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Introduction: Computational studies were performed to investigate the unknown status of endosomal and cell surface receptors in SARS-CoV-2 infection. The interactions between Toll-like receptors (TLRs)-4/7/8/9 or ACE2 receptor and different SARS-CoV-2 variants were investigated. Methods: The RNA motifs for TLR7, TLR8 and a CpG motif for TLR9 were analyzed in different variants. Molecular docking and molecular dynamics (MD) simulations were performed to investigate receptor-ligand interactions. Results: The number of motifs recognized by TLR7/8/9 in the Alpha, Delta and Iranian variants was lower than in the wild type (WT). Docking analysis revealed that the Alpha, Delta and some Iranian spike variants had a higher affinity for ACE2 and TLR4 than the WT, which may account for their higher transmission rate. The MD simulation also showed differences in stability and structure size between the variants and the WT, indicating potential variations in viral load. Conclusion: It appears that Alpha and some Iranian isolates are the variants of concern due to their higher transmissibility and rapid spread. The Delta mutant is also a variant of concern, not only because of its closer interaction with ACE2, but also with TLR4. Our results emphasize the importance of ACE2 and TLR4, rather than endosomal TLRs, in mediating the effects of different viral mutations and suggest their potential therapeutic applications. [ABSTRACT FROM AUTHOR]

Chicken gut microbiota is affected by factors such as diets, environmental,and bird age. In the current study, the effects of age and region on the ileum bacterial population of broiler chickens were investigated. A total of 2679 chickens in four geographical regions of Iran were sacrificed in the first to eighth week of age. Stool samples were collected and DNA was extracted and analyzed for the detection of Lactobacillus, Enterococcaceae, Clostridiaceae, Streptococcaceae, and Actinobacteria, using specific primers and probes. Purified amplicons were quantified by QuantiFluor® and pooled for sequencing. Findings showed that L. acidophilus was the dominant bacterium during the first four weeks, and was substituted with L. crispatus and L. salivarius in the next four weeks. So, the Lactobacillus family was the most dominant bacteria at all ages showing its essential role in chicken physiology. The age of chickens significantly affected the percentage of L. crispatus, L. acidophilus, L. salivarius, Clostridiaceae, Enterococcaceae, Streptococcaceae, and Actinobacteria. The breeding region influenced Streptococcaceae, with the highest percentage in the hot region. Chicken weight had a significant effect on Enterococcaceae. Broiler breeder age and distance to the nearest farm had no effect on ileum bacterial populations. This study showed there are several factors during the broiler breeding period that have an impact on microbial population changes at different ages. [ABSTRACT FROM AUTHOR]

The Clostridium genus is composed of a large spectrum of heterogeneous bacteria. They are Gram-positive, mostly mesophilic, and anaerobic spore-forming strains. Clostridia are widely distributed in oxygen-free habitats. They are found principally in the soil and intestines of ruminants as normal flora, but also are the cause of several infections in humans. The infections produced by important species in humans include botulism, tetanus, pseudomembranous colitis, antibiotics-associated diarrhea, and gas gangrene. Immunization with toxoid or bacterin-toxoid or genetically modified or other vaccines is a protective way against clostridial infection. Several experimental or commercial vaccines have been developed worldwide. Although conventional vaccines including toxoid vaccines are very important, the new generation of vaccines is an effective alternative to conventional vaccines. Recent advances have made it possible for new vaccines to increase immunogenicity. This review discusses briefly the important species of clostridia in humans, their toxins structure, and vaccine development and usage throughout the world. [ABSTRACT FROM AUTHOR]

The aim of this study was to identify the production capacity of six commercial silkworm Bombyx mori L. (Lepidoptera: Bombycidae) lines (numbers 31, 32, 103, 104, 153, and 154), which were contaminated with Nosema bombycis Nageli spores (103 or 105 per ml), the fungus responsible for pebrine (pepper disease) in silkworms. Effects of the two fungal Microsporidia concentrations were tested on silkworm feeding on either treated (with 1.0% Lactobacillus rhamnosus probiotics or 0.5% Echinacea purpurea plant extract) or non‐treated (control) mulberry leaves (Moraceae). Traits included larval weight, cocoon weight, cocoon shell weight, pupal rate, silk shell percentage, total cocoon number, cocoon yield per 10 000 larvae, larval mortality, and incidence of infection in moth. Results showed that lines, feed supplements, and level of Microsporidia spore infection had significant effects on most estimated parameters. Both concentrations of Microsporidia spores led to a decrease in the silkworm performance and an increase in mortality. Supplementation of mulberry leaves with probiotics or plant extract resulted in improvement of economic traits in B. mori. Commercial line 153 performed the best in most of the studied traits and was highlighted as the optimum line for breeding in Iranian silkworm industry. Also, line 104 was the most resistant line to N. bombycis. This information will be helpful for selection of the best line for breeders according to the rearing conditions and desirable economic traits. [ABSTRACT FROM AUTHOR]

