Pathophysiology of enteropathogenic Escherichia coli during a host infection

Enteropathogenic Escherichia coli (EPEC) is a major cause of infantile diarrhea in developing countries. However, sporadic outbreaks caused by this microorganism in developed countries are frequently reported recently. As an important zoonotic pathogen, EPEC is being monitored annually in several countries. Hallmark of EPEC infection is formation of attaching and effacing (A/E) lesions on the small intestine. To establish A/E lesions during a gastrointestinal tract (GIT) infeciton, EPEC must thrive in diverse GIT environments. A variety of stress responses by EPEC have been reported. These responses play significant roles in helping E. coli pass through GIT environments and establishing E. coli infection. Stringent response is one of those responses. It is mediated by guanosine tetraphosphate. Interestingly, previous studies have demonstrated that stringent response is a universal virulence regulatory mechanism present in many bacterial pathogens including EPEC. However, biological signficance of a bacterial stringent response in both EPEC and its interaction with the host during a GIT infection is unclear. It needs to be elucidated to broaden our insight to EPEC pathogenesis. In this review, diverse responses, including stringent response, of EPEC during a GIT infection are discussed to provide a new insight into EPEC pathophysiology in the GIT.     

Potential of indigenous Bacillus spp. as probiotic feed supplements in an extruded low‐fish‐meal diet for juvenile olive flounder,Paralichthys olivaceus

A 12‐week feeding trial was designed to assess the probiotic potential of indigenous Bacillus amyloliquefaciens and/or Bacillus subtilis singly or in combination with Bacillus licheniformis in an extruded feed for olive flounder (Paralichthys olivaceus) juveniles. A high fish meal (FM) diet (control) and a low‐FM diet containing an alternative protein blend (30% FM replacement, FM30) were formulated. Three other experimental diets were prepared by inclusion of B. amyloliquefaciens (BA), B. subtilis (BS), or a mixture of B. amyloliquefaciens, B. subtilis, and B. licheniformis (BASL) into FM30 diet, with a final concentration of 10⁶ CFU/g diet. Results indicated that the FM30 diet was well tolerated by flounder, and the overall performance was not affected by dietary treatments. Lysozyme activity and total immunoglobulin level were significantly reduced in flounders when fed with the FM30 diet compared with the BASL and BA diets, respectively. The Bacillus additives neither enriched the relative abundance of the corresponding Bacillus spp. in the relevant gut microbiota of olive flounder nor modulated the presumptive gene functions of the gut microbiome. Despite the absence of growth‐promoting effect, the tested probiotics could still be economically viable for use as immunostimulants in commercial flounder diets with partial FM replacement.     

Analysis of Variation in Anthocyanin Composition in Korean Coloured Potato Cultivars by LC-DAD-ESI-MS and PLS-DA

Anthocyanin concentration and composition and the effect of steaming and baking on these were evaluated in tubers of Korean red- and purple-fleshed potato cultivars and breeding clones using liquid chromatography with diode array detection and electrospray ionization-mass spectrometry (LC-DAD-ESI-MS). Twenty-six anthocyanins were isolated, of which 24 were identified. Remarkably, five cis isomers were identified, of which four, viz., cis-petanin, cis-peonanin, petunidin 3-cis-caffeoylrutinoside-5-glucoside, and petunidin 3-cis-feruloylrutinoside-5-glucoside, are reported for the first time. Moreover, pelargonidin 3-p-coumaroylrutinoside-5-glucoside (pelanin), peonidin 3-p-coumaroylrutinoside-5-glucoside (peonanin) and petunidin 3-p-coumaroylrutinoside-5-glucoside (petanin) were identified as the principal anthocyanins. We found that the total anthocyanin content of coloured potatoes was decreased by steaming and baking compared with the raw state. In addition, we performed partial least square discriminant analysis (PLS-DA) to discriminate between the analyzed anthocyanins. Cis isomers seemed to play a vital role as a biomarker in the PLS-DA model based on the type of processing and colour of the tubers.     

