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This study explored whether zinc oxide (ZnO) supplementation could alleviate weanling-induced intestinal injury through TLR and NOD-like receptor signaling pathways. Twelve early-weanling piglets were allotted to two dietary treatments (control vs 2200 mg Zn/kg from ZnO) for 1 wk. The results showed that supplemental ZnO improved daily gain and feed intake, decreased post weaning scour scores, increased villus height and villus height:crypt depth ratio at the jejunal mucosa, and decreased diamine oxidase activity and endotoxin concentration in plasma. The intestinal mRNA levels of TLR4 and its downstream signals, including MyD88, IL-1 receptor-associated kinase 1 and TNF-α receptor-associated factor 6, were decreased, and the expressions of intestinal pro-inflammatory cytokines and chemokines were decreased simultaneously in the ZnO-supplemented piglets. Although NF-κB p65 mRNA abundance was not affected by ZnO supplementation, NF-κB p65 protein expression was down-regulated by ZnO. However, ZnO supplementation had no effect on intestinal expressions of NOD1 and NOD2, and their adaptor molecule receptor-interacting serine/threonine-protein kinase 2, as well as protein expressions of caspase-3 and heat shock protein 70. The results indicated that the protective effects of ZnO on intestinal integrity were closely related to decreasing the expressions of genes associated with inflammation through inhibiting the TLR4-MyD88 signaling pathways., (© The Author(s) 2013 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.)

A full sequence of TaHSP70 (heat shock protein 70 of Tanichthys albonubes) was amplified which was 2398 bp, including an open reading frame (ORF) of 1392 bp encoding a polypeptide of 643 amino acids with all three HSP70 family signatures and cytosolic motif of EEVD. Genomic DNA structure analysis revealed that the TaHSP70 gene contained one intron in 5'UTR. BLAST analysis revealed that the TaHSP70 gene shared high similarity with other known HSP70 genes. The alignment of inferred amino acid sequences also showed high degrees of similarity among the homologues. There was a basal mRNA expression of TaHSP70 in the different tissues from the non-exposed T. albonubes and the highest expression level in the liver. To investigated the time- and dose-dependent relationship of the expression of TaHSP70 following exposure to heavy metals, T. albonubes were exposed to 2(-1) 96 h-LC50 (0.027 mg L(-1)), 4(-1) 96 h-LC50 (0.0135 mg L(-1)) of copper and 2(-1) 96 h-LC50 (2.31 mg L(-1)), 4(-1) 96 h-LC50 (1.15 mg L(-1)) of cadmium for 96 h. Hsp70 expression relative to the control was analyzed by Quantitative real-time PCR and Western blotting. The results indicated that there were a dose-dependent expression pattern and an exposure time effect in the liver responded to heavy metal stress. Interestingly, TaHSP70 gene expressions did not show consistent changes between transcription and translation levels. Taken together, the dynamics of TaHSP70 expression observed provided important insight into heavy metal stress, heat shock protein activity, and the potential ways to monitor the chronic stressors in T. albonubes culture environments., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Simple Summary: The elongation of very-long-chain fatty acids gene 2 (elovl2) of Chinese sturgeon was identified in acipenseriformes species for the first time. Elovl2 was highly conserved in molecular evolution among vertebrates. Functional characterization in yeast demonstrated that Chinese sturgeon Elovl2 could efficiently elongate C20 (20:4n-6 and 20:5n-3) and C22 (22:4n-6 and 22:5n-3) substrates, confirming its critical roles in long-chain polyunsaturated fatty acid (LC-PUFA) biosynthesis. Diets with different types of PUFAs can affect the transcription levels of elovl2 in Chinese sturgeon. This study enhances our understanding about the evolutionary and functional characteristics of elovl2 and also provides novel insights into the mechanism of LC-PUFA biosynthesis in vertebrates. Elongases of very-long-chain fatty acids (Elovls) are critical rate-limiting enzymes that are involved in LC-PUFA biosynthesis through catalyzing the two-carbon elongation of a pre-existing fatty acyl chain. Thus far, several Elovls have been extensively studied in teleost. However, the functional and physiological roles of Elovls in chondrichthyans have rarely been reported. In this study, we identified and characterized elovl2 from the endangered Chinese sturgeon (Acipenser sinensis) by whole genome scanning. The results show that the coding sequence of elovl2 was 894 bp in length, for a putative protein of 297 amnio acids. Comparative genomic analyses indicated that Chinese sturgeon elovl2 was evolutionarily conserved. Functional characterization in yeast demonstrated that the Chinese sturgeon Elovl2 could efficiently elongate C20 (ARA and EPA) and C22 (22:4n-6 and 22:5n-3) substrates, confirming its critical roles in LC-PUFA biosynthesis. Spatial and temporal expression analyses showed high elovl2 mRNA levels were detected in the liver and brain and showed an increase trend both in embryonic and post-hatching stages. Interestingly, diets with vegetable oils as lipid sources could significantly induce the high expression of elovl2 in Chinese sturgeon, implying that the endogenous LC-PUFA biosynthesis pathway was stimulated by lack of LC-PUFA in their diets. Our findings will enhance our understanding about the evolutionary and functional roles of elovl2 and provide novel insights into the LC-PUFA biosynthesis mechanism in vertebrates. [ABSTRACT FROM AUTHOR]

