Differential expression of microRNA between normally developed and underdeveloped female worms of Schistosoma japonicum.

Eggs produced by bisexual infected mature female worms (MF) of Schistosoma japonicum are important in the transmission of the parasite and responsible for the pathogenesis of schistosomiasis. The single-sex infected female worms (SF) cannot mature and do not produce normal eggs; also they do not induce severe damage to the host. In this study, the microRNA (miRNA) expression profiles of 25d MF and 25d SF were investigated through Solexa deep-sequencing technology to explore the developmental mechanisms of schistosome female worms. There were 36 differentially expressed miRNA, 20 up-regulated and 16 down-regulated found in MF/SF worms, including some development related miRNA such as bantam (ban), let-7, miR-124, miR-8, miR-1, miR-7. There were 166 target genes of up-regulated miRNA and 201 target genes of down-regulated miRNA after comparing the target gene prediction software results with RNA-Seq transcriptome results. Analysis of the target genes shows that different ones are involved in MF and SF worms in Gene Ontology terms, with a similar situation in KEGG. This observation indicates that different genes regulated by differentially expressed miRNA take part in MF and SF and lead to differential sexual status. This means that the sexual status of female worms is regulated by miRNA.

Temporal modulation of host aerobic glycolysis determines the outcome of Mycobacterium marinum infection.

Macrophages are the first-line host defense that the invading Mycobacterium tuberculosis (Mtb) encounters. It has been recently reported that host aerobic glycolysis was elevated post the infection by a couple of virulent mycobacterial species. However, whether this metabolic transition is required for host defense against intracellular pathogens and the underlying mechanisms remain to be further investigated. A pathogenic mycobacterial species, M. marinum, is genetically close to Mtb and was utilized in this study. Through analyzing cellular carbon metabolism of RAW 264.7 (a murine macrophage-like cell line) post M. marinum infection, a strong elevation of glycolysis was observed. Next, three glycolysis inhibitors were examined for their ability to inhibit mycobacterial proliferation inside RAW264.7 macrophages. Among them, a glucose analog, 2-deoxyglucose (2-DG) displayed a protective role against mycobacterial infection. Treatment with 2-DG at concentrations of 0.5 or 1 mM significantly induced autophagy and decreased the phagocytosis of M. marinum by macrophages. Moreover, 2-DG pre-treatment exerted a significantly protective effect on zebrafish larvae by limiting the proliferation of M. marinum, and such effect was correlated to tumor necrosis factor alpha (TNF-α) as the 2-DG pre-treatment increased the expression of TNF-α in both mouse peritoneal macrophages and zebrafish. On the contrary, the 2-DG treatment post infection did not restrain proliferation of M. marinum in WT zebrafish, and even accelerated bacterial replication in TNF-α-/- zebrafish. Together, modulation of glycolysis prior to infection boosts host immunity against M. marinum infection, indicating a potential intervention strategy to control mycobacterial infection.

Transcriptome analysis of low-temperature-affected ripening revealed MYB transcription factors-mediated regulatory network in banana fruit.

Low temperature leads to abnormal ripening and poor quality of the harvested banana fruit, which is an urgent problem limiting the development of industry in China. To comprehensively understand the mechanism underlying low-temperature-affected ripening, we performed comparative RNA-Seq analysis of ethylene-induced ripening of banana fruit after 3 days of pre-storage at 7 °C and 22 °C. A total of 986 differentially expressed genes (DEGs) were identified in both RT-0 d versus RT-3 d, LT-0 d versus LT-3 d, RT-0 d versus LT-0 d and RT-3 d versus LT-3 d, and the RNA-Seq outputs of 15 randomly selective DEGs were verified using qRT-PCR. Among the 986 DEGs obtained in the four groups, 9 MYB genes (MaMYB75/281/219/4/151/156/3/37 and MaMYB3R1) and 32 genes related to carotenoid biosynthesis (MaPSY1/2a), flavor formation (MaLOX6, MaADH7, MaAAT1), sucrose transport (MaSUS2/4), ethylene production (MaSAM1, MaACO9/10/12, MaACS1/12), starch degradation (MaAMY1A/1B, MaPHS1/2, MaMEX2, MapGlcT1) and cell wall degradation (MaPG3/X1, MaPME25/41, MaXTH5/7/22/23/25, MaEXP2/20/A1/A15) were characterized. Combining transcription factor binding site (TFBS) analysis as well as cis-acting element analysis, the regulatory network of low-temperature-affected ripening mediated by MYBs were constructed. The data generated in this study may unravel the transcriptional regulatory network of MYBs associated with low-temperature-affected ripening and provide a solid foundation for future studies.

A host lipase prevents lipopolysaccharide-induced foam cell formation.

