Identification and virulence test of a new pathogen that causes verticillium striping on rapeseed in northwestern China

Five stems of rapeseed with abundant black microsclerotia were collected from Huangyuan County of Qinghai Province,China,and fungal isolates were obtained from the stems.They were identified based on morphology,molecular features and specific PCR detection.The results showed that the 10 fungal isolates belonged to Verticillium longisporum lineage A1/D3.One of the 10 isolates(HW7-1)was tested for virulence on three species of rapeseed,including B.napus Zhongshuang 9,B.rapa Qingyou 9 and B.juncea Tayou 2 by conidia inoculation of HW7-1 on roots of young seedlings.Control seedlings were inoculated with V.dahliae conidia or water alone.The seedlings of these treatments were transplanted in culture mix and incubated in a growth chamber(20℃).Results suggested that the control seedlings of three cultivars appeared quite healthy,while the seedlings inoculated with HW7-1 turned yellowing leaves,seedling stunting or even death after 22 days post-inoculation.V.longisporum was re-isolated from he yellow leaves,thus fulfilling Koch's postulates.Moreover,compared to the control treatments,inoculation with HW7-1 caused flowering delay and seed yield reduction on Tayou 2 with production of microsclerotia on the stems.To our knowledge,this is the first report of V.longisporum lineage A1/D3 on rapeseed in northwestern China.

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.

A potential hyphal fusion protein complex with an important role in development and virulence interacts with autophagy-related proteins in Fusarium pseudograminearum

Hyphal fusion(anastomosis)is a common process serving many important functions at various developmental stages in the life cycle of ascomycetous fungi.However,the biological roles and molecular mechanisms in plant pathogenic fungi were widely unknown.In this study,a hyphal fusion protein FpHam-2 was screened from a T-DNA insertion mutant library of Fusarium pseudograminearum,and FpHam-2 interacts with another 2 hyphal fusion protein homologues FpHam-3 and FpHam-4.Each of these 3 genes deletion mutant revealed in similar defective phenotypes compared with the WT and complemented strains,including reduction in growth rate,defects in hyphal fusion and conidiation,more sensitive for cell membrane,cell wall and oxidative stress responses,and decreased in virulence.The yeast two-hybrid assay was used to identify that FpHam-2 interacts with 3 autophagy-related proteins,including FpAtg3,FpAtg28 and FpAtg33.Furthermore,FpHam-2-deletion mutant showed decreased accumulation of autophagic bodies in hypha.In conclusion,FpHam-2,FpHam-3 and FpHam-4 have an essential role for hyphal fusion and regulating the growth,conidiation and virulence in F.pseudograminearum.

Molecular Detection of Resistance and Virulence Genes in Coagulase Negative Staphylococci Isolated from Blood Cultures at the University Teaching Hospital of Bouake

Introduction: Coagulase-negative staphylococci (CoNS) are currently recognized as genuine pathogens. However, little is known about the resistance and virulence genes that explain their pathogenicity in hospitals in Cte d'Ivoire. The aim of this study was to contribute to the genotypic identification of resistance and virulence genes in CoNS isolated from blood cultures at the University Teaching Hospital (CHU) of Bouak, in order to improve patient management. Material and Methods: This was a descriptive study conducted from September to December 2023. The CoNS isolates studied came from the collection of strains isolated from blood cultures of febrile patients hospitalized or attending consultations at the CHU of Bouak. The strains were analyzed using conventional simplex PCR. Results: Of the 45 isolates analyzed, 46.7% carried both the aacA-aphD and tetK genes and 40% carried the mecA gene. With regard to virulence genes, only the LukS-PV gene was observed in S. epidermidis and S. haemolyticus isolates. Conclusion: The high prevalence of CoNS isolates carrying the mecA gene and the presence of virulence genes observed in this study give cause for concern in hospitals. It is important to develop comprehensive surveillance strategies against nosocomial and multi-resistant infections at the CHU of Bouak.