Simple Summary: Tachykinin1 (tac1) gene products participate in the regulation of hormone secretion in the pituitary of Dabry's sturgeon. Two products of tac1 gene, Substance P (SP) and neurokinin A (NKA), were initially confirmed to regulate the gene expression of two gonadotropins (lh and fsh) in the Dabry's sturgeon pituitary tissue by intraperitoneal injection in vivo and co-culture of primary cells in vitro. This research presents the initial investigation into the reproductive role of SP and NKA in Dabry's sturgeon, offering a valuable insights for advancing efficient ripening techniques in artificial breeding of sturgeon. As an ancient and endangered species unique to the Yangtze River in China, the wild population of the Dabry's sturgeon has become scarce. Due to the long time till the first sexual maturity of Dabry's sturgeon, the population of artificially bred Dabry's sturgeon recovered slowly. As a member of the tachykinin family, TAC1 has been reported to have a variety of functions in mammals such as pain control, smooth muscle contraction and reproductive cycle regulation, but the function of Tac1 in fish has been rarely reported. In this study, we synthesized two tac1 gene products, Substance P (SP) and neurokinin A (NKA), and further verified the effect of two tac1 gene products on the secretion of related hormones in the pituitary of Dabry's Sturgeon by intraperitoneal injection and co-culture of primary cells. Expression studies revealed that the newly cloned tac1 were mainly distributed in the hypothalamus and pituitary tissue of the brain. In prepubertal Dabry's sturgeon, this study showed that the two gonadotropins' mRNA levels in pituitary tissue can be significantly increased by SP and NKA through intraperitoneal injection, and the LH protein level in serum was also increased. Further study showed that both NKA and SP could promote the two gonadotropins' mRNA expression in pituitary cells of Dabry's sturgeon. In addition, we explored the optimal dose and time of SP and NKA on pituitary cells is 24 h and over 10 nM. These results, as a whole, suggested that tac1 gene products play an important role in gonadotropin release and gonadal development in prepubertal Dabry's sturgeon. [ABSTRACT FROM AUTHOR]

