Animal Gene最新文献

Bombyx mori bidensovirus (BmBDV) replicates in the midgut cells of the silkworm to cause fatal flacherie disease. Flacherie disease causes a major impact in silk production and severe economic loss in the silk industries of India. BmBDV possesses two single stranded DNA that encodes for putative protein-primed DNA polymerase and adopts its own mechanism for replication. Development of rapid, sensitive and simple-to-use novel technology such as real-time polymerase chain reaction (RT-qPCR) will be useful in detection of BmBDV in silkworms. This RT-PCR assay will help to understand the significant pathology of BmBDV. The primers in specific to VD2 ORF1 region of BmBDV were designed to study their sensitivity and specificity. The detection limit of qPCR was found to be 3.01 × 10¹ copy number in the plasmid DNA with 96.3% efficiency in RT-PCR. In accordance, results shown in our study are 1000 times more sensitive than the conventional PCR with a detection limit of 3 × 10⁴ in PCR assay. Apart from displaying an increased sensitivity, these primers show high specificity towards BmBDV pathogenicity. On detection of flacherie infection studies with the specific primers shows gradual increase in detection of infectivity of BmBDV from 12 h to post infection for 48 h. In this case, the copy number reported at 48 h was found to be >10⁷ copies/reaction through qPCR analysis. The validation of this method was conducted using 90 diseased samples collected from 10 different sericulture farms. Here, detected virus copy numbers range from 1.24 × 10³ to 1.08 × 10¹⁰ where the optimization of RT-PCR could be used as a valuable tool in detection of BmBDV virus. Further aid in formulating appropriate disease control strategies, screening silkworm breeds for resistance during early infection stages are initiating reasons for this study. This particular analysis will provide a platform for developing silkworm hybrids to prevent further crop loss at the field level.

Understanding the effects of antibiotics on immunogen expression and the genetic consequences of antibiotic overuse in rabbit farms are becoming increasingly important as customers become more interested in healthy and safe products. The present study was destined to evaluate the effect of Neomycin, as a common antibiotic used in rabbit farms, on immune genes, some hematological parameters, and small intestinal histology in two rabbit lines: V and Alexandria. A total of 160 rabbits were orally provided 50 mg of Neomycin in 5 ml of water daily. The immunity genes have been studied using IL-2, IL-4 and CD1. The results showed an independent relationship between Neomycin-treated groups. At 21 days of age, V-line rabbits had the lowest levels of hemoglobin (Hb), red blood cells (RBCs), and lymphocytes compared to the Alexandria line. The responder rabbits showed the highest value in Hb, RBCs, and lymphocytes while the non-responder rabbits showed the highest value in WBCs. Several cytokines were tested, and no significant differences were found between the two lines. While CD1A1 (T cell surface glycoprotein CD1) increased the gene expression in the V-line innate-immune group and the responder group to the Neomycin. Based on the findings, Neomycin enhanced rabbits' immune responses.

Crabs are one of the most diversified crustaceans among metazoans, from both morphological and ecological point of view. Among them, ghost crabs (Genus Ocypode; Family Ocypodidae) play an important role in the tropical and sub-tropical intertidal ecosystems. Due to a lack of comprehensive data, their cryptic behaviour, mimicry or camouflage, and a lack of formal taxonomic documentation, identification of this group is often difficult. Molecular taxonomy can help to overcome these barriers by assisting in species identification. The present study is the first large scale attempt to identify ghost crabs using DNA barcodes, which are now available globally, for 16 morphospecies in the genus Ocypode. A total of 164 barcodes were analysed for species delimitation that are available globally including specimens collected from the Indian Sundarbans. Various species delimitation methods (K2P, NJ, Bayesian supported PTP and Bayesian tree) revealed a total of 19 MOTUs for 16 distinct species where multiple MOTUs were observed in 3 species with inconsistencies in genetic distances and tree topologies. We recognised cryptic diversity in Ocypode macrocera, O. platytarsis and O. quadrata with intraspecies distances greater than the species delimitation threshold of 0.053 ± 0.01. The current study also concluded that O. platytarsis and O. brevicornis are two distinct species, putting an end to the debate over their species synonymy. Furthermore, our Bayesian analysis confirmed the retraction of the genus Hoplocypode specified only for O. occidentalis. As a result, the current research added a new dimension to ghost crab taxonomy by contributing to the detection of cryptic species, thus speculating the possibility of new species and reaffirming the effectiveness of the DNA barcoding while demanding enrichment of the global database for comprehensive species identification and extensive approach of molecular taxonomy as well as morphological interventions in solving taxonomic puzzles.