Although microbe-associated molecular pattern (MAMP) molecules can promote cholesterol accumulation in macrophages, the existence of a host-derived MAMP inactivation mechanism that prevents foam cell formation has not been described. Here, we tested the ability of acyloxyacyl hydrolase (AOAH), the host lipase that inactivates gram-negative bacterial lipopolysaccharides (LPSs), to prevent foam cell formation in mice. Following exposure to small intraperitoneal dose(s) of LPSs, Aoah -/- macrophages produced more low-density lipoprotein receptor and less apolipoprotein E and accumulated more cholesterol than did Aoah +/+ macrophages. The Aoah -/- macrophages also maintained several pro-inflammatory features. Using a perivascular collar placement model, we found that Aoah -/- mice developed more carotid artery foam cells than did Aoah +/+ mice after they had been fed a high fat, high cholesterol diet, and received small doses of LPSs. This is the first demonstration that an enzyme that inactivates a stimulatory MAMP in vivo can reduce cholesterol accumulation and inflammation in arterial macrophages.

Seasonal Dynamics of Gastrointestinal Nematode Infections of Goats and Emergence of Ivermectin Resistance in Haemonchus contortus in Hubei Province, China.

INTRODUCTION: Gastrointestinal nematodes (GINs) are a major constraint to the survival and productivity of animals. In southern China, goats are the most important small domestic ruminants. METHODS: From May 2013 to May 2017, we conducted a longitudinal study of hircine GIN infections in Huangshantou Town, Gongan County, Hubei Province, China, using fecal egg counts. RESULTS: Our investigation revealed that the GINs of goats in Hubei Province have changed significantly. Over 90% of eggs detected in the first month of investigation, May 2013, belonged to the species Haemonchus contortus and Chabertia sp. There was no seasonal variation in positive rates (PRs) of GINs, but the mean eggs per gram (EPG) of GINs were higher between April and July than between September and November (P < 0.05). The gradual increase in the percentage of H. contortus eggs among all detected eggs during our research and the low cure rate of IVM mass treatment revealed the emergence of IVM resistance in H. contortus. After the implementation of an integrated GIN control strategy, which included two mass treatments (one in April/May with ABZ and another in September/October with IVM + ABZ), in 2016 and 2017, both the PRs and EPG of GINs were significantly reduced. CONCLUSION: The results presented here reveal that controlling GINs of small ruminants in small farms in southern China requires an integrated control strategy that should include monitoring of infection and anthelmintic resistance, and increased farmer education on the importance of using the appropriate drugs at the correct dose.

LPS inactivation by a host lipase allows lung epithelial cell sensitization for allergic asthma.

Allergic asthma is a chronic inflammatory disease primarily mediated by Th2 immune mechanisms. Numerous studies have suggested that early life exposure to lipopolysaccharide (LPS) is negatively associated with allergic asthma. One proposed mechanism invokes desensitization of lung epithelial cells by LPS. We report here that acyloxyacyl hydrolase (AOAH), a host lipase that degrades and inactivates LPS, renders mice more susceptible to house dust mite (HDM)-induced allergic asthma. Lung epithelial cells from Aoah-/- mice are refractory to HDM stimulation, decreasing dendritic cell activation and Th2 responses. Antibiotic treatment that diminished commensal LPS-producing bacteria normalized Aoah-/- responses to HDM, while giving LPS intrarectally ameliorated asthma. Aoah-/- mouse feces, plasma, and lungs contained more bioactive LPS than did those of Aoah+/+ mice. By inactivating commensal LPS, AOAH thus prevents desensitization of lung epithelial cells. An enzyme that prevents severe lung inflammation/injury in Gram-negative bacterial pneumonia has the seemingly paradoxical effect of predisposing to a Th2-mediated airway disease.

A candidate recombinant antigen for diagnosis of schistosomiasis japonica in domestic animals.

Domestic animals infected with Schistosoma japonicum are a major source of infection and play an important role in transmission to humans. A key strategy for the elimination of schistosomiasis is to control the sources of infection. In the present study, we identified a candidate diagnostic antigen-encoding gene, SjMRP1, the putative multidrug resistance protein 1 gene, by screening a cDNA phage display library from 44-day-old S. japonicum worms using IgGs from goat, cattle, and buffalo infected with S. japonicum. We cloned and expressed the fragment of SjMRP1 and subsequently evaluated the diagnostic potential of the recombinant protein rSjMRP1. In the enzyme-linked immunosorbent assay of rSjMRP1 (rSjMRP1-ELISA), the sensitivity in goat, cattle, and buffalo was 95.6% (86/90), 100% (22/22), and 90% (81/90), respectively, and the specificity was 100% (30/30) in goat and cattle and 96.67% (29/30) in buffalo. These results were not significantly different from soluble egg antigen (SEA)-ELISA results. Notably, rSjMRP1-ELISA has no cross reaction with Haemonchus contortus, a most common nematode seen in goat and bovine in China, in 13 infected goats, and with Orientobilhazia turkestanica, which is genetically under Schistosoma, in 36 infected goats; whereas SEA-ELISA showed false positive rate of 15.38% and 83.33% in the two respective animal groups. The results obtained here suggest that rSjMRP1 may be used for diagnosis of S. japonicum infection of domestic animals.

