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Abstract Giant freshwater prawn Macrobrachium rosenbergii is an economically important species. However, its growth retardant have brought serious economic losses in recent years. Antibiotics abuse is suggested as a reason for M. rosenbergii 's growth retardant, while few studies focused on the toxic effect of antibiotics on M. rosenbergii. To investigate the effect of enrofloxacin, a widely used antibiotic, on juvenile M. rosenbergii , a 14 days exposure study was carried out within 0.2, 1 and 5 mg/L enrofloxacin and followed by 7 days decontamination. Results showed that during the test period, enrofloxacin had the largest accumulation in juvenile shrimp at day 3, and gradually decreased at day 7 and 14, and almost all the drugs are cleared after 3 days decontamination. Short-term exposure to low dose enrofloxacin can promote the growth of juveniles. High dose enrofloxacin inhibited the growth of juvenile shrimp, to gill and liver damage, and induced apoptosis of the hepatopancreatic cells. These adverse effects was possibly caused by enrofloxacin-induced oxidative stress. Moreover, we also found the damage caused by high concentrations of enrofloxacin was irreversible in the short term. Collectively, these data indicated that enrofloxacin did affect the juvenile shrimp growth and development, and high level enrofloxacin abuse may contributed to M. rosenbergii 's growth retardant. Highlights • Low dose enrofloxacin can promote the growth of M. rosenbergii. • High dose enrofloxacin inhibited the growth of M. rosenbergii. • The damaging effects of enrofloxacin is related to oxidative stress. • Damage caused by high dose enrofloxacin was irreversible in the short term. • Enrofloxacin abuse may contributed to M. rosenbergii 's growth retardant. [ABSTRACT FROM AUTHOR]

Macrobrachium rosenbergii is an important cultural species in China and other Southeast Asian countries. However, Enterobacter cloacae infection has caused a great economic loss in M. rosenbergii culture industry. The immune responses of M. rosenbergii to the E. cloacae infection is not fully characterized. To investigate the immune response of M. rosenbergii against E. cloacae , we performed transcriptome analysis of the M. rosenbergii hepatopancreas with and without E. cloacae infection using RNA-seq. After assembly and annotation, 29,731 high quality unigenes were obtained from RNA-seq data. Differential expression analysis revealed the existence of 2498 significantly differently expressed genes (DEGs) at 12 h post infection, with 1365 up-regulated and 1133 down-regulated genes. Among these DEGs, some well-known immune-related genes were up-regulated significantly, including C-type lectin 1, lectin 3, anti-lipopolysaccharide factor 2, Cu/Zn superoxide dismutase and heat shock protein 70. GO analysis demonstrated 24 biological process subcategories, 14 cellular component subcategories, and 12 molecular function subcategories that were enriched among these DEGs, and some DEGs were clustered into immune related subcategories such as immune system process, response to stimulus, biological adhesion, and antioxidant activity. These DEGs were enriched into 216 KEGG pathways including a core set of immune correlated pathways notably in phagosome and lysosome. In addition, 5 up-regulated and 5 down-regulated immune-related DEGs were selected for further validation by quantitative real-time PCR and the results showed consistence with the RNA-seq data. Additionally, the expression level of six selected immune-related genes (ALF2 , CLEC1 , LEC3 , hemocyanin1 , HSP70 and SOD) based on the transcriptomic data were monitored at different point of time in hepatopancreas, gill, hemolymph and intestine. Results revealed these immune-related genes were significantly up-regulated in different tissues from 6 to 24 h after E. cloacae infection. Overall, these results provided valuable information for further studying the immune response of M. rosenbergii against E. cloacae infection. • The immune response of M. rosenbergii against E. cloacae was determined by transcriptome analysis. • DEGs were enriched in many immune-related pathways. • E. cloacae triggers a wide defensive response of various immune related genes in M. rosenbergii. [ABSTRACT FROM AUTHOR]