Toll-like receptors (TLRs) are a class of pattern recognition receptors (PRRs) that detect pathogen associated molecular patterns and activate innate and adaptive immune system. Coronaviruses can be detected via TLRs through their biological materials such as ribonucleic acids, glycoproteins and CpG motifs. During COVID-19 pandemic, different strategies have been used for combating SARS-CoV-2 to initiate a proper and balanced immune response through TLRs or other PRRS. Understanding the trigerred TLR signaling pathways during coronavirus infections would assist to understand the control and defense mechanisms against these viral diseases. In this review, we summarize different studies on the use and function of of TLRs and their signaling pathways as vaccines/adjuvants or therapeutic agents against coronavirus infections. Since the pandemic is ongoing and there still many unknowns with respect to COVID-19 immunology, we highlight the role of TLRs and their agonists/antagonists in previous coronavirus infections, and show their potential role in the current SARS-CoV-2 immunopathology. [ABSTRACT FROM AUTHOR]

Clostridium is a Gram-positive, rod-shaped, anaerobic, and spore-forming bacterium, which is found in the surrounding environments throughout the world. Clostridium species cause botulism, tetanus, enterotoxaemia, gas gangrene, necrotic enteritis, pseudomembranous colitis, blackleg, and black disease. Clostridium infection causes severe economic losses in livestock and poultry industries. Vaccination seems to be an effective way to control Clostridial diseases. This review discusses the toxins and vaccine development of the most common pathogenic Clostridium species in animals, including Clostridium perfringens, Clostridium novyi, Clostridium chauvoei, and Clostridium septicum. In this comprehensive study, we will review different kinds of clostridial toxins and the vaccines that are experimentally or practically available and will give a short description on each vaccine focusing on its applications, advantages, and disadvantages. [ABSTRACT FROM AUTHOR]

An efficient method for detection of foot and mouth disease (FMD) and, particularly, differentiation of vaccinated from infected animals is the use of nonstructural (NS) proteins as antigens in Enzyme-Linked Immunosorbent Assay (ELISA) Kits. In this study, only epitopic regions of 3AB and 3D NS proteins were used for recombinant protein production, as a cost-effective method instead of peptide synthesis, for application in in-house ELISA diagnostic kits. Specific primers were designed according to the antigenic regions of 3AB (C-terminus of 3A and the whole 3B) and 3D (N-terminus) proteins, and the polymerase chain reaction (PCR) amplification was performed. Purified amplicons were cloned into pET21a (+) vectors and then transformed into Escherichia coli (BL21). Thereafter, bacteria were induced with 1 mM isopropyl β-d-1-thiogalactopyranoside (IPTG) for expression of antigenic proteins. Antigenic 3AB protein was expressed in soluble form, but 3D protein was extracted from the bacterial lysate. Protein expression was confirmed using sodium dodecyl sulfate–polyacrylamide gel electrophoresis and Western blot analyses. An indirect ELISA was developed for each protein, and the diagnostic sensitivity and specificity were determined. The 3AB-ELISA showed higher sensitivity and specificity than 3D-ELISA (95.24% and 100%, compared with 90.48% and 88.71%, respectively). The epitopic 3AB-ELISA developed here can be used for detection and differentiation of FMD infected from vaccinated animals, but the epitopic 3D-ELISA showed lower efficiency in screening for FMD status. [ABSTRACT FROM AUTHOR]

SlC11A1 gene is a potential candidate gene related to susceptibility to a wide range of intracellular pathogens. In this research, we used logistic regression analysis in a casecontrol study to find the relationship between SLC11A1 gene and estimated breeding values of production traits (estimated breeding values of protein and fat yield, and protein and fat percentage) in Holstein dairy cattle in a selective genotyping method. Samples have been genotyped based on clinical mastitis data using PCR-SSCP method, previously. The contrast between heterozygous genotype and both homozygous genotypes showed that despite of significant relationship between clinical mastitis and SLC11A1, there was no negative effect of the gene of interest on production traits. Hence, considering the unfavorable relationship between clinical mastitis and production traits, selection of SLC11A1 gene will have no unfavorable effect on production traits, and it will not disorganize selection breeding programs based on health data. [ABSTRACT FROM AUTHOR]

