The selection pressure analysis of chicken anemia virus structural protein gene VP1

Chicken anemia virus (CAV) is the pathogen of chicken infectious anemia. To clarify the driving force in CAV evolution, we have detected positive selection in the structural protein gene VP1 by using maximum-likelihood models. Strong evidence was found that VP1 proteins were subject to the high rates of positive selection, and eight sites were identified to be under positive selection using the Bayes Empirical Bayesian method. Interestingly, four selected sites (amino acids 75, 125, 141, and 144) might be responsible for the attenuation exhibited. One selected site (amino acid 287) was connected with the virulence of CAV. This study provided some implication for the evolution of CAV, development of vaccines, and investigation into the structural and functional profiles of the VP1 protein. Dong Wang and Wei Fan contributed equally to this work.     

A new anti-infective strategy to reduce the spreading of antibiotic resistance by the action on adhesion-mediated virulence factors in Staphylococcus aureus

Staphylococcus aureus is a flexible microbial pathogen frequently isolated from community-acquired and nosocomial infections. S. aureus expresses a wide array of secreted and cell surface-associated virulence factors, including proteins that promote adhesion to damaged tissue and to the surface of host cells, and that bind proteins in blood to help evade immune responses. Furthermore, surface proteins have a fundamental role in virulence related properties of S. aureus, including biofilm formation. The present study evaluates the anti-infective capabilities of a secreted protein of Serratia marcescens (serratiopeptidase, SPEP), in impairing some staphylococcal virulence-related properties, such as attachment to inert surfaces and adhesion/invasion on eukaryotic cells. SPEP seems to exert its action by modulating specific proteins. It is not assessed if this action is due to the proteolytic activity of SPEP or to a specific mechanism which triggers an out/inside signal. Proteomic studies performed on surface proteins extracted from SPEP treated S. aureus cultures revealed that a number of proteins are affected by the treatment. Among these we found the adhesin/autolysin Atl, SdrD, Sbi, EF-Tu and EF-G. EF-Tu and EF-G are known to perform a variety of function, depending on their cytoplasmic or surface localization. All these factors can facilitate bacterial colonization, persistence and invasion of host tissues. Our results suggest that SPEP could be developed as a potential “anti-infective agent” capable to hinder the entry of S. aureus into human tissues, and also impairs the ability of this pathogen to adhere to prostheses, catheters and medical devices.     

Characterization of Listeria monocytogenes isolated from Ganges water,human clinical and milk samples at Varanasi,India

Listeria monocytogenes isolated from Ganges water, human clinical and milk samples were characterized by antibiotic susceptibility, serotype identification, detection of virulence genes and ERIC- and REP-PCR fingerprint analyses. All isolates were uniformly resistant to ampicillin, except two isolates, and showed variable resistance to gentamicin, cotrimoxazole, ofloxacin, rifampicin and tetracycline. Of the 20 isolates found positive for pathogens, seven (four human and three water isolates) belong to serogroups 4b, 4d and 4e; six (one human and five water isolates) belong to serogroups 1/2c and 3c; four milk isolates belong to serogroups 1/2b and 3b; and three milk isolates belong to serogroups 1/2a and 3a. Two water isolates, all human isolates, except one (Pb1) lacking inlJ gene, and three milk isolates possess inlA, inlC, plcA, prfA, actA, hlyA and iap genes. The remaining water and milk isolates showed variable presence of inlJ, plcA, prfA, and iap genes. ERIC- and REP-PCR based analyses collectively indicated that isolates of human clinical samples belong to identical or similar clone and isolates of water and milk samples belong to different clones. Overall study demonstrates the prevalence of pathogenic L. monocytogenes species in the environmental and clinical samples. Most of the isolates were resistant to commonly used antibiotics.     

A single nucleotide mutation results in loss of enzymatic activity in the hyaluronate lyase gene of Streptococcus pyogenes

Group A streptococci produce a variety of extracellular proteins, many of which are considered to be virulence factors. One of these is hyaluronate lyase (HylA), an enzyme capable of degrading the extracellular matrix of the host as well as the bacterial capsule. The current study examined three genotypes of hylA (full, truncated and deleted). Only isolates containing a full-length gene produced an enzymatically active hyaluronate lyase; however, truncation of the protein was not the reason for loss of activity. A single nucleotide substitution, resulting in an amino acid change at position 199 of the lyase was present in a highly-conserved region of the protein in isolates not producing active enzyme. In serotypes 4 and 22, those producing active enzymes, this residue was an aspartic acid, in serotypes not showing hyaluronate lyase activity, it was a valine. Site-directed mutagenesis indicated the loss of enzymatic activity of the hyaluronate lyase is in part determined by the mutation resulting in an amino acid residue change. This mutation results in an inactive form of the enzyme and is found in the more virulent serotypes of Streptococcus pyogenes, suggesting that hyaluronate lyase could interfere with the disease process, in essence being an anti-virulence factor.     