Polyphenols sourced from Ilex latifolia Thunb. (PIT) contain high levels of phenolic acids, tannic acids, triterpenoids and so on, which play important roles in antioxidant function. This study was conducted to investigate the effects of PIT against intestinal injury in piglets under oxidative stress. Thirty-two weanling piglets were arranged by a 2 × 2 factorial experiment with diets (basal diet vs. PIT diet) and oxidative stress (saline vs. diquat). All piglets were injected with saline or diquat on d 21, respectively. After 7 days, all pigs were slaughtered and intestinal samples were collected. PIT enhanced jejunal villus heights and crypt depth in the piglets under oxidative stress. PIT increased the activities of intestinal mucosal lactase, sucrase and maltase in the challenged piglets. PIT also increased the jejunal ratio of protein to DNA and ileal protein content. PIT increased the jejunal activities of GSH-PX and GSH content and reduced the ileal MDA amounts. Furthermore, PIT regulated the expression of ferroptosis mediators, such as TFR1, HSPB1, SLC7A11 and GPX4. These results indicate that dietary PIT supplementation enhances the histological structure and function of the intestinal mucosa, which is involved in modulating antioxidant capacity and ferroptosis. [ABSTRACT FROM AUTHOR]

Enhancing the synthesis of microbicidal and immunomodulatory host defense peptides (HDP) is a promising host-directed antimicrobial strategy to combat a growing threat of antimicrobial resistance. Here we investigated the effect of several natural cyclooxygenase-2 (COX-2) inhibitors on chicken HDP gene regulation. Our results indicated that phenolic COX-2 inhibitors such as quercetin, resveratrol, epigallocatechin gallate, anacardic acid, and garcinol enhanced HDP gene expression in chicken HTC macrophage cell line and peripheral blood mononuclear cells (PBMCs). Moreover, these natural COX-2 inhibitors showed a strong synergy with butyrate in augmenting the expressions of multiple HDP genes in HTC cells and PBMCs. Additionally, quercetin and butyrate synergistically promoted the expressions of mucin-2 and claudin-1, two major genes involved in barrier function, while suppressing lipopolysaccharide-triggered interleukin-1β expression in HTC macrophages. Mechanistically, we revealed that NF-κB, p38 mitogen-activated protein kinase, and cyclic adenosine monophosphate signaling pathways were all involved in the avian β-defensin 9 gene induction, but histone H4 was not hyperacetylated in response to a combination of butyrate and quercetin. Because of their HDP-inducing, barrier-protective, and antiinflammatory activities, these natural COX-2 inhibitors, when combined with butyrate, may be developed as novel host-directed antimicrobial therapeutics. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance is a major concern to public health demanding effective alternative strategies to disease control and prevention. Modulation of endogenous host defense peptide (HDP) synthesis has emerged as a promising antibiotic alternative approach. This study investigated a potential synergy between sugars and butyrate in inducing HDP gene expression in chickens. Our results revealed that sugars differentially regulated HDP expression in both gene- and sugar-specific manners in chicken HD11 macrophage cells. Among eight mono- and disaccharides tested, all were potent inducers of avian β-defensin 9 (AvBD9) gene (p <0.05), but only galactose, trehalose, and lactose obviously upregulated cathelicidin-B1 (CATHB1) gene expression. The expression of AvBD14 gene, on the other hand, was minimally influenced by sugars. Moreover, all sugars exhibited a strong synergy with butyrate in enhancing AvBD9 expression, while only galactose, trehalose, and lactose were synergistic with butyrate in CATHB1 induction. No synergy in AvBD14 induction was observed between sugars and butyrate. Although lactose augmented the expression of nearly all HDP genes, its synergy with butyrate was only seen with several, but not all, HDP genes. Mucin-2 gene was also synergistically induced by a combination of lactose and butyrate. Furthermore, lactose synergized with butyrate to induce AvBD9 expression in chicken jejunal explants (p <0.05). Mechanistically, hyper-acetylation of histones was observed in response to both butyrate and lactose, relative to individual compounds. Mitogen-activated protein kinase, NF-κB, and cyclic adenosine monophosphate signaling pathways were also found to be involved in butyrate- and lactose-mediated synergy in AvBD9 induction. Collectively, a combination of butyrate and a sugar with both HDP-inducing and barrier protective activities holds the promise to be developed as an alternative to antibiotics for disease control and prevention. [ABSTRACT FROM AUTHOR]