Toll-like receptors (TLRs) are known for their essential roles in promotion of innate immunity and induction of adaptive immunity through recognition of pathogen-associated molecular patterns (PAMPs) or danger associated molecular patterns (DAMPs). TLR genes are excellent models for the study of the selective pressure enforced by microorganisms on the host genome. Phylogenetic analyses have shown that interactions between pathogens and immune systems have changed during evolution. Selective pressure for maintenance of specific pathogen recognition has led to evolution of TLRs under both positive and purifying selection. However, intracellular and cell-surface TLRs have been affected differently due to their variation in conservational constraints. In this review, we summarize some of the main studies on the influence of selection on shaping the evolution of several TLR gene families in different terrestrial vertebrate species. We also describe the effect of evolution on the function of different TLRs and their specific conservations in these species and show similarities and differences in evolutionary patterns of TLR orthologs among species as well as among TLR gene families. [ABSTRACT FROM AUTHOR]

Maternal effect genes have highly impressive effects on pre-implantation development.Filia/Ecat1/Khdc3lis a maternal effect gene found in mouse oocytes and embryos, loss of which causes a 50% decrease in fertility. In the present study, we investigatedFiliamRNA expression in bovine oviduct, 30- to 40-day fetus, liver, heart, lung, and oocytes (as a positive control), by RT-PCR and detected it only in oocytes. A 443 bp fragment was amplified only in oocytes and was sequenced as a part of bovine predictedFiliamRNA. We analyzed bovine and ovine Filia N-terminal peptide sequence in PHYRE2, and a KH domain was predicted. Protein alignment using ClustalW indicated a highly identical N-terminal extention between the 2 species. Immunohistochemical analysis using anti-bovine Filia antibody showed the expression of Filia protein in the zone surrounding the nuclear membrane, and in the subcortex of ovine oocytes of primary and antral follicles. However, in the bovine, Filia has been found through the oocyte cytoplasm of antral follicles, and here it is further confirmed in the primary follicles. Our data suggests a difference in Filia expression pattern between cow and sheep, although the sequence is highly conserved. [ABSTRACT FROM AUTHOR]

Sonic Hedgehog (SHH) is a signaling pathway mediated through a receptor system which seems to have effects on oocyte maturation and embryonic development. Purmorphamine is an SHH agonist that performs a crucial role in the regulation of the activity of SHH receptors and downstream transcription factors. The aim of this study was to analyze the effect of purmorphamine on the mRNA expression of SHH signaling downstream molecules (Patched1, Glioma-Associated Oncogene1, Smoothened, Histone Deacetylase1, Histone Deacetylase2 and Histone Deacetylase3) in ovine two-cell embryo. Ovaries were obtained from a slaughterhouse, and cumulus-oocyte complexes were aspirated and cultured in maturation media containing 0, 250 or 500 ngmL-1 purmorphamine. Then, oocytes were fertilized and cultured in a CR1 culture medium and after 24 h, two-cell embryos were collected for RNA extraction. Gene expression was evaluated by real-time polymerase chain reaction (PCR). Results indicated that in 250 ng mL-1 purmorphamine, Smo, Ptch1 and Hdac3 expression reduced, Hdac1 expression increased, and GUI and Hdac2 expression levels did not change. In 500 ng mL-1 purmorphamine, GUI and Smo transcripts increased, while Ptch1, Hdac2 and Hdac3 transcripts decreased. Regarding to the presence of SHH signaling molecules in two-cell embryos and their response to purmorphamine, it can be suggested that SHH signaling is probably active before embryonic genome activation in ovine embryos. [ABSTRACT FROM AUTHOR]

Different culture system conditions can affect cleavage, zygote genome activation, embryo development, and viability. The purpose of the present study was to compare a monoculture synthetic oviductal fluid medium with modified sequential G1/G2 media and also to investigate the effect of epidermal growth factor (EGF) on bovine embryo development. Oocytes were aspirated from 2- to 6-mm ovarian follicles and matured in maturation media. After in vitro fertilization of matured oocytes, presumptive zygotes were cultured in test media with or without additional exogenous EGF. Results indicated that EGF significantly increased blastocyst formation but not cleavage rate in both media. There was no significant difference between the sequential and the monoculture media. [ABSTRACT FROM AUTHOR]

