Distribution and association of antimicrobial resistance and virulence characteristics in Enterococcus spp. isolates from captive Asian elephants in China.

Enterococcus spp., as an opportunistic pathogen, are widely distributed in the environment and the gastrointestinal tracts of both humans and animals. Captive Asian elephants, popular animals at tourist attractions, have frequent contact with humans. However, there is limited information on whether captive Asian elephants can serve as a reservoir of antimicrobial resistance (AMR). The aim of this study was to characterize AMR, antibiotic resistance genes (ARGs), virulence-associated genes (VAGs), gelatinase activity, hemolysis activity, and biofilm formation of Enterococcus spp. isolated from captive Asian elephants, and to analyze the potential correlations among these factors. A total of 62 Enterococcus spp. strains were isolated from fecal samples of captive Asian elephants, comprising 17 Enterococcus hirae (27.4%), 12 Enterococcus faecalis (19.4%), 8 Enterococcus faecium (12.9%), 7 Enterococcus avium (11.3%), 7 Enterococcus mundtii (11.3%), and 11 other Enterococcus spp. (17.7%). Isolates exhibited high resistance to rifampin (51.6%) and streptomycin (37.1%). 50% of Enterococcus spp. isolates exhibited multidrug resistance (MDR), with all E. faecium strains demonstrating MDR. Additionally, nine ARGs were identified, with tet(M) (51.6%), erm(B) (24.2%), and cfr (21.0%) showing relatively higher detection rates. Biofilm formation, gelatinase activity, and α-hemolysin activity were observed in 79.0, 24.2, and 14.5% of the isolates, respectively. A total of 18 VAGs were detected, with gelE being the most prevalent (69.4%). Correlation analysis revealed 229 significant positive correlations and 12 significant negative correlations. The strongest intra-group correlations were observed among VAGs. Notably, we found that vancomycin resistance showed a significant positive correlation with ciprofloxacin resistance, cfr, and gelatinase activity, respectively. In conclusion, captive Asian elephants could serve as significant reservoirs for the dissemination of AMR to humans.

Biliopancreatic Limb Length of Small Intestinal Bypass in Non-obese Goto-Kakizaki (GK) Rats Correlates with Gastrointestinal Hormones, Adipokines, and Improvement in Type 2 Diabetes.

BACKGROUND: The purpose of this study was to explore the effects on type 2 diabetes, gastrointestinal hormones, and adipokines after the small intestinal bypass of different biliopancreatic limb (BPL) lengths in non-obese type 2 diabetic rats. METHOD: Small intestinal bypass with the BPL length at 10cm, 20cm, 30cm, and 40cm, respectively, and sham surgery were performed in non-obese GK rats. Fasting serum was collected at 2 days preoperatively and 1, 3, 6, and 9 weeks postoperatively. Body weight and fasting blood glucose (FBG) were measured during the experiment. Glycated hemoglobin (GHb), fasting insulin (FINS), C-peptide, ghrelin, leptin, adiponectin, and somatostatin were measured postoperatively. RESULT: Rats with a bypassed length of 40cm died within 5-9 weeks. No statistically significant was observed in body weight between the sham group and the bypass groups at the 9th week postoperatively. FBG, GHb, FINS, C-peptide, and HOMA-IR in the bypass groups were lower than those in the sham group postoperatively and were negatively correlated with BPL length. Ghrelin and leptin declined compared with preoperative but were not associated with BPL length. Adiponectin of the bypass groups increased after operation and was positively correlated with BPL length. Somatostatin remained stable among groups during the experiment. CONCLUSION: Ghrelin and leptin of non-obese GK rats decreased postoperatively without a linear relationship with the BPL length, while adiponectin increased with positively correlation with the BPL length. In addition, somatostatin remained steady after small intestinal bypass. Further studies are expected to confirm the effect of the BPL length of small intestinal bypass on gastrointestinal hormones and adipokines.

miR-4999-5p Predicts Colorectal Cancer Survival Outcome and Reprograms Glucose Metabolism by Targeting PRKAA2.

