Solar photocatalytic pathogenic disinfection:Fundamentals to state-of-the-art

It is necessary to treat pathogen-infected water before its utilisation.Of conventionally used treatment methods,solar photocatalysis has gained considerable momentum owing to its operational simplicity and capacity to use freely and abundantly available solar energy.This article systematically reviewed the disinfection of water with photocatalysis.It addressed the concerns of microbial infection of water and the fundamentals behind its treatment with photocatalysis.It presented an in-depth description of pathogenic deactivation with powerful reactive oxygen species.Special emphasis was given to process intensification as it is an attractive technique that provides multifunctionality and/or equipment miniaturisation.Solar reactor design regarding mobilised/immobilised photocatalysts and compound parabolic concentrators were elucidated.Finally,key parameters governing photoperformance,corresponding trade-offs,and the need for their optimisation were discussed.Overall,this article is a single point of reference for researchers,environmentalists,and industrialists who address the ever-severing challenge of providing clean water whilst also maintaining energy sustainability.

Isolation and Identification of a Pathogenic E.coli Strain Causing Diarrhea in Foxes

[Objectives] The study aimed to identify the pathogenic E. coli strain that caused diarrhea in foxes and to analyze its drug sensitivity.[Methods] A pathogenic E. coli strain was isolated from dead foxes with diarrhea. By conventional bacterial isolation and culture, morphological observation, pathogenicity test and K-B disc method, the isolated strain was identified as pathogenic E. coli .[Results] The isolated pathogen was highly sensitive to ceftriaxone, cefotaxime, ciprofloxacin and lincomycin, moderately sensitive to enrofloxacin, neomycin, gentamycin, spectinomycin, florfenicol, amikacin and polymyxin, and resistant to ampicillin, amoxicillin and doxycycline.[Conclusions] This study provided reference for the prevention and control of diarrheal diseases in foxes in Qinhuangdao region.

Detection of High Pathogenicity Island(HPI) in Pathogenic Escherichia coli of Mink by PCR

[Objective] The paper was to analyze the carrying status of high pathogenicity island （HPI） in pathogenic Escherichia coli of mink. [Method] Eight strains of E. coli were isolated from dead mink, and conducted pathogenicity test of artificial infection. The carrying status of HPI （irp2, fyuA） was detected by PCR. [Result] Eight strains of E. coli were pathogenic E. coli, and the carrying rate of HPI （irp2, fyuA） was 100%, positively correlated with the pathogenicity. [Conclusion] The results lay a foundation for further exploring the pathogenic mechanism of E. coli..

Pathogenicity of Escherichia coli O_(123) from Rex Rabbit on White Mice

[Objective]The paper was to analyze the pathogenesis of Escherichia coli O123 from rex rabbit. [Method]E. coli O123 isolated from rabbit liver with diarrhea symptom in scale rex rabbit farm was intraperitoneally injected into 18- 22 g Kunming mice,and its pathogenicity was determined by clinical symptoms and pathological examination. [Result]When the inoculation concentration was about 8. 5 × 107 CFU /mL,Kunming mice appeared the clinical symptoms of drooping spirit,diarrhea and gathering,and the mortality reached 50%. Anatomical examination found that intestinal wall was thinning and intestinal mucosa was bleeding. [Conclusion]E. coli from rex rabbit has strong pathogenicity,and establishing animal model with Kunming mice to study its pathogenesis is of great reference significance for diagnosis and prevention of E. coli disease of rex rabbit.