Enterobacter cloacae is a member of the Enterobacter family, which could prevent Macrobrachium rosenbergii from growing and cause mass mortality. However, no research has focused on microRNA immunity in M. rosenbergii infected with E. cloacae. To clarify the immune response mechanisms, transcriptomic analysis was performed on the miRNAs of M. rosenbergii infected with E. cloacae YZ3 strain. Following quality screening, 10,616,712 clean reads were obtained from the control group and 12,726,421 from the infected group. Among 899 known miRNAs, 446 differentially expressed miRNAs (DEMs) were identified. Meanwhile, 59 novel miRNAs were predicted, along with 39 DEMs. Target genes of DEMs have been predicted in order to gain a deeper understanding of the immune-related functions. GO and KEGG pathway analysis revealed the biological functions and signaling pathways of target genes. The results indicated that E. cloacae significantly affected the NOD-like receptor, RIG-I-like receptor and Toll-like receptor pathways. Ten DEMs were randomly selected, and their expression level was verified by Quantitative Real-time PCR technology. Overall, this study highlights the influential role of miRNAs in the innate immune system of M. rosenbergii , which has important implications for developing new strategies to prevent and treat related diseases in the future. • MiRNA transcriptome analysis of M. rosenbergii infected with E. cloacae was first performed. • 446 known and 39 novel miRNAs were differentially expressed during E. cloacae infection. • The target genes of DEMs were enriched in many immune-related pathways. [ABSTRACT FROM AUTHOR]

Outbreaks of mass mortalities occurred in Macrobrachium rosenbergii farms in Gaoyou county, Jiangsu Province of China. The bacterial isolates from M. rosenbergii exhibited the same phenotypic traits and biochemical characteristics, and were identified as non-O1/O139 Vibrio cholerae according to biochemical characteristics and molecular identification. In challenge test, M. rosenbergii infected with non-O1/O139 V. cholerae GXFL1-4 developed similar pathological signs to the naturally diseased prawns, and LD 50 of the strain to M. rosenbergii was 4.5 × 106 CFU/mL at 96 h post-infection. Histopathological analysis revealed that hepatopancreas and intestines of diseased M. rosenbergii exhibited obvious inflammatory responses to non-O1/O139 V. cholerae infection. Detection virulence factors of the strain GXFL1-4 showed that the bacteria produced caseinase, lipase, amylase, lecithinase and hemolysin, and carried toxR , hlyA , ompW , ompU , hap , rtxA and rtxC virulence related genes, supporting the strong virulence to M. rosenbergii. Additionally, the immune related gene expression in M. rosenbergii evaluated by qRT-PCR analysis showed that HSP70 , Crustin , Lysozyme , TRL1 , ALF1 , Lectin , Peroxinectin , proPO and SOD immune related genes were significantly up-regulated at 6 and 12 h after infection with GXFL1-4. The results of our study suggested that non-O1/O139 V. cholerae was an etiological element in the mass mortalities of M. rosenbergii and this study provided preliminary insights into the diversity in the immune response of M. rosenbergii to the bacterial invasion. • Mass mortalities of cultured M. rosenbergii from several farms in Gaoyou country of China was reported. • Pathogenesis and histopathological analysis related to M. rosenbergii with non-O1/O139 V. cholerae infection were studied. • Non-O1/O139 V. cholerae infection induced a wide defensive response of various immune related genes in M. rosenbergii. [ABSTRACT FROM AUTHOR]

Abstract Outbreaks of mass mortality among cultured zoea of M. rosenbergii occurred in different hatcheries of Gaoyou country of China in April 2017. Of the isolates obtained from the diseased zoea, XL3–1 was pathogenic to the zoea and postlarva of M. rosenbergii in the virulence test, with a LD50 value 3.2 × 106 CFU/mL and 3.5 × 106 CFU/mL, respectively. XL3–1 was identified as Enterobacter cloacae by biochemical characteristics and molecular tools. Detection of virulence-associated genes by PCR indicated that XL3–1 was positive for irp2 , fhuA , sodB , sltA , flaD and OmpX. According to the antibiotic resistant assay, the isolate was sensitive to quinolones, while they were resistant to cephalosporins and penicillins. This study also investigated the protection of specific chicken egg yolk antibodies (IgY) in E. cloacae -infected M. rosenbergii. The highest titer against E. cloacae of the obtained specific IgY by ELISA was 1: 213. M. rosenbergii treated with specific egg yolk powder could achieve the survival rate of 67% after E. cloacae infection, while M. rosenbergii treated with non-specific egg yolk powder or no egg yolk powder could achieve survival rates of 20% and 17%, respectively. As well, the bacterial burden in hepatopancreas was significantly lower in M. rosenbergii treated with specific egg yolk powder than those treated with non-specific egg yolk powder. This study clarified the virulence characteristics and antibiotic resistance of E. cloacae , and provided a strategy to control the disease by using specific IgY. Highlights • Mass mortality of cultured zoea of M. rosenbergii from different farms in Gaoyou country of China is reported. • E.cloacae was identified as a pathogen to M. rosenbergii. • Oral administration of specific IgY significantly improved the survival rate of E. cloacae -infected M. rosenbergii. [ABSTRACT FROM AUTHOR]

