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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.

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: 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.

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.

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.

The objective of this study was to investigate the effect of whey protein concentrate (WPC) and selenium (Se) supplementation on sperm quality, antioxidant enzymes activity and lipid peroxidation in seminal plasma, liver and testis of roosters. Forty-five Ross-308 broiler breeder roosters aged 60 weeks were used for an eight-week period in a 3 × 3 factorial arrangement of dietary treatments. Three levels of WPC (0.0, 1.5 and 3.0% of diet) and selenium supplementation (0.0, 0.2 and 0.4 mg/kg of diet) with five replications were tested. Total and progressive sperm motility, sperm concentration, plasma membrane integrity, and sperm viability were significantly lower in birds treated with Se supplementation-free diet (P < 0.05). Also, abnormal sperms were significantly higher in Se supplementation-free diets when compared to the diets supplemented with 0.4 mg/kg Se (P < 0.05). The use of 1.5% of WPC resulted in significantly increased total and progressive sperm motility compared to the WPC-free diet (P < 0.05). Selenium at the level of 0.4 mg/kg along with 3.0% WPC were associated with significantly increased Glutathione peroxidase and superoxide dismutase activity in seminal plasma as compared to other levels (P < 0.05). The highest level of total antioxidant capacity (TAC) in seminal plasma was observed at the level of 0.2 mg Se (P < 0.05). Further, 3.0% WPC resulted in significantly increased TAC concentration in seminal plasma compared to the WPC-free diet (P < 0.05). Moreover, the Malondialdehyde (MDA) level of seminal plasma in selenium supplementation-free diets was significantly higher than those of other levels (P < 0.05). Glutathione peroxidase activity, TAC, and MDA levels in the testis and liver were not affected by the WPC and Se levels. It can be concluded that dietary inclusion of WPC and Se improved the semen quality, increased antioxidant enzymes activity and decreased lipid peroxidation in seminal plasma of broiler breeder roosters.

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

Abstract KRAS mutation is responsible for 40–50% of colorectal cancers (CRCs). RNA-seq data and bioinformatics methods were used to analyze the transcriptional profiles of KRAS mutant (mtKRAS) in comparison with the wild-type (wtKRAS) cell lines, followed by in-silico and quantitative real-time PCR (qPCR) validations. Gene set enrichment analysis showed overrepresentation of KRAS signaling as an oncogenic signature in mtKRAS. Gene ontology and pathway analyses on 600 differentially-expressed genes (DEGs) indicated their major involvement in the cancer-associated signal transduction pathways. Significant hub genes were identified through analyzing PPI network, with the highest node degree for PTPRC. The evaluation of the interaction between co-expressed DEGs and lncRNAs revealed 12 differentially-expressed lncRNAs which potentially regulate the genes majorly enriched in Rap1 and RAS signaling pathways. The results of the qPCR showed the overexpression of PPARG and PTGS2, and downregulation of PTPRC in mtKRAS cells compared to the wtKRAS one, which confirming the outputs of RNA-seq analysis. Further, significant upregualtion of miR-23b was observed in wtKRAS cells. The comparison between the expression level of hub genes and TFs with expression data of CRC tissue samples deposited in TCGA databank confirmed them as distinct biomarkers for the discrimination of normal and tumor patient samples. Survival analysis revealed the significant prognostic value for some of the hub genes, TFs, and lncRNAs. The results of the present study can extend the vision on the molecular mechanisms involved in KRAS-driven CRC pathogenesis.