Effect of dietary fiber and threonine content on intestinal barrier function in pigs challenged with either systemic E.coli lipopolysaccharide or enteric Salmonella Typhimurium

Background:The independent and interactive effects of dietary fiber(DF)and threonine(Thr)were investigated in growing pigs challenged with either systemic E.coli lipopolysaccharide(LPS)or enteric Salmonella Typhimurium(ST)to characterise their effect on intestinal barrier function.Results:In experiment 1,intestinal barrier function was assessed via oral lactulose and mannitol(L:M)gavage and fecal mucin analysis in pigs challenged with E.coli LPS and fed low fiber(LF)or high fiber(HF)diets with graded dietary Thr.Urinary lactulose recovery and L:M ratio increased(P<0.05)during the LPS inoculation period in LF fed pigs but not in HF fed pigs.Fecal mucin output was increased(P<0.05)in pigs fed HF compared to LF fed pigs.In experiment 2,RT-qPCR,ileal morphology,digesta volatile fatty acid(VFA)content,and fecal mucin output were measured in Salmonella Typhimurium challenged pigs,fed LF or HF diets with standard or supplemented dietary Thr.Salmonella inoculation increased(P<0.05)fecal mucin output compared to the unchallenged period.Supplemental Thr increased fecal mucin output in the HF-fed pigs(Fib×Thr;P<0.05).Feeding HF increased(P<0.05)VFA concentration in cecum and colon.No effect of either Thr or fiber on expression of gene markers was observed except a tendency(P=0.06)for increased MUC2 expression with the HF diet.Feeding HF increased goblet cell numbers(P<0.05).Conclusion:Dietary fiber appears to improve barrier function through increased mucin production capacity(i.e.,goblet cell numbers,MUC2 gene expression)and secretion(i.e.,fecal mucin output).The lack of effect of dietary Thr in Salmonella-challenged pigs provides further evidence that mucin secretion in the gut is conserved and,therefore,Thr may be limiting for growth under conditions of increased mucin production.

Isolation and Characterization of E. Coli O157 : H7 from Infected Newborn Calves in Northeast China

Escherichia coli O157 : H7 is a foodborne pathogen that poses a major threat to public health. Epidemiologic investigations have identified dairy cows, especially calves, are the principal reservoir of E. coli O157 : H7. In this study, based on the results, E. coli O157 : H7 was the main cause of E. coli disease outbreak in late October, 2015, and more than 90% of newborn calves died of serious diarrhea. Through further experiments, the drug sensitivity and resistance of the strain, the expression of the virulence gene and virulence pathogenicity were studied. E. coli O157 : H7 isolates were resistant to 12 antibiotics including penicillin, tetracycline and ampicillin, and were sensitive to eight antibiotics including cefoperazone, ceftazidime and amikacin. Resistance genes included tetB, strB, aadB, aphA, floR, TEM and virulence genes included stx1, eaeA and hlyA. Using specific pathogen free mice, the result showed that the isolate was pathogenic with a median lethal dose of 7.9×107 CFU · mL-1. This study described the pathogenesis and clinical manifestations of E. coli O157 : H7 infection. These results guided the use of antibiotics in prevent and control of bacterial infections in the future.

UV-TiO_2 photocatalytic disinfection and photoreactivation of pathogenic bacterium in municipal wastewater

The disinfected bacteria will be a photoreactivation under the irradiation of the sunlight,and the light intensity plays an important role in the bacteria resurrection.The effect of light intensity on photoreactivation of Escherichia coli(E.coli) and Enterococcus faecalis(E.faecalis) in secondary effluents which were disinfected respectively by pure UV and UV-TiO_2 was investigated.The results show that the disinfection efficiency of UV-TiO_2 is much higher than that of the pure UV disinfection.The photoreactivation rate of E.coli is much higher in pure UV disinfection than in UV-TiO_2 photocatalytic disinfection.Under high light intensity in UV-TiO_2 disinfection,high resurrection rate can be induced.However,a higher resurrection rate can be introduced even under low light intensity in pure UV disinfection alone.Meanwhile,UV-TiO_2 disinfection has a strong inhibition effect on E.faecalis photoreactivation.When the light intensity is lower than 21 μW/cm^2,nearly no resurrection of E.faecalis occurs after 72 h resurrection irradiation,and a little resurrection rate is observed only under a strong photoreactivating light intensity.