Mechanistic target of rapamycin (mTOR), an evolutionarily conserved serine/threonine kinase with diverse functions, is expressed ubiquitously in central and peripheral tissues. Recent data strongly implicated mTOR signaling in the regulation of food intake, but the relevant information is still limited in teleost. To reveal the effects of mTOR on food intake in an endemic economic fish in the upper reaches of the Yangtze River, this study firstly cloned the full length cDNA of mTOR in Schizothorax prenanti (S. prenanti) and found its mRNA was widely distributed in various tissues. Rapamycin (1 mg/kg body weight), the specific inhibitor of mTOR, significantly increased the food intake of S. prenanti (P < 0.05), which was accompanied by significantly elevated Ghrelin mRNA levels and had no influence on NUCB2 mRNA levels in the hepatopancreas and intestine (P < 0.05). Whereas L-leucine (10 mg/kg body weight), an activator of mTOR, inhibited the expression of Ghrelin mRNA and stimulated the transcription level of NUCB2 in hepatopancreas and intestine at 4 h post intraperitoneal (i.p.) injection (P < 0.05), but failed to induce a significant decrease in food intake. Collectively, this study provides a novel information on mTOR in the regulation of food intake, which is linked to Ghrelin but not NUCB2 in S. prenanti.

Keratin-associated proteins (KAPs) are constituents of wool and play an important role in determining the characteristics of wool. Variation in many ovine KAP genes are associated with wool traits and are useful as markers for improving wool traits in sheep. The identification of promoters in a gene is an important part of the recognition of a gene's complete structure. In silico study was done with aim of analyzing the promoter regions and transcription factors with the aid of transcription start site (TSSs), regulatory elements and motifs of sheep KAP genes. Most of predicted TSSs (64%) found below -500 bp away from the start codon of the gene being expressed, and the rest were found from −515 to −1502 upstream of start codon. MKAP3 was identified as the most common motif for sheep KAP genes, which serves as binding sites for Zinc finger transcription factor to regulate the expression of the genes. Poor CpG Islands observed might suggest that their gene expression regulation pattern is tissue specific. In general, in silico analysis of KAP genes of sheep could be helpful to add knowledge about gene regulatory elements in the promoter regions. Furthermore, this finding will help for wool traits improvement of sheep.

Diaphorina citri (Kuwayama) (Homoptera: Liviidae) is an important pest of Rutaceae crops. D. citri is the vector of citrus Huanglong disease. This pest often causes outbreaks of citrus Huanglong disease and has a devastating impact on the citrus industry. To date, there has been no systematic study of genes related to oviposition and spermatogenesis in D. citri. Based on the transcriptome and proteome sequencing of D. citri abdominal tissues, genetic information on multiple gene families related to oviposition and spermatogenesis in male and female adults was obtained. The gene and protein expression differences of 16 gene family members related to oviposition and spermatogenesis were analyzed in male and female adults. The transcriptome and proteome sequencing data of the Vg, VgR, and Tssk gene families were analyzed. In this study, we analyzed the differences in the gene and protein expression of 16 gene families related to abdominal oviposition and spermatogenesis in female and male D. citri adults. Some of the data indicated that the differences in abdominal oviposition- and spermatogenesis-related gene expression between male and female adults can affect reproductive behavior. However, how the expression of related genes affects the reproductive behavior of male and female adults, including courtship, mating and oviposition, requires further study. The results provide an important research direction for the control of D. citri by using genetic engineering technology.

Genome-wide association study (GWAS) is a powerful tool for identifying loci and individual polymorphisms associated with economically important traits in various species of productive animals. The objective of this study was a genome-wide association search in sheep of the North-Caucasian meat and wool breed to identify single nucleotide polymorphisms (SNPs) associated with a class assessment determined by a set of productive parameters. Sixty rams of the North-Caucasian sheep breed were selected based on the criteria of phenotype and assigned to one of the groups - elite (control, 50 rams) and super-elite (case, 10 rams). They were genotyped using Ovine Infinium HD BeadChip 600 K DNA array and search of SNPs associations with productivity class were conducted. Candidate genes was annotated using ENSEMBLE database. Twelve SNPs were identified that have highly significant differences in the frequency of occurrence in animals with a super-elite score. From these twelve substitutions, five are in introns, seven are located at different distances from the genes. Most of the genes located next to the detected SNPs regulate development and functions of neurons. The greatest reliability in the associations was shown by the substitutions rs410503867 (p = 2 × 10⁻¹³) and rs428223899 (p = 1.2 × 10⁻¹⁰) located on chromosomes 4 and 21. Only in animals with a super-elite score three substitutions - rs404739757, rs402746571 and rs413668028 were found, they are presented together next to NALCN gene. The studies revealed a number of new candidate genes located near of SNPs associated with a general assessment of the complex of sheep traits (DGKB, PAK1, CHL1, CTTNBP2, NALCN and NFATC2) and proposed new molecular genetic markers in the form of single nucleotide polymorphisms. We are promising to use the identified genetic markers of super-elite animals in marker-associated breeding to improve the productive qualities of some sheep breed.