Risk factors in enterococci isolated from foods in Morocco: Determination of antimicrobial resistance and incidence of virulence traits

A collection of enterococci isolated from meat, dairy and vegetable foods from Morocco including 23 Enterococus faecalis and 15 Enterococcus faecium isolates was studied. All isolates were sensitive to ampicillin, penicillin, and gentamicin. Many E. faecalis isolates were resistant to tetracycline (86.95%), followed by rifampicin (78.26% ciprofloxacin (60.87%), quinupristin/dalfopristin (56.52%), nitrofurantoin (43.47%), levofloxacin (39.13%), erythromycin (21.73%), streptomycin (17.39%), chloramphenicol (8.69%), vancomycin (8.69%), and teicoplanin (4.34%). E. faecium isolates showed a different antibiotic resistance profile: a high percentage were resistant to nitrofurantoin (73.33%), followed by erythromycin (66.60%), ciprofloxacin (66.66%), levofloxacin (60.00%), and rifampicin (26.66%), and only a very low percentage were resistant to tetracycline (6.66%). One isolate was resistant to vancomycin and teicoplanin. The incidence of virulence factors was much higher among E. faecalis isolates, especially for genes encoding for sex pheromones, collagen adhesin, enterococcal endocarditis antigen, and enterococcal surface protein. Isolates with multiple factors (both antibiotic resistance and virulence traits) were also more frequent among E. faecalis isolates, in which one isolate cumulated up to 15 traits. By contrast, several isolates of E. faecium had only very few unwanted traits as compared to only two isolates in E. faecalis. The high abundance of isolates carrying virulence factors and antibiotic resistance traits suggests that the sanitary quality of foods should be improved in order to decrease the incidence of enterococci.     

黑龙江地区幽门螺杆菌毒力基因型与胃十二指肠疾病关系的研究

幽门螺杆菌（Hp）感染除与宿主和环境因素有关外，菌株的毒力差异对临床结果的影响也是不可忽视的因素。Hp基因亚型的分布具有明显的地域性，地理差异在导致不同临床结局中所起的作用仍是当前研究的热点。本研究旨在阐明Hp主要毒力基因亚型在黑龙江地区的分布情况及其与胃、十二指肠疾病的关系。     

磷酸肌酸佐治病毒性脑炎

目的：观察磷酸肌酸治疗病毒性脑炎的临床疗效。方法：对符合诊断标准的病毒性脑炎，按入院先后分为治疗组和对照组，治疗组在常规西医治疗基础上加用磷酸肌酸1g/d，通过观察临床主要症状持续天数及疗程判断疗效。结果：治疗组的发热、头痛、呕吐、抽搐及意识障碍持续天数较对照组缩短，两者有显著差异。结论：磷酸肌酸佐治病毒性脑炎疗效显著。     

中国香港地区幽门螺杆菌毒力基因型与十二指肠溃疡关系的研究

目的：研究幽门螺杆菌（Hp）的重要毒力因子vacA，cagA，iceA及插入序列IS在中国香港地区分离菌株中的分布特征及其与十二指肠溃疡的关系。方法：采用聚合酶链反应（PCR）和Southern blot方法，对72例证实为Hp感染的胃十二指肠疾病患者的胃黏膜标本直接进行检测。结果：72例患者中，69例（95.8％）感染的Hp菌株为vacA sIc型，3例（4.2％）为sIa型；23例（31.9％）为vacAm1b型，46例（63.9％）为vacAm2型；6例（8.3％）为混合型。63.9％（46/72）的患者感染菌株为iceA1型，29.2％（21/72）为iceA2型。CagA的阳性率为88.9％（64/72）。结论：Hp毒力因子vacA，cagA和iceA在香港菌株中的分布有自己的特点；未发现特定cagA，vacA和iceA基因型别与DU相关。     

Increased virulence using engineered protease-chitin binding domain hybrid expressed in the entomopathogenic fungus Beauveria bassiana

Insect cuticles consist mainly of interlinked networks of proteins and the highly insoluble polysaccharide, chitin. Entomopathogenic fungi, such as Beauveria bassiana, invade insects by direct penetration of host cuticles via the action of diverse hydrolases including proteases and chitinases coupled to mechanical pressure. In order to better target cuticle protein-chitin structures and accelerate penetration speed, a hybrid protease (CDEP-BmChBD) was constructed by fusion of a chitin binding domain BmChBD from Bombyx mori chitinase to the C-terminal of CDEP-1, a subtilisin-like protease from B. bassiana. Compared to the wild-type, the hybrid protease was able to bind chitin and released greater amounts of peptides/proteins from insect cuticles. The insecticidal activity of B. bassiana was enhanced by including proteases, CDEP-1 or CDEP:BmChBD produced in Pichia pastoris, as an additive, however, the augment effect of CDEP:BmChBD was significantly higher than that of CDEP-1. Expression of the hybrid protease in B. bassiana also significantly increased fungal virulence compared to wild-type and strains overexpressing the native protease. These results demonstrate that rational design virulence factor is a potential strategy for strain improvement by genetic engineering.     

Candida commensalism and virulence: the evolution of phenotypic plasticity.

Candida albicans and related species live as benign commensals in one or more body locations in a majority of healthy individuals. As opportunistic pathogens, they are poised to overgrow cavities and penetrate tissue in response to an alteration in host physiology that presumably compromises the immune functions that normally suppress their growth. There is little evidence of the emergence of successful drug-resistant or hypervirulent strains that predominate in either the commensal or disease states. It appears more likely that all strains possess similar capabilities for rapid adaptation to drug therapy, the immune response and changes in body location and in host physiology. It is suggested that the mechanisms for rapid adaptation lie in the developmental programs of the bud-hypha transition and high frequency phenotypic switching, and in the modulation of the expression of virulence genes in response to environmental cues.