A Comparative Investigation on Growth of Three Food Born Pathogenic Bacteria Inoculated with Withania somnifera:an Invitro Experimental Study

Background:Withania somnifera(WS)is proposed as one of the alternatives instead of the antibiotic.This study is aimed to evaluate the inhibitory potency of enzymatic extract of the fruits of the WS.Methods:As an invitro experimental study,the growth rate of Shigella dysenteriae,Salmonella typhimurium,and Escherichia coli inoculated in different concentrations(25%,12.5%,6.25%and 3.125%)of the extract were assessed.A microtitre plate method was conducted.ANOVA was applied to identify statistical differences with p-value<0.05.Results:Different concentrations of extract,in comparison with control,declined the growth rate of all tested bacteria.All concentrations inhibited the growth of S.typhimurium(p<0.05).Compared to the microorganism control,effective concentration of the extract inhibiting the growth of E.coli was 12.5%,and 6.25%,while it was 12.5%,and 6.25%for Sh.dysenteriae(p<0.05).A dose-dependent response of E.coli was observed.The antibacterial activity of the extract tested was found mainly against E.coli and Sh.dysenteriae.The most resistant microorganism compared to E.coli and Sh.dysenteriae was S.typhimurium(p<0.05).25%of the concentration of the extract showed the different inhibitory effect among three tested bacteria(p<0.05).Conclusions:The extract was labeled as an antibacterial agent against the representative of three foodborne bacteria,Invitro.The common effective concentrations of the extract(12.5,and 6.25%)is recommended for further research,as food additive,to remedy digestive ailments related to E.coli,S.typhimurium and Sh.dysenteriae.

The Wheat Pathogenesis Related Protein (TdPR1.2) Ensures Contrasting Behaviors to <i>E. coli</i>Transformant Cells under Stress Conditions

The pathogenesis-related proteins 1 (PR-1) gene family play important roles in the plant metabolism in response to biotic and abiotic stresses. The wheat TdPR1.2 has been previously isolated and characterized. Here we showed by bio-informatic analysis that TdPR1.2 contains six cysteine residues that are conserved between all PR-1 proteins tested. Using ScanProsite tool, we found that TdPR1.2 structure has a CRISP family signature 1 and 2 located at the C-terminal part of the protein. Those two domains are conserved in many identified PR1.2 proteins in plants. Moreover, SignalIP-5.0 analysis revealed that TdPR1.2 contains a putative signal peptide formed by 25 amino acids at the N-terminal extremity. The presence of this signal peptide suggested that the mature proteins will be secreted after the cleavage of the signal sequence. Further, we investigate the role of the TdPR1.2 proteins in the growth of Escherichia coli transformants cells under different abiotic stresses. Our results showed that the full-length form of TdPR1.2 enhanced tolerance of E. coli against salt and osmotic stress but not to KCl. Moreover, TdPR1.2 protein confers bacterial tolerance to heavy metals in solid and liquid mediums. Based on these results, we suggest that the TdPR1.2 protein could play an important role in response to abiotic stress conditions.

贵州部分地区猪源大肠杆菌耐药性分析及ESBLs基因型检测

为了解贵州地区规模养猪场大肠杆菌菌株耐药情况和ESBLs基因型的流行情况,试验采用CLSI推荐的方法对采自贵州省4个地区规模养猪场的164株大肠杆菌进行药物敏感性试验和产ESBLs菌的检测,并用PCR方法对TEM、SHV、OXA-1和CTX-M-1 4种常见ESBLs基因进行检测。结果显示,164株大肠杆菌对10种常用抗菌药物耐药率分别是头孢噻呋93.29%、氨苄西林87.19%、四环素86.59%、庆大霉素81.10%、链霉素53.66%、多黏菌素51.83%、环丙沙星53.05%、卡那霉素47.56%、金霉素34.76%和氟苯尼考21.95%,且大多为多重耐药,其中检测出ESBLs阳性菌株137株,阳性率为83.54%,各个地区的检出率不同;137株产ESBLs大肠杆菌中,TEM、SHV、OXA-1和CTX-M-1基因的检出率分别为90.51%、70.07%、51.82%和43.07%,且多为复合基因型耐药菌株,各地区的各种基因检出率不同。试验结果表明,贵州部分地区的猪源大肠杆菌耐药现象严重,ESBLs菌株的检出率很高,耐药基因的检出率也极高,且多为复合基因型耐药菌株,应加强当地产酶耐药菌的监测和研究,有效防制此类细菌引发的疾病。