The present study was aimed at investigating whether dietary anemonin could alleviate LPS-induced intestinal injury and improve intestinal barrier restoration in a piglet model. Eighteen 35-d-old pigs were randomly assigned to three treatment groups (control, LPS and LPS+anemonin). The control and LPS groups were fed a basal diet, and the LPS + anemonin group received the basal diet + 100 mg anemonin/kg diet. After 21 d of feeding, the LPS- and anemonin-treated piglets received i.p. administration of LPS; the control group received saline. At 4 h post-injection, jejunum samples were collected. The results showed that supplemental anemonin increased villus height and transepithelial electrical resistance, and decreased crypt depth and paracellular flux of dextran (4 kDa) compared with the LPS group. Moreover, anemonin increased tight junction claudin-1, occludin and ZO-1 expression in the jejunal mucosa, compared with LPS group. Anemonin also decreased TNF-α, IL-6, IL-8 and IL-1β mRNA expression. Supplementation with anemonin also increased TGF-β1 mRNA and protein expression, Smad4 and Smad7 mRNA expressions, and epidermal growth factor and epidermal growth factor receptor (EGFR) mRNA expression in the jejunal mucosa. These findings suggest that dietary anemonin attenuates LPS-induced intestinal injury by improving mucosa restoration, alleviating intestinal inflammation and influencing TGF-β1 canonical Smads and EGFR signaling pathways. [ABSTRACT FROM AUTHOR]

Weaning stress caused marked changes in intestinal structure and function. Transforming growth factor-β1 (TGF-β1) and canonical Smads signaling pathway are suspected to play an important regulatory role in post-weaning adaptation of the small intestine. In the present study, the intestinal morphology and permeability, developmental expressions of tight junction proteins and TGF-β1 in the intestine of piglets during the 2 weeks after weaning were assessed. The expressions of TGF-β receptor I/II (TβRI, TβRII), smad2/3, smad4 and smad7 were determined to investigate whether canonical smads signaling pathways were involved in early weaning adaption process. The results showed that a shorter villus and deeper crypt were observed on d 3 and d 7 postweaning and intestinal morphology recovered to preweaning values on d 14 postweaning. Early weaning increased (P<0.05) plasma level of diamine oxidase (DAO) and decreased DAO activities (P<0.05) in intestinal mucosa on d 3 and d 7 post-weaning. Compared with the pre-weaning stage (d 0), tight junction proteins level of occludin and claudin-1 were reduced (P<0.05) on d 3, 7 and 14 post-weaning, and ZO-1 protein was reduced (P<0.05) on d 3 and d 7 post-weaning. An increase (P<0.05) of TGF-β1 in intestinal mucosa was observed on d 3 and d 7 and then level down on d 14 post-weaning. Although there was an increase (P<0.05) of TβR II protein expression in the intestinal mucosa on d3 and d 7, no significant increase of mRNA of TβRI, TβRII, smad2/3, smad4 and smad7 was observed during postweaning. The results indicated that TGF-β1 was associated with the restoration of intestinal morphology and barrier function following weaning stress. The increased intestinal endogenous TGF-β1 didn't activate the canonical Smads signaling pathway. [ABSTRACT FROM AUTHOR]