A novel colloidal gold immunochromatography assay strip for the diagnosis of schistosomiasis japonica in domestic animals.

BACKGROUND: Schistosomiasis remains a major public health concern in China and an epidemiological survey has revealed that schistosome-infected bovines and goats are the main transmission sources for the disease. Therefore, development of a sensitive technique for the diagnosis of schistosomiasis in domestic animals is necessary. METHOD: A novel colloidal gold immunochromatography assay (GICA) strip was developed for detecting Schistosoma japonicum in domestic animals. The colloidal gold was conjugated with recombinant streptococcal protein G (rSPG). As the test and control lines, the schistosome soluble egg antigen and rSPG, respectively, were blotted on nitrocellulose membrane. RESULTS: The lowest detectable serum dilution was 1∶640 for schistosome-infected buffaloes. The cross-reaction rate of GICA was 14.29% with Paramphistomum sp. in buffaloes, 16.67% with Haemonchus sp. in goats, and 33.33% with Orientobilharzia sp. in goats. These results were slightly lower and similar to those obtained through ELISA. Moreover, the strips for detecting S. japonicum in mice, rabbits, buffaloes, and goats showed high sensitivity (100.00%, 100.00%, 100.00%, and 100.00%, respectively) and specificity (100.00%, 100.00%, 94.23%, and 88.64%, respectively). And the sensitivity or specificity of the GICA strips did not present any significant differences after storage for 12 months at room temperature. When compared with ELISA, the GICA strips exhibited similar sensitivity and specificity in the diagnosis of schistosomiasis in mice, rabbits, buffaloes, and goats. Besides, only 5 µl of serum are required for the test and the detection can be completed within 5 min. CONCLUSION: This study is the first to develop a GICA strip using gold-rSPG conjugate for the diagnosing of schistosomiasis in domestic animals, and preliminary results showed that the developed strip may be suitable for large-scale screening of schistosomiasis in endemic areas.

Evaluation of recombinant multi-epitope proteins for diagnosis of goat schistosomiasis by enzyme-linked immunosorbent assay.

BACKGROUND: Schistosomiasis is a huge threat to human and animal health. Apart from bovines, goats play an important role in the transmission of schistosomiasis in some endemic areas of China. An accessible, quality-assured goat schistosomiasis diagnostic technique is needed. Recently, our laboratory identified two recombinant diagnostic antigens, SjPGM and SjRAD23 via an immuno-proteomic method. The application of these two recombinant antigens to develop a higher sensitivity and specificity technique for the sheep schistosomiasis diagnosis is urgently needed. METHODS: Epitopes of SjPGM and SjRAD23 were predicted and three polypeptides, two from SjRAD23 and one from SjPGM, were selected. Recombinant plasmids containing two to three DNA sequences encoding predicted polypeptides or large hydrophilic region of Sj23 (LHD-Sj23) were constructed and expressed. Eight recombinant schistosome antigens including four multi-epitope proteins and four recombinant single-molecule antigens as well as SEA, were assessed by ELISA in 91 sera from schistosome-infected goats, 44 sera from non-infected goats, 37 sera from Orientobilharzia-infected goats, and 12 from Haemonchus contortus-infected goats. RESULTS: ELISA tests showed that three multi-epitope proteins had higher sensitivity than the four single-molecule antigens (rSjRAD23, rSjPGM, rBSjRAD23-1, rBSj23) and the multi-epitope protein rBSjPGM-BSjRAD23-1-BSj23 had the highest sensitivity (97.8 %, 89/91) and maintained good specificity (100 %, 44/44) as well as low cross-reactivity with haemonchosis (8.33 %, 3/12) and orientobilharziasis (13.51 %, 5/37) in the diagnosis of goat schistosomiasis. In contrast, when SEA was applied as a diagnosis antigen, it had 100 % (91/91) sensitivity, 75 % (33/44) specificity, 25 and 83.78 % cross-reactivity with haemonchosis (3/12) and orientobilharziasis (31/37), respectively. CONCLUSIONS: The application of recombinant multi-epitope proteins may increase the sensitivity of diagnosis technique and retain high specificity of single-molecule antigens for schistosomiasis, and the recombinant antigen rBSjPGM-BSjRAD23-1-BSj23 has the potential to be used as a diagnosis antigen for goat schistosomiasis.