Mechanistic research:Selenium regulates virulence factors,reducing adhesion ability and inflammatory damage of Helicobacter pylori

BACKGROUND The pathogenicity of Helicobacter pylori is dependent on factors including the environment and the host.Although selenium is closely related to pathogenicity as an environmental factor,the specific correlation between them remains unclear.AIM To investigate how selenium acts on virulence factors and reduces their toxicity.METHODS H.pylori strains were induced by sodium selenite.The expression of cytotoxin-associated protein A(CagA)and vacuolating cytotoxin gene A(VacA)was determined by quantitative PCR and Western blotting.Transcriptomics was used to analyze CagA,CagM,CagE,Cag1,Cag3,and CagT.C57BL/6A mice were infected with the attenuated strains subjected to sodium selenite induction,and H.pylori colonization,inflammatory reactions,and the cell adhesion ability of H.pylori were assessed.RESULTS CagA and VacA expression was upregulated at first and then downregulated in the H.pylori strains after sodium selenite treatment.Their expression was significantly and steadily downregulated after the 5th cycle(10 d).Transcriptome analysis revealed that sodium selenite altered the levels affect H.pylori virulence factors such as CagA,CagM,CagE,Cag1,Cag3,and CagT.Of these factors,CagM and CagE expression was continuously downregulated and further downregulated after 2 h of induction with sodium selenite.Moreover,CagT expression was upregulated before the 3rd cycle(6 d)and significantly downregulated after the 5th cycle.Cag1 and Cag3 expression was upregulated and downregulated,respectively,but no significant change was observed by the 5th cycle.C57BL/6A mice were infected with the attenuated strains subjected to sodium selenite induction.The extent of H.pylori colonization in the stomach increased;however,sodium selenite also induced a mild inflammatory reaction in the gastric mucosa of H.pylori-infected mice,and the cell adhesion ability of H.pylori was significantly weakened.CONCLUSION These results demonstrate that H.pylori displayed virulence attenuation after the 10th d of sodium selenite treatment.Sodium selenite is a low toxicity compound with strong stability that can reduce the cell adhesion ability of H.pylori,thus mitigating the inflammatory damage to the gastric mucosa.

Molecular investigation of exoU and exoY virulence genes in Pseudomonas aeruginosa collected from hospitalized patients in North of Iran:A descriptive-analytical study

Objective:To investigate the frequency of exoU and exoY genes in patients with Pseudomonas aeruginosa infection.Methods:In this study,100 clinical isolates of Pseudomonas aeruginosa were collected from patients hospitalized in educational-therapeutic hospitals and were identified using standard microbiological tests.Then,the antibiotic resistance pattern of the isolates was determined by the disk agar diffusion method.The bacterial DNAs were extracted by the alkaline lysis method.Finally,the presence of exoU and exoY genes was evaluated by the PCR test.Results:In this study,47%,72%,29%,39%,40%,and 44%of the isolates were non-susceptible to piperacillin,aztreonam,ceftazidime,imipenem,tobramycin,and ciprofloxacin,respectively.In addition,95%and 93%of the clinical isolates carried the exoU and exoY genes.Blood and fecal isolates had both virulence genes,while only one wound isolate had neither genes.Meanwhile,all urinary isolates contained the exoY gene and only one isolate lacked the exoU gene.Also,88 isolates simultaneously had both exoU and exoY genes.Conclusions:High prevalence of exoU and exoY genes in this region indicates a significant role of typeⅢsecretion system in pathogenesis of Pseudomonas aeruginosa.The typeⅢsecretion system may be a suitable target to reduce the pathogenicity of this bacterium.

psk1 virulence gene-induced pulmonary and systemic tuberculosis in a young woman with normal immune function:A case report

BACKGROUND Tuberculosis is a chronic infectious disease and an important public health pro-blem.Despite progress in controlling tuberculosis,the incidence of tuberculosis in China is still very high,with 895000 new cases annually.This case report des-cribes the investigation of a case of severe disseminated tuberculosis in a young adult with normal immune function,conducted to ascertain why a Mycobacterium tuberculosis(M.tuberculosis)strain caused such severe disease.CASE SUMMARY A previously healthy 28-year-old woman presented to our hospital with a 1-mo-nth history of fever and fatigue.She was diagnosed with severe disseminated pulmonary tuberculosis,spinal tuberculosis with paravertebral abscesses,and tuberculous meningitis.M.tuberculosis was isolated from bronchoal-veolar lavage fluid.She was treated with standard antituberculous therapy and underwent debridement,bone graft,and internal fixation surgery for spinal tuberculosis.She responded to therapy and regained her ability to walk following the surgery.We analysed the whole-genome sequence of the strain and designated it BLM-A21.Additional M.tuberculosis genomes were selected from the Virulence Factor Database(http://www.mgc.ac.cn/cgi-bin/VFs/genus.cgi?Genus=Mycobacterium)for comparison.An evolutionary tree of the BLM-A21 strain was built using PhyML maximum likelihood software.Further gene analysis revealed that,except for the pks1 gene,BLM-A21 had similar virulence genes to the CDC 1551 and H37Rv strains,which have lower dissemination.CONCLUSION We speculate that the pks1 virulence gene in BLM-A21 may be the key virulence gene responsible for the wide-spread dissemination of M.tuberculosis infection in this previously healthy adult with normal immune function.

