Isolation,identification,and virulence gene analysis of pathogenic Aeromonas dhakensis in Macrobrachium rosenbergii and histopathological observation

To identify the cause of mass mortality of adult Macrobrachium rosenbergii in a farm in Gaoyou City,Jiangsu Province,China,a dominant strain named DKQ-1 was isolated from the hepatopancreas of dying M.rosenbergii and identified as Aeromonas dhakensis by purification culture,biochemical characterization,and 16S rRNA and gyrB gene sequence analysis.The results of the challenge test revealed that the strain was highly pathogenic and the 50%lethal dose(LD_(50))in 72 h to M.rosenbergii was 1.54×10^(5)CFU/mL.The amplification results of virulence genes show that strain DKQ-1 carried 9 virulence genes,including ascV,aexT,aer,act,lip,ompAI,gcaT,acg,and exu,supporting the strong virulence of strain DKQ-1 to M.rosenbergii.Histopathological observation of the hepatopancreas,gills,and intestines indicated that DKQ-1 injection into M.rosenbergii could cause serious tissue damage,which further supported the strong virulence of this strain.In addition,a drug susceptibility test revealed that strain DKQ-1 was sensitive to 16 kinds of antibiotics,resistant to 9 kinds of antibiotics,and had intermediate resistance to spectinomycin and kanamycin.This study is the first report of A.dhakensis isolated from M.rosenbergii and provided a reference for the pathogen identification of bacterial diseases in M.rosenbergii,and for the prevention and treatment caused by A.dhakensis.

Genetic parameter evaluation for growth and cold resistance traits of the giant freshwater prawn Macrobrachium rosenbergii

The giant freshwater prawn Macrobrachium rosenbergii distributed from tropical to subtropical regions,is a warm-water species,and its survival temperature is 14-35°C,which greatly limits its culture cycle and culture area in China.Therefore,it is urgent to cultivate a new high quality,high yield variety with improved cold-resistance,but the genetic parameters for cold-resistance traits are unknown in M.rosenbergii.In this study,the cold-resistance of adult M.rosenbergii populations was tested using the indoor artificial cooling method.Individuals were selected from 139 families of Shufeng G3 generation and cultured for 200 days.A linear mixed model was constructed by ASReml-R to evaluate the genetic parameters of the cold-resistance trait(cooling degree hours,CDH)and growth traits(body weight,BW,and body length,BL)based on the restricted maximum likelihood(REML)method.The results show that the heritability of CDH was low(0.12±0.05),while the growth traits(BW and BL)had low to moderate heritability,with 0.20±0.06 for BW and 0.06±0.04 for BL.The phenotypic and genetic correlation between BW and BL was significantly positive,but significantly negative phenotypic and genetic correlations were detected between CDH and BW and between CDH and BL.Furthermore,the analysis of the differences between cold-resistance and phenotypic traits showed that the female reproductive status,exoskeleton hardness and claw number of adult prawns had a great influence on the cold-resistance of M.rosenbergii(P<0.05),indicating that adults with claws and hard exoskeletons are preferred as parents in subsequent breeding selection.The present results can be attributed to the selection and breeding of a new cold-resistant variety of M.rosenbergii.

Isolation,identification,and evaluation of intestinal bacteria in Macrobrachium rosenbergii

Probiotics are live microbial food supplements that have been shown to have beneficial effects on animal health.Endogenous probiotic bacteria have long been used for their proposed health promoting properties and have become a hot research topic in growth improvement in aquaculture.The endogenous probiotic bacteria from intestines of Macrobrachium rosenbergii(giant river prawn)was explored for their probiotic potential,from which 367 bacterial strains were isolated from the intestine of M.rosenbergii.After 16 S rDNA sequence analysis,234 isolates were identified as Lactococcus garvieae,which accounted for 63.76%of the total number of culturable intestinal bacteria,suggesting that this bacterium was the main component of the microbiota.Furthermore,to reveal the probiotic properties of L.garvieae,this isolated bacterial strain was characterized morphologically,physiologically,and biochemically.Its enzyme production capacity,bacteriostatic activity,and resistance to acid,high temperature,and pH,were assessed.In vitro experiments showed that the L.garvieae(No.C 6 a 2)had a fast growth rate and entered the logarithmic phase rapidly.Besides,it had characteristics of acid-production and resistance,enzyme-producing capacity,and strong antibacterial activity against pathogenic Staphylococc us aureus,Aeromonas hydrophila,and Aeromonas veronii.However,it lacked the ability to tolerate high temperature.Our results provide novel data to deepen our understanding of the intestinal bacteria structure of M.rosenbergii and valuable information for probiotic screening and the application for M.rosenbergii.

Heterosis and combining ability evaluation for growth traits of blunt snout bream(Megalobrama amblycephala)when crossbreeding three strains

The blunt snout bream(Megalobrama amblycephala)is a major aquaculture species in the Chinese freshwater polyculture system.In this study,complete diallel crossing of three strains(i.e.,Liangzi[LZ],Poyang[PY]and Yuni[YN])was used to evaluate the combining ability and heterosis effect of intraspecific crossbreeding on the growth performance(i.e.,body weight,total length,body length,body height,and Fulton’s coefficient of condition).The offspring produced from the three strains were reared in a communal pond for 20 months,and nine microsatellites were used to assess their pedigree.Of the749 offspring,708(94.53%)could be assigned directly to a single parental pair.Significant differences were observed in body weight(BW)among the different combinations,indicating that the combining ability of BW from each strain was significantly different.The general combining ability(GCA)of BW from the sire was much higher than that from the dam.The GCA for the dam and sire of the LZ and YN were the largest and the smallest,respectively.The special combining ability of YN$9 PY#was the largest,and YN$9 PY#showed significantly positive heterosis effects on all F1growth traits,except for Fulton’s condition coefficient(K)(P\0.05).In addition,significant positive linear correlations were found between the mean BW of corresponding progeny and the H E of the parental generation(y=600.7x-29.472,r=0.8651,P=0.0026),H O(y=1206.9x-518.14,r=0.7436,P=0.0216),and the polymorphism information content(y=1021.3x-331.31,r=0.8245,P=0.0063)as determined by a correlative test.The findings obtained from the study will be important for future considerations of M.amblycephala stock improvement programs.