The Chinese sturgeon (Acipenser sinensis) is critically endangered in China. Owing to the lack of visible sexual dimorphism and effective DNA markers for sexing in this species, a novel non-invasive method for sex identification in Chinese sturgeons would be helpful for conservation management. The zona pellucida (ZP) gene and sex determining region Y-box 9 (Sox9) gene were considered as candidate genes involved in gonadal differentiation. Here, we first detected the differences in expression of the ZP3.2 and Sox9 genes in the gonads and ventral fins of 107 Chinese sturgeons aged 2, 3, 6, 7, and 9–11 years old, using quantitative real-time polymerase chain reaction (qRT-PCR) to develop non-invasive expression markers. The accuracy of the developed marker was confirmed using endoscopy in another 240 individuals in four different age groups (aged 5–8 years old). The expression levels of the ZP3.2 gene in the ovaries and ventral fins at different ages were significantly (p < 0.05; p < 0.05, respectively) higher than those in the counterparts of males. The expression levels of the Sox9 gene in the testes at different ages were significantly (p < 0.05) higher than those in the ovaries, whereas the expression levels of the Sox9 gene in the male ventral fins were low, similar to those observed in the female ventral fins. Furthermore, a sex identification accuracy of 78.33% to 86.66% for the ZP3.2 gene in the ventral fin was confirmed by conducting endoscopies on individuals of four different age groups. The differential expression of ZP3.2 in the ventral fins of female and male Chinese sturgeon can be used as an effective non-invasive sex indicator in individuals over 2 years of age. This work also provides a novel solution for non-invasive sex identification in other surgeon species. • In this study, quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression level of ZP3.2 gene and Sox9 gene in 2, 3, 6, 7 and 9-11 years old Chinese sturgeons. The results show that the relative expression of ZP3.2 instead of Sox9 in ventral fin could be an effective indicator used to non-invasive sex identification of Chinese sturgeon in early stage of gonadal development. • Furthermore, the accuracy of 78.33% to 86.66% of ZP3.2 gene in ventral fin for sex identification was confirmed by endoscopy in another four different age groups Compared to traditional methods，the sex determination of Chinese sturgeon could be advanced to two years old. • This study explored a new technology for early sex identification of Chinese sturgeons, which can advance the time of sex identification to the 2 to 3 years old , withing a high accuracy rate (about 80%). [ABSTRACT FROM AUTHOR]

The Chinese sturgeon ( Acipenser sinensis ) is an anadromous fish distributed in the Yangtze River and the East China Sea. In this study, we report the novel finding of cleft palates in Chinese sturgeons derived from artificial fertilization. To explore the genetic basis of palate malformation in A. sinensis , Illumina RNA-seq technology was used to analyze the transcriptome data of farmed Chinese sturgeons with normal palates and cleft-palates. Raw reads were obtained and assembled into 808,612 unigenes, with an average length of 509.33 bp and an N50 of 574 bp. Sequence similarity analyses against four public databases (Nr, UniProt, KEGG, and COGs) found 158,642 unigenes that could be annotated. GABAergic synapses and TGF-β signal pathways were the two most enriched pathways with high Rich Factors in the analyses of differentially expressed genes. In these two signal pathways, six genes ( GABRA4 , GS , GNS , S6K , PITX2 , and BMP8 ) were found as candidate cleft-palate genes in Chinese sturgeon. These findings contribute to our understanding of cleft palate genetics in sturgeon, while simultaneously adding to our knowledge about craniofacial development. [ABSTRACT FROM AUTHOR]

Citrobacter freundii is one of the important bacterial diseases responsible for disease outbreaks to wild and cultured fishes globally. However, no known empirical research has focused on exploring relationships between immune response after C. freundii infection in sturgeons. In this study, C. freundii was isolated and identified from artificially breeding Chinese sturgeon, and global measurement of transcriptome response to C. freundii infection in head-kidney and spleen of A. sinensis were conducted to the acknowledgement of the potential mechanisms of pathogen-host interaction triggered by the bacterial infection. In total, differentially expressed genes which significantly associated with immune responses were found to be participated in antigen processing and presentation (MHC I, MHC II, HspA1, Hsp90A, Hsp70, CTSL, and CTSE), and acute phase response (serotransferrin and CP), as well as changing of other immune-related cytokine, such as chemokine and interferon, which proving their reacting and regulatory role during the response of thehost against C. freundii infection in fish. C. freundii can cause serious disease in sturgeon species was first reported in this study, and innate immune responses to C. freundii infection in this study will be conducive to understand the defense mechanisms and making appropriate prevention strategies in A. sinensis aquaculture operations. • Citrobacter freundii was isolated for the first time from Chinese sturgeon (Acipenser sinensis). • Transcriptome sequencing was performed to explore the alteration in immune response following bacterial infection. • DEGs profiles provides insights for understanding of host defense mechanisms against C. freundii infection. [ABSTRACT FROM AUTHOR]