Genomic Sequencing and Molecular Characteristics of A Very Virulent Strain of Infectious Bursal Disease Virus Isolated in China

[Objective] The paper was to determine the genomic sequence of a very virulent strain of infectious bursal disease virus（IBDV）,and study its molecular characteristics.[Method] A very virulent strain（vvIBDV）（HLJ-0504） of infectious bursal disease virus（IBDV） with special characters was isolated in China and its genome was sequenced.[Result] Sequence analysis showed that segment A of HLJ-0504 was derived from vvIBDV,while segment B was from a distinct ancestor.The morbidity and mortality of HLJ-0504 was 100% and 86.7%to SPF chickens,respectively.[Conclusion] vvIBDV with distinct segment B were still circulating and the evolution of IBDV was diversified in China.Besides,it is hard to imagine that the virulence of IBDV is determined solely by segment A or B.

Pathogenic Antigenic and Molecular Characterization of the Very Virulent Strain(Gx) of Infectious Bursal Disease Virus Isolated in China

The very virulent infectious bursal disease virus (vvIBDV) strain Gx was isolated from a poutl-try farm in Guangxi Province, China, during 1996. The mortality in the infected flock was 80% and occurred 5 days after immunization with serotype I IBD vaccine. The results of antigen-capture ELISA (AC-ELISA), pathogenicity testing, cloning and sequence analysis of the VP2 gene showed that the deduced amino acid sequence of strain Gx VP2 was the same as vvIBDV UK661, which is considered as a reference strain for European vvIBDVs. The antigenicity of the Gx strain was the same as an European vvIBDV strain 849. The EID50 of Gx virus was 10-8.25/0. 2 ml, and the mortality was 64% when 4 week-old SPF chickens were challenged at dosage of 2×10~3EID50. We have demonstrated that the IBDV strain Gx isolated in China is vvIBDV according to European standards.

Screening of High Virulent Isolate of Beauveria spp. against Helicoverpa assulta and Changes in Protective Enzymes Activities of Infected Larvae

Eleven isolates of Beauveria spp., including B. bassiana(Bb062) newly isolated from Helicoverpa assulta, and other seven B. bassiana isolates and three B. brongniartii isolates that were originally isolated from different geographic origins and various hosts, were tested against the 3^(rd) instar larvae of H. assulta. The protective enzyme activity in the 3^(rd) larvae of H. assulta infected by highly virulent isolate was also assayed. The results showed that the isolate Bb062 had the highest virulence to the 3^(rd) instar larvae among eleven isolates of Beauveria spp., with a corrected mortality reaching 91.07% within 10 d post treatment, and the LT_(50) was 4.67 d. After inoculated with three concentrations(1.0 ×10~6, 1.0 ×10~7 and 1.0×10~8 conidia/mL) of Bb062 conidial suspension, the accumulative mortality of H. assulta larvae increased with the increase of concentration and observation time, and the LC_(50) was 1.82×10~7 conidia/mL. Activities of superoxide dismutase(SOD), peroxidase(POD) and catalase(CAT) in H. assulta larvae first increased rapidly then dropped sharply within 72 h. Bb062 showed high virulence to H. assulta and could inhibit activities of protective enzymes. Therefore, it will be a promising biocontrol agent against H. assulta.