Abstract Macrobrachium rosenbergii is one of the most economically important freshwater shimp, with fast growth and high nutrient content in the agricultural development of China. However, it had been suffering diseases infection, causing mass death and great economic losses. In the present study, a bacteria strain was isolated from the diseased zoea of M. rosenbergii and was identified as Vibrio vulnificus by biochemical characteristics and 16S rRNA homologous analysis. The infection test showed that the strain GXFL1-3 was pathogenic to zoea and postlarva of M. rosenbergii , and the half lethal dose (LD 50) were 1.16 × 106 CFU/mL and 1.45 × 106 CFU/mL, respectively. Detection of virulence-associated genes by PCR indicated that GXFL1-3 was positive for fur, OmpU, acfA, flaA, vvhA, vvp and tcp , the detection of extracellular enzymes and hemolysin showed that GXFL1-3 was positive for protease, amylase, lecithin, urease and hemolysin activity, further supporting its pathogenicity. A duplex PCR for rapid detection of V. vulnificus was established. Only V. vulnificus could amplify two specific bands of flaA and fur , while the other six strains of Vibrio were negative. The minimum detectable amount of template was 2.4 × 103 CFU/mL through sensitivity test. Highlights • Mass mortality from cultured zoea of M. rosenbergii from different farms in Gaoyou country of China is reported. • V. vulnificus was identified as a pathogen to M. rosenbergii. • V. vulnificus had virulence to the zoea and postlarva of M. rosenbergii , with a LD 50 value of 1.16 × 106 CFU/mL and 1.45 × 106 CFU/mL. • A double PCR for rapid detection of V. vulnificus was established. [ABSTRACT FROM AUTHOR]

Non-O1 Vibrio cholerae , a member of the Vibrio family, could cause gastrointestinal infection of Macrobrachium rosenbergii and result in significant economic losses. However, few studies on microRNA immunity related to non-O1 V. cholerae infection of M. rosenbergii. The aim of this study was to elucidate the mechanism of miRNA in the potential immune response of M. rosenbergii. to non-O1 V. cholerae MSVC-GY01 infection by transcriptome sequencing. Following quality screening, the control group received 10, 616, 712 clean reads, whereas the infected group received 9,727,616. The miRNA sequences in the two samples are extremely consistent and have a length of roughly 23 nt. In all, 871 known miRNAs were discovered, with 279 differentially expressed miRNAs (DEMs). Meanwhile, 62 novel miRNAs were predicted, including 43 DEMs. In order to understand the immune-related biological functions of DEMs, target genes were predicted. Pathway function annotation analysis showed that non-O1 V. cholerae affected the NOD-like receptor signaling pathway, RIG-I-like receptor signaling pathway, and Toll-like receptor signaling pathway, suggesting that miRNAs in the hepatopancreas play a key role in immune responses to pattern recognition receptors. Twelve DEMs were randomly selected for Quantitative Real-time PCR (qRT-PCR). Overall, the expression trends of qRT-PCR were consistent with the sequencing results. These findings corroborate the immunomodulatory function of miRNA in M. rosenbergii against non-O1 V. cholerae infection and provide guidance for the prevention and treatment of related illnesses. • MiRNA transcriptome analysis of M. rosenbergii under non-O1 V. cholerae infection was firstly performed. • A total of 871 known miRNAs were identified, among which 279 differentially expressed miRNAs were associated with infection. • The target genes of differentially expressed miRNAs were significantly enriched in 19 immune-related pathways. [ABSTRACT FROM AUTHOR]