PURPOSE: Colorectal cancer (CRC) is the third most common cancer, and the second leading cause of cancer death worldwide. Dysregulation of microRNAs has been shown to modulate glucose metabolic reprogramming in CRC. However, the functional role of miR-4999-5p in the CRC glucose metabolic shift has not been characterized. PATIENTS AND METHODS: The levels of miR-4999-5p and PRKAA2 were evaluated by RT-qPCR. Univariate and multivariate survival analyses were conducted to evaluate the prognostic value of miR-4999-5p. Cell proliferation was assessed using the CCK-8 and colony formation assays. Extracellular acidification rate, glucose uptake, cellular glucose-6-phosphate level, and lactate production were evaluated to assess the effects of miR-4999-5p on CRC glycolysis. Dual-luciferase reporter assay was conducted to investigate the direct interaction between miR-4999-5p and PRKAA2. Mouse xenograft models were established to assess the functions of miR-4999-5p in vivo. RESULTS: miR-4999-5p was highly expressed in CRC tissues and cell lines. In addition, miR-4999-5p was associated with tumor differentiation and TNM stage, and elevated expression of miR-4999-5p was an independent predictor of poorer overall survival. Furthermore, miR-4999-5p promoted cell proliferation and glycolysis in CRC. miR-4999-5p targeted PRKAA2 to exert its tumor-promoting functions, and PRKAA2 knockdown rescued decreased cell proliferation and glycolysis in miR-4999-5p-silenced CRC cells. In vivo experiments showed that miR-4999-5p promoted CRC growth. CONCLUSION: miR-4999-5p facilitated cell growth and glucose metabolic reprogramming through direct targeting of PRKAA2. Our results showed that miR-4999-5p may be a novel prognostic marker and therapeutic target for CRC.

The evidence of apelin has the bidirectional effects on feeding regulation in Siberian sturgeon (Acipenser baerii).

Apelin is a peptide, mainly produced in the brain, which participates in several physiologic effects. However, knowledge about the mechanism of appetite regulation in teleosts, including the role of apelin is not well understood. The aim of this study is to explore the effect of feeding status on the expression of apelin mRNA in the whole brain and the effects of injection of apelin on food intake in Siberian sturgeon (Acipenser baerii). In this study, we first cloned the apelin cDNA sequence of the Siberian sturgeon. We obtained a 1046-bp cDNA fragment, including a 237-bp open reading frame (ORF) that encoded 78 amino acids. Apelin was widely distributed in 11 tissues related to feeding regulation, with the highest expression in thewhole brain, followed by the spleen and trunk kidney. In addition, we measured the effects of periprandial (preprandial and postprandial) change, fasting and re-feeding on apelin mRNA expression in whole brain. The level of apelin mRNA was significantly decreased 1h after feeding. The results of the fasting experiment showed that the expression of apelin mRNA in the brain was significantly reduced after 1day of fasting but consistently increased throughout the 15-day food deprivation period. When the 15-day fasted fish were re-fed, apelin mRNA expression in the brain was significantly increased as compared to that of the control. These results suggest that apelin may play a bidirectional role in the regulation of food intake in the Siberian sturgeon. In order to further examine the effect of apelin on feeding regulation in Siberian sturgeons, acute and chronic intraperitoneal (i.p.) injection experiments were performed and food intakes were recorded. Results showed that acute i.p. injection of apelin-13 reduced food intake, however, chronic i.p. injection apelin-13 increased the food intake for 7days in Siberian sturgeons. In conclusion, our results show that apelin has a bidirectional effect on feeding regulation in Siberian sturgeons by acting as a satiety factor in short-term feeding regulation and a starvation factor in long-term feeding regulation.

Identification and characterization of a ß-defensin gene involved in the immune defense response of channel catfish, Ictalurus punctatus.

Antimicrobial peptides are small peptides that play important roles in a host's innate immune response. As an important antimicrobial peptide, ß-defensin widely distribute in mammals, insects and plants with broad-spectrum antimicrobial activity. In this study, the ß-defensin gene of the channel catfish, Ictalurus punctatus, was cloned, sequenced, and subjected to a bioinformatic analysis. The ß-defensin gene of the channel catfish contains three exons and two introns, and encodes a precursor peptide consisting of two domains: a signal peptide of 24 amino acid residues and a mature peptide of 43 amino acid residues. The mature peptide is estimated to have a molecular mass of 7.1kDa and a theoretical isoelectric point of 8.21. Channel catfish ß-defensin (ccBD) has six conserved cysteine residues, forming three disulfide bridges at C1-C5, C2-C4, and C3-C6, and a ß-sheet in the predicted three-dimensional structure. A phylogenetic analysis suggests that ccBD belongs to the type 1 ß-defensins. Real-time quantitative PCR showed that channel catfish ß-defensin transcripts are constitutively expressed in various tissues in healthy fish, with highest expression in the skin. The expression of ccBD in vivo increased significantly in the head kidney (2.9-fold), gill (2.2-fold), and skin (6.6-fold) at 48h after bacterial (Edwardsiella ictaluri) challenge. In vitro, lipopolysaccharide (LPS), a bacterial mimic, induced significant changes in ccBD expression in leukocytes from the spleen (3.4-fold) and head kidney (3.9-fold) 24h after stimulation. Chemically synthesized ccBD displayed marked inhibitory activity against a broad range of bacteria. These results suggest that ccBD is involved in the innate antibacterial defenses of the channel catfish.

