Research Progress in Phage

With excessive utilization of antibiotics in recent years,bacterial drug resistance problem is serious increasingly,and it is more and more difficult to develop anti-infective drug,while it does not have these problems to use phage controlling disease.Phage is a kind of prokaryotic virus,widely exists in the nature and includes bacteriophage,cyanophage and actinophage.Due to its potential of replacing antibiotics to treat disease,phage receives more and more attention.In this paper,based on development status of phage research at home and abroad,discovery process,naming method and classification basis of phage are introduced comprehensively,and advantages and limitations of phage applying in prevention and control of bacterial diseases are analyzed.We introduce application status of phage in human medicine,prevention and control of diseases for terrestrial animals and aquaculture,and the effects of phage in sewage treatment,prevention and control of microbial contamination of food and detection technology,and point out the shortages of phage in the above application.Meanwhile,we also discuss application prospects of phage in disease prevention and control,environmental protection and food safety.

Monoclonal antibody based Dot-ELISA and indirect fluorescence antibody technique for detecting Edwardsiella ictaluri infection in yellow catfish (Pelteobagrus fulvidraco)

Edwardsiella ictaluri is known as the etiological agent of Red-head disease of yellow catfish(Pelteobagrus fulvidraco),and is the cause of heavy economic losses in the aquaculture industry.In this study,a DotEnzyme Linked Immunosorbent Assay(Dot-ELISA)and an Indirect Fluorescence Antibody technique(IFAT)for detecting Edwardsiella ictaluri were developed by using a monoclonal antibody,5D11.For DotELISA,the working dilutions of 5D11 and the secondary antisera(enzyme-labeled goat anti-mouse Ig)were 1:320 dilution and 1:3000 dilution,respectively.For IFAT,the working dilutions of 5D11 and goatanti-mouse Ig conjugated with fluorescein isothiocyanate were as respectively 1:80 and 1:256.Both of the methods established were highly sensitivity(Minimum detectable concentration,5
107 CFU/mL)and had a high degree of accuracy(positive rate was 100%for both artificial infected fish and spontaneous diseased Pelteobagrus fulvidraco).The two reliable methods developed have high potential for quick and efficient detection of Edwardsiella ictaluri in aquaculture production units.

Identification and histopathological and pathogenicity analysis of Aeromonas salmonicida salmonicida from goldfish (Carassius auratus) in North China

In this study,bacterial pathogens were isolated from the liver,spleen,kidney,heart,and muscle of diseased goldfish(Carassius auratus)after a massive outbreak of disease in an aquaculture farm in Anshan,Liaoning Province,North China(N41°05′58.42″E122°53′31.89″).Based on physiological and biochemical characterization of the bacteria and 16S rDNA sequence analysis,the bacterial isolates were identified as Aeromonas salmonicida salmonicida.Virulence gene multilocus sequence typing analysis showed that the bacterial isolates from all the organs collected were of the same type,and clustered in the same phylogenetic group as A.salmonicida salmonicida.The pathogen,was named AS.17 and was resistant to rifampicin,bacitracin,vancomycin,penicillin and compound sulfamethoxazole,but was highly sensitive to nalidixic acid,lomefloxacin,spectinomycin,ofloxacin,and ciprofloxacin.Goldfish experimentally infected with AS.17 had similar symptoms to naturally infected fish,including gill filament anemia,intestinal bleeding,intra-abdominal fluid,and jaw bleeding.Histological analyses showed that the liver,the spleen,the kidney and the intestines of fish infected with AS.17 had severe pathological alterations.In summary,this study reports the isolation of A.salmonicida salmonicida from C.auratus in North China and provides a reference strain and a research foundation for further studies on A.salmonicida disease control and epidemiology.

Enhancing the immune response in the sea cucumber Apostichopus japonicus by addition of Chinese herbs Houttuynia cordata Thunb as a food supplement

The Chinese herb Houttuynia cordata Thunb(abbreviated as HCT hereafter)has been widely used in human and livestock medical research.However,few studies have tested the effects of HCT in aquaculture systems,particularly in echinoderms.In this study,the impact of HCT was evaluated when used as a feed additive to enhance the immune response of the sea cucumber,Apostichopus japonicus.Two concentrations,1.5%and 3%,of HCT powder were tested by adding them to the feed.Feeding experiments had a four week duration and each week,the humoral and cellular immunity index of the intestine,tentacles,peristome,and coelomic fluid were analyzed.The results indicated that the percentage of A.japonicus coelomocytes significantly increased after supplementing feed with HCT,and reached a peak after one week of feeding with the 3%HCT supplementation.Analysis of reactive oxygen species(ROS)indicated that HCT in feed caused a notable increase in the coelomocyte ROS concentrations over the experimental period.In both the 1.5%and 3%HCT addition groups,the ROS peaked in the third week and then remained stable.In addition,alkaline phosphatase,acid phosphatase,superoxide dismutase,lysozyme activity of the intestine,tentacles,peristome and coelomic fluid were significantly improved during the four-week test period.In conclusion,1.5%and 3%HCT added to feed boosts the immune responses of A.japonicus.HCT has potential as an immune enhancer for mariculture of sea cucumbers.