Conidia of one Fusarium solani isolate from a soybean-production field enable to be virulent to soybean and make soybean seedlings wilted

Fusarium is usually thought to cause soybean root rot, which results in a large quantity of annual yield loss in soybean production, by its secretions including Fusarium toxins and cell wall degrading enzymes, but not by the conidia themselves that do not underlie any virulence so far. Here we report that the conidia of one Fusarium solani isolate are able to be virulent to soybean and make soybean seedlings wilted alone. We isolated them from the wilted plants in a soybean-production field and molecularly identified 17 Fusarium isolates through phylogenetic analysis. Of them, except for one isolate that showed diversity of virulence to different soybeans （virulent to one soybean whereas avirulent to another soybean）, the others were all virulent to the two tested soybeans： both conidia cultures and secretions could make soybean seedlings wilted at 5 days post infection, and their virulence had dosage effects that only conidia cultures of at least 5×10^6 conidia mL-1 could show virulence to soybean; however, the sole conidia of the F. solani isolate #4 also exhibited virulence to soybean and could make soybean seedlings wilted. Finally, we developed the specific cleaved amplified polymorphic sequences （CAPS） markers to easily differentiate Fusarium isolates. The isolate #4 in this work will likely be used to investigate the new mechanism of virulence of Fusarium to soybean.

Identification and Characterization of Putative Virulent Genes in Streptococcus equi ssp. zooepidemicus

Suppression subtractive hybridization （SSH） was performed with virulent strain ATCC35246 and avirulent strain ST171 to identify novel genes associated with virulence in Streptococcus equi ssp. zooepidemicus （SEZ）. There were fourteen genomic regions that only presented in virulent strain ATCC35246. These regions encoded 14 proteins, some of them were homologous to proteins associated with cellular surface structure, molecular synthesis, energy metabolism, regulation, transport systems, and other unknown functions. Primers for 6 particular regions were designed from the already published SEZ sequence. Then, we used PCR to evaluate the distribution and conservation of these 6 DNA fragments in various SEZ strains collected from different sources, regions, groups, and times. The results showed that these 6 DNA fragments were widely distributed in SEZ strains, yet they were not existence in the avirulent strain ST171. Moreover, these fragments could not be detected in other Streptococcus groups.

The prevalence of virulent clonal strains of mutans streptococci in vivo and co-culture succession of the strains in vitro—Virulence potential of mutans streptococci

Purpose:To examine selected putative virulent prop-erties of mutans streptococci in two groups with dif-ferent caries activity and to examine co-culture hy-bridization of the strains in vitro. Methods: A set of strains from caries-free subjects (115) and another set from caries-active subjects (165) were isolated. Each strain was characterized for three virulence determi-nants. The clinical bacteria were then cocultured by three strains exhibiting the highest levels of virulence. Isolate colonies of last filial generation bacteria were enrichment-incubated and estimated for virulence again. RAPD-PCR and MLEE analyses were processed for parental bacteria and last filial genera-tion one. Results: No difference associated with caries activity of the subjects from whom the isolates origi-nated. Virulent properties of a filial generation strains was not different in the same generation, but was very different from their parent strains. Conclu-sion: The coexist properties of virulent polyclonal strain of MS may hold in a very general conditional sense in a dental plaque ecosystem in vivo, however, one of the co-culture strains may became dominant and displa- ced the others as the result of continuous ecological succession in vitro.

Immunological Changes of Chicks Immunized with March＇s Disease Vaccines and Challenged with Virulent MD Virus

The experiment was conducted to inoculate one-day old chicks with March's disease (MD) trivalent and herpesvirus of turkey (HVT) vaccines separately, and then to challenge them with virulent MD virus (vMDV) at the age of 15 days. and 5, 25, 45 and 75 days after the challenge with vMDV, comparing with the control-challenged chicks without immunization, to detect the immunoprotetive efficacy and dynamic changes of the inductive activity of interleukin-2(IL-2), expression of IL-2 receptor and proliferative function of T cells in thymus and spleen; the number of ANAE+T, AP+T cells and IgG, IgM, IgA antibody-producing cells in Bursa Fabricius, spleen,thymus, cecal tonsil and Harder gland; as the amount of T cells and IgG, IgM, IgA in peripheral blood as well as the content of IgG, IgM and IgA in the tear, trachea washings, bile and intestinal fluids of the experimental chicks. The experimental results firstly demonstrate that the immunorcgulation of IL-2, and IL-2 receptor, the cellualr and humoral immune responses were significantly enhanced in the central and peripheral immune organs; the local mucosal immune function were markedly amplified in the respiratory and digestive tracts of the immunized-challenged,chicks, which were closely correlated with the immunoprotection against MD; the immune response and immunoprotective effect of the trivalent vaccine-immunized chicks were much better than those of HVT vaccine-immunized chicks:

Susceptibility patterns and virulence genotypes of Helicobacter pylori affecting eradication therapy outcomes among Egyptian patients with gastroduodenal diseases