Epidermal growth factor receptor (EGFR) gene plays a pivotal role in cell communication and proliferation in both vertebrates and invertebrates. The objectives of this study were to monitor for single nucleotide polymorphisms (SNPs) in the 3′untranslated region (3′-UTR) of EGFR in geese and evaluate the relationship between the polymorphisms of these SNPs and egg production. Using the direct sequencing method, a single nucleotide polymorphism c.*7750G > A was detected in 3′-UTR region of EGFR gene. Two alleles (G and A) and three genotypes (GG, AG and AA) were identified. Association analysis results showed that c.*7750G > A SNP was significantly associated with egg production at 34-week egg-laying period (P < 0.01). Geese with GG genotype (mutant) significantly produced more eggs compared to those with AA genotype (wildtype). Quantitative real-time PCR indicated that the expression levels of geese EGFR mRNA was highly expressed in kidney and granulosa cells and also expressed in follicles granulosa cells at different egg production stages with high expression in the large white follicle. Also, the mRNA expression level of EGFR in geese ovaries indicated that the geese with GG genotype recorded significantly higher expression levels compared to the geese with AA genotype. Luciferase activity assays showed that dme-miR-5-3p could significantly decrease luciferase activity in the presence of allele A compared with negative control and allele G (P < 0.05). These results suggest that c.*7750G > A in the 3′-UTR of EGFR gene may play a significant role in egg production in Yangzhou geese.

Two most recently described macaque species; the white-cheeked macaque (Macaca leucogenys) and Arunachal macaque (Macaca munzala) were discovered from a single biodiversity hotspot, the Eastern Himalaya. We conducted surveys in the West Siang, Arunachal Pradesh, India and collected five faeces and two skin samples of macaques that on DNA analysis identified as white cheeked macaques. Subsequently, we undertook intensive field surveys and successfully captured white cheeked macaques in camera traps as well as found a captive juvenile individual during questionnaires from the same region. We report white-cheeked macaque from West Siang about 197 km away from China. Unfortunately, white-cheeked macaque has not been yet included in the Wildlife (Protection) Act, 1972 of India and therefore, the present study laid foundation to promote field studies in Central Arunachal Pradesh to delineate distribution boundary and population size of white-cheeked macaque in Arunachal Pradesh.

Infestation of commercially important silkworm Bombyx mori by dipteran endoparasitoid Exorista bombycis causes ~20% revenue loss. Though invasion has induced hemocyte encapsulation and melanisation of invaded larva, strategies to compromise with host-defense system by dipteran endoparasitoids is unknown. In the present study variation in genome wide gene expression was analyzed by utilizing microarray data acquired from hemocytes collected at 6 h after invasion that showed significant (P < 0.05) up-regulation of 423 genes and down-regulation of 587 genes. Among them 11 detoxification genes and 34 immune genes down-regulated showing repressed defense reactions substantiated by reduced activity of detoxification enzymes esterase and FMO in early stage of infestation. The down-regulated genes were enriched after comparing with control and by filtering unreliable probes. GO annotation of significantly (P < 0.05) down-regulated genes was used for clustering and categorization of functionally related defense processes. Networking of interacting partners of down-regulated defense genes showed relatedness among the down- regulated biological processes viz., Wnt signalling, DNA repair, transcription, catabolism, glutamine metabolism, components of cytoskeletal structure and transport. These clusters perform immune responses, signalling, stress responses, cell redox homeostasis, cytoskeletal organization, glutamine, glycogen and chitin metabolism, protein, lipid and carbohydrate metabolism and transmembrane transport. Autophagy, apoptosis, cell adhesion, cell growth and cellular lipid metabolism also showed down-regulation independently. Down-regulation of defense genes, interacting components and low detoxification enzyme activity in early infestation revealed active suppression of host defense at genic, enzymic and interactome level which is an unknown anti-defense mechanism of dipteran endoparasitoids.