Intraperitoneal injection urocortin-3 reduces the food intake of Siberian sturgeon (Acipenser baerii).

Urocortin-3 (UCN3), one of the corticotropin releasing factor (CRF) family peptides, which was discovered in 2001, has a variety of biological functions. However, the researches of UCN3 in fish were scarce. In order to understand whether UCN3 play a role in regulating food intake in fish, we first cloned the ucn3 cDNAs sequence of Siberian sturgeon (Acipenser baerii Brandt), and investigated the ucn3 mRNA levels in 11 tissues. The Siberian sturgeon ucn3 cDNA sequence was 1044bp, including an open reading frame (ORF) of 447bp that encoded 148 amino acids with a mature peptide of 40 amino acids, a 5'-terminal untranslated region (5'-UTR) of 162bp and a 3'-terminal untranslated region (3'-UTR) of 435bp. The result of tissue distribution showed that ucn3 widely distributed in 11 tissues with highest expression in brain. We also assessed the effects of periprandial (pre- and post-feeding), fasting and re-feeding on ucn3 mRNAs abundance in brain. The results showed the expression of ucn3 mRNA in brain was significantly elevated after feeding, decreased after fasting 17 days and increased after re-feeding. To further investigate the food intake role of UCN3 in Siberian sturgeon, we performed intraperitoneal (i.p.) injection of Siberian sturgeon UCN3 (SsUCN3) with three doses (60, 120 or 240ng/g) and recorded the food intake. Acute and chronic i.p. injection SsUCN3 reduced the food intake in a dose-dependent pattern. In conclusion, this study indicates that SsUCN3 acts as a satiety factor to inhibit the food intake of Siberian sturgeon.

Molecular characterization and expression analysis of Hsp90 in Schizothorax prenanti.

Aquatic animals suffer from various environmental stresses because the aquatic environment is a very complex system. To monitor the health status of fish, Hsp90 a potential early warning marker was determined in Schizothorax prenanti after infection with a bacterium. In this study, we cloned Hsp90 from S. prenanti for the first time. The full-length cDNA sequence of SpHsp90 was 2663 bp, contains an open reading frame of 2181 bp, and has a gene encoding 726 amino acids, an estimated molecular mass of 83.38 kDa, and a theoretical isoelectric point of 4.91. The SpHsp90 amino acid sequence has five conserved HSP90 family signatures and shares 87.0-95.5 % identity with other vertebrates. Phylogenetic analysis and structure comparison indicated that SpHsp90 should be a ß isoform of the HSP90 family. SpHsp90 was ubiquitously expressed in all examined tissues, and the highest level of expression was in the kidney. After Streptococcus agalactiae infection, the level of SpHsp90 expression had significant changes (P < 0.05) in the hepatopancreas, spleen, kidney, and blood. The expression increased to the highest level at 6 h in the blood and at 24 h in the hepatopancreas, spleen, and kidney. The results suggested that the SpHsp90 gene could be induced by S. agalactiae in S. prenanti and that SpHsp90 may be involved in resistance to bacterial infection and provide an early warning information. The kidney is the most suitable for detecting SpHsp90 after bacterial infection.

Molecular cloning, expression analysis, and appetite regulatory effect of peptide YY in Siberian sturgeon (Acipenser baerii).

Peptide YY (PYY) is an anorectic brain-gut peptide involved in feeding regulation and well characterized in mammals. However, the functional role of PYY in the appetite regulatory of fish is not clear. In this study, we characterized a high conservation of PYY cDNA and found high expression levels of PYY mRNA in the brain and digestive tract of Siberian sturgeon. Then, we examined preprandial (pre- and post-feeding) changes of PYY mRNA expression in the brain that showed a significantly increased in 3h post-feeding, suggesting an anorectic possible function of PYY in Siberian sturgeon. Next, we examined the expression of PYY mRNA during 15 days fasting and refeed after fasting. The SsPYY mRNA expression of unfed fish had a significant 2.4, 1.7, 2.0, 2.2, and 2.1-fold decrease compared to 1-, 3-, 6-, 10- and 15-day ad libitum fed animals, respectively. After refeed, SsPYY mRNA significantly increased 1.9 and 4.1-fold above that of the 15-day fed and unfed fish control group (P<0.01). Furthermore, a single intraperitoneal injection of 10, 100 and 200 ng/g BW SsPYY1-36 caused a reduction in the next feeding and no significant reduction in food intake was observed in fish injected with a 1 ng/g BW. Overall, PYY has a potentially role in food intake attenuation of Siberian sturgeon.