Purmorphamine is a Sonic Hedgehog agonist that plays an important role in activity regulation of receptors and transcription factors. This study was carried out to assess the effects of Purmorphamine on nuclear and cytoplasmic maturation, epigenetic changes of in vitro matured oocytes and development of parthenogenetic ovine embryos. Ovine ovaries were randomly collected from industrial slaughterhouse, and cumulus-oocyte complexes were cultured in media containing 250 or 500 ng/ml Purmorphamine or without Purmorphamine as a control group. Then, after in vitro maturation (IVM), Hoechst stain, cell tracker blue and histone H4K12 antibody were used, respectively to assess the nuclear and cytoplasmic maturation, and histone acetylation rate of matured oocytes. Matured oocytes were activated and cultured to the blastocysts stage in order to determine the parthenogenetic embryo cleavage rate. Expression of Histone Deacetylase 1, 2 and 3 (Hdac1 , Hdac2 and Hdac3 ) genes were evaluated in matured oocytes by quantitative Real Time PCR. Results showed that although the concentration of 500 ng/ml Purmorphamine had no significant influence on the oocyte nuclear maturation, it led to an increase in oocyte cytoplasmic maturation. Also, no significant difference in the rates of cleavage and blastocysts per cleavage was observed (P > 0.05) in the groups treated with Purmorphamine compared to the control. Quantitative PCR analysis indicated that in 500 ng/ml Purmorphamine treatment, Hdac 2 and 3 transcripts decreased and the H4K12 acetylation increased significantly (P < 0.05). Consequently, this study showed that 500 ng/ml Purmorphamine can induce the cytoplasmic maturation of oocytes, and may possibly be a good addative for improvement of ovine oocyte cytoplasmic maturation. [ABSTRACT FROM AUTHOR]

Enterotoxaemia, a disease that affects domestic ruminants, is caused by the epsilon toxin of Clostridium perfringens type D and B. Control and prophylaxis are based on systemic vaccination of small ruminant herds with epsilon toxoid. Purified epsilon toxin is an essential material for vaccine evaluation. It is also necessary for diagnosis of enterotoxaemia disease in the field by in vitro tests including ELISA. The aim of this study was to set up a method for preparation of functional purified epsilon toxin of C. perfringens type D to be used in serum neutralization test. In this study, epsilon toxin was prepared from C. perfringens type D culture precipitated with ammonium sulfate, dialyzed against phosphate buffered saline (PBS) buffer and then, purified using chromatography system. Then, the purified epsilon toxin was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Toxin function was confirmed by cell culture and minimum lethal dose (MLD) assays. Also rabbits were immunized by vaccine in two turns with a 28-day interval. Then, blood samples were collected, and serum neutralization (SN) test was carried out. Results showed that the purified toxin was suitable for SN assay. Our purification method was simple, fast and cost-effective for preparation of epsilon toxin. • A simple method for purification of Clostridium perfringens epsilon toxin was developed. • Epsilon was purified by (NH₄)₂SO₄ precipitation, dialysis and chromatography. • Toxin function was confirmed by cell culture and minimum lethal dose assays. • The amount of neutralizing epsilon antibodies was 5.5 IU/mL. [ABSTRACT FROM AUTHOR]

Melatonin is a hormone mainly secreted by the pineal gland during the circadian cycle, with low levels during the daytime and prominent levels during the night. It is involved in numerous physiological functions including the immune system, circadian rhythm, reproduction, fertilization, and embryo development. In addition, melatonin exerts anti-inflammatory and antioxidant effects inside the body by scavenging reactive oxygen and reactive nitrogen species, increasing antioxidant defenses, and blocking the transcription factors of pro-inflammatory cytokines. Its protective activity has been reported to be effective in various reproductive biotechnological processes, including in vitro maturation, embryo development, and survival rates. In this comprehensive review, our objective is to summarize and debate the potential mechanism and impact of melatonin on oocyte maturation and embryo development through various developmental routes in different mammalian species., (© The Author(s) 2024. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The Clostridium genus is composed of a large spectrum of heterogeneous bacteria. They are Gram-positive, mostly mesophilic, and anaerobic spore-forming strains. Clostridia are widely distributed in oxygen-free habitats. They are found principally in the soil and intestines of ruminants as normal flora, but also are the cause of several infections in humans. The infections produced by important species in humans include botulism, tetanus, pseudomembranous colitis, antibiotics-associated diarrhea, and gas gangrene. Immunization with toxoid or bacterin-toxoid or genetically modified or other vaccines is a protective way against clostridial infection. Several experimental or commercial vaccines have been developed worldwide. Although conventional vaccines including toxoid vaccines are very important, the new generation of vaccines is an effective alternative to conventional vaccines. Recent advances have made it possible for new vaccines to increase immunogenicity. This review discusses briefly the important species of clostridia in humans, their toxins structure, and vaccine development and usage throughout the world., Competing Interests: The authors declare no competing interests.