BACKGROUND Helicobacter pylori(H.pylori)is a significant human pathogen that is responsible for a variety of illnesses,including mucosa-associated lymphoid tissue lymphoma,gastric cancer,peptic ulcers,and gastritis.AIM To investigate the frequency of H.pylori infection and its resistance patterns among Egyptian patients and to determine the influence of H.pylori virulence genetic determinants on the eradication success of 14-d triple therapy regimen.METHODS H.pylori infections were investigated in 72 patients with gastroduodenal complications suggestive of H.pylori infection.The cagA and vacA genotypes of cultured strains were studied using polymerase chain reaction.The patients underwent 14 d of triple-therapy treatment.The treatment response was examined using histology and a rapid urease test 6 wk after therapy discontinuation.RESULTS The intention-to-treat eradication rate was 59.2%(95%CI:48.2%-70.3%).Rates of H.pylori resistance to clarithromycin,amoxicillin,and metronidazole were 52.8%,81.9%,and 100%,respectively.Successful eradication of H.pylori was more significantly associated with vacA s1-positive strains[adjusted odds ratio(aOR)=0.507,95%CI:0.175-0.822].A significant association was found between failed eradication rate and H.pylori strains resistant to clarithromycin(aOR=0.204,95%CI:-0.005 to 0.412)and amoxicillin(aOR=0.223,95%CI:0.026-0.537).CONCLUSION This study’s low H.pylori eradication rate following 14-d triple therapy is concerning and worrying.H.pylori pan-resistance to metronidazole followed by the high resistance to ciprofloxacin,amoxicillin,and clarithromycin in this research is challenging and of great concern.

Molecular epidemiology, characterization of virulence factors and antibiotic resistance profile of Streptococcus agalactiae isolated from dairy farms in China and Pakistan

Streptococcus agalactiae is one of the most common pathogens that cause bovine mastitis worldwide. Identifying pathogen prevalence and virulence factors is critical for developing prevention and control approaches. Herein, 1 161 milk samples from various dairy farms in China(n=558) and Pakistan(n=603) were collected between 2019–2021 and were subjected to S. agalactiae isolation. Prevalence, serotyping, virulence genes, and antibiotic-resistant genes of S. agalactiae were evaluated by PCR assay. All isolates were characterized for haemolysis, biofilm production, cytotoxicity, adhesion, and invasion on bovine mammary epithelial cells. The prevalence of S. agalactiae-induced mastitis in cattle was found to be considerably higher in Pakistan than in China. Jiangsu and Sindh provinces had the highest area-wise prevalence in China and Pakistan, respectively. Serotypes Ia and II were prevalent in both countries, whereas serotype III was found only in Pakistan. Moreover, all isolates tested positive for PI-2b gene but negative for PI-1 and PI-2a genes. All isolates harboured cfb, cylE, hylB, and fbsB virulent genes, whereas many of them lacked bibA, rib and bca. However, the absence of bac and scp genes in Chinese isolates and cspA in Pakistani isolates was noted, while spb1 and lmb were not detected in isolates of both countries. Pakistani isolates, particularly serotype Iapositive, had a considerably higher ability to produce biofilm, haemolysis, cytotoxicity, adhesion, and invasion than Chinese isolates. Most of the isolates were phenotypically resistant to tetracycline, erythromycin, and clindamycin and genotypic resistance was confirmed by the presence of ermA, ermB, tetM and tetO genes. Our study highlights the antimicrobial resistance profile and virulence-related factors contributing to the epidemiological spread of mastitis-causing S. agalactiae in China and Pakistan. The findings may facilitate future studies designed to develop improved treatment and control strategies against this pathogen.

The vital role of CovS in the establishment of Streptococcus equi subsp.zooepidemicus virulence