产毒性大肠杆菌(ETEC)中毒力岛分布的研究

目的 了解小肠结肠炎耶尔森菌强毒力岛在产毒性大肠杆菌中的分布，以便于进一步深入研究毒力岛在大肠杆菌中的结构的完整性和功能。方法ＰＣＲ扩增和原位杂交及基因序列分析。结果 在检测的９３株产毒性大肠杆菌（ＥＴＥＣ）的毒力岛的检出率为 ３２．２５％（３２／９３），而且这些阳性菌株中的毒力岛大部分连接到 ａｓｎｔＲＮＡ（天门冬氨酸ｔＲＮＡ）位点。结论 产毒性大肠杆菌是致病性较强的病原菌之一，而小肠结肠炎耶尔森菌强毒力岛在产毒性大肠杆菌中阳性率较高的分布．

肠聚集性大肠杆菌ybtE基因缺失突变株的构建

目的构建肠聚集性大肠杆菌ybtE缺失突变株,为下一步ybtE基因功能的研究奠定基础。方法应用PCR扩增肠聚集性大肠杆菌的ybtE基因,将其克隆至pUC18载体,酶切缺失988bp ybtE片段,并插入998bp的卡那霉素抗性基因,利用定向克隆技术将突变的ybtE片段克隆至自杀质粒pKTN701,形成重组自杀质粒。将携带重组自杀质粒的SM10λpir与EAEC17-2进行接合转移,利用同源重组,根据卡那霉素抗性筛选并鉴定接合子。结果经PCR、酶切和探针杂交鉴定,得到2株肠聚集性大肠杆菌ybtE缺失突变株。结论成功构建肠聚集性大肠杆菌ybtE基因缺失突变株。

Department of Etiology and Department of Pharmacology

In order to understand the role of transmembrane signal transduction of host cells in the early steps of infection,the adherence of E. coli to HEp 2 cells and the change of activity of phospholipase C γ (PLC γ) induced by the adherence were investigated.The adherence of enteropathogenic E.coli (EPEC), strain E.7, induced a significant increase of inositol triphosphat (IP 3) level in HEp 2 cells. The adherence of the bacteria and the increase of IP 3 was kinetically correlated. Whereas the increase of IP 3 level induced by the adherence of the control strain EPEC (H511), a non piliated strain, was much meager than that by E7, a piliated strain. The results highlighted an important role of transmembrane signals like IP 3 in the pathogenesis of EPEC.

Effects of Preparation and Storage of Agar Media on the Sensitivity of Bacterial Forward Scattering Patterns

Recent worldwide foodborne outbreaks emphasize the need for the development of rapid and accurate method for pathogen detection. To address such issues, a new colony based label-free detection method working on the principles of elastic light scattering was introduced. In order to build libraries of scattering images for bacterial pathogens, it is pertinent to determine the effect of preparation and storage of the agar media on the scatter patterns. Scatter patterns of three Escherichia coli serovars (O26, O111 and O157) were studied and used in a model system, after growth on Sorbitol-MacConkey agar plates that were prepared and stored at different conditions in the laboratory. Quantitative image processing software was used to analyze variation in scatter patterns of the same serovar on media prepared under various standard laboratory conditions and to generate a cross-validation matrix for comparison. Based on the results, it was determined that attention should be given during preparation of media so that the agar plates are not air-dried more than 10 - 20 min after solidification at room temperature. The plates could be stored in sealed bags in cold room (4oC - 10oC) for up to a month before use. The findings of this study should provide guidelines in preparation, storage, and handling of media for generation of reproducible scatter patterns of bacterial colonies with the light scattering sensor for pathogen detection.