Introduction: Identifying 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). Methods: Genomic 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. Results: The 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. Discussion: These 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. [ABSTRACT FROM AUTHOR]

Introduction: Cow's milk is one of the most important animal products for any country and known as an important source for fat and protein originated from animals (Fenelon and Guinee, 1999, Martini et al., 2016). Milk fat has a high impact on taste of milk and other dairy products (Spelman et al., 1996). The liver in ruminant animals including dairy cows plays an important role in the metabolism of carbohydrates, fats, vitamins, hormones, and etc. The absorbed nutrients pass through the liver from the gastrointestinal tract and enter the blood circulation system, and eventually enter the mammary glands of dairy cows. Therefore, the liver plays an essential role in cow lactation (Graber et al., 2010 and Schlegel et al. 2012). All of the components that determine milk quality can be considered as quantitative traits that are controlled by many genes and are influenced by environmental factors. If genetic markers can explain a significant part of the variation, they can be considered as ideal candidates for genomic selection (Shi etal. 2019). Previously, microsatellite markers were frequently used to identify quantitative trait locus (QTL). With the progress of science, the advent of the single-nucleotide polymorphisms (SNP) are used in genome-wide association studies (GWAS) to identify QTL. In dairy cows, some of the major genes with significant effects on milk fat have been identified in previous GWAS studies. Therefore, with the existence of a large number of GWAS in dairy cows, these studies can be combined using meta-analysis to achieve higher power results. These studies contribute to our current understanding of the genetic regulation of milk fat yield traits. This approach provides a better understanding of the genetic architecture of complex traits. The network clustering algorithm and cluster identification are important tools in the structural analysis of networks. Various types of clustering algorithms are used for protein-protein interactions (PPIs) networks analysis. In this study, we used an algorithm known as MCODE to identify dense regions in the PPIs diagram (Bader and Hogue, 2003). The overall purpose of PPIs network clustering is grouping of genes or proteins that according to various scales are related to each other. The network of PPIs contains different proteins important in different pathways. These genes or proteins are clustered based on the similarity of metric and are known as matrix distances. It is also important to predict molecular assemblies of protein interaction data because it provides another level of functional annotation (Gollapalli et al., 2015). The purpose of this study was to conduct a meta-analysis of GWAS in cluster analysis to identify genes that are effective in milk fat yield in dairy cows. Material and methods: In this study, the data used were GWAS summary data. All data were collected from 19 published studies from 2010 to 2019. This research included main papers and dissertation (valid dissertations with published papers). All available genes were combined, synthesized, and evaluated using a meta-analysis method. The Cytoscape v3.7.2 software was used to analyze and visualize the genes examined by the STRING v1.5.0 plugin and to extract clusters from the MCODE v1.5.1 algorithm. Therefore, the results of the GWAS summary data were combined in molecular networks with PPIs, which have a significant role in increasing the association studies power to identify genes affecting milk fat trait. Also, the DAVID server was used to identify the gene ontology (GO) term enrichment in order to detect enriched biological terms associated with genomic regions and to identify gene networks using functional annotation clustering tools based on enriched pathways analysis. Results and discussion: In this study, we analyzed 223 genes using the STRING plugin in Cytoscape software. These genes were associated with at least another gene and had a direct and partial correlation with each other. In the gene network, the correlation created for milk fat yield trait included 213 genes or nodes and 219 edges (gene connection). The P-value calculated in the STRING network was statistically significant for enriched pathways in PPIs ( ). The collection of important and popular genes were evaluated using the MCODE plugin. Seven clusters were identified and grouped in this network. For instance, proteins in cluster 1 included: ARHGAP39, CPSF1, CYHR1, PPP1R16A, GRINA, MROH1, and SMPD5 genes. As shown in Table 2, cluster 1 (score=7) was connected with 7 node density to 21 nodes. This cluster showed proteins that play important roles in the internal space of the endoplasmic reticulum (cellular components), metal ion binding (molecular function), and integral to the membrane (cellular components). CPSF1, CYHR1, and GRINA were the major genes involved in the internal space of endoplasmic reticulum, metal ion binding, and membrane integral, respectively. It was found that clusters 1 and 2 have the highest score between all reported clusters. Conclusion: These results show that using data from different sources can increase the reliability and accuracy of GWAS studies. We were able to identify the most important genes in the network pathways using the GWAS summary data in cluster analysis. This method determines the quality of proteins involved in fat yield while facilitating our understanding of the molecular structures of proteins. The most important genes with high scores were identified in cluster one (ARHGAP39, CPSF1, CYHR1, PPP1R16A, GRINA, MROH1, and SMPD5) and two (HERC1, UBR4, ASB17, TRIM9, KLHL2, and BTRC). These clusters based on existing biological knowledge can help data mining and system models understand network interactions and pathways. These protein clusters provide a deep insight into how genes interact with each other in network analysis for fat yield. Moreover, it was observed that meta-analysis of GWAS summary data can play an important role in the wide understanding of network visualization and cluster analysis of identified genes in enriched pathways. Therefore, cluster analysis can improve the identified genes power for economically important traits such as milk fat yield in a population of dairy cows and can be used in future genomic evaluations and breeding programs.