Genetic analysis of ORF5 of recent Korean porcine reproductive and respiratory syndrome viruses (PRRSVs) in viremic sera collected from MLV-vaccinating or non-vaccinating farms

The 23 open reading frame (ORF) 5 sequences of Korean type II porcine reproductive and respiratory syndrome virus (PRRSV) were collected from viremic sera from the (modified live vaccine) MLV-vaccinating and non-vaccinating farms from 2007 to 2008. The samples were phylogenetically analyzed with previous ORF5 sequences, including type I Korean PRRSV, and previously reported or collected sequences from 1997 to 2008. A MN184-like subgroup of type II Korean PRRSV was newly identified in the viremic sera collected from 2007 to 2008. And of the type I PRRSVs, one subgroup had 87.2~88.9% similarity with the Lelystad virus, showing a close relationship with the 27~2003 strain of Spain. The maximum parsimony tree of type II PRRSV from 1997 to 2008 showed that they had evolved to four lineages, subgroups 1, 2, 3 and 4. Most of the recently collected type II PRRSVs belonged to subgroup 4 (48%). The region of three B-cell epitopes and two T-cell epitopes of ORF5 amino acids sequences was considerably different from the MLV in subgroups 3 and 4. In conclusion, the existence of type I PRRSV, which was genetically different from Lelystad virus (Prototype of type I PRRSV), and heterologous type II PRRSVs of viremic pigs detected even in the MLV-vaccinating farms indicated the need for new vaccine approaches for the control of PRRSV in Korea.     

溶菌酶在食品工业中的应用

综述了溶茵酶的基本性质,着重讨论了溶菌酶抑制的微生物种类、机理,系统地阐述了溶菌酶的制备及其在食品工业中的应用进展.     

Exogenous DNA Uptake of Boar Spermatozoa by a Magnetic Nanoparticle Vector System

The sperm‐mediated gene transfer method is applicable to transgenesis in many species that use spermatozoa for reproduction recently, which has been shown various results. In the current study, we show that transgenic porcine embryos can be efficiently produced by employing a simple transfection method that uses magnetic nanoparticles (MNPs). The complexes formed between plasmid DNA and MNPs were bounded on ejaculated boar spermatozoa at a higher efficiency compared to methods using DNA alone or lipofection. Using confocal microscopy, rhodamine fluorophore‐labelled MNPs were detected on external surfaces of the spermatozoa membrane, which were bounded on zona pellucida of in vitro maturated oocyte during in vitro fertilization. Electron microscopy revealed that clusters of MNPs were detected in inside of plasma membrane and nucleus of the spermatozoa head. Additionally, we found that magnetofected boar spermatozoa could be fertilized with oocytes in vitro and that the resulting gene of green fluorescent protein was detected in fertilized eggs by genomic PCR analysis. Taken together, these results suggest that MNPs can be used to efficiently introduce a transgene into embryo via spermatozoa.     

One year's study of dynamic and evolution of types I and II PRRSV in a swine farm

This study was to investigate dynamic and evolution of PRRSV in a seed-stock farm by monitoring PRRSV status from 11 June 2009 to 4 August 2010. For laboratory test, around 18-24 umbilical cords from farrowed sows and 5-95 sera from nursery and grow/finish pigs were submitted around every 2 weeks interval during the study. The submitted samples were tested for PRRSV using IDEXX PRRS 2XR ELISA kit, RT-nested PCR. The PRRSV-positive samples were further sequences based on ORF5 and analyzed using MEGA 3.1 program and Beast 1.5.4 package. The surveyed farm was first infected with type II PRRSV but it was infected newly with type I PRRSV of unknown origin, showing rapid substitution to type I PRRSV as a dominant strain in 2 weeks. The type I PRRSV was first detected from umbilical cord of a farrowed sow in 12 January 2010, and secondly from nursery pigs in 26 January 2010. Although sudden increase of mean S/P ratio was found in grow/finish pigs around 2 months earlier than first type I PRRSV detection, no type I PRRSV viremia was found. Thirty three ORF5 full sequences from 14 type II to 19 type I PRRSVs were obtained chronologically in this farm and the genetic characteristics and evolution rates of those sequences were analyzed. The substitution rates (/site/day) of two types were 4.03×10(-5) (type I), 3.09×10(-5) (type II), respectively, which was more frequent than previous reports. The calculated divergence time of type I PRRSV was consistent with the time when the sudden elevation of serum IgG in grow/finish barn was first observed. This study provided fundamental data for type I PRRSV dynamic in a previously type II PRRSV-infected farm and suggested grow/finisher barn could be a primary site for PRRSV introduction.