Streptococcus equi subsp.zooepidemicus(SEZ)is an important zoonotic agent.Here,a virulence-attenuated strain M35246 derived from natural variation of wild-type SEZ ATCC35246 was found.M35246 showed a deletion of 25contiguous genes as well as a loss-of-function mutation in covS.Subsequently,a 25-gene-deleted strain(ΔPI),a covS-mutant strain(Mcov S),and relevant complementary strains were constructed and investigated.M35246 and Mcov S were significantly less encapsulated and exhibited poorer anti-phagocytic capacity compared to wild-type SEZ.McovS was significantly more sensitive toβ-lactams,aminoglycosides,macrolides,and lincosamides than wild-type SEZ.M35246,McovS,andΔPI exhibited an increase in median lethal dose(LD_(50))in mice by 10~5,10~5,and 5 times when compared to wild-type SEZ,respectively.Neither M35246 nor McovS were isolated from mice 48 h after being challenged with approximately 2000 times the LD_(50)of wild-type SEZ.Transcriptome analysis showed that 668 significantly differentially expressed genes existed between McovS and wild-type SEZ.Numerous virulence factor-encoding genes and anabolicrelated genes in McovS that were involved in anti-phagocytosis,capsule formation,pathogenicity,and antibiotic resistance were downregulated significantly relative to the wild-type strain.This study revealed that the CovS plays a vital role in the establishment of SEZ virulence.

Impact of Helicobacter pylori virulence markers on clinical outcomes in adult populations

BACKGROUND In recent years,associations between specific virulence markers of Helicobacter pylori(H.pylori)and gastrointestinal disorders have been suggested.AIM To investigate the presence of virulence factors including vacuolating cytotoxin A genotypes(s1m1,s1m2,s2m1,and s2m2),cytotoxin-associated gene A(CagA),and urease activity in H.pylori strains isolated from Arab and Jewish populations in northern Israel and to assess associations between these factors and patients’demographics and clinical outcomes.METHODS Patients(n=108)who underwent gastroscopy at the Baruch Padeh Medical Center,Poriya due to symptomatic gastroduodenal pathologies as part of H.pylori diagnosis were enrolled in the study.Gastric biopsy specimens were collected from the antrum of the stomach.Clinical condition was assessed by clinical pathology tests.Bacteria were isolated on modified BD Helicobacter Agar(BD Diagnostics,Sparks,MD,United States).Bacterial DNA was extracted,and PCR was performed to detect CagA and vacuolating cytotoxin A genes.Urease activity was assessed using a rapid urease test.RESULTS A significant correlation was found between disease severity and patient ethnicity(P=0.002).A significant correlation was found between CagA presence and the s1m1 genotype(P=0.02),which is considered the most virulent genotype.Further,a higher level of urease activity was associated with isolates originating from the Jewish population.Moreover,higher urease activity levels were measured among CagA-/s1m1 and CagA-/s2m2 isolates.CONCLUSION Our study highlights the importance of incorporating molecular methods for detection of virulence markers of H.pylori in order to tailor optimal treatments for each patient.Further investigation should be performed regarding associations between H.pylori virulence factors and ethnicity.

Virulence Factors and Biofilm Formation in Vancomycin Resistant Enterococcus faecalis and Enterococcus faecium Isolates in Brazil

In this work, we evaluated biofilm formation of Vancomycin Resistant of E. faecalis and E. faecium (VRE) in different culture media and adhesion substrate, as well as cellular hydrophobicity and presence of virulence genes. For this, 35 isolates were collected from a public hospital in Recife, Pernambuco, Brazil and identified by the Matrix-Assisted Laser Desorption Ionization - Time-of-flight - Mass Spectrometry (MALDI-TOF-MS) technique. Biofilm formation was analyzed by the Crystal Violet (CV) method and fluorescence microscopy, cellular hydrophobicity by hydrocarbon interaction and the presence of gelE, esp and asa1 genes by Polymerase Chain Reaction (PCR). 12 isolates were identified as E. faecalis and 23 as E. faecium. Most were obtained in Coronary Units (40.0%) and Intensive Care Unit (31.4%). E. faecium isolates were more resistant to the antibiotics tested than E. faecalis;however, E. faecalis stood out as a biofilm producer. Regarding the presence and gene frequency, it was observed that gelE (54.3%) and esp (54.3%) were the most prevalent, followed by asa1 (22.9%). When comparing the gene frequency, it was observed that gelE and esp were predominant (48.6% for both species), while asa1 was more frequent in E. faecalis (20.0%). The data presented here are worrying, because they reveal the virulence potential of isolates VRE, which contributes to the dissemination and persistence of these pathogens in the hospital environment.

Genotypic Resistance of F_1 Cotton Hybrids by Inoculation with Different Virulent Isolates of the Fungus Verticillium Dahliae Klebahn

The plant pathogen Verticillium dahliae causes severe cotton losses in Uzbekistan. To create cotton varieties that are resistant to the more virulent races of V.dahliae we wanted to determine