation.

Objective Salmonella is a Gram-negative, anaerobic, flagellated bacterium of Salmonella type in Enterobacteriaceae family, which is known as a zoonosis infectious agent. In recent years, different types of antibiotics have been used to overcome the Salmonella infection; however, the increasing number of antibiotic-resistant bacteria, the provision of alternative methods seems be necessary. Specific immunoglobulin Y (IgY) originated from egg yolk causes passive immunity in the fetus, and extracting these antibodies from egg yolk is cheaper and more feasible than other antibodies. Numerous studies have shown that the egg yolk IgY has immunogenic function against a wide range of bacterial and viral infections in mammals. The purpose of this study was to produce specific immunoglobulin Y against Salmonella typhimurium and Salmonella enteritidis bacteria in laying hens. Materials and Methods In the current study, six pullets were inoculated and boosted with one milliliter of killed Salmonella typhimurium and Salmonella enteritidis. Injections were done fortnightly until one week before egg production. After breast muscle injection of chickens, the eggs were collected, and the IgY was extracted from yolk by the Polson method with polyethylene glycol 6000. The produced IgY was approved by SDS-PAGE method. The specific indirect ELISA done by specificity test for purified IgY also approved the specific IgY against both Salmonella typhimurium and Salmonella enteritidis. Results The results of SDS - PAGE demonstrated the presence of two protein bands of 27 and 67 kDa representing light and heavy chains of IgY, respectively. The specific indirect ELISA method proved that the specific IgY against Salmonella typhimurium and Salmonella enteritidis was produced in the egg yolk. Conclusions It might be possible to administer the egg yolk containing specific IgY against both Salmonella typhimurium and Salmonella enteritidis, to induce passive immunity for preventing some pathogenic diseases in calves such as diarrhea.

ABSTRACT: Clostridium perfringens-induced necrotic enteritis (NE) is an economically important disease of broiler chickens. The present study evaluated the effect of C. perfringens on the intestinal histomorphometry, enteric microbial colonization, and host immune responses using 3 experimental NE reproduction methods. The experimental groups consisted of 1) unchallenged Control diet (corn-soybean meal), 2) Control diet + Eimera inoculation at d 11 followed by C. perfringens challenge at d 15 (ECp), 3) Wheat-based diet + C. perfringens challenge (WCp), and 4) Wheat-based diet + Eimeria inoculation followed by C. perfringens challenge (WECp). The results showed that chickens receiving ECp and WECp had reduced (P < 0.05) bird performance coupled with enteric gross lesions and epithelial damage at d 17 and 24 of age compared to unchallenged control birds. These ECp and WECp administered birds also had increased (P < 0.05) ileal colonization by clostridia and E. coli at d 17 and 24, while the resident Lactobacillus counts were reduced (P < 0.05) at d 24 of age. Furthermore, at d 24, jejunal transcription of IL-6, IL-10, annexin-A1 and IL-2 genes was upregulated (P < 0.05) in the ECp group, whereas the transcription of TNF receptor associated factor (TRAF)-3 gene was increased (P < 0.05) in WECp treated birds when compared to unchallenged control group. Additionally, stimulation of chicken splenocytes and cecal tonsilocytes with virulent C. perfringens bacilli or their secretory proteins resulted in a higher (P < 0.05) frequency of T cells and their upregulation of MHC-II molecule, as determined by flow cytometry. These findings suggest that C. perfringens, while inducing epithelial damage and changes in microbiota, can also trigger host immune responses. Furthermore, NE reproduction methods using coccidia with or without the wheat-based dietary predisposition seem to facilitate an optimal NE reproduction in broiler chickens and thus, may provide better avenues for future C. perfringens research.

Hilsa species are broadly described as largely anadromous fish with a large valuable population size. It is well-known as one of the most critical commercial and occurs in marine, estuarine and riverine environments Hilsa shad, Tenualosa ilisha locally known as Sbour, migrates to the freshwater environment of the Shatt Al-Arab River systems for breeding. It was determined the genetic variation in 70 specimens of T. ilisha from four sites at the Shatt Al-Arab River, the mtDNA cytochrome-b gene was selected as a molecular marker for detecting genetic diversity, origin, and divergence of the population through comparing them with 6 samples from different locations at Indo-Pacific. The AMOVA analysis showed that the variation between groups is 60.97% and 39.02% within population indicating statistically significant P value (P

This study aimed to monitor the effect of including rumen-protected L-carnitine (Carneon 20 Rumin-Pro, Kaesler Nutrition GmbH, Cuxhaven, Germany) in the transition diet on the productive and metabolic responses of multiparous high-producing Holstein dairy cows. Thirty-two multiparous cows were allocated in a completely randomized design to receive the same diet plus 60 g fat prill containing 85% palmitic acid (control, n = 16) or 100 g rumen-protected L-carnitine (RLC, n = 16); at 28 days before expected calving until 28 days in milk (DIM). Fat prill was included in the control diet to balance the palmitic acid content of both experimental diets. Milk production over the 28 DIM for the control and RLC groups was 46.5 and 47.7 kg, respectively. Milk fat content tended to increase upon rumen-protected L-carnitine inclusion (p = 0.1). Cows fed rumen-protected L-carnitine had higher fat- and energy-corrected milk compared with the control group. Pre- and post-partum administration of L-carnitine decreased both high- and low-density lipoprotein concentrations in peripheral blood of post-partum cows. The results of this study indicated that the concentration of triglycerides and beta-hydroxybutyrate was not significantly different between the groups, whereas the blood non-esterified fatty acid concentration was markedly decreased in cows supplemented with L-carnitine. Animals in the RLC group had a significant (p < 0.05) lower blood haptoglobin concentration at 7 and 14 DIM than the control. Animals in the RLC group had a lower concentration of blood enzymes than those of the control group. The mRNA abundance of Toll-like receptors 4, cluster of differentiation 14, and myeloid differential protein 2 did not significantly change upon the supplementation of L-carnitine in the transition diet. In summary, the dietary inclusion of RLC improved dairy cow's performance during the early lactation period. Greater production, at least in part, is driven by improved energy utilization efficiency and enhanced metabolic status in animals during the periparturient period.