一株仔猪产肠毒素大肠杆菌分离鉴定、耐药性分析及其卵黄抗体的制备

为诊断某规模化养猪场仔猪腹泻病原,并制备其特异性卵黄抗体,采用细菌分离培养、生化试验、16SrRNA PCR鉴定及系统进化树分析、药敏试验等方法对病料进行研究,并利用全菌体灭活和超声波破碎法制备抗原,将其分别与不同佐剂(ISA 71VG、ISA201VG和15AVG)配伍乳化制备疫苗免疫蛋鸡、聚乙二醇法提取高免卵黄抗体、SDS-PAGE电泳和Bradford法鉴定抗体质量及浓度、双向琼脂扩散试验测定抗体效价。结果表明,分离菌株的形状、大小及生化特性均与大肠杆菌相符。分离株的主要毒力基因E.coli-estb和elt基因分别为113、272 bp;分离株elt和estb基因与美国流行毒株如UMNK88处于同一分支,亲缘关系较近;与丹麦株CFSAN018748、瑞士株14OD0056处于不同分支,亲缘关系较远。分离菌株对头孢噻肟、诺氟沙星和复方新诺明敏感,对头孢曲松和多黏菌素B中介,对多西环素、呋喃唑酮以及多种氨基糖苷类和喹诺酮类抗菌药物耐药。各组卵黄抗体的重链(约70 ku)和轻链(约25 ku)均明显,纯度高。首免后56 d 0.2%甲醛处理的菌液与ISA 15AVG配伍所致疫苗的卵黄抗体蛋白含量最高(1.462 mg/g卵黄),超声波破碎处理的菌液与ISA 71VG配合所制疫苗的卵黄抗体蛋白含量次之(1.459 mg/g卵黄)。血清抗体与菌体抗原反应性良好,超声波破碎抗原与ISA 71VG配伍所制疫苗的血清抗体、卵黄抗体效价分别高达1∶16、1∶4。

猪致病性大肠杆菌耐药质粒检测及其中药消除作用研究

为探讨中药对河南省规模化养猪场致病性大肠杆菌(E.coli)耐药质粒消除的效果,本研究在对87个猪致病性E.coli分离株进行耐药质粒检测的基础上,将各分离株分别于含有不同浓度的石榴皮、黄芩、黄连等中药提取液的培养液中培养,进行耐药质粒消除试验;并采用影印培养法和纸片扩散法进行耐药性消除菌落筛选及其对抗菌药物敏感性恢复的检测。结果表明:各分离株均含有2个～8个质粒,大小约为1.0 kb～87.0 kb;经石榴皮、黄芩、黄连作用后,各分离株均可以获得数量不等的耐药性消除菌落,消除率为0.34%～16.2%,以黄连的平均消除率最高,为9.33%;在耐药性消除的各分离株中,有25个分离株未发生质粒消除,其余均消除了1条～3条质粒,并且对7种抗菌药物的平均敏感率由消除前的5.39%分别提高至60.4%、75.2%和76.5%。表明石榴皮、黄芩、黄连对猪致病性E.coli具有质粒消除和耐药性逆转作用,其中黄连作用效果最好。