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.

To investigate the effects of dietary supplementation with industrial vinegar (IV) and waste date vinegar (WDV) on growth performance, intestinal morphology, and immune response of broilers, five hundred Ross 308 chickens were randomly divided into 50 cages in a 42-day breeding period. The experiment consisted of ten treatments with five replications in each treatment including; control 1 (standard feed without WDV or IV), control 2 (control 1 + 2% water into the feed), 1, 2, and 3% of WDV and 2% industrial vinegar (IV) into the feed and 0.5, 1, and 1.5% of WDV and 1% industrial vinegar (IV) into the water. One chicken from each replicate was slaughtered on days 24 and 42 to investigate gastrointestinal tissue growth as well as intestinal morphology. The results showed that growth performance was not affected by treatments in any period. The height and width of the villus in the treatments containing 1% of WDV into the water and 2% of WDV into the feed increased with time compared to the control treatments. On day 42 of the experiment, the crypt depth was also higher in the treatment containing 1% of WDV into the feed compared to the other treatments. The ileal coliforms were also affected by WDV addition into the feed at 42 days of age compared to other groups (P < 0.05). The highest amount of IgM and IgG were found to levels of 2% and 1% of WDV into the diet, respectively (P < 0.05). However, SRBC, ND titter was not affected by treatments. Also, no difference was found between industrial vinegar in water or feed in most of the studied parameters. The results of this study showed that supplementation of the diet with WDV had a positive effect on intestinal morphology and immune system of broilers compared to industrial vinegar.

In the present study, kashk samples were collected from two regions of Iran, the Fars (Abadeh) and Razavi Khorasan (Kalat) provinces. Fifteen bacteria were isolated and physiological and biochemical assays were performed. After identification to the genus level, eight isolates were identified as lactic acid bacteria (LAB) and subjected to molecular identification and probiotic properties assays. The results revealed that the isolates were Enterococcus faecium KKP 3772 (KF1), Enterococcus faecium C1 (KF2), Pediococcus pentosaceus H11 (KF3), Pediococcus pentosaceus VNK-1 (KK4), Lactococcus lactis RSg (KK1), Enterococcus faecalis P190052 (KK2), Enterococcus mundtii CECT972T (KK3), and Lactiplantibacillus plantarum PM411 (KK5). Only the numbers of L. lactis RSg (KK1) and Lpb. Plantarum PM411 (KK5) decreased to below 9 Log CFU/mL after acidic conditions (pH = 3) and showed weak antibacterial activity. Enterococcus mundtii CECT972T (KK3) and E. faecium C1(KF2) were highly susceptible to bile salts, while P. pentosaceus VNK-1 (KK4) and P. pentosaceus H11 (KF3) showed the highest resistance. All of the isolates were resistant to tetracycline and sensitive to chloramphenicol and gentamicin. The antimicrobial activity of P. pentosaceus VNK-1 (KK4) and P. pentosaceus H11 (KF3) was higher than other isolates and consequently, their inhibition zones were larger. The adhesion capabilities of LAB isolates to intestinal epithelial cells were evaluated by examining the auto-aggregation factor and cell surface hydrophobicity. The highest and lowest cell surface hydrophobicity and auto-aggregation were obtained from P. pentosaceus VNK-1 (KK4) and E. mundtii CECT972T (KK3), respectively. In general, P. pentosaceus VNK-1 (KK4) and P. pentosaceus H11 (KF3) have shown better probiotic properties as compared to other isolates.