规模化猪场大肠杆菌β-内酰胺类药物耐药性检测及其超广谱β-内酰胺酶调查

为阐明我国规模化猪场大肠杆菌(E.coli)对抗菌药物的耐药现状和耐药机制,本研究于2006年2月至2008年12月从19个省市97个规模化猪场分离鉴定602株E.coli,进行了4大类16种抗菌药物耐药性检测和产超广谱β-内酰胺酶(ESBLs)菌株的筛选,采用PCR方法检测了9种常见β-内酰胺酶耐药基因,对产ESLBs菌株进行了脉冲电场凝胶电泳(PFGE)分析。结果显示:E.coli分离株对抗菌药物产生了不同程度的耐药性,602株E.coli中共有57株产ESBLs,产ESBLs菌株普遍耐受第二及第三代头孢药物。分别有35株和20株E.coli为blaCTX-M型和blaSHV型ESBLs,尚未发现blaOXA型和blaKPC型。PFGE结果显示,产ESBLs菌株间同源性低,推测我国猪源E.coli产ESBLs多为零星发生或以质粒介导的形式传播。本研究为了解我国猪源E.coli对β-内酰胺类药物的耐药现状和ESBLs变化趋势提供了实验依据。

规模化养鸭场鸭源大肠杆菌分离株的致病性及系统发育分析

为研究鸭源大肠杆菌(E.coli)的致病性,明确鸭病原性E.coli与人和其他动物E.coli的亲缘关系及O血清型与菌种间遗传关系的相关性,本研究从我国西南地区规模化养鸭场患典型E.coli败血症的雏鸭体内分离37个血清型共82个E.coli分离株(其中优势血清型32株,其他血清型50株),以每株0.2 mL(l09 cfu/mL)腿部肌肉接种7日龄健康鸭进行致病性试验;并对其中14个鸭源E.coli代表株进行16S rRNA基因克隆、测序及系统发育分析。结果表明:所有分离株全部为致病性E.coli,其中高致病株、中等致病株和低致病株分别占受试菌株的80.5%(66/82)、17.1%(14/82)和2.4%(2/82),4个优势血清型全部为高致病株和中等致病株,分别占受试菌株的84.4%(27/32)和15.6%(5/32);14个代表株鸭源E.coli的16S rRNA形成2个主要分支,3株新发现的血清型单独形成一个较远的分支,11株常见血清型与人及其他动物源E.coli形成另一个大的分支,其中有6株与人的O157亲缘关系较近,2株与出血性E.coli O157∶H7 sakai聚为一个小分支;从遗传进化的关系分析,这些菌株有可能是人类的潜在病原菌;同时还发现不同血清型的分离株可以聚为一支,而同一血清型的分离株可以处于不同的分支,常规的O血清学分型不能体现菌种间遗传关系的远近。

规模化猪场致病性大肠杆菌、沙门氏菌药敏区系调查

从四川省 1 6个和广西 1个规模化猪场临诊发生典型仔猪黄、白痢猪只的肛试粪样及死亡猪只的心血和肝中分离出致病性大肠杆菌 2 45株 ,从四川省 1 2个规模化猪场临诊发生典型仔猪副伤寒猪只中分离出致病性沙门氏菌 2 7株。用 1 6种常用抗菌药物对分离菌株进行了药敏测定 ,结果表明 ,大肠杆菌、沙门氏菌存在广泛的耐药性 ,以多重耐药性为主。大肠杆菌菌株最为敏感的药物是头孢唑啉、新霉素 ,而对林可霉素、青霉素钾的敏感率极低。对沙门氏菌菌株最为敏感的药物是新霉素、磺胺、庆大霉素、妥布霉素等 ,而对红霉素、林可霉素几乎全部耐药。

猪致病性大肠杆菌的分离鉴定及耐药性分析

从湖北省襄樊地区7家猪场采集病料进行大肠杆菌分离培养,获得36株菌株。通过菌落和细菌形态学观察、生化试验和动物试验确定分离细菌为致病性大肠杆菌。应用纸片扩散法对分离的大肠杆菌进行药物敏感试验,结果表明,对丁胺卡那霉素敏感的菌株有81%,氟苯尼考42%,庆大霉素39%,痢菌净33%,新霉素31%。对阿莫西林耐药的菌株为86%,氯霉素72%,土霉素56%,卡那霉素47%、庆大霉素47%。通过本次试验可知丁胺卡那霉素可作为襄樊地区防治致病性大肠杆菌的首选药物,氟